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autoMACS® Pro Separator User manual

1. EEE 03 3 Figure 6 20 A reagent vial containing CD4 MicroBeads human was placed into the Reagent Rack position R1 1 Inthe Separation menu select the desired position s in the sample rack template field by touching it 2 Assign a reagent for autolabeling from the Labeling submenu to each position The recommended separation and wash programs will be automatically displayed in the Separation menu 3 Optional The pre selected separation and wash programs can be changed in the corresponding submenus 4 Toassign a sample volume click on the Volume submenu and enter the corresponding value using the numeric keypad Select Enter 6 Cell separation using the autoMACS Pro Separator 133 Note option under Labeling denotes that NO autolabeling will be performed Only a cell separation will be performed manual magnetic labeling must be performed prior to placing the sample in the Chill Rack Note If the Clean program has been enabled it will also appear in the Wash submenu HE 03 25 Figure 6 21 Performing a cell separation with CD4 MicroBeads human using the autolabeling feature The cell separation program Possel and wash program Qrinse were selected 5 Select Run 6 2 5 2 Cell separation after manual labeling After completing the priming process or a wash program the autoMACS Pro Separator is ready for separation The status of the instrument is displayed in the Status menu
2. Automated arm Front door Fluid bottle basket Bottom cover Figure 10 1 Front view of autoMACS Pro Separator Automated arm with ports for sample uptake and release of cell fractions The automated arm figure 10 1 is a computer controlled part of the autoMACS Pro Separator It holds two ports one for sample uptake and release of the magnetically labeled positive cell fraction and one for release of the non labeled negative cell fraction The automated arm moves in y and z directions The ports are automatically washed in the autoMACS Pro washing station during and after the separation process to prevent cross contamination between samples Access covers The front door figure 10 1 is opened sideways to allow access to the parts of the fluidic system that require periodic maintenance by the user This includes the autoMACS Columns pump syringe and upper valves The bottom cover figure 10 1 gives access to the lower valves and can be removed by pulling gently The washing station cover is opened sidewards giving access to the washing station the peristaltic pump and the tubing of the autoMACS Pro washing station 10 Technical data and specifications 189 Fluid bottles and fluid bottle baskets Two baskets holding two fluid bottles each are located at each side of the instrument Fluid bottles are connected to the autoMACS Pro system with color coded tubing and sensors at the bottle closures for fluid level cont
3. Cancel to abort 7028 Barcode on Chill Rack could not be read MACS Check electrical connection of MACS MiniSampler If MiniSampler is not connected properly detected the MACS MiniSampler symbol would be displayed in the status screen 7029 Unable to read rack bar code correctly Check bar code on rack or try different rack Make sure ambient or direct sun light does not hit sensor Touch Retry to try again touch Select to set Chill Rack type without automatic bare code reading 7030 The instrument has been shut down without Always use Sleep to shut down the instrument using Sleep 9 Troubleshooting 187 188 Error code 7031 7032 7033 7034 7035 7036 7037 7038 7039 7048 Cause The given whole blood sample size exceeds the maximum volume specified The sample cannot be diluted sufficiently Air in system during calibration of tubing Date and time is outdated Air intake during sample uptake although needle did not yet hit bottom of tube Leakage of air into system in front of bubble sensor liquid level has been overestimated or needle did not move to bottom as fast as necessary Air intake during sample uptake although needle did not yet hit bottom of tube Leakage of air into system in front of bubble sensor liquid level has been overestimated or needle did not move to bottom as fast as necessary Uptaken sample has been processed Portion of
4. MicroBead Kits Chill 15 Rack Direct MicroBeads human rat non human primate Direct MicroBeads mouse Whole Blood MicroBeads Cell Isolation Kits Cell Isolation Kits MicroBead Kits Chill 50 Rack Whole Blood MicroBeads Number of Reagents Strategy Positive 1 selection or depletion Positive 1 selection or depletion Whole blood or 1 bone marrow Untouched 2 isolation Untouched 3 isolation Positive 2 selection or depletion Positive 1 selection or depletion Positive 1 selection or depletion Whole blood or 1 bone marrow Untouched 2 isolation Untouched 3 isolation Positive 2 selection or depletion Whole blood or 1 bone marrow Dilution volume 10 cells per 80 uL 10 cells per 90 uL Original volume 10 cells per 40 uL 10 cells per 30 uL 10 cells per 60 uL 107 cells per 80 uL 107 cells per 90 uL Original volume 10 cells per 40 uL 10 cells per 30 uL 10 cells per 60 uL Original volume Minimal volume 160 uL 180 pL 0 25 mL 160 uL 120 uL 120 uL 160 uL 180 uL 1mL 160 uL 120 uL 120 uL 4 mL Autolabeling Maximal volume Minimal total cell number 2x10 1600 uL 2x10 1800 uL 1mL 4x10 800 uL 4x10 600 uL 2x10 1200 uL 2x10 5200 pL 2x10 5850 uL 4mL 4x107 2600 uL 4x10 1950 uL 2x107 3900 uL 8mL 1 Max number of samples 6 min first incubatio
5. 1 Verify that the washing station is not clogged with salt deposits Take out the washing station and clean as indicated in section 7 3 3 Reassemble washing station and run a Rinse program Select Wash Now from the lower menu bar in the Separation menu Highlight Rinse and click Run If the problem persists follow the steps below Make sure that the peristaltic waste pump works properly Run the Check_up program Select User settings from the Option menu Highlight Check_up and click Run If the Check_up program reports a problem with the waste pump remove the pump head and clean the pin that drives the pump for details refer to section 7 4 3 Clean the washing station refer to section 7 3 3 Reassemble the unit and check whether the problem persists If the problem persists replace the pump head refer to section 7 4 3 9 Troubleshooting 9 173 174 Note If initial wash Rinse program is enabled press Stop as soon as the Status menu appears on the screen A window opens displaying that the process is paused Press Cancel 9 1 6 Outlet port is clogged 1 Ifthe outlet ports are clogged e g due to salt deposits the elution process might be affected 2 Wipe the outlet port with a tissue soaked with 70 ethanol or double distilled water 3 Flush the outlet port manually using a syringe filled with 70 ethanol or double distilled water 9 1 7 MACS MiniSampler does not move properly 1 Check whet
6. 5 1 2 Hardware autoMACS Columns 5x2 columns 130 021 101 autoMACS Columns can be used for up to 14 days or 100 separations within these 14 days Their capacity reaches up to 2x108 magnetically labeled cells from up to 4x10 nucleated total cells or up to 15 mL of whole blood 5 Materials required 5 2 autoMACS Pro Separator 5 accessories Accessory Description Order no Chill 5 Rack Chill Rack for 5 mL tubes 130 092 951 Chill 5 Rack Box of 3 3x Chill Racks for 5 mL tubes 130 097 041 Chill 15 Rack Chill Rack for 5 and 15 mL 130 092 952 tubes Chill 15 Rack Box of 3 3x Chill Racks for 5 and 15 mL 130 097 036 tubes Chill 50 Rack Chill Rack for 5 15 and 50 mL 130 092 953 tubes Chill 50 Rack Box of 3 3x Chill Racks for 5 15 and 130 097 037 50 mL tubes Chill 5 15 50 Rack Set Set of three Chill Racks 130 097 038 MACS Reagent Rack 4 Accommodates 4 vials of 130 094 574 MACS Reagents Air Filter Extension Set Extension tube and 130 091 339 hydrophobic air filter for the autoMACS Fluid Bottles autoMACS Pro Laminar Metal plate for operating the 130 093 246 Hood Plate autoMACS Pro Separator ina laminar flow hood autoMACS Pro Angle Space saving angle adaptors 130 093 245 Connector Set for cables connecting to the autoMACS Pro Separator autoMACS Pro Protection foil for long term 130 093 532 Protection Cover storage of the autoMACS Pro Separator Table 5 2 List of accessories for the autoMACS Pro Separator
7. Calibr_2 4 Select Calibrate The calibration is performed automatically 5 Press Save to finish Calibr_2 11 Hardware and software installation Technical support 12 For technical support please contact your local Miltenyi Biotec representative or the Technical Support team at Miltenyi Biotec headquarters Miltenyi Biotec GmbH Friedrich Ebert Stra e 68 51429 Bergisch Gladbach Germany Phone 49 2204 8306 830 Fax 49 2204 8306 89 macstec miltenyibiotec de www miltenyibiotec com Visit www miltenyibiotec com local to find your nearest Miltenyi Biotec contact 12 Technical support 203 204 12 Technical support Limited warranty 13 Except as stated in a specific warranty statement which may accompany your autoMACS Pro Separator the Product or unless otherwise agreed in writing by an authorized representative of Miltenyi Biotec Miltenyi Biotec s warranty if any with respect to this Product is subject to the terms and conditions of sale the Terms of the company within the Miltenyi Biotec group which supplied the Product The Terms may vary by country and region Copies of these Terms are available on request or at www miltenyibiotec com Nothing in this document should be construed as constituting an additional warranty Miltenyi Biotec s product warranty only covers Product issues caused by defects in material or workmanship encountered during ordinary use as described in the user manual or othe
8. FITC Fluorescein isothiocyanate 14 Glossary Fluid bottle 1 5 L bottles holding the fluids for operational use of 1 the autoMACS Pro Separator Fluid sensors monitor the fluid levels in the bottles for Running Buffer Washing Solution and waste via electrolyte conductivity The fluid level in the bottle for the storage solution cannot be monitored as no electrolytes are present in the solvent Fluid sensors This sensor type measures electrolyte conductivity and is an integral part of the bottle closures of the fluid bottles for waste Running Buffer and Washing Solution Fluid sensor cable Cable connecting the fluid sensor to the autoMACS Pro Separator The sensor cable connectors on the bottle closures are color coded red for waste green for Washing Solution and blue for Running Buffer Fraction collection tube 5 mL 15 mL or 50 mL plastic sample tubes to collect the positive and negative fractions The process has been optimized using tubes from BD Falcon Front door The front door opens sidewards giving access to the autoMACS Columns pumps valves washing station and tubing MACS Technology Technology developed by Miltenyi Biotec for immunomagnetic labeling and subsequent separation of cells or biomolecules in a high gradient magnetic field MACS MicroBeads Super paramagnetic particles conjugated to antibodies for magnetic labeling of cells or biomolecules eee Magnet cover Black cover surrounding th
9. For information on spare parts please refer to section 7 8 5 Materials required 113 114 5 Materials required Cell separation using the 6 autoMACS Pro Separator The following chapter describes how to perform cell separations using the autoMACS Pro Separator IMPORTANT The autoMACS Pro Separator is intended for research applications only and not for diagnostic or therapeutic use essi 6 1 General considerations The autoMACS Pro Separator can perform fully automated cell separations This includes all steps of the separation procedure from cell labeling to sample loading the separation over autoMACS Columns and the collection of unlabeled and labeled cell fractions Cell labeling can be performed manually or using the autolabeling feature of the instrument The pre set separation programs of the autoMACS Pro Separator have been developed for optimal separation results independent from the applied labeling and separation strategy 6 1 1 Cell labeling strategies There are two basic approaches for magnetic labeling of cells with MACS MicroBeads direct and indirect magnetic labeling For direct labeling MACS MicroBeads bind directly to a specific cell surface marker protein For indirect labeling MACS MicroBeads recognize a primary antibody its conjugates or a ligand binding to a specific cell surface marker protein Direct magnetic labeling Direct labeling with MACS MicroBeads is the fastest way of magnetic l
10. Miltenyi Biotec autoMACS Pro Separator User manual Version 4 Original instructions Copyright 2013 Miltenyi Biotec All rights reserved No part of this user manual may be reproduced stored in a retrieval system transmitted published or distributed in any form or by any means electronically mechanically by photocopying microfilming recording or otherwise without the prior written consent of Miltenyi Biotec however notwithstanding the foregoing the owners of the autoMACS Pro Separator may make copies solely for purposes of training personnel in the use and servicing of the unit within their business or organization Maximal care has been taken by Miltenyi Biotec in the preparation of this user manual However Miltenyi Biotec shall not be liable for any technical or editorial errors or omissions contained herein or for incidental or consequential damages in connection with the furnishing performance or use of this document The information in this document is provided as is without warranty of any kind and is subject to change without notice MACS autoMACS and VioBlue are registered trademarks of Miltenyi Biotec GmbH Ficoll and Ficoll Paque are trademarks of GE Healthcare companies All other trademarks mentioned in this document are the property of their respective owners and are used for identification purposes only Unless otherwise specifically indicated Miltenyi Biotec products and services are for research
11. Miltenyi Biotec B V Schipholweg 68 H 2316 XE Leiden The Netherlands macs miltenyibiotec nl Customer service The Netherlands Phone 0800 4020120 Fax 0800 4020100 Customer service Belgium Phone 0800 94016 Fax 0800 99626 Customer service Luxembourg Phone 800 24971 Fax 800 24984 China Miltenyi Biotec GmbH Shanghai Office Rm 2309 No 319 Xianxia Rd Shanghai 200051 P R China Phone 86 21 62351005 Fax 86 21 62350953 macs miltenyibiotec com cn France Miltenyi Biotec SAS 10 rue Mercoeur 75011 Paris France Phone 33 1 56 98 16 16 Fax 33 156 98 16 17 macs miltenyibiotec fr Italy Miltenyi Biotec S r l Via Persicetana 2 D 40012 Calderara di Reno BO Italy Phone 39 051 6 460 411 Fax 39 051 6 460 499 macs miltenyibiotec it Japan Miltenyi Biotec K K Nittsu Eitai Building 5F 16 10 Fuyuki Koto ku Tokyo 135 0041 Japan Phone 81 3 5646 8910 Fax 81 3 5646 8911 macs miltenyibiotec jp Nordics and Baltics Miltenyi Biotec Norden AB Scheelev gen 17 223 70 Lund Sweden macs miltenyibiotec se Customer service Sweden Phone 0200 111 800 Fax 046 280 72 99 Customer service Denmark Phone 80 20 30 10 Fax 46 46 280 72 99 Customer service Norway Finland Iceland and Baltic countries Phone 46 46 280 72 80 Fax 46 46 280 72 99 Singapore Miltenyi Biotec Asia Pacific Pte Ltd 100 Beach Road 28 06 to 28 08 Shaw Tower Singapore 189702 Phone 65 6238 8183 Fax 65 6238 0302 macs
12. Place the bottle in its appropriate position remove the cap and fasten the bottle closure Note the color coding and connect each sensor cable to the respective bottle closure 4 Attach the sensor cable plug to the socket for sensor cables at the back of the autoMACS Pro Separator labeled Bottle Sensor and fasten securely 5 Connect the hydrophobic air filters 0 2 um to the appropriate connectors on the bottle closures AN x d ie y Figure 3 13 Positioning of the hydrophobic air filters on the bottle closures 3 Assembly and installation Note Ensure that the autoMACS Pro Separator has been correctly installed section 3 1 and that the empty fluid bottles used for transport purposes have been replaced by fresh fluid bottles section 3 2 3 3 Running the autoMACS Pro 3 Separator for the first time performing a test calibration 3 3 1 Switch ON the autoMACS Pro Separator The main power switch is located on the right side of the instrument in front of the bottle baskets I indicates ON O indicates OFF Switch on the autoMACS Pro Separator ON OFF switch Figure 3 14 Location of the ON OFF switch 3 3 2 Check the date and time 1 Toset the time and date perform the following steps Select Option User settings and Set_time Select Run Highlight either the time or date fields by touching the display Use the keypad to change the date or time accordingly u Sel new li
13. groove 2 hole for adjustment pins 7 Make sure that the bracket of the new valve is positioned horizontally 7 Maintenance 157 158 Note In case the valve exchange program Valve_ex cannot be performed e g valve does not turn any more switch OFF instrument and turn the valve bracket manually to the position corresponding to the groove in the valve drive Figure 7 19 Rear view of the autoMACS Pro Separator valve 1 bracket 2 adjustment pins 8 Carefully insert the new valve allowing the bracket to find the groove in the drive At first the bracket will slide in only halfway Figure 7 20 Inserting the new valve 9 10 11 12 13 14 15 Gently rotate the valve The two adjustment pins will slide into their corresponding holes in the valve plate Make sure that the valve is fully inserted into the driving station Fasten valve screws using the screwdriver Connect the tubing with the installed new valve and fasten tubing by hand until finger tight Plug in and switch ON the autoMACS Pro Separator Perform a Rinse program and check the valves visually for leakage and air inlet Refasten tubing connectors using caution If leakage persists call Technical Support Take care not to pinch the tubings at the bottom left of the instrument when closing the bottom cover 7 Maintenance 7 4 2 Exchange of dilutor valve 1 Switch OFF and unplug the instrument 2 Remove the pump
14. t utilis e ou s est d vers e prenez les pr cautions appropri es pour d contaminer soigneusement le syst me Toujours v rifier le syst me fluidique le jeu complet de tubulures les bouteilles et leurs bouchons les valves les colonnes la valve diluteur et les aiguilles avant d allumer l instrument Si des fuites sont d tect es remplacer les pi ces endommag es avant d allumer l instrument Si les pi ces ne peuvent pas tre remplac es d brancher l instrument et ne pas l utiliser Les pieces endommag es contenant du mat riel biologique infectieux ou des liquides qui ont t en contact avec un tel mat riel peuvent tre potentiellement dangereuses 1 Informations importantes Les colonnes les plaques les tubes et tous les autres consommables 1 qui ont t en contact avec des chantillons nocifs pour l organisme doivent tre trait s l autoclave avant l limination Les d chets liquides doivent tre trait s l autoclave ou d contamin s l aide d un d sinfectant adapt l agent pathog ne sp cifique par ex Ga 10 eau de Javel alcool isopropylique ou 70 d thanol L limination des d chets doit tre effectu e conform ment aux r glementations locales a utoMACS Pro Separator SN 0563 Order na 190 097 545 Power Supply 100 240 VAL 50 00 Hz 200 VA Fuser 2014 2304 Figure 1 4 Les symboles d alerte pour les dangers biologiques sont localis s sur la part
15. 58 1 Informazioni importanti Informacion importante 1 Lea la siguiente informaci n antes de usar el instrumento equipo Por favor lea ntegramente la informaci n contenida en este manual de usuario antes de utilizar el instrumento equipo La lectura y o seguimiento incorrectos de estas directrices pueden llevar a un uso manejo o mantenimiento incorrectos del equipo lo cual puede poner en peligro a sus usuarios producir resultados impredecibles derivar en un mal funcionamiento del aparato o que ste sufra da os as como causar un desgaste prematuro y reducir el tiempo de vida del instrumento pudiendo anular su garant a Mantenga este manual de usuario en un lugar seguro al alcance de todo aquel que est usando la Unidad de Separaci n autoMACS Pro Separator separador autoMACS Pro El presente cap tulo describe las instrucciones de seguridad y los requisitos de emplazamiento de su separador autoMACS Pro Las siguientes advertencias y precauciones tienen como objetivo evitar da os personales o materiales 1 1 S mbolos y niveles de peligro Disposici n de advertencias de seguridad Ejemplo CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard Las advertencias de seguridad informan al usuario sobre riesgos potenciales en caso de que no se sigan las advertencias y precauciones resumidas a continuaci n El s mbolo que aparece a la izquie
16. Calibr 1 7021 A separation program has been started but Please rinse system by touching Wash or abort with system had not been rinsed properly Cancel 7022 Columns are overdue Please install new columns Touch Cancel to abort and PEA then install columns touch Continue to ignore and use old columns not recommended 7023 Plunger movement blocked because of column Touch Cont to discard the remaining negative cells into clogging blocked tubing set or any other the waste bottle To output negative fraction again at cross section constriction during output of the lower speed exchange negative tube with an empty negative fraction The negative fraction has not tube and touch Retry been eluted completely Negative cells are still remaining in the system 7024 The number of programmed sample positions Exchange rack with rack holding more samples and exceeds the actual positions of the rack on the touch OK or touch Cancel to abort and reprogram MACS MiniSampler 7026 The protective cover of the MACS MiniSampler Check configuration and connection of the protective seems to be opened by the moving needle arm cover the MACS MiniSampler and the front support at hitting the cover the autoMACS Pro Separator then touch Continue Touch Cancel to abort 7027 The protective cover of the MACS MiniSampler Please close protective cover and touch Continue needs to be closed Touch
17. Check whether the uptake needle is positioned in the middle of the sample tube during the mixing procedure If not limited mixing could lead to low purity Refer to the respective data sheet for the recommended separation program Recovery of target cells is low Refer to section 9 3 1 Overall recovery is low Refer to section 9 3 1 9 Troubleshooting 9 3 3 MACS MicroBeads for indirect labeling 9 Positive cells have not been retained on the column 1 The unbound primary antibody was not completely removed and inhibits magnetic labeling with MACS MicroBeads for indirect labeling Wash cells carefully optionally wash twice by adding 1 2 mL of buffer per 10 cells i e 10 20 times the incubation volume after incubation with primary antibody Centrifuge and remove supernatant completely preferably with a pipette or by vacuum 2 The concentration of the primary antibody was too low Therefore magnetic labeling was not sufficient to retain the desired cells on the column To avoid this the primary antibody should be titrated carefully For tips and hints on titration refer to the FAQs under Customer Support at www miltenyibiotec com 3 The isotype of the primary antibody was not recognized by the Anti Immunoglobulin MicroBeads Make sure to use appropriate MicroBeads 4 Check the expression of the antigen epitope with flow cytometry 5 Antibodies may have degraded Check the antibody for its function Storage o
18. During depletion the unwanted cells are labeled and depleted from the target cells The target cells are collected as the negative fraction Furthermore sequential sorting allows the performance of two consecutive separations Positive selection Positive selection means that a particular target cell type is magnetically labeled During separation the magnetically labeled cells are retained within the column Unlabeled cells flow through and are collected as the negative fraction After automated retraction of the magnet the magnetically labeled target cells are eluted from the autoMACS Column as the positive fraction Positive selection can be performed after direct or indirect magnetic labeling using various MACS MicroBeads or MicroBead Kits MACS Whole Blood MicroBeads are specially developed for the positive selection of leukocyte subsets directly from human whole blood or bone marrow Depletion To remove a certain cell type from a mixture of cells the unwanted cell type is magnetically labeled During separation the unwanted labeled cell type is retained within the column The unlabeled target cells flow through the column and are collected as the negative fraction After retraction of the magnet the retained cells are eluted from the autoMACS Column 6 Cell separation using the autoMACS Pro Separator Untouched isolation 6 To isolate a particular target cell type in an unlabeled i e untouched form non target cells are magne
19. Esistono potenziali rischi in caso di utilizzo di uno strumento AutoMACS Pro Separator aperto caduto o danneggiato in caso di infiltrazione di liquidi all interno dello strumento in caso di penetrazione di oggetti nello strumento attraverso le fessure di ventilazione o in caso di inserimento accidentale di oggetti nello strumento In caso di presenza di fiamme o fumo spegnere immediatamente il autoMACS Pro Separator staccare lo strumento dalla presa di corrente e contattare un rivenditore Miltenyi Biotec autorizzato o il servizio di assistenza clienti Miltenyi Biotec E severamente vietato utilizzare uno strumento danneggiato o provvisto di cavo dell alimentazione danneggiato 1 3 2 Forte campo magnetico WARNING Extremely strong magnetic fields Risk of severe personal injury to persons carrying pacemakers or electronical medical implants stay clear Keep all magnetically sensitive objects away from device AVVERTENZA autoMACS Pro Separator provvisto di un magnete estremamente potente Tenere i supporti magnetici come carte di credito nastri magnetici e supporti di memorizzazione e i dispositivi elettronici come apparecchi acustici pacemaker strumenti di misurazione e controllo computer e orologi ad una distanza minima 1 Informazioni importanti di 20 cm dal coperchio del magnete Questi oggetti potrebbero 1 essere danneggiati oil loro funzionamento essere compromesso dal campo magnetico Figura 1 1 Segna
20. It is necessary to configure the reagent rack before starting the run _ I Ey Figure 4 22 Selecting a separation template Expt_2a was selected 2 Select and highlight the desired template in this case Expt_2a 3 Select Ok LES Figure 4 23 The template Expt_2a was successfully loaded more information Figure 4 24 The Reagent Rack has to be configured before starting the run To delete a separation template 1 Select Load Template from the lower navigation bar 2 Scroll through the list of saved templates use the navigation arrows A V 3 Select the template for deletion 4 An overview of the autoMACS Pro Separator user interface 103 104 En Figure 4 25 Expt_3 template was selected for deletion 4 Select Delete Template 4 5 Reagent menu The Reagent menu is used to program any reagent vials that are required for automated magnetic labeling and subsequent cell sorting Reagents can be entered using the 2D code reader or manually using the Enter Reagent input panel For convenience it is possible to load and save reagent templates For more information on the Reagent menu please refer to section 6 2 4 Define reagent rack N 11 31 Figure 4 26 The Reagent menu 4 6 Log list menu RE 03 11 Figure 4 27 The Log list display 4 An overview of the autoMACS Pro Separator user interface The log list records a complete log of actions performed by
21. To select a particular sample position in the programming field touch it once To deselect it touch it once again If the same combination of separation program and washing program is required for more than one sample highlight all the desired sample positions first and then select the programs Alternatively define the program combination for one position and then highlight the other desired positions to adopt the program combination To erase selected program combinations from the programming field first mark the respective positions and choose Delete selected from the lower navigation bar If positions are not marked the button can be used to delete the entire template In this case the option Delete selected will switch to Delete all Alternatively start the separation from a previously saved separation template 1 Select Load template from the lower navigation bar 2 Select the desired template by using the arrows 6 Cell separation using the autoMACS Pro Separator 135 136 Note The touchscreen displays the type of tube rack upon starting the separation 3 Select OK 4 Select Run to start the separation 5 Check that there is enough buffer for the number of programmed separations as instructed by a pop up dialog box Click Continue to proceed The autoMACS Pro Separator automatically detects the type of tube rack in use and allows the user to utilize only the number of samples and sample positions the tube
22. and source of the hazard as well as potential consequences prohibitions and measures are pointed as follows 1 Important information 11 WARNING CAUTION o o Op gt gt gt gt Symbols and hazard levels The following chart is an illustrated glossary depicting the symbols that are used in this user manual and on the autoMACS Pro Separator WARNING indicates a potentially hazardous situation which if not avoided could result in death or serious injury CAUTION indicates a potentially hazardous situation which if not avoided could result in minor or moderate injury Attention consult the user manual for further instructions and proceed with caution Warnings include the risk of damage to the equipment severe personal injury or loss of life Hazard of crushing and shearing Risk of crushing and shearing of bodily parts due to mechanical hazards Laser radiation Risk of serious eye and skin injuries Strong magnetic field The magnetic field can interfere with magnetizable objects and electronic devices or damage magnetic information carriers Risk of severe personal injury to persons carrying pacemakers or electronic medical implants Risk of contamination if biohazardous material is used Indicates the risk of death severe injury to the instrument operator or equipment damage due to potentially dangerous biological material Indicates the risk of death or severe injury to the instrument operator due to ha
23. cole E 9 un ua DG 59 un a lo o O S OS gt lu S mu a a amp 3 SEA mio 21 7 a6 ma ES ES mo DO vw mr fh Wg D Hi Or me tn n 2 n gt AS a A N58 sia 28 3 2 83 82 3 2 MM ES TE o fey tel v v amp ae aa Ta amp AE a aa aa Purity will slightly decrease Recovery will slightly decrease 144 6 Cell separation using the autoMACS Pro Separator Maintenance 7 Appropriate maintenance of the autoMACS Pro Separator helps to maintain excellent reproducibility of the cell separation results The following section gives you an overview on the procedures required for efficient maintenance of the instrument NOTICE Insufficient or improper maintenance of your autoMACS Pro Separator can cause unpredictable results avoidable malfunction and premature failure of the instrument and may void ES your warranty NOTICE Please do not perform any maintenance procedures other than specifically described in this user manual Any other maintenance procedures must be performed by qualified service personnel Inquire with your local Miltenyi Biotec representative about Miltenyi Biotec s extensive instrument service and support arrangements or refer to www miltenyibiotec com support NOTICE When replacement or spare parts are required for maintenance only use genuine Miltenyi Biotec parts or third party parts specified and recommended by Miltenyi Biotec Using unauthorized replacement or spare
24. device after spilling of biohazardous material Wird oder wurde mit biologischen Gefahrenstoffen gearbeitet muss der Bediener des Ger tes entsprechend den f r die verwendeten Substanzen geltenden Warnhinweisen und Schutzbestimmungen eine pers nliche Schutzausr stung tragen Tragen Sie Schutz handschuhe Schutzkleidung und Schutzbrille um Ber hrung der Gefahrenstoffe mit Haut und Augen zu vermeiden Sch tzen Sie auch Ihren Mund und Nase da Aerosole aus undichten Stellen des Systems austreten k nnten Mangelhafte oder unzureichende Schutzausr stung kann den Bediener des Instrumentes gef hrden Werden biologische Gefahrenstoffe oder unbekannte Substanzen eingesetzt sollten Sie mit dem autoMACS Pro Separator in einer Sterilbank arbeiten Falls Gefahrstoffe verwendet wurden oder ausgetreten sind achten Sie auf eine sorgf ltige Dekontaminierung des Ger tes Vor Inbetriebnahme des Ger tes berpr fen Sie das fluidische System das Schlauchsystem Flaschen und deren Verschl sse Ventile S ulen Verd nnerventil und Nadeln und ersetzen Sie beim Feststellen einer undichten Stelle alle besch digten Teile K nnen besch digte Teile nicht ersetzt werden ziehen Sie den Netzstecker und benutzen Sie das Ger t nicht Besch digte Teile die mit biologischen Gefahrenstoffen in Kontakt waren sind potentiell gef hrlich S ulen Auffanggef e und alle weiteren Verbrauchsmaterialien die in Kontakt mit biologischen Gefahrenstoffe
25. gauche d finit le risque En haut le danger mentionn pr c demment est affect un niveau de danger Le niveau le type et la source du danger ainsi que les ventuelles cons quences les interdictions et les mesures prendre sont mentionn s conform ment au tableau ci dessous 1 Informations importantes 23 24 WARNING CAUTION 00o Ob Db PPP gt Al Symboles et niveaux de danger Le tableau suivant est un glossaire illustr expliquant les symboles utilis s dans ce mode d emploi et sur le s parateur autoMACS Pro Separator AVERTISSEMENT indique une situation dangereuse qui si elle n est pas vit e peut entrainer la mort ou des blessures graves ATTENTION indique une situation dangereuse qui si elle n est pas vit e peut entra ner des blessures mineures ou mod r es Attention consultez le mode d emploi pour obtenir plus de consignes et agissez avec prudence Les avertissements incluent le risque de d t rioration de l quipement de blessure corporelle grave et le danger de mort Risque d crasement et de cisaillement Risque d crasement et de cisaillement des membres corporels d des dangers m caniques Rayonnement laser Danger de graves l sions oculaires et cutan es Champ magn tique puissant Le champ magn tique interf re avec les objets magn tisables et les appareils lectroniques ou endommage les supports d information magn tiques Risque de grave blessure corporelle pour
26. if the instrument cannot be restarted follow procedure B CAUTION Depending on the nature of your sample it is recommended to wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes Dispose tissues swabs and vials appropriately 7 Maintenance Note In order to perform a cell rescue procedure the O_ init programm must be disabled If initial wash O_init program is enabled press Stop as soon as the Status menu appears on the screen A window opens displaying that the process is paused Press Cancel 7 7 1 Rescue procedure A 1 Restart the instrument by switching it OFF and ON again 2 Undo the tubing connector at the negative port and place into a 50 mL tube 3 Take out the uptake port needle from the needle holder and place it into a 50 mL tube 4 Undo the tubing connector of the waste tube at the waste bottle and place itinto a 50 mL tube place a second 50mL tube beside this one 5 Run the program Qrinse This will rinse the complete fluidic system with autoMACS Pro Running Buffer eluting the cells into the 50 mL tubes Depending on which step of the separation program that the interruption occurred the cells will be found in any one of the vials 6 Combine all fractions and centrifuge at 350xg for 10 minutes 7 Discard the supernatant and apply cells to a reseparation as soon as possible Keep cells on ice until the separation 8 Reconnect all tubing at the a
27. in order to reduce formation of cell aggregates Dead cells in the cell suspension may non specifically bind to MicroBeads and will then be co enriched in the positive fraction Remove dead cells before separation by using the MACS Dead Cell Removal Kit 130 090 101 or by Ficoll Paque density gradient The concentration of MicroBeads or the temperature used for magnetic labeling were too high It is recommended to dilute the MicroBeads according to the data sheet Cells should be incubated with MicroBeads in the refrigerator 2 8 C The total number of cells may have been miscounted leading to an inappropriate ratio of MicroBeads and cells Use a ratio of MicroBeads and cells as stated in the respective MACS Cell Separation Reagent data sheet Incubation time with MicroBeads was too long leading to background labeling Reduce the incubation time to the recommended values in the respective MACS Cell Separation Reagent data sheet Typically an incubation time of 15 minutes is required when incubating in the refrigerator 2 8 C For exceptions refer to the respective data sheet When target cells are extremely rare lt 5 of total cells few non labeled cells which are retained non specifically may constitute a high portion compared to the target cells Performa second separation without re labeling to remove non specifically retained cells or choose a double column separation like PosselD PosselD2 or PosselDS When us
28. staining reagents For tips and hints on titration refer to the FAQs under Customer Support at www miltenyibiotec com Note To prevent contamination run a Sleep program before turning OFF the instrument 9 4 Fluorescent staining 9 4 1 Cells are poorly stained 1 Fluorochrome conjugated antibodies may have deteriorated Store MACS Fluorochrome conjugated Antibodies at 2 8 C 2 Fluorochrome conjugated antibodies and MACS MicroBeads might compete for epitopes Check the efficiency of the fluorescent staining on cell samples that are not magnetically labeled Especially for weakly expressed antigens it might be necessary to use a fluorochrome conjugated antibody that is directed against an epitope different from that recognized by the MicroBeads 3 For indirect staining of biotinylated antibodies with streptavidin fluorochromes please note that buffers supplemented with fetal bovine serum FBS or BSA may contain free biotin which can inhibit streptavidin binding to the biotinylated antibody Alternatively use MACS Anti Biotin VioBlue FITC PE or APC MACS Anti Biotin antibodies do not bind to free biotin 9 4 2 Cells are excessively stained 1 High background staining may occur when the concentration of the staining reagents is too high and the incubation time for the staining procedure is too long The use of the pre titrated MACS Fluorochrome conjugated Antibodies is recommended to achieve optimal results 2 Chec
29. 1 located at the front of the instrument Tilt the MACS MiniSampler and slide the guiding into the receiving slot until resistance is met lower the rack to a horizontal position i e the rack is locked in the position illustrated by the figure below Figure 3 4 The receiving slot 1 and guide 2 of the MACS MiniSampler 5 Ensure that the MACS MiniSampler is completely inserted and secure the connection by fastening the MiniSampler screw as shown in the figure below A Figure 3 5 Securing the MACS MiniSampler with its screw 6 Attach the sensor cables to the cable guide at the back of the autoMACS Pro Separator 3 Assembly and installation 77 78 Figure 3 6 Sensor cables are attached to the back of the instrument as shown above 7 Place the MACS MiniSampler cable underneath the autoMACS Pro Separator and connect it to the socket labeled External CAN at the rear panel of the instrument Use the screw driver to fasten the screws of the connecter 8 Guide the 2D code reader cable underneath the instrument Connect the 2D code reader plug with the corresponding outlet at the back of the instrument labeled RS232 BCR Use the screw driver to fasten the screws of the connecter 9 Attach the sensor cable plug to the socket for sensor cables at the back of the autoMACS Pro Separator labeled Bottle Sensor and fasten securely Use the screw driver to fasten the screws of the connecter Sensor c
30. 12 Figure 6 14 CD4 MicroBeads human assigned to reagent rack position R4 was selected for deletion Deleting the entire reagent list 1 Select an unassigned position on the reagent rack In this example position R4 was selected If there are no unassigned positions delete an individual reagent as described above 6 Cell separation using the autoMACS Pro Separator nl o Figure 6 15 To delete all reagents from the reagent rack the unassigned position R4 was selected 2 Select Delete All 6 2 4 3 Working with reagent templates For convenience it is possible to load and save reagent templates In this example the following reagents were scanned into the following reagent rack positions R1 CD4 MicroBeads human 130 045 101 R2 CD8a Ly 2 MicroBeads mouse 130 049 401 R3 No reagent R4 No reagent Saving a reagent template 1 Assign reagent vials to reagent rack positions as outlined above refer to section 6 2 4 1 YES Figure 6 16 Using the 2D code reader two reagent vials were assigned to the reagent rack positions R1 and R2 2 Select Save Template 3 Allocate a name to the template In this example the template was saved as EXPT_2 experiment 2 6 Cell separation using the autoMACS Pro Separator 131 132 IA 035 Figure 6 17 Using the alphanumeric keypad assign a name to the template 4 Select Ok Loading a reagent template 1 Select Load
31. 60825 1 Utilizzare unicamente cavi di connessione inferiori ai 3 metri di lunghezza 1 5 Funzionamento manutenzione trasporto e smaltimento sicuri Seguire le istruzioni di seguito riportate onde assicurare che il funzionamento la manutenzione il trasporto e lo smaltimento del vostro autoMACS Pro Separator avvengano in modo corretto e sicuro 1 5 1 Funzionamento sicuro Qualora lo strumento non funzioni correttamente e le istruzioni o i messaggi visualizzati sullo schermo invitino a contattare l assistenza tecnica la sicurezza di impiego non pi garantita Spegnere immediatamente il autoMACS Pro Separator staccare lo strumento dalla presa dell alimentazione e contattare un rivenditore Miltenyi Biotec autorizzato o il servizio di assistenza clienti Miltenyi Biotec 1 5 2 Manutenzione IMPORTANTE Se non diversamente indicato nel presente manuale d uso o in altri documenti forniti da Miltenyi Biotec non eseguire autonomamente la manutenzione del vostro autoMACS Pro Separator Gli interventi di manutenzione e riparazione devono essere eseguiti da personale qualificato Interventi di manutenzione e riparazione scorretti o impropri del autoMACS Pro Separator possono rappresentare un pericolo per l incolumit degli operatori avere conseguenze imprevedibili causare malfunzionamenti o danni provocare l usura prematura e una minore durata di esercizio dello strumento nonch invalidare la garanzia Per i contratti di assistenza e
32. 9 2 3 steps 1 to 4 2 Low recoveries can also be caused by partial column blockage Run the Safe program and exchange the autoMACS Columns ESS refer to sections 4 7 and 3 3 4 3 Performa test run using PBS as a mock sample and check output volumes For correct volumes refer to table 6 1 in section 6 1 3 1 If volumes are not correct go to section 9 2 1 4 Ifthe problem persists contact Miltenyi Biotec Technical Support When using the autolabeling feature 1 Check whether the uptake needle is positioned in the middle of the sample tube during the mixing procedure If not mixing will be inadequate 2 Check whether the reagent volume contents of reagent vial is sufficient for labeling 3 Check that the recommended maximal sample processing time of 90 minutes was not exceeded In the event of an elevated room temperature the cooling capacity of the Chill Racks may be compromised Higher labeling temperatures could cause unspecific binding which may lead to lower purities when using positive selection strategies 9 2 5 Sample not or only partly taken up 1 If the sample has been taken up only partly check if the sample contains clumps larger than 1 mm Continue with the separation of this sample As soon as the separation is finished continue with step 2 Before processing any remaining samples use a Pre Separation Filter 30 um 130 041 407 to remove the clumps from the sample 9 Troubleshooting 177 178 For separati
33. Cells were not labeled with MicroBeads because the MicroBeads were degraded Check the expiration date Check for sterility of the MicroBeads if the vial has been opened before Cells were labeled e g with fluorochrome conjugated antibodies prior to magnetic labeling with direct MicroBeads When antibodies recognizing the same epitope are used for fluorescent and magnetic labeling fluorochrome conjugated antibodies and MicroBeads might compete for the binding sites This can result in insufficient magnetic labeling It is recommended to perform magnetic labeling prior to staining with fluorochrome conjugated antibodies Alternatively use indirect MicroBeads A non optimal separation program was chosen e g Possel instead of Possel S for dimly labeled cells When using the autolabeling feature 1 Check whether the uptake needle is positioned in the middle of the sample tube during the mixing procedure If not inadequate mixing will be performed Check whether the reagent volume contents of reagent vial is sufficient for labeling Verify whether the sample volume was programmed correctly A reduced input volume will result in insufficient labeling of cells 9 Troubleshooting 179 180 Low purity of magnetically labeled cell fraction 1 Check for cell aggregates Negative cells may be retained when forming clusters with positive cells and thus contaminate the positive fraction Use buffers devoid of Ca and Mg
34. Inbetriebnahme und Betrieb des Ger ts ist die Dokumentation zu beachten 1 Wichtige Informationen 1 2 Warn und Sicherheitshinweise 1 Der autoMACS Pro Separator ist ein hochmodernes computergesteuertes Ger t zur automatischen Separation magnetisch markierter Zellen mittels MACS Technologie Der MACS MiniSampler ist an den autoMACS Pro Separator angeschlossen und damit Teil der Zellseparationsvorrichtung autoMACS Pro Separator und MACS MiniSampler gelten bei Einbau und Bedienung durch fachkundiges Personal und Beachtung der allgemein blichen Sicherheitspraktiken und Hinweise in dieser Bedienungsanleitung als sicher Dieses Kapitel erkl rt die im Umgang mit dem Ger t m glichen Gefahren und gibt wichtige Sicherheitshinweise zur Vermeidung dieser Gefahren Halten Sie sich genau an diese Hinweise um sich selbst und das Ger t vor m glichen Gefahren zu sch tzen und ein sicheres Arbeitsumfeld zu garantieren Bei Verwendung des Ger ts unter Missachtung der Herstelleranweisungen ist die Sicherheit nicht gew hrleistet ACHTUNG Bitte lesen und befolgen Sie alle in diesem Benutzerhandbuch gegebenen Hinweise zum Betrieb des Ger tes Beachten Sie auch alle auf dem Display erscheinenden Warnhinweise Bewahren Sie dieses Benutzerhandbuch sowie alle weiteren mit diesem Ger t erhaltenen Sicherheits und Betriebsanleitungen an einem f r alle Nutzer des Ger ts zug nglichen Ort auf so dass diese k nftig jederzeit darauf zur ckgreifen
35. Pro Separator automatically records the date of the last column exchange and displays the date for column replacement only if the column exchange program Col_ex has been used for column installation 6 2 3 Prime the autoMACS Pro Separator 1 Switch ON the autoMACS Pro Separator and wait for the instrument to complete initialization 2 After initialization the autoMACS Pro Separator will display the Status menu For more details on the Status menu please refer to section 4 3 EE Sample status Fluid bottle filling status Column status Tube rack status MiniSampler status Figure 6 1 Overview of the instrument s status panel 3 Ensure that the symbols for the fluid bottles are colored green If they are red check whether the fluid bottles are filled the waste bottle is empty and the color coded bottle sensors are connected to the appropriate bottle If symbols are gray check the bottle sensors for proper connection Figure 6 2 Fluid bottle status symbols Left Fluid bottles are ready Right Fluid bottles need to be replaced 4 Ensure that the symbol for the columns is colored green For more details refer to section 4 3 2 If the symbol for the columns is red the columns must be exchanged The fill level on the symbol is an indicator for the remaining operation life of the columns Figure 6 3 The column status graphic Left The columns are ready Right The columns must be changed 6 Cell
36. The fluid bottles are illuminated green Select the Separation menu 1 Select the desired position s in the sample rack template field by touching it 2 Select from the Labeling submenu for manual labeling 3 Optional It is not mandatory to assign a volume for manually labeled samples However the autoMACS Pro Separator requires this information to calculate and display the total sample processing time Enter the sample volume in the Volume submenu using the numeric keypad Select Enter 4 Select a separation program and a washing program for each sample position The selected programs will be displayed in the programming field 6 Cell separation using the autoMACS Pro Separator 1osRion s Select RUN lo start I 25 Fig 6 22 Performing a cell separation after manual labeling with CD4 MicroBeads human The cell separation program Possel and wash program Qrinse were selected 5 Optional Autolabeling and manual labeling can be assigned in parallel to two independent samples To assign a reagent for autolabeling to a sample select the position of interest in the sample rack template field and assign the respective reagent from the Labeling submenu Note The reagent has to be entered in the Reagent menu to be displayed in the Labeling submenu 6 Optional Templates can be saved by selecting Save template from the lower navigation bar Follow the prompt to enter a template name 7 Select Ok and Run
37. User settings 11 23 35 47 59 71 71 72 75 76 81 83 83 83 84 86 88 90 91 93 93 93 94 95 96 97 97 98 104 104 106 107 107 5 1 5 1 1 5 1 2 5 2 6 1 6 1 1 6 1 2 6 1 3 6 1 3 1 6 1 3 2 6 2 6 2 1 6 2 1 1 6 2 1 2 6 2 2 6 2 3 6 2 4 6 2 4 1 6 2 4 2 6 2 4 3 6 2 5 6 2 5 1 6 2 5 2 6 2 5 3 6 2 5 4 6 2 5 4 1 6 2 5 4 2 6 2 6 6 2 6 1 6 2 6 2 6 2 6 3 6 3 7 7 1 7 2 7 2 1 7 2 2 7 2 3 7 3 7 3 1 7 3 2 7 3 3 7 3 4 7 3 5 74 74 1 74 2 Content Materials required Materials required for operation Buffers and Solutions Hardware autoMACS Pro Separator accessories Cell separation using the autoMACS Pro Separator General considerations Cell labeling strategies Cell separation strategies autoMACS Pro separation wash and maintenance programs Separation programs Wash and maintenance programs Experimental considerations Preparation of samples Prepare single cell suspensions Adjusting sample volumes Select appropriate tube rack Prime the autoMACS Pro Separator Labeling of cells Entry of reagents for autolabeling Deleting reagents or reagent lists Working with reagent templates Cell separation Cell separation after autolabeling Cell separation after manual labeling Example of application entry of sample separation instructions Monitoring the cell separation process Status menu before separation Status menu during the separation process Shut down o
38. Washing Solution Sterile and ready to use solution for washing and special rinsing programs The tubing connector is color coded green Bottle closure Vented screw on closure with fluid uptake ports canules and distribution tubes The bottle closures contain fluid sensors and are equipped with sensor cable connectors 14 Glossary 14 207 208 Chill Racks Three different acrylic tube racks are provided with the instrument They are designed for optimal positioning of sample tubes and fraction collection tubes at the ports of the automated arm They contain a coolant allowing to pre cool the racks in the refrigerator for cooling of the cells during the separation process The racks have four tube positions for each sample Position A holds the sample tube containing the starting material Position B holds the tube for the non labeled fraction Position C holds the tube for the magnetically labeled fraction Position D is auxiliary For details refer to the table in section 4 1 2 Column connector Luer to thread connector connecting the autoMACS Columns to the fluidic system Column substitute Column without spheres that replace the autoMACS Columns for long term storage and shipment Column substitutes cannot be used for cell separations Depletion Isolation of target cells by labeling all cells other than the target cells with MACS MicroBeads and subsequently performing a MACS Separation The non labeled fraction contain
39. decontaminate the fluidic system every 3 to 6 months using the Safe program If predominantly whole blood bone marrow or tissue samples are separated the safe procedure can be performed more often without harming the instrument e g every four weeks instead of performing a column exchange 7 Maintenance 153 154 CAUTION If sample tubes and fraction collection tubes have been in contact with biohazardous material they should be autoclaved after use Bottle closures can be autoclaved In case of spillage it is recommended to use a disinfectant that is appropriate for the potential pathogen e g bleach isopropyl alcohol or 70 ethanol to decontaminate surfaces with tissue or swabs Switch OFF and unplug the instrument before decontamination Dispose tissues and swabs appropriately It is recommended to wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes Perform instrument decontamination according to the following instructions 1 Select menu Option Special and the Safe program RE 15 05 Figure 7 12 Safe program is underway 2 Press Run Follow the screen prompts 3 Disconnect the tubings from storage solution and buffer bottles Select OK 4 Place the ends of the tubing in a minimum of 15 mL of disinfectant solution e g MACS Bleach Solution Select OK 5 Fillasample tube with 25 mL of disinfectant solution Select Done 6 Optional Wash fluid bottle
40. has been hindered 263 Unable to read rack bar code correctly 264 Unable to read rack bar code correctly 769 Diluter plunger could not be initialized Syringe not mounted correctly syringe damaged or diluter valve not positioned correctly 775 Diluteris not initialized After a diluter malfunction an initialization of the module is necessary This might also be a subsequent error if another error has been displayed shortly before 777 Plunger movement blocked because of column clogging blocked tubing set or any other cross section constriction Possible remedies Restart instrument Please call Technical Support if error is displayed again Switch OFF the instrument wait for 5 seconds and switch the instrument ON again If error persists touch the button Details Exchange displayed valve If error is displayed again call Technical Support Touch button Details Exchange displayed valve if applicable If error is displayed again or magnet is displayed call Technical Support Switch OFF the instrument for 5 seconds and turn it ON again If error persists switch OFF instrument loosen air filters on buffer waste bottles by a half turn exchange columns for dummy columns verify that washing station is free of clogs and salt deposits eventually clean washing station verify that uptake needle is not clogged Switch on instrument if error persists touch Details Exchange displayed valve if applic
41. labeling please refer to the Cell Separation Reagent data sheet Autolabeling may be performed prior to cell separation Miltenyi Biotec continually develops and optimizes autolabeling protocols For a list of Cell Separation Reagents optimized for autolabeling please contact Miltenyi Biotec Technical Support 6 2 4 1 Entry of reagents for autolabeling The Reagent menu is used to program any reagent vials that are required for automated magnetic labeling and subsequent cell sorting Reagents can be entered using the 2D code reader or manually using the Enter Reagent input panel 6 Cell separation using the autoMACS Pro Separator Note If the reagent vial cannot be identified by the 2D code reader please enter the reagent information manually refer to Entering reagents manually below Define reagent rack N Figure 6 6 The Reagent menu is displayed Entry of reagents with 2D code reader 1 Select Reagent tab and highlight the position where the vial will be placed in the reagent rack Four positions are available R1 R2 R3 and R4 EE Figure 6 7 Position R1 was selected for the reagent vial 2 Activate the reader by selecting Read Reagent and present a reagent vial in front of the 2D code reader Ensure the 2D code is facing the blinking code reader light The optimal reading distance is 0 5 2 5 cm from the code reader cover tilt the vial as depicted in figure 6 8 Figure 6 8 Scannin
42. les personnes portant un pacemaker ou des implants m dicaux lectroniques Risque de contamination en cas d utilisation de mati res nocives pour l organisme Indique le danger de mort le risque de blessure grave pour l op rateur de l appareil ou le risque de d t rioration de l quipement du a des mati res biologiques potentiellement dangereuses Indique le danger de mort le risque de blessure grave pour l op rateur de l appareil en raison de la tension dangereuse Borne pour conducteur de protection Le symbole est appos a l int rieur de l quipement Avertissement destin au personnel de maintenance Alimentation lectrique activ e Alimentation lectrique d sactiv e La notice d utilisation doit tre consult e avant de proc der l installation et l utilisation du syst me 1 Informations importantes 1 2 Avertissements et pr cautions 1 Le s parateur autoMACS Pro fonctionne avec une technologie de pointe Il s agit d un appareil command par ordinateur con u pour la s paration automatique des cellules marquage magn tique las l aide de la technologie MACS Le mini chantillonneur MACS est branch au s parateur autoMACS Pro et fait donc partie du dispositif de s paration des cellules Le s parateur autoMACS Pro et le mini chantillonneur MACS sont congus pour fonctionner en toute s curit apr s leur installation s ils sont utilis s par des personnes qualifi es selon les pratiques
43. miltenyibiotec com sg Spain Miltenyi Biotec S L C Luis Bu uel 2 Ciudad de la Imagen 28223 Pozuelo de Alarcon Madrid Spain Phone 34 91 512 12 90 Fax 34 91 512 12 91 macs miltenyibiotec es United Kingdom Miltenyi Biotec Ltd Almac House Church Lane Bisley Surrey GU24 9DR UK Phone 44 1483 799 800 Fax 44 1483 799 811 macs miltenyibiotec co uk Miltenyi Biotec provides products and services worldwide Visit www miltenyibiotec com local to find your nearest Miltenyi Biotec contact Unless otherwise specifically indicated Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use MACS autoMACS and VioBlue are registered trademarks of Miltenyi Biotec GmbH All other trademarks mentioned in this document are the property of their respective owners and are used for identification purposes only Copyright 2013 Miltenyi Biotec GmbH All rights reserved 140 002 016 06
44. n al l ser puede ocasionar lesiones oculares Utilizar un pa o suave y seco para limpiar cualquier sustancia del esc ner No utilizar alcohol u otra sustancia de limpieza El autoMACS Pro Separator est clasificado como producto l ser 1M de est ndar IEC 60825 1 1993 A1 1997 A2 2001 1 Informaci n importante ATENCI N El uso de controles el ajuste o la realizaci n de 1 procedimientos con excepci n de los aqui especificados puede dar lugar a una peligrosa exposici n de radiaci n wu N Figura 1 3 Posici n de los l seres El l ser de detecci n del rack es invisible y se localiza dentro del rea rectangular El lector de c digos de barra visble est localizado dentro del c rculo abierto 1 4 Instalaci n segura El presente cap tulo describe los requisitos que debe reunir el lugar donde desea instalar y operar el separador autoMACS Pro Lea las instrucciones de este cap tulo y aseg rese de que el emplazamiento est correctamente preparado antes de conectar el instrumento a la corriente el ctrica Cuando planee la distribuci n del emplazamiento y la ubicaci n del equipo tenga en cuenta las precauciones recogidas en este cap tulo para evitar aver as del instrumento y reducir la posibilidad del cierre de la empresa por razones medioambientales 1 4 1 Accesorios de montaje No coloque el separador autoMACS Pro sobre una mesa carrito estante tripode o soporte inestable puesto que pod
45. of the washing station 11 Carefully remove the waste distributor from its position by pulling in an upward direction and remove the draining tube from the old diluter valve Figure 7 27 The waste distributor 12 Mount the new draining tube and finger tighten Take care to clean spilled fluids with ethanol or disinfectant TN r Figure 7 28 The draining tube is attached 13 Install the waste distributor back to its former position and reinstall the washing station 14 Install the syringe 15 Switch ON the autoMACS Pro Separator 7 Maintenance 161 162 Note The peristaltic pump head should be exchanged once a year 16 Check for correct function by running the Rinse program 17 Run the program Calibr_2 to calibrate the fluidic volume control of the instrument 7 4 3 Exchange of the peristaltic pump head 1 Switch OFF and unplug the instrument 2 Remove the washing station as described in section 7 3 3 steps 1 6 3 Press the tubing clamp on the left hand side of the waste distributor and remove the tubing Figure 7 29 Disconnecting the tubing from the waste distributor 4 If necessary pull out the bottom cover as depicted in figure 7 14 5 Press clamps on both sides of the pump and pull out the pump head Figure 7 30 Detachment of the pump head 6 This will uncover a pin that protrudes from the instrument Clean the pin using 70 ethanol but do not attempt to pull it out The pin dr
46. on screen menus Tubing connector Plastic threaded connector with square nut used to connect the tubings to the bottle closures the columns the pump or valves Tubing system Permanent set of Teflon tubing through which fluids circulate in the autoMACS Pro Separator fluidic system Washing Solution bottle Bottle for autoMACS Pro Washing Solution The bottle closure is equipped with a fluid sensor The closure the fluid sensor cable and the tubing connector are color coded green Waste bottle Bottle for waste fluid The closure is equipped with a fluid sensor The closure the fluid sensor cable and the tubing connector are color coded red Whole Blood MicroBeads MACS MicroBeads developed for isolating target cells directly from human whole blood Wrench Black wrench used to tighten and loosen tubing connections 14 Glossary 211 Miltenyi Biotec Germany Austria Switzerland Miltenyi Biotec GmbH Friedrich Ebert Stra e 68 51429 Bergisch Gladbach Germany Phone 49 2204 8306 0 Fax 49 2204 85197 macs miltenyibiotec de USA Canada Miltenyi Biotec Inc 2303 Lindbergh Street Auburn CA 95602 USA Phone 800 FOR MACS Phone 1 530 888 8871 Fax 1 530 888 8925 macs miltenyibiotec com www miltenyibiotec com Australia Miltenyi Biotec Australia Pty Ltd Unit 16A 2 Eden Park Drive North Ryde NSW 2113 Australia Phone 61 2 8877 7400 Fax 61 2 9889 5044 macs miltenyibiotec com au Benelux
47. or damp location Avoid high humidity or condensation and protect the machine against water splashes 1 4 4 Grounded earthed product The instrument is equipped with a three wire electrical grounding type plug that has a third pin for grounding This plug only fits into a grounded power outlet This is a safety feature Do not try to insert the plug into a non grounded power outlet If you cannot insert the plug into the outlet contact your local electrician to replace the outlet 1 4 5 Power sources The instrument should only be operated from a power source indicated on the product s electrical ratings label If you have questions about the type of power source to use contact your authorized Miltenyi Biotec service provider or local power company Do not use extension cords or power strips Do not overload an electrical outlet The overall system load must not exceed 80 of the branch circuit rating 1 4 6 Accessibility Make sure that the main switch as well as the connector for the power cable are easily accessible and located as close to the operator of the instrument as possible If it is necessary to disconnect the power supply unplug the cable from the power outlet 1 4 7 Peripheral devices Only peripheral devices that comply to UL 60950 are allowed to be connected to the RS232 connector labeled COM The connector labeled RS232 AUX is not in use In addition only original autoMACS Pro Separator equipment should be
48. or modify a cell separation program 1 Highlight the appropriate sample s using the sample template 2 Scroll through the Separation submenu using the arrows M W 3 Select and highlight a Separation program Figure 4 18 Positive cell separation sensitive mode Possel_s was selected Assigning a wash program The Wash submenu 5 is used to instruct the autoMACS Pro Separator which wash program should be applied to each sample Four wash programs are available to choose from Qrinse quick rinse Recommended to save time between sample separation steps The tubing and column receive a quick rinse Rinse The system is primed receiving an extensive wash Rinse is recommended between rare cell whole blood cord blood and bone marrow separations 4 An overview of the autoMACS Pro Separator user interface Note If the Clean program has been enabled it will also appear in the Wash submenu Sleep A rinse is performed before the system is shutdown A Clean An optional very extensive rinsing program Clean may be used after whole blood cord blood and bone marrow separations To assign or modify a wash program 1 Highlight the appropriate sample s using the sample template 2 Scroll through the Wash submenu using the arrows A V 3 Select highlight a Wash program ES Figure 4 19 Top Qrinse program was selected Bottom The Clean program has been enabled and was selecte
49. parts can cause malfunction of the instrument and impair cell separation results Miltenyi Biotec does not honor any warranty or accept any responsibility for instrument failure or damages resulting from the use of inappropriate replacement or spare parts WARNING During a maintenance procedure potentially contaminated liquid may spill out of the orifice of the washing station and the tubing Therefore wear protective gloves protective clothing and safety glasses to avoid contact with skin and eyes Dispose used gloves and clothing appropriately 7 Maintenance 145 146 Note Do not use a dishwasher to clean and do not autoclave any of the removable parts unless indicated otherwise Note In case of malfunction during cell separation the O_ init program must be disabled in order to perform a cell rescue procedure The instrument automatically prompts the user to perform a rinse before performing a cell separation 7 1 General considerations Please consider that the sample quality has a significant influence on the system s performance It is crucial to use single cell suspensions for cell separation Furthermore dead cells and other small particles potentially derived from the sample preparation should be removed prior to cell separation For further information on sample preparation refer to section 6 2 1 7 2 Daily maintenance The following maintenance procedures must be performed on a daily basis If the inst
50. positions A1 B1 and C1 onto the MACS MiniSampler Select Done 3 Place the MACS Reagent Rack 4 onto the MACS MiniSampler Select Done 4 Test position Washing station will be displayed on the screen Select Test position The instrument will automatically place the needle in the washing station Ensure that the needle is positioned directly above the center of the rear opening of the washing station Click Done The next position will be addressed automatically running Test position washing station Select action Verify washing station position N 11 32 Fig 3 20 Test calibration of the washing station 5 Inthe following steps further calibration settings regarding the needle arm can be tested including the tube rack positions Al Ori B1 Neg and C1 Pos the x position and vials one to four in the Reagent Rack To test a position select Test position Check all approached positions by using the needle navigation buttons Move up and Move down then select Done To skip a position select Next The test calibration can be canceled at any time by selecting Test end 3 Assembly and installation 85 86 Note When delivered the autoMACS Pro fluidic system is filled with double distilled water The autoMACS Columns are reusable for two weeks or up to 100 separations whichever comes first The instrument auto matically records the date of the column exchange and can display the due date for the
51. rack During double positive selection programs Posseld Posseld2 Posselds and Posselwb the magnetically labeled target cells are first retained in the autoMACS Column 1 The negative fraction containing the non labeled cells is retrieved in the negative fraction collection tube i e row B of the tube rack Then the magnetically labeled cells are held in a reservoir and loaded onto the autoMACS Column 2 Again the unlabeled cells are released into the negative fraction collection tube i e row B of the tube rack Finally the magnetically labeled cells are eluted into the positive fraction collection tube i e row C of the tube rack When running any depletion program the magnetically labeled non target cells are retained in the autoMACS Column 1 The non labeled target cells pass through the column and are released into the negative fraction collection tube i e row B of the tube rack The magnetically labeled fraction containing the unwanted cells is eluted into the positive fraction collection tube i e row C of the tube rack 6 Cell separation using the autoMACS Pro Separator 119 120 Program Volume of non Volume of Loading labeled fraction labeled fraction rate i e negative fraction i e positive fraction Possel 2mL sample volume 2mL 4 mL min Possel_s 2 mL sample volume 2mL 1 mL min Posseld 2mL sample volume 0 5mL 4mL min column 1 1mL min column 2 Posseld2 2 mL s
52. rer la fiche dans une prise de courant sans contact de mise la terre Si vous ne pouvez pas ins rer la fiche dans la prise de courant veuillez vous adresser votre lectricien local qui remplacera la prise de courant 1 4 5 Sources lectriques Ne faites fonctionner l appareil qu partir d une source lectrique indiqu e sur la plaque signal tique du produit Si vous avez des questions sur le type de courant lectrique que vous pouvez utiliser contactez votre prestataire de services Miltenyi Biotec agr ou votre fournisseur d lectricit local N utilisez pas de rallonges ni de barrettes de connexion Ne surchargez pas la prise lectrique La charge totale du syst me ne doit pas d passer 80 de la valeur nominale du circuit de d rivation 1 4 6 Accessibilit Assurez vous que l interrupteur principal et le connecteur du cable lectrique soient facilement accessibles et plac s aussi pr s que possible de l op rateur de l appareil S il s av re n cessaire de d connecter l alimentation lectrique d branchez le cable 1 4 7 Appareils p riph riques Seuls les appareils p riph riques conformes a la norme UL 60950 peuvent tre branch s au connecteur RS232 tiquet COM Le connecteur tiquet RS232 AUX n est pas utilis De plus seuls des appareils autoMACS Pro originaux doivent tre raccord s aux connecteurs tiquet s External CAN CAN1 et CAN2 1 Informations important
53. row C of the sample rack Running Buffer bottle Bottle for Running Buffer The bottle closure is equipped with a fluid sensor The closure the fluid sensor cable and the tubing connector are color coded blue 14 Glossary Safe solution Solution of 1 w v sodium hypochlorite in 1 distilled water used to decontaminate the autoMACS Pro fluidic system with the Safe program The safe solution is fed into the system from a 50 mL tube Upon completion of the Safe program the fluidic system contains Running Buffer Storage solution Solvent used during the Sleep Store Safe Clean and Column Exchange Col_ex programs to minimize the contamination risk The fluidic system of the autoMACS Pro Separator is filled with storage solution 70 ethanol Storage solution bottle Bottle for 70 ethanol The bottle closure the fluid sensor cable and the tubing connector are color coded black Store program Prior to long term storage the Store program should be applied During this procedure the autoMACS Columns are replaced with substitutes Upon completion of the Store program the fluidic system contains 70 ethanol Syringe pump Computer controlled high precision syringe pump with Teflon seal plunger that drives fluids through the autoMACS Pro fluidic system Touchscreen High resolution TFT color touchscreen located on top of the autoMACS Pro Separator The touchscreen is used to operate and monitor the instrument through
54. safety and operating instructions provided with the instrument in a place accessible to all users for future reference IMPORTANT The autoMACS Pro Separator is intended for indoor use only Do not use the instrument in areas classified as hazardous locations such as oxygen laden environments At all times local working area safety instructions laboratory policies and standards regarding laboratory health and safety and prevention of accidents must be adhered to Contact your local authority governing electrical power supply building constructions maintenance or safety for more information regarding the installation of the equipment If you have a serious concern regarding the safe use of your insrument please contact your authorized Miltenyi Biotec service provider or call Miltenyi Biotec Customer Service 1 Important information 13 1 3 General precautions To reduce potential risks associated with operating the autoMACS Pro Separator please observe the following general precautions Failure to observe these precautions could result in fire bodily harm and or damage to the instrument 1 3 1 Hazard of electric shock and spread of fire WARNING Hazardous voltages Risk of loss of live severe personal injury Unplug before cleaning Do not use if device is opened or damaged if liquids have been spilled into device if objects device through ventilation slots WARNING Electrical devices pose the risk
55. sample volumes exceeding the maximum volume specified for rack or program type If volumes are in specified range but error is displayed anyway please call Technical Support Please clean outside of needle manually If this error is displayed frequently please call Technical Support Check if tubes are correctly positioned in rack corresponding to template programming Check MACS MiniSampler connection in front of autoMACS Pro Separator Error Cause Possible remedies code 7010 Not certain if liquid surface of sample hasbeen Touch Ignore to continue without liquid detection detected correctly Needle will be moved to bottom directly This might result in a subsequent warning 7008 if the liquid level is higher than 60 mm above the tube bottom see above Touch Retry to continue with liquid detection Ensure that the tubing from the needle arm to bubble sensor can move freely Adjust the tubing if necessary Otherwise call Technical Support 7011 Restart of the instrument is required Please restart instrument 7012 Calibration data not found Please calibrate needle arm axes Run program Calibr_1 7013 Calibration data not found Please calibrate needle arm axes Run program Calibr_1 7014 Calibration data not found Please calibrate tubing Run program Calibr_2 7015 Columns are not installed Please install columns 7018 Calibration data not found Please calibrate MACS MiniSampler using program
56. separation using the autoMACS Pro Separator 6 125 126 Note If priming of the instrument has not been performed the autoMACS Pro Separator will automatically ask to run a Rinse program before starting the separation The process can be monitored in the Status menu At this point the fluid bottles are illuminated yellow It is possible to include an initial Rinse program that will be performed automatically upon switching ON the instrument The setting of this option is described in section 4 7 O_init Optional priming of the instrument at startup Note For autolabeling the reagent rack configuration must be programmed using the Reagent tab menu Refer to section 4 5 for an overview of the Reagent tabbed menu 5 Ensure that the MACS MiniSampler is installed correctly For more details concerning correct installation of the MACS MiniSampler refer to section 3 Figure 6 4 MACS MiniSampler status graphic Left The MiniSampler was successfully installed Right No MiniSampler was detected 6 The autoMACS Pro Separator is now ready for priming Select Separation and Wash Now from the lower navigation bar Figure 6 5 The Wash Now command displays two options Qrinse for a quick rinse and Rinse A full rinse is required for instrument priming 7 Select Rinse and Run 6 2 4 Labeling of cells Cells can be labeled manually with MACS MicroBeads For detailed information on manual
57. syringe as described in section 7 3 2 Note The pump syringe should be exchanged once 3 Unscrew the tubing a year A Of mets Figure 7 21 Unscrewing the tubing connected to the dilutor valve 4 Unscrew the two hexagonal socket screws using the key that was delivered with the new or exchange valve a bd Figure 7 22 Unscrewing the dilutor valve s socket srews 5 Pull the valve out of the coupling Take care to note the orientation of the valve shaft as shown below 6 When inserting the new valve into the coupling make sure that the short side of the trapezoid shaped valve shaft points towards the triangle in the coupling Note The profile of the valve shaft is shaped asymmetrically similar to a trapezoid refer to figure 7 23 Figure 7 23 Inserting a new valve take care to note the orientation of the valve 7 Maintenance 159 160 7 When the valve is properly inserted mount the hexagonal socket screws 8 Connect the tubing according to the positioning of the diluter valve e Figure 7 24 Connect tubing to the dilutor value as shown above 9 Guide the draining tube towards the washing station as shown below Figure 7 25 Red colored line indicates the positioning of the draining tube The draining tube connects the valve to the washing station 10 Remove the washing station Take care to clean spilled fluids with ethanol or disinfectant 7 Maintenance Figure 7 26 Disassembly
58. target cells are eluted in row C of the sample rack This program is used for cells with normal to high frequency and with normal antigen expression Possel_s Positive selection program sensitive mode using one autoMACS Column The target cells are eluted in row C of the sample rack This program is used for cells with normal to high frequency which weakly express the antigens used for magnetic labeling or to achieve the highest recovery of the target cells Posseld Positive selection program normal mode using both autoMACS Columns The target cells are eluted in row C of the sample rack in a volume of 0 5 mL This program is used to isolate rare cells or to achieve a higher purity of the target cells Posseld2 Special positive selection program normal mode using both autoMACS Columns The target cells are eluted in row C of the sample rack in a volume of 2 mL This program is used to isolate rare cells from whole blood cord blood or large cell samples and to achieve a higher recovery of the target cells Posselds Positive selection program sensitive mode using both autoMACS Columns The target cells are eluted in row C of the sample rack in a volume of 2 mL This program is used to isolate rare cells that weakly express antigens used for magnetic labeling e g CD133 Posselwb Special positive selection program using both autoMACS Columns for the isolation of cell subsets from of whole blood The target cells are eluted in
59. the autoMACS Pro Separator is regulated by five valves The valve that is connected with the pump syringe is referred to as dilutor valve If a valve of the autoMACS Pro Separator starts to leak an exchange of the valve should be performed Exchange one valve at a time according to the following instructions 1 Select User settings from the Options menu Highlight Valve _ ex and press Run to start the program Select valves which have to be exchanged Valves will automatically be turned to exchange position 2 Switch OFF and unplug the autoMACS Pro Separator Open the front cover For exchange of the lower valve remove the bottom cover by pulling firmly 7 Maintenance 155 156 gt a Figure 7 14 The bottom cover must be removed for access to the lower valve 3 Using the supplied wrench unscrew the valve cover screw which releases the tubing Figure 7 15 The valve cower screw is unscrewed 4 Detach all tubing and valve port locks or blind screws from the respective valve Figure 7 16 The valve cover screw is unsrewed Tubing and valve port locks or blind screws are detached 5 Loosen the two valve screws using the screwdriver from the autoMACS Pro Separator Starter Kit and pull out the valve 7 Maintenance NI 6 Check if the groove in the valve drive is positioned horizontally and in the lower half of the axis Figure 7 17 Detachment of the valve Figure 7 18 Valve plate driving section 1
60. to start 9 Check buffer levels as instructed by a pop up dialog box Click Continue to proceed 10 Itis recommended to monitor the process from the Status menu Click the Status menu 6 Cell separation using the autoMACS Pro Separator Autolabeling IA 12 7 Figure 6 29 Sample processing is underway as instructed 6 2 5 4 Monitoring the cell separation process The autoMACS Pro Separator is a sensor controlled instrument that allows easy monitoring during operation 6 2 5 4 1 Status menu before separation Refer to section 4 3 for an overview of how to monitor the instrument status using the Status menu prior to performing a cell separation However a brief summary is included below Status of fluid bottles The status of fluid bottles is indicated by color coded graphic symbols and by a text table Bottle Symbol Symbol color and user action Green no action required Red refill bottle Gray connect bottle sensor Running Buffer Green no action required Red refill bottle Gray connect bottle sensor Washing Solution visually check volume Waste Green no action required Red empty waste Gray connect bottle sensor Storage solution Gray no liquid detection Table 6 5 Status of fluid bottles displayed in the Status menu 6 Cell separation using the autoMACS Pro Separator 139 140 Note Interrupting the process after labeling will prolong the incubation period Column status If
61. tube rack Select Done to continue Figure 11 7 Calibrating the position of the needle arm in the sample tube 12 Select Save to store the new settings Calibration 1 Calibration of the x axis step 3 4 13 Select Calibrate to proceed with calibration 14 Place a Chill Rack and a MACS Reagent Rack 4 onto the MACS MiniSampler It is recommended to use a Chill 15 Rack 15 Select Done 11 Hardware and software installation Note It is possible that the stored calibration value is zero In this event step 16 will be skipped Continue with step 17 Note Due to mechanical limits of the MiniSampler the correction potential is limited Exceeding these limits is reported by the message standard range exceeded Calibration outside the standard range is not recom mended The outer positions reagent vial and sample row 6 will not be reached properly Labeling in sample position 6 will be prohibited 16 Select Use to start the calibration at current position recommended Select Reset to use factory settings 17 If necessary adjust the central positioning of the uptake needle by using the needle navigation buttons Move left Move right Height The uptake needle must be positioned above the center of the tube in row A of the tube rack Select Done to continue Process running Calibration 1 Step 3 4 x axis Adiust uptake needle centrically to sample tube row 4 using buttons Position 23 standard
62. usare contattare il rivenditore Miltenyi Biotec autorizzato o la societ elettrica locale Non utilizzare prolunghe o ciabatte multi presa Non sovraccaricare la presa di corrente Il carico complessivo del sistema non deve superare 80 della potenza del circuito 1 4 6 Accessibilit Accertarsi che l interruttore principale e l attacco del cavo dell alimentazione siano facilmente accessibili e posti quanto pi vicino possibile all operatore dello strumento Qualora si renda necessario interrompere l alimentazione staccare il cavo dalla presa di corrente 1 Informazioni importanti 1 4 7 Periferiche 1 Soltanto le periferiche conformi a UL 60950 possono essere collegate al connettore RS232 contrassegnato con COM Il connettore contrassegnato come RS232 AUX non in uso inoltre possibile collegare soltanto apparecchi autoMACS Pro Ga originali ai connettori contrassegnati con External CAN CAN1 e CAN 2 livelli di tensione su questi connettori non devono superare i livelli di tensione ammessi pari a 30 V rms e 42 4 V di picco o 60 V cc Al connettore del sensore flacone pu essere esclusivamente collegato il cavo del sensore del flacone autoMACS Pro Al connettore RS232 BCR pu essere collegato soltanto un lettore per codice 2D raccomandato da Miltenyi Biotec dispositivi laser esterni collegati al connettore contrassegnato come RS232 BCR devono essere conformi alla norma IEC
63. w gt Figure 1 3 Position des lasers Les lasers invisibles de d tection de r teliers sont situ s dans la zone rectangulaire Le lecteur de code 2D visible est entour d un cercle blanc 1 4 S curit de l installation Ce paragraphe d crit les exigences auxquelles votre site doit r pondre pour garantir l installation et la s curit d utilisation de votre s parateur autoMACS Pro Lisez les consignes de ce paragraphe et assurez vous que votre site a t am nag en cons quence avant de brancher l appareil sa source lectrique Lors de la planification de l am nagement de votre site et de l emplacement de l quipement tenez compte des pr cautions d crites dans ce paragraphe afin d viter que l appareil ne tombe en panne et de r duire le nombre d arr ts pouvant tre caus s par l environnement de l appareil 1 4 1 Accessoires de montage Ne placez pas le s parateur autoMACS Pro sur une table un chariot un pied un tr pied ou un support L appareil pourrait sinon chuter provoquant des blessures corporelles graves et ou la d t rioration consid rable l appareil Utilisez le s parateur uniquement sur une table un chariot un pied un tr pied ou un support recommand par Miltenyi Biotec ou vendu avec l appareil Ne placez pas le s parateur autoMACS Pro dans un appareil int gr ou un espace restreint comme un casier sauf si le dispositif a t sp cialement con u pour accueillir l ap
64. 3 368 Cover MiniSampler 130 093 669 Magnet Lock front cover 130 093 370 4 port 3 way Valve 130 093 371 autoMACS Pro Tube t31 130 093 372 Uptake Port Needle Guiding needle arm 130 093 397 Thumb Screw M5x40 MiniSampler 130 093 407 Pump Syringe Hamilton 5 mL Cavro XP 130 094 682 Dilutor Valve 6 port distribution V2 130 094 729 Table 7 1 List of available spare parts 7 Maintenance 167 168 7 Maintenance Quality control of separations performed with the autoMACS Pro Separator Note To count the cells of the original fraction collect an aliquot of the cell sample after magnetic labeling directly before the magnetic separation to analyze whether cell losses are due to centrifugation steps OR to magnetic separation Also take counting statistics into consideration The standard deviation when counting cells is N N1 2 Therefore cell counting might be associated with large statistical errors To evaluate any MACS Cell Separation the separated cells can be analyzed with regard to purity recovery and viability Using MACS MicroBeads the magnetically labeled cells can be simultaneously stained with fluorochrome conjugated antibodies Antibodies of the same specificity can be used in most cases MACS Fluorochrome conjugated Antibodies are standardized to evaluate MACS Cell Separations The stained cells can subsequently be analyzed by flow cytometry fluorescence microscopy or other techniques 8 1 Recovery of
65. 4 Laser radiation WARNING The instrument is equipped with four vertical cavity surface emitting lasers VCSELs for automated rack detection Class 1M The radiation is not visible Do not view directly with optical instruments e g lenses magnifying glasses and microscopes Viewing the VCSEL port within 100 mm distance with optical instruments could be hazardous to the eye The instrument is also equipped with a 2D Code Reader which uses a visible semiconductor laser as a target pointer for adjusting the reading position and powerful light emitting diodes LEDs for illuminating the reading area and the fluid bottles as well as the waste bottle Do not remove the bottle holders According to the international standard IEC 62471 this lamp system has an exposure hazard value EHV of 0 91 and is in excess of the Exempt Risk Group The hazard distance HD for the Exempt Risk Group is 61 cm The hazard distance for Risk Group 1 is 20 cm Do not look directly at laser or LED radiation or reflected laser or LED radiation from a mirrored surface Otherwise eye injury may result Do not intentionally direct the laser beam at others Do not disassemble modify or remove the installed laser or LED radiation sources or their mounting rackets The laser or LED radiation sources do not automatically stop emitting when disassembled Radiation of disassembled units may lead to eye injuries Be careful of the path of the laser beam or reflectio
66. 81 182 Non labeled fraction shows low purity 1 Magnetic labeling was insufficient due to a low expression level of the surface marker It is recommended to use MACS MicroBeads for indirect labeling e g MACS Anti Biotin MicroBeads in order to increase magnetic labeling of the cells The number of magnetically labeled cells exceeds the column capacity Calculate the number of expected magnetically labeled cells The number of cells should not exceed 2x108 per sample If necessary split the sample Labeling of cells was ineffective due to an incubation temperature lower than recommended Incubate the cells in the refrigerator 2 8 C for labeling Labeling of cells was ineffective due to too much debris and or dead cells in the sample Debris and dead cells will non specifically bind to all other components present Antibodies and MicroBeads will be captured non specifically and not be able to label the cells in the positive fraction sufficiently any more Cells were not labeled with MicroBeads because the MicroBeads were degraded Check the expiration date Check for sterility of the MicroBeads if the vial has been opened before Labeling of cells was performed for a shorter time than recommended The typical incubation time is 15 min Please refer to the respective data sheet for more details When using the autolabeling feature 1 Check whether the reagent vial has enough reagent to perform magnetic cell labeling
67. 90 835 Valve Blind Screw 5 pcs 130 091 996 autoMACS Pro Tube t27 130 093 284 autoMACS Pro Tube t2 130 093 285 autoMACS Pro Tube t7 130 093 286 autoMACS Pro Tube t21 130 093 287 autoMACS Pro Tube t5 130 093 288 autoMACS Pro Tube reservoir t9 t24 130 093 289 autoMACS Pro Uptake Port Needle 130 093 290 autoMACS Pro Bubble Sensor 130 093 291 MACS Fluid Container 1 5 L 130 093 292 autoMACS Pro Tube t4 130 093 293 Fuse 5x20 T4A set 130 093 302 autoMACS Pro Sensor Cable 130 093 303 autoMACS Pro Bottle Closure green 130 093 304 166 7 Maintenance NI Spare part Order no autoMACS Pro Bottle Closure blue 130 093 305 autoMACS Pro Bottle Closure black 130 093 306 autoMACS Pro Bottle Closure red 130 093 307 Dilutor Valve 6 port distribution 130 093 308 autoMACS Pro Tube t32 130 093 309 autoMACS Pro Tube t26 130 093 310 autoMACS Pro Tube t20 130 093 311 autoMACS Pro Tube t12 t14 130 093 312 autoMACS Pro Tube t3 130 093 313 autoMACS Pro Tube t18 130 093 314 autoMACS Pro Tube t11 t25 130 093 315 autoMACS Pro Tube t6 t8 t23 130 093 316 autoMACS Pro Tube t10 t16 t17 t19 t22 130 093 317 Washing Station needle arm 130 093 349 autoMACS Pro Tube t1 t13 130 093 350 Tube Connector 2x2 Ports 1 4 28 UNF 130 093 362 Tube Guiding Ball needle arm 130 093 364 Peristaltic Pump Head incl Tube 130 093 365 Outlet Port Unit needle arm 130 093 366 Guard needle arm 130 093 367 Thumb Screw M5x16 130 09
68. Antes de encender el instrumento siempre inspeccione todo el sistema de fluidos todas las conexiones y tubos v lvulas columnas jeringas y agujas Si detectase alguna rotura o fuga sustituya todas las piezas da adas antes de encender el instrumento Si alguna parte da ada no puede ser sustituida desconecte y no use el equipo La falta de piezas por donde pasa material de riesgo biol gico o el contacto con l quidos que han estado en dichas piezas podr a ser peligroso Las columnas placas tubos de ensayo y cualquier otro objeto que haya estado en contacto con las muestras peligrosas deber n ser autoclavados antes de poder volver a ser utilizados Los residuos l quidos deber n ser autoclavados o descontaminados usando un desinfectante industrial apropiado para el pat geno espec fico por ejemplo hipoclorito de sodio al 10 alcohol isoprop lico o etanol al 70 La eliminaci n de los residuos debe cumplir la normativa nacional 1 Informaci n importante autoMACS Pro Separator SN 0563 Didier no 120 092 545 Power Supply 100 240 VAL 50 60 Hz 200 VA Fuser 2141 2504 JUS USTED LAR OP SILA Figura 1 4 Se al de cuidado de riesgo biol gico localizada en la parte inferior del panel delantero del separador autoMACS Pro izquierda y en la parte superior de la botella de Fluido del separador autoMACS Pro derecha 1 5 5 Transporte El separador autoMACS Pro debe ser transportado con cuidado en el em
69. Patna oeo Figure 6 11 Reagent information is manually entered using the reagent order number 3 Select Ok 4 Ifa correct number is inserted the software will immediately recognize the reagent or kit To confirm select the reagent from the list by using the touch screen Detine reagent rack Define ES C5 Figure 6 12 CD4 MicroBeads human 130 045 101 was manually entered 5 Select Ok to confirm the identified reagent and reagent vial position R2 6 Cell separation using the autoMACS Pro Separator 129 130 IA 05 03 Figure 6 13 Reagent vial CD4 MicroBeads human was assigned to the reagent rack position R2 6 2 4 2 Deleting reagents or reagent lists In this example the following reagents were scanned and assigned to the following reagent rack positions e R1 FcR Blocking Reagent human component 1 3 of product Monocyte Isolation Kit Il human 130 091 153 e R2 Monocyte Antibody Cocktail human component 2 3 of product Monocyte Isolation Kit II human 130 091 153 e R3 Anti Biotin MicroBeads component 3 3 of product Monocyte Isolation Kit Il human 130 091 153 e R4 CD4 MicroBeads human 130 045 101 Deleting an individual reagent from the reagent rack lists 1 Select the reagent name that must be deleted In this example CD4 MicroBeads human on position R4 must be removed 2 Select Delete Selected to remove the highlighted reagent Define reagent rack BR 03
70. S MiniSampler is automatically recognized by the rack detector after starting the separation process During operation the tube rack should be covered with the MACS MiniSampler lid that is connected to the lid guiding The MACS MiniSampler can be disconnected from the autoMACS Pro Separator by pulling it up on the front side and pulling it towards the user A MACS MiniSampler lid Chill 5 Rack MACS MiniSampler Figure 10 5 Rear view of MACS MiniSampler with MACS Reagent Rack 4 and Chill 5 Rack The cell separation unit The central part of the cell separation unit consists of a magnet and two autoMACS Columns figure 10 6 Once installed the autoMACS Columns become part of a closed fluidic system and can be used for up to two weeks OR 100 separations whichever comes first Fluids are put through the fluidic system with the help of a syringe pump and four valves 10 Technical data and specifications 10 191 Valves 1 3 Diluter valve Syringe pump Colum 1 and 2 Magnet Valve 4 Figure 10 6 Front of autoMACS Pro Separator with access and bottom covers removed 2D code reader barcode reader The autoMACS Pro Separator is equipped with a 2D code reader that uses lasers and powerful light emitting diodes LEDs for illuminating the reading area The 2D code reader is classified as a Class 1 laser product per standard IEC 60825 1 1993 A1 1997 A2 2001 maximum output 116 pW wavelength 655 nm pulse duration 1 ms P
71. S232 AUX Adicionalmente deben conectarse exclusivamente equipos originales de autoMACS Pro con los conectores rotulados con External CAN CAN1 y CAN2 Los niveles de voltaje de estos conectores no debe sobrepasar los niveles de voltaje peligroso de 30 V CA y 42 4 V punta o 60 V CC Se debe conectar solamente el cable de sensor de botella MACS Pro con el conector del sensor de botella Con 1 Informaci n importante el conector RS232 BCR debe conectarse solamente un lector de 1 c digo 2D recomendado por Miltenyi Biotec Los dispositivos de l ser externos conectados con el conector rotulado con RS232 BCR deben cumplir con la norma IEC 60825 1 Utilice exclusivamente cables de conexi n con un largo menor a 3 m ee 1 5 Manejo mantenimiento transporte y eliminaci n seguros Observe las siguientes instrucciones para asegurar un manejo mantenimiento transporte y eliminaci n de su separador autoMACS Pro seguros 1 5 1 Manejo seguro Si el instrumento no funciona correctamente y las instrucciones o mensajes que aparecen en la pantalla aconsejan ponerse en contacto con el servicio t cnico el uso del instrumento ya no es seguro Apague inmediatamente el separador autoMACS Pro desench felo y p ngase en contacto con el proveedor de servicios de Miltenyi Biotec autorizado o con el servicio de atenci n al cliente de Miltenyi Biotec 1 5 2 Revisi n t cnica IMPORTANTE Salvo que el presente manual de u
72. Template from the lower navigation bar To scroll through the list of saved templates use the navigation arrows A V The corresponding template is displayed in the adjacent panel lil Figure 6 18 Selecting a reagent template Expt_2 was selected which comprises CD4 h and CD8 m on rack positions R1 and R2 respectively 2 Select and highlight the desired template in this case Expt_2 3 Select Ok DE 0 53 Figure 6 19 The template Expt_2 was successfully loaded 6 Cell separation using the autoMACS Pro Separator Deleting a reagent template 6 1 Select Load Template from the lower navigation bar 2 Scroll through the list of saved templates using the navigation arrows A V 3 Select the template for deletion 4 Select Delete Template 6 2 5 Cell separation 6 2 5 1 Cell separation after autolabeling Note To obtain brief It is an option to use the autolabeling feature to label cells with information on the MACS MicroBeads After entering the desired reagent s in the A separation and wash Reagent menu assign the MicroBead reagent vials to the respective programs highlight the positions in the MACS Reagent Rack and go to the Separation menu program of interest and press the information button located between the arrows A pop up window will be displayed Alternatively refer to sections 4 4 and 6 1 3 for more information on the Separation menu and separation programs Define reagent rack
73. abeling MACS MicroBeads specifically bind to antigens on the cell surface Only one incubation step is necessary Direct magnetic labeling requires a minimal number of washing steps and therefore minimizes cell loss Highly specific cell separation reagents for direct labeling of numerous cell types with MACS MicroBeads are available for human mouse rat and non human primate cells 6 Cell separation using the autoMACS Pro Separator 115 116 Indirect magnetic labeling Indirect magnetic labeling is based on a two step procedure In a first step cells are labeled with a primary antibody directed against a cell surface marker The primary antibody can either be unconjugated biotinylated or fluorochrome conjugated In a second step cells are magnetically labeled with MACS MicroBeads which either bind to the primary antibody or to a molecule conjugated to the primary antibody Accordingly magnetic labeling is achieved with Anti Immunoglobulin MicroBeads Anti Biotin MicroBeads or Anti Fluorochrome MicroBeads Indirect labeling can also be performed using a cocktail of primary antibodies to concurrently label a number of unwanted cell types e g for the untouched isolation of target cells 6 1 2 Cell separation strategies There are two basic strategies for separating specific cell populations positive selection and depletion During positive selection the target cells are magnetically labeled and collected as the positive fraction
74. able Make sure to prepare samples as specified If error is displayed again call Technical Support If object blocked movement remove object Switch OFF instrument Wait 5 seconds Switch ON instrument Otherwise or if error is displayed again call Technical Support Check bar code on rack or try different rack Make sure ambient or direct sun light does not hit sensor Clean sensor to remove salt or dust using a cloth soaked in 70 ethanol or double distilled water and wipe dry Check bar code on rack or try different rack Make sure ambient or direct sun light does not hit sensor Check if syringe is fastened correctly Exchange if broken or damaged If error is displayed again call Technical Support Restart instrument Please call Technical Support if error is displayed again Restart instrument and try a Qrinse to wash out clogged material If error persists run Safe program Otherwise exchange diluter valve or standard valves depending on where a constriction is suspected Please call Technical Support if error is displayed again 9 Troubleshooting 9 185 186 Error code 6006 6009 6216 7001 7002 7003 7004 7005 7006 7007 7008 7009 Cause Air intake during sample uptake although needle did not yet hit bottom of tube Leakage of air into system in front of bubble sensor liquid level has been overestimated or needle did not move to bottom as fast as nece
75. able plug Code reader plug MACS MiniSampler plug ell z Power socket Figure 3 7 Location of the MACS MiniSampler sensor cable code reader and power sockets at the back of the instrument 10 Carefully remove the uptake port needle from the foam packaging 3 Assembly and installation and underside of the touch screen are supported by foam 11 Place the uptake port needle into its guiding at the needle arm g CAM d Figure 3 9 The needle port is positioned as illustrated above 12 Note the position of the power socket on the rear panel of the autoMACS Pro Separator figure 3 7 Ensure that the main power switch is in position 0 13 Connect the power cord 14 Elevate the touchscreen and insert the memory card as shown below D 3 Assembly and installation 79 80 gt Figure 3 10 Inserting the software memory card 15 Replace the fluid bottles Refer to section 3 2 for full details 16 Switch ON the autoMACS Pro Separator 17 Secure the MACS Reagent Rack 4 onto the MACS MiniSampler into the left recess The engagement hook has to snap into the undercut i fl Y va Figure 3 11 Securing the MACS Reagent Rack 4 onto the MACS MiniSampler 3 Assembly and installation Note Racks can be pre cooled for 3 4 hours at 2 8 C Do not cool below 0 C since samples may freeze If recognition of the tube rack fails the instrument will display a screen for manual s
76. about 20 uL please be sure to overfill the vial with reagent accordingly 9 3 Magnetic labeling and separation 9 3 1 Positive selection Magnetically labeled cells have not been retained on the column 1 Magnetic labeling of the cells was insufficient because the MicroBead concentration was too low This may be due to the 9 Troubleshooting fact that either too much buffer was added to the cells or too much buffer was left on the cell pellet after centrifugation before adding the MicroBeads Furthermore the total number of cells may have been miscounted Use a ratio of MicroBeads and cells as stated in the respective MACS Cell Separation Reagent data sheet Labeling of cells was ineffective due to too much debris and or dead cells in the sample Debris and dead cells will non specifically bind to all other components present Antibodies and MicroBeads will be captured non specifically and not be able to label the cells in the positive fraction sufficiently any more Labeling of cells was ineffective due to an incubation temperature lower than recommended It is recommended to incubate cells in the refrigerator 2 8 C for labeling The number of magnetically labeled cells exceeded the column capacity Calculate the number of magnetically labeled cells e g by staining with flurochrome conjugated antibodies and subsequent fluorescence analysis The number should not exceed 2x108 cells per sample If necessary split the sample
77. akage Pump syringe leakage Pump syringe is filled with air during operation Washing station overflow Outlet port is clogged MACS MiniSampler does not move properly Touchscreen remains dark Disruption of power supply during cell separation Performance errors not indicated by a warning or error screen Output volumes are not correct Low cell viability in final fraction Low purity of isolated cells Low recovery of isolated cells Sample not or only partly taken up Reagent vial runs dry Magnetic labeling and separation Positive selection Depletion MACS MicroBeads for indirect labeling Fluorescent staining Cells are poorly stained Cells are excessively stained Contamination of tubing system Problems indicated by error or warning screens Technical data and specifications Schematics of the autoMACS Pro Separator Technical data and specifications of the autoMACS Pro Separator 162 163 163 164 164 165 165 166 169 169 170 170 171 171 171 172 172 173 173 174 174 174 174 175 175 175 176 177 177 178 178 178 181 183 184 184 184 184 184 189 189 193 11 11 1 11 1 1 11 2 12 13 14 Content Hardware and software installation Installation of new software Exchanging the software cards Calibration of the autoMACS Pro Separator Technical support Limited warranty Glossary 197 197 197 198 203 205 207 Thank you for choosing a Miltenyi Biotec product The a
78. al Liquid waste shall be autoclaved or decontaminated using a disinfectant that is appropriate for the specific pathogen e g 10 bleach isopropyl alcohol or 70 ethanol Waste disposal must be in accordance with any local regulations 1 Important information autoMACS Pro Separator SN 0563 Dider na 130 097 545 Power Supply 100 240 VAL 5060 Hz 200 VA Fuser TAANS c JUS USTED LMR CP SILA Figure 1 4 Warning signs for biohazard located on lower facing panel of the autoMACS Pro Separator left and on top of autoMACS Pro Separator fluid bottle right Flammable 70 ethanol is used in Sleep and Store programs The solvent is flammable Therefore keep the instrument away from fire 1 5 5 Transport The autoMACS Pro Separator should be transported with care in packaging specified by Miltenyi Biotec Internal damage can occur if it is subjected to excessive vibration or if it is dropped If the instrument needs to be shipped back to the manufacturer for service decontaminate the instrument from any hazardous material prior to shipment If you have questions regarding proper decontamination or shipment please contact Technical Support for assistance Refer to section 7 3 4 for further information on instrument decontamination 1 5 6 Instrument disposal Please contact Technical Support for assistance if you wish to dispose of your instrument 1 Important information 21 22 1 Important informat
79. ample volume 2mL 4mL min columns 1and 2 Posselds 2mL sample volume 2mL 1 mL min columns 1 and 2 Posselwb 2mL 3x sample 2mL 4mL min volume predilution columns 1and2 Deplete 2mL sample volume 2mL 4mL min Depletes 2mL sample volume 2mL 1mL min Depl05 2mL sample volume 2mL 0 5 mL min Depl025 2mL sample volume 2mL 0 25 mL min A_Depl07 3 mL per 1 mL sample 2 mL per 1 mL 4mL min volume sample volume A_Depls7 3 mL per 1 mL sample 2mLper1mL 1 mL min volume sample volume This program must only be used with manual labeling Predilution 3 fold of the sample volume Table 6 1 Output volumes and loading rates of separation programs 6 1 3 2 Wash and maintenance programs The autoMACS Pro Separator is equipped with reusable autoMACS Columns Therefore after each cell separation a thorough washing procedure rinses the columns of the autoMACS Pro Separator After the wash program is completed columns and tubing system are filled with Running Buffer Moreover there are obligatory and optional wash programs which have to be performed daily or at times to ensure proper operation and maintenance of the instrument Please find below a list of the available wash and maintenance programs 6 Cell separation using the autoMACS Pro Separator Note The autoMACS Pro Separator will not start a separation program before a wash program has been completed Wash programs Qrinse is the standard short wash program that only uses Run
80. and installation 3 3 7 Test of barcode scanner 3 1 Select Reagent 2 Press Read Reagent 3 Place the barcode e g the barcode below in front of barcode scanner also refer to figure 1 3 Ki Se RE 15 19 Figure 3 31 The name ofthe scanned reagent here CD4 MicroBeads human 130 045 101 appears in the display 3 Assembly and installation 91 92 3 Assembly and installation An overview of the autoMACS Pro 4 Separator user interface The autoMACS Pro Separator is operated through a TFT color touchscreen After switching ON the instrument the startup screen ie will appear i 12 15 Figure 4 1 The autoMACS Pro Separator startup screen 4 1 Working with the lower menu bar Figure 4 2 The lower menu bar of the Separation menu A lower menu bar is accessible from all menu screens Depending on the status of the instrument and the selected upper tabbed menu option the lower menu buttons switches between an inactive grey background and active white background state For example Stop can only be selected when the instrument is actively performing a process such as cell separation Figure 4 3 Left Stop button is active and can be selected Right Stop button is inactive and cannot be selected 4 2 The main menu screen Five main menus allow easy interaction with the instrument They are accessed through the upper tabbed menu 4 An overview of the autoMACS Pro Separator u
81. anol analytical reagent The storage solution consists of 70 ethanol and has to be prepared grade without additive from absolute ethanol p a grade and distilled water MACS Bleach Solution is used in combination with the Safe program 5 Materials required 111 112 Note Program Safe can also be used for decontaminating the autoMACS Pro Separator fluidic system For special decontamination procedures contact Miltenyi Biotec Technical Support Depending on the level of use it is recommended to run a Safe program at least every 3 to 6 months If material like whole blood or tissue is primarily used it is recommended to run the Safe program once a month Buffers amp Description Capacity Order no Solutions autoMACS Cell separation buffer 6x1 5L 130 091 221 Running Buffer containing azide as preservative autoMACS For rinsing of the autoMACS 6x1 45 L 130 091 222 Rinsing Pro Separator s fluidic Solution system and preparation of preservative free cell separation buffer MACS BSA For preparation of 6x0 75 L 130 091 376 Stock Solution preservative free cell separation buffer autoMACS Pro For rinsing of the autoMACS 6x1 5L 130 092 987 Washing Pro Separator s fluidic Solution system Storage 70 v v ethanol in distilled solution water make sure to use absolute ethanol p a MACS Bleach Used in combination with 1x1 L 130 093 663 Solution Safe program Table 5 1 Buffers and solutions required for operation
82. any magnetic information carriers such as credit cards magnetic tapes and floppy disks any electronic equipment such as hearing aids pacemakers measuring and control instruments computers and watches and magnetizable tools and objects at a distance of at least 20 cm from the magnet cover These items may be affected or damaged by the magnetic field 1 Important information Figure 1 1 Location of warning sign for strong permanent magnet 1 3 3 Hazard of crushing and shearing CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard CAUTION Do not open the front access covers while the instrument is in operation Do not obstruct the movement of the automated arm and accessory hardware during operation Keep fingers etc away from all moving parts of the autoMACS Pro Separator and accessory hardware to avoid crushing or shearing injuries or damage to the instrument Do not touch fluid pumps or adjust the tubing while the instrument is in operation Always switch OFF the instrument before adjusting any part of the fluidic system Always stop or abort a procedure before handling accessory hardware e g MACS MiniSampler or loading removing tubes from the tube rack placed on the sampler Do not circumvent any safety measures or devices Figure 1 2 Open circle shows warning sign for hazard of crushing and shearing 1 Important information 15 1 3
83. ardous samples it is recommended to fill the waste bottle with 100 mL of disinfectant before use e g MACS Bleach Solution 130 093 663 For proper disposal please follow local regulations and carefully read the chapter 1 3 Assembly and installation 81 82 Note When removing the waste bottle unscrew the bottle closure and then fasten a standard lid i e without sensor cables onto the bottle before removing the waste bottle from the basket This is to ensure that the content of waste bottle is not accidentally spilt over the autoMACS Pro Separator during removal from the basket Note The correct positioning of each solution bottle recognizable by the color code and the symbols is crucial in order to perform successful procedures using the autoMACS Pro Separator To keep buffers sterile each bottle closure should be equipped with a hydro phobic air filter Avoid any contact of hydrophobic air filters with fluids as this may cause clogging of the filter Bottle Symbol Bottle Running Storage Buffer solution blue black Washing Waste Solution red green Table 3 1 Symbols and color coding of fluid bottles 1 Install one fluid bottle at a time Please note the corresponding color coding refer to table 3 1 2 Unscrew bottle closures counter clockwise and remove the empty bottle Do not disconnect the color coded tubing 3 Donotopena fresh bottle until it is placed in the basket
84. as como la botella de desechos Por favor no extraiga las cestas de las botellas De acuerdo con la normativa internacional IEC 62471 este sistema de l mpara tiene un valor de riesgo por exposici n EHV de 0 91 siendo en exceso para el grupo exento de riesgos Exempt Risk Group La distancia de riesgo HD para el grupo exento de riesgos es de 61 cm La distancia de riesgo para el Grupo de Riesgo 1 Risk Group 1 es de 20 cm No mirar directamente el l ser radiaci n LED l ser reflejado o radiaci n del LED de una superficie reflejada De otra manera podr a ocasionarse una lesi n ocular No dirigir intencionalmente el rayo l ser a otras personas No desmontar modificar ni quitar el l ser o la fuente de radiaci n LED o sus soportes de montaje El l ser o las fuentes de radiaci n LED no interrumpen la emisi n cuando est n desmontados La radiaci n de unidades desmontadas puede ocasionar lesiones oculares Tener cuidado con la trayectoria del rayo o la reflexi n l ser de una superficie reflejada Durante la instalaci n del autoMACS Pro Separator tener cuidado de que durante la operaci n la trayectoria del rayo l ser no est a la misma altura que el ojo humano No permitir que agua aceite polvo u otras sustancias ajenas se peguen a la ventana de abertura del lector 2D Code Esto puede causar errores de lectura Aseg rese de parar la emisi n del l ser antes de limpiar el esc ner De no ser as la exposici
85. ate the instrument as instructed below The autoMACS Pro Separator contains various control boards During initialization of the instrument with a software card all of these components are automatically checked for the currently installed software 11 1 1 Exchanging the software cards 1 Ensure the instrument is switched OFF before proceeding 2 Note the memory card slot at the right hand side of the touch screen 2 ne at Figure 11 1 Location of the memory card slot 3 Remove the old memory card by pressing the black release button beneath the card slot 4 Insert the new memory card 5 Switch the instrument ON A dialog box will appear prompting the operator to proceed if desired 6 Select Update to confirm that you wish to proceed 11 Hardware and software installation 197 198 Note During the panel software update process the display may turn black or white or may freeze or flicker This process may take several minutes The progress will be indicated by an acoustic signal If warning 7030 is reported confirm by selecting OK If warning 7016 is reported select Ignore Note If warning 7016 Columns are not installed Start column exchange program appears select Ignore Panel software version conflict Version AMI_01 _02_5 required ER 154 Figure 11 2 The user must confirm any update The current software panel software version will be overwritten 11 2 Calibration of th
86. ationen 41 42 1 4 2 Bel ftung Das Instrument sollte nicht in der N he von Radiatoren Hei l fter fen oder anderen W rme erzeugenden Ger ten stehen Verst rker eingeschlossen Erm glichen Sie im Betrieb eine ausreichende Luftzirkulation im Abstand von mindestens 15 cm in allen Richtungen um den autoMACS Pro Separator um eine ausreichende K hlung zu gew hrleisten Vermeiden Sie das Ger t direkter Sonneneinstrahlung auszusetzen ffnungen und Schlitze am Ger t sind zur Bel ftung gedacht und sollten niemals bedeckt oder blockiert werden da sie das Ger t vor berhitzung sch tzen und sicheren Betrieb erm glichen F hren Sie niemals Fremdk rper durch ffnungen in das Ger t ein 1 4 3 Wassereintritt und N sse Setzen Sie das Instrument niemals in einer feuchten oder nasskalten Umgebung ein Setzen Sie das Ger t nie hoher Feuchtigkeit oder Kondensation aus und sch tzen Sie es gegen Spritzwasser 1 4 4 Geerdetes Produkt Das Ger t ist ausgestattet mit einem Schutzkontaktstecker und einer dreiadrigen Anschlussleitung F hren Sie den Stecker nicht in eine Steckdose ohne Schutzkontakt ein Wenn Ihre Steckdose keinen Schutzkontakt aufweist bitten Sie einen ortsans ssigen Elektriker diese zu ersetzen 1 4 5 Stromquellen Das Ger t sollte nur von einer Stromquelle aus betrieben werden die den elektrischen Angaben auf dem Typschild entsprechen Sollten Sie Fragen zur Art der Stromversorgung haben wenden Sie sic
87. attached to the connectors labeled External CAN CAN1 and CAN2 The voltage levels on these connectors shall not exceed hazardous voltage levels of 30 V RMS and 42 4 V peak or 60 Vdc Only the autoMACS Pro Bottle Sensor Cable should be attached to the Bottle Sensor connector 1 Important information Only a 2D code reader recommended by Miltenyi Biotec should be 1 connected to the RS232 BCR connector External laser devices connected to the connector labeled RS232 BCR have to comply with the standard IEC 60825 1 Only use connector cables less than 3m in length 1 5 Secure operation maintenance transport and disposal Observe the following instructions to ensure secure operation maintenance transport and disposal of your autoMACS Pro Separator 1 5 1 Safe operation If the instrument is not working properly and instructions or messages on the display screen advise to contact technical service secure operation is no longer possible Immediately switch OFF the autoMACS Pro Separator unplug the instrument from the electrical outlet and contact an authorized Miltenyi Biotec service provider or the Miltenyi Biotec Customer Support team 1 5 2 Servicing IMPORTANT Unless otherwise specifically noted in this user manual or other Miltenyi Biotec documentation do not service the autoMACS Pro Separator yourself Servicing and repair must be performed by qualified service personnel Improper or inc
88. balaje especificado por Miltenyi Biotec Se pueden producir da os internos si es expuesto a vibraciones excesivas o si se cae En caso de que sea necesario devolver el instrumento a su fabricante para su revisi n limpielo y desinf ctelo de cualquier sustancia peligrosa antes de realizar el env o Si tiene preguntas relativas a la descontaminaci n o el env o p ngase en contacto con el servicio de asistencia t cnica 1 5 6 Eliminaci n del instrumento equipo P ngase en contacto con el servicio de asistencia t cnica si desea desprenderse de su instrumento 1 Informaci n importante 69 70 1 Informaci n importante Introduction 2 2 1 MACS Technology the gold standard in cell separation MACS Technology has become the standard method in cell separation It is based on i MACS MicroBeads i e highly specific antibodies coupled to superparamagnetic 50 nm particles ii MACS Columns containing a matrix of ferromagnetic spheres and iii MACS Separators providing a strong magnetic field When a MACS Column is placed in a MACS Separator the ferromagnetic spheres amplify the magnetic field by 10 000 fold thus inducing a high gradient within the column This magnetic field is strong enough to retain cells that are labeled with minimal amounts of MACS MicroBeads Columns rapidly demagnetize when the column is removed from the separator or in case of the autoMACS Pro Separator the magnet is automatically retract
89. cell sample will be rinsed out at the waste port 9 Troubleshooting 3 Select the Separation menu and Wash now Select Qrinse and 9 press Run The cells will be rinsed out with Running Buffer 4 Reconnect waste closure and waste bottle 5 Perform the Rinse program to prime the autoMACS Pro Separator 6 The rescued cell sample can now be centrifuged and subsequently be reprocessed with the autoMACS Pro Separator 9 2 Performance errors not indicated by a warning or error screen 9 2 1 Output volumes are not correct 1 Check for air inlet and leakage in the fluidic system by running the Rinse program Select Wash Now from the lower menu bar in the Separation menu Highlight Rinse and click Run 2 Ifthe tubing is leaking go to section 9 1 2 3 If the pump syringe is leaking go to section 9 1 3 4 Ifthe column is leaking go to section 9 1 1 5 If one or more valves are leaking go to section 7 4 1 6 Check whether all tubing connections including column connections bottle connections and connections at the pump syringe are fastened Loose connections may allow air to enter the system and therefore affect the performance 7 If the problem persists run the Calibr_2 program Select User settings from the Option menu Highlight Calibr_2 and click Run 8 If the problem persists contact Miltenyi Biotec Technical Support 9 2 2 Low cell viability in final fraction 1 Low cell viability can result from problems during both the c
90. cells In most cases the number of isolated cells will be compared to the number of cells theoretically expected from the heterogeneous starting population To calculate the target cell recovery take an aliquot from the magnetically labeled fraction just before starting the cell separation The target cell recovery e g positive cells in the magnetically labeled cell fraction can be calculated as follows No of cells in pos fraction x positive cells in pos fraction Target cell recovery 100 x No of cells in orig sample x positive cells in orig sample The overall cell recovery can be calculated as follows No of cells in pos fraction No of cells in neg fraction Overall cell recovery 100 x No of cells in orig sample 8 Quality control of separations performed with the autoMACS Pro Separator 169 8 2 Purity of isolated cell population For most experiments that follow the cell separation it is necessary to document the purity of the isolated cell subset It is recommended to analyze the cells by flow cytometry Alternatively fluorescence microscopy or immunocytochemistry can be used Purity of the positively selected cell fraction Purity positive cells in positive magnetically labeled fraction Purity of the depleted cell fraction Purity negative cells in negative non labeled fraction 8 3 Viability of the cells Different dyes are available to discriminate betwee
91. ch are directly attached to the instrument and ready to use sterile MACS Buffers are available for maximum convenience The autoMACS Pro Separator is supplied with the MACS MiniSampler and therefore offers an additional sampling option This feature allows the sequential processing of multiple samples without further manual handling The MiniSampler is supplied with three different tube racks and an additional reagent rack The MiniSampler and tube racks are automatically detected by the autoMACS Pro Separator adding to its operational efficiency 2 Introduction The user manual aims at explaining the principles of cell sorting with the autoMACS Pro Separator and assists you by providing step by step protocols for the separation and quality control Tips and hints in the troubleshooting section are intended to help optimize your magnetic cell separations 2 Figure 2 1 Front image of the autoMACS Pro Separator the access cover was made transparent for the purpose of illustration 2 Introduction 73 74 2 Introduction Assembly and installation 3 The autoMACS Pro Separator is a benchtop instrument that fits in laminar flow hoods or safety cabinets If the instrument is placed in a laminar flow hood the following accessories might be required autoMACS Pro Laminar Hood Plate 130 093 246 and optionally autoMACS Pro Angle Connector Kit 130 093 245 The laminar hood plate provides a stable and even surface even
92. chlusskabel mit einer pe Lange von max 3 Metern verwendet werden 1 5 Sicherer Betrieb Wartung Transport und Entsorgung Beachten Sie die folgenden die folgenden Hinweise bez glich Betriebssicherheit Wartung Transport und Entsorgung Ihres autoMACS Pro Separators 1 5 1 Sicherer Betrieb des Ger tes Falls Ihr Ger t nicht einwandfrei arbeitet und Anzeigen auf dem Display Sie dazu auffordern den technischen Kundendienst zu kontaktieren ist die Betriebssicherheit nicht mehr l nger gew hrleistet Schalten Sie das Ger t sofort aus unterbrechen die Stromzufuhr und kontaktieren einen autorisierten Miltenyi Biotec Service Provider oder das Miltenyi Biotec Customer Support Team 1 5 2 Wartung WICHTIG Versuchen Sie nicht den autoMACS Pro Separator selbst zu warten oder zu reparieren es sei denn es ist in diesem Benutzerhandbuch oder anderen technischen Unterlagen der Miltenyi Biotec GmbH ausdr cklich vermerkt Wartung und Reparaturen m ssen durch geschulte Fachkr fte ausgef hrt werden Falsche oder unsachgem e Wartung oder Reparatur an Ihrem Ger t kann zur Gef hrdung des Anwenders unvorhersehbaren Resultaten Fehlfunktionen Ger tesch den vorzeitigem Verschlei und verringerter Lebensdauer f hren und kann den Verlust Ihrer Garantieanspr che zur Folge haben Fragen Sie Ihren rtlichen Miltenyi Biotec Vertriebsmitarbeiter nach unseren weit reichenden Vereinbarungen zum Ger teservice und Kundendienst oder besuchen
93. cht absichtlich auf andere Personen Demontieren wechseln oder entfernen Sie nicht eingebaute Laser oder LED Strahlungsquellen oder deren Halterung Die Laser oder LED Strahlungsquellen h ren bei Demontage nicht automatisch auf Strahlung zu emittieren Strahlung von demontierten Ger ten kann zu Augenverletzungen f hren Achten Sie auf den optischen Weg des Laserstrahls oder der Reflektion durch eine Spiegeloberfl che Achten Sie bei der Einrichtung des autoMACS Pro Separator darauf dass sich der optische Weg des Laserstrahls beim Betrieb nicht auf gleicher H he mit dem menschlichen Auge befindet Vermeiden Sie dass Wasser Ol Staub oder andere Fremdk rper dem Offnungsfenster des 2D Code Lesers anhaften Dies kann zu Lesefehlern f hren Stellen Sie vor der Reinigung des Scanners sicher dass keine Laserstrahlen mehr emittiert werden Der Umgang mit dem Laser kann sonst zu Augenverletzungen f hren Verwenden Sie zum Abwischen von Substanzen auf dem Scanner ein weiches trockenes Tuch Verwenden Sie keinen Alkohol oder andere Reinigungssubstanzen Der autoMACS Pro Separator ist nach Norm IEC 60825 1 1993 Al 1997 A2 2001 als Klasse 1M Laserprodukt eingestuft 1 Wichtige Informationen VORSICHT Die Verwendung von anderen als hierin genannten 1 Bedienungselementen sowie die Anpassung oder Durchf hrung von anderen als hier genannten Verfahren kann gef hrliche Strahlung freisetzen Abbildung 1 3 Position von Las
94. chten 1 Wichtige Informationen 45 46 1 Wichtige Information Informazioni importanti 1 Leggere prima dell uso Si prega di leggere tutte le informazioni riportate nel presente manuale d uso prima dell utilizzo La mancata lettura e l inosservanza delle istruzioni possono condurre ad un impiego improprio o scorretto dello strumento a manipolazione e manutenzione inadeguate o errate e potrebbero rappresentare un pericolo per l operatore avere conseguenze imprevedibili causare il malfunzionamento o danni allo strumento condurre ad un usura prematura e ad una minore durata di esercizio dello strumento nonch invalidarne la garanzia Conservare il presente manuale in un luogo sicuro accessibile a tutti gi addetti che utilizzano l unita di separazione autoMACS Pro autoMACS Pro Separator Il presente capitolo illustra le istruzioni di sicurezza e i requisiti ambientali per l installazione dello strumento autoMACS Pro Separator seguenti simboli di avvertenza e attenzione servono quale ausilio alla prevenzione di lesioni personali e danni al dispositivo 1 1 Simbolie livelli di pericolo Indicazioni di sicurezza Esempio CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard Le indicazioni di sicurezza informano l operatore dei rischi potenziali derivanti dal mancato rispetto delle avvertenze e precauzioni descritte di seguito L icona su
95. ck if the leakage persists If so unscrew the tubing and check if the screw thread is damaged If this is the case order and install new tubing 6 Ifthe problem persists contact Miltenyi Biotec Technical Support 9 1 3 Pump syringe leakage Verify that the Running Buffer has equilibrated to room temperature before performing a washing or separation program Cold buffer will make the plunger seal constrict more than usual and may lead to leakage Salt crystals may also damage the pump syringe seal To clean the pump syringe refer to section 7 3 2 and retry If the problem persists follow the guidelines below 1 Run the Sleep program Press the shutdown button on the upper right hand corner of the screen W Click Yes to shutdown the instrument 2 Switch OFF the instrument 3 Wash the pump syringe as described in section 7 3 2 4 Switch ON the instrument and run a Qrinse program to ensure that the problem is solved Select Wash Now from the lower menu bar in the Separation menu Highlight Qrinse and click Run 5 If the leakage persists order and install either a new Pump Syringe 130 090 339 or a new Pump Seal 130 022 101 For details on the installation refer to section 7 3 2 9 Troubleshooting Note Depending on the level of use the pump syringe should be cleaned every 1 3 months Appropriate maintenance and long term storage assures that no salt deposits accumulate in the pump syringe Salt deposits may ca
96. d C Details of each instrument action are listed 4 7 Option menu nil Figure 4 29 The Option display The Option menu allows maintenance procedures such as exchange of autoMACS Columns or decontamination of the system The menu is divided into two main categories Special and User settings An overview of the functions available under each category are given below 4 Anoverview of the autoMACS Pro Separator user interface 4 7 1 Special A Special options comprise three special programs for column exchange Col_ex instrument decontamination Safe and cleaning of the instrument for long term storage Store Safe This is a disinfectant procedure which uses MACS Bleach Solution for cleaning and decontamination of the autoMACS Pro Separator Depending on the level of use and general instrument maintenance it is recommended to decontaminate the fluidic system every 3 to 6 months using the Safe program asa Store The program Store should be applied to prepare the instrument for long term storage Upon completion of the Store program the fluidic system contains 70 ethanol To store the autoMACS Pro Separator for a period longer than two weeks the tubing system should be cleaned and the columns should be replaced with column substitutes The Store program automatically performs the cleaning procedure and prompts the user to install column substitutes 4 7 2 User settings The User settings are for maintena
97. d as the last wash program Working with separation templates For convenience it is possible to load and save separation templates Separation templates can be used in combination with reagent templates refer to above To save a separation template 1 Configure the sample template an example follows 4 An overview of the autoMACS Pro Separator user interface 101 102 YA Figure 4 20 A sample template was setup to process two samples Sample 1 Isolation of untouched human monocytes using the Monocyte Isolation Kit Il human 130 091 153 autolabeling program MO IK h separation program Deplete Sample 2 Direct labeling and positive selection of CD4 cells using CD4 MicroBeads human 130 045 101 autolabeling program CD4 h separation program Possel 2 Select Save Template 3 Allocate a name to the template In this example the template was saved as EXPT_2A experiment 2a piale name and press OK ee 95 11 Figure 4 21 Using the alphanumeric keypad assign a name to the template 4 Select Ok To load a reagent template 1 Select Load Template from the lower navigation bar To scroll through the list of saved templates use the navigation arrows A V The corresponding template is displayed on the sample template panel 4 An overview of the autoMACS Pro Separator user interface Note The sample separation template was loaded however the reagent rack has not yet been configured
98. de s curit g n rales et les consignes mentionn es dans ce mode d emploi Les instructions de ce chapitre expliquent les risques potentiels li s l utilisation de l appareil et fournissent des informations de s curit importantes afin de r duire ces risques Respectez strictement ces consignes pour prot ger l quipement et vous m me des risques potentiels et cr er un environnement de travail s r La protection n est pas garantie si cet appareil n est pas utilis conform ment aux instructions donn es par le fabricant IMPORTANT Veuillez lire et respecter toutes les instructions d utilisation fournies dans ce mode d emploi et observez tous les avertissements affich s sur l appareil Conservez ce mode d emploi et les autres consignes de s curit et d utilisation fournies avec l appareil un endroit accessible pour tous les utilisateurs en vue de les consulter ult rieurement IMPORTANT Le s parateur autoMACS Pro est destin une utilisation int rieure uniquement N utilisez pas l appareil dans des zones class es dangereuses telles que des environnements teneur lev e en oxyg ne Les consignes de s curit concernant la zone de travail locale les bonnes pratiques de laboratoire ainsi que les directives relatives la sant la s curit et la pr vention des accidents en laboratoire doivent tre observ es en permanence Pour de plus amples d informations concernant l installation de l quipemen
99. deionized water Remove the finger guard that is attached underneath the outlet port holder by pulling gently Clean the finger guard as described for the ports Figure 7 2 Removing the finger guard for cleaning 4 Wipe the outlet port for the negative fraction as indicated above The port can be flushed by using a syringe To clean the uptake port ensure the instrument is switched OFF Move the needle holder up and down to get access to the entire surface of the needle Push the finger guard back into position Switch ON the instrument Run the Sleep program 7 Maintenance 147 148 Note Ensure enough storage solution is available 7 2 3 Instrument shutdown Make sure to run the Sleep program prior to switching the instrument OFF This program ensures most efficient preservation of the fluidic system by rinsing with storage solution Shutdown button 1 Press the shutdown button on the upper right hand corner of the screen 2 Select Yes from the pop up dialog box Do you really want to shutdown the device FE Figure 7 3 Shutting down the autoMACS Pro Separator 3 The autoMACS Pro Separator will automatically perform a Sleep program 4 Upon completion of the Sleep program you will be prompted to shut down the instrument The fluid bottle illumination is purple at this point Switch OFF the autoMACS Pro Separator by using the main power switch Store the program for long term storage To st
100. document de Miltenyi Biotec L entretien et la r paration doivent tre effectu s par des r parateurs qualifi s Tout entretien ou toute r paration inappropri e ou incorrecte de votre s parateur autoMACS Pro peut mettre en danger les utilisateurs fournir des r sultats impr visibles entra ner le dysfonctionnement ou la d t rioration de l appareil son usure pr matur e et r duire la dur e de vie de l appareil ainsi qu annuler votre garantie Renseignez vous aupr s de votre repr sentant Miltenyi Biotec local sur les conditions d entretien et de support compl mentaires propos es par Miltenyi Biotec ou consultez le site www miltenyibiotec com support IMPORTANT Si des pi ces de remplacement ou de rechange sont requises assurez vous que le prestataire de services utilise exclusivement des pi ces Miltenyi Biotec d origine ou des pi ces de fabricants tiers sp cifi es et recommand es par Miltenyi Biotec L utilisation de pi ces de remplacement ou de rechange non autoris es peut entra ner le dysfonctionnement de l appareil et fausser les r sultats de la s paration des cellules Miltenyi Biotec n accorde pas de prestation de garantie ou d cline toute 1 Informations importantes 31 32 responsabilit pour les pannes et les dommages de l appareil r sultant de l utilisation de pi ces de remplacement ou de rechange inappropri es Une fois les travaux d entretien ou de r paration achev s demandez a votre prestata
101. e autoMACS Pro Separator The autoMACS Pro Separator is calibrated by using the two programs Calibr_1 and Calibr_2 Calibration 1 is recommended after installation of new software Calibration 2 is always necessary when the pump syringe the Diluter Valve or the tubing are changed Calibr_1 is used to calibrate the settings of the needle arm x y and z axis including the alignment of the needle arm with the washing station and the Chill Racks Calibr_2 automatically calibrates the liquid volume control This is crucial for the correct measurement and processing of the sample volumes The fluidic system must be filled with buffer before commencing this step i e the Rinse program must be performed Calibration 1 Beginning the calibration sequence 1 Select menu Option 2 Highlight User settings and select Calibr_1 3 Select Run 4 Select Calibrate to proceed The first of four 1 4 calibration steps will begin namely calibration of the washing station 11 Hardware and software installation RE 1 01 Figure 11 3 Performing calibration of the needle arm a 4 step calibration process Calibration 1 Calibration of the washing station step 1 4 5 Select Calibrate and then Use to proceed with calibration The needle arm will automatically move towards the washing station and should be located directly above the center of the rear opening of the washing station Calibration 1 Step 1 4 Washing station Use current cal
102. e magnets The magnet cover is located in the center of the fluidic system and has slots for the autoMACS Columns Memory card Removable compact flash SD RAM card containing the autoMACS Pro Separator programs Memory card slot Slot located on the right hand side of the autoMACS Pro touchscreen giving access to the memory card The memory card should be removed by trained personnel only Negative fraction Sample fraction containing the non labeled cells that pass through the autoMACS Column while the column is placed in the magnetic field PE Phycoerythrin 14 Glossary 209 210 Ports The automated arm carries two ports the port located proximal to the instrument is designed for computer controlled fluid detection and distribution such as magnetic labeling sample mixing sample uptake and release of the magnetically labeled fraction while the port in the front releases the non labeled fraction Positive fraction Sample fraction containing the cells labeled with MACS MicroBeads These cells are retained on the column while the column is placed in the magnetic field The cells are eluted from the column after the magnet has been removed from the column Positive selection Process of isolating cells by labeling the target cells with MACS MicroBeads and performing a MACS Separation The labeled target cells are eluted in row C of the tube rack Possel Positive selection program in standard mode using one autoMACS Column The
103. e nominal voltage Transient overvoltages present on the mains supply category Il The instrument is suitable for rated pollution degree 2 The autoMACS Pro Separator is not specified for use in the cold room WARNING Read the chapter 1 for important safety information before installation and use The emission sound pressure level at the workstation is lt 70 dB A This equipment has been tested and found to comply with the limits for a Class B digital device pursuant to Part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference in a residential installation This equipment generates uses and can radiate radio frequency energy and if not installed and 10 Technical data and specifications used in accordance with the instructions may cause harmful 1 0 interference to radio communications However there is no guarantee that interference will not occur in a particular installation If this equipment does cause harmful interference to radio or television reception which can be determined by turning the equipment off and on the user is encouraged to try to correct the interference by one or more of the following measures Reorient or relocate the receiving antenna e Increase the separation between the equipment and receiver Connect the equipment into an outlet on a circuit different from that to which the receiver is connected Consult the dealer or an experienced radio TV t
104. eagent human 130 059 901 or FcR Blocking Reagent mouse 130 092 575 before labeling 2 The concentration of MicroBeads and or the temperature used for magnetic labeling were too high It is recommended to dilute the MicroBeads according to the data sheet Cells should be incubated with MicroBeads in the refrigerator 2 8 C 3 Incubation time with MicroBeads was too long leading to background labeling Reduce the time for incubation to the recommended values in the respective MicroBeads data sheet 4 Dead cells in the cell suspension may bind non specifically to MicroBeads and will then be co enriched in the positive fraction Remove dead cells before separation by using the MACS Dead Cell Removal Kit 130 090 101 or by Ficoll Paque density gradient centrifugation 5 Incubation temperatures were higher than recommended typically 2 8 C leading to non specific binding 6 Inadequately stringent separation programs were chosen e g DepleteS instead of Deplete Please refer to the respective data sheet for the recommended program When using the autolabeling feature 1 Check that the recommended maximal sample processing time of 90 minutes was not exceeded If the room temperature is too high the cooling capacity of the Chill Racks may be compromised Higher labeling temperatures could cause unspecific binding which may lead to lower purities when using positive selection strategies 9 Troubleshooting 9 1
105. echnician for help Changes or modifications of the equipment unless expressly approved by Miltenyi Biotec may void your authority to operate the equipment pursuant to FCC 47 CFR The autoMACS Pro Separator in combination with the MACS MiniSampler complies with all essential requirements of the following directives 2004 108 EC electromagnetic compatibility 2006 42 EC machinery and is in conformity with the following harmonized standards EN 61010 1 EN 61010 2 081 EN 61326 1 The autoMACS Pro Separator has been investigated by Underwriter Laboratories in accordance with the standards UL 61010 1 c US CAN CSA C22 2 No 61010 1 Listen EC 61010 2 081 LABORATORY EQUIPMENT 52ZA The MACS MiniSampler has been investigated by Underwriter Laboratories in accordance with the standards Al UL 61010 1 C US CAN CSA C22 2 No 61010 1 For other safety considerations refer to the product label or visit www miltenyibiotec com Design and specifications are subject to change without notice 10 Technical data and specifications 195 196 10 Technical data and specifications Hardware and software installation 11 Note Do not remove or insert a memory card while the instrument is turned ON The memory card must remain in the unit Removing the memory card during operation will abort all running processes 11 1 Installation of new software After installing an autoMACS Pro software update it is necessary to calibr
106. ecovery 2 Introduction 71 72 2 2 The autoMACS Pro Separator The autoMACS Pro Separator is a benchtop magnetic cell sorter that allows gentle sorting of more than ten million cells per second from a sample of up to 4x10 total cells The instrument is designed for use with more than 250 MicroBead based cell separation reagents for research applications Thus it is possible to choose between different cell separation strategies according to the respective experimental design from positive selection of abundant or rare cells to the isolation of untouched cells by depletion of non target cells or to the isolation of sophisticated cell subsets by sequential sorting Twelve preset separation programs simplify and standardize the application The autoMACS Pro Separator features automated sample labeling autolabeling sample loading elution of the non labeled negative as well as the labeled positive cell fractions Up to six samples can be processed in one programming step Furthermore automated procedures for maintenance of the system are included Different wash programs are available to rinse the columns before a new separation is performed One pair of columns can be used for up to 100 cell separations or for two weeks whichever comes first The thin film transistor TFT color touchscreen with intuitive screen menus makes operation and monitoring of the instrument user friendly and easy Finally standard MACS Fluid Bottles whi
107. ed from the autoMACS Column to elute the labeled cells The colloidal suspensions of MACS MicroBeads allow easy handling and short incubation times Cells can be separated in less than 30 minutes As minimal amounts of MicroBeads are required for magnetic cell labeling a large number of epitopes remains available for fluorescent labeling Thus staining of cells with fluorochrome conjugated antibodies for subsequent cell analysis can be performed simultaneously MACS MicroBeads are non toxic biodegradable and do not need to be removed from cells after the separation process MicroBeads do not alter structure function or activity status of labeled cells Isolated cells can be immediately used for downstream applications including flow cytometric analysis cell culture and molecular analysis It takes only a few simple steps to get pure cell populations Cells in a single cell suspension are magnetically labeled with MACS MicroBeads The sample is applied to the autoMACS Column placed in the autoMACS Pro Separator The unlabeled cells pass through while the magnetically labeled cells are retained within the column The unlabeled cells are collected as the negative fraction After a short washing step the magnet is retracted from the autoMACS Column and the magnetically labeled cells are eluted from the column as the positive fraction Thus with MACS Technology both labeled and unlabeled cells can easily be isolated with high purity and r
108. election of the tube rack Before confirming the choice ensure that the rack is placed correctly into the recess The MACS logo should face towards the user Note The 70 ethanol does not contain electrolytes Therefore the filling status of the storage solution bottle cannot be determined 18 Seta tube rack e g Chill 5 Rack onto the MACS MiniSampler 3 into the right recess ensuring that the rack barcode is facing the autoMACS Pro Separator A Alli a te y f fl gt 5 ve s U Figure 3 12 Positioning the Chill 5 Rack adjacent to aMACS Reagent Rack 4 on the MACS MiniSampler 19 Perform a system calibration test refer to section 3 3 for more details 3 2 Replacing fluid bottles and the connection of fluid sensor cables Operating the autoMACS Pro Separator requires Running Buffer Washing Solution and storage solution absolute ethanol diluted to 70 with double distilled water It is recommended to operate the instrument with ready to use MACS Buffers The autoMACS Pro Separator is delivered with four empty fluid bottles which are connected to the instrument with specifically designed bottle closures The bottle closures consist of a fluid uptake port or a fluid outlet port waste bottle as well as a sensor for measuring electrolyte conductivity The fluid bottles bottle closures and fluid sensor cables are color coded for easy handling refer to table 3 1 below WARNING When working with biohaz
109. ell preparation and the cell separation Hence also check section 9 3 2 Check the 70 ethanol Make sure that analytical reagent grade ethanol without additives is used to prepare the solvent 3 Check the Running Buffer Make sure that the appropriate buffer has been used and that no contamination has occurred 9 Troubleshooting 175 176 8 Check Washing Solution Make sure that the appropriate solution has been used and no contamination has occurred Check the pump syringe for contamination If contamination is obvious refer to section 9 1 3 and change columns afterwards Check for contamination of the tubing system and refer to the appropriate section Run the Safe program to decontaminate the fluidic system Check the column status The autoMACS Columns should be exchanged after 100 separations or two weeks after the last column exchange whichever comes first The procedure is described in detail in section 3 3 4 If the problem persists contact Miltenyi Biotec Technical Support 9 2 3 Low purity of isolated cells 1 Low purities can be caused by problems during both the cell preparation and the cell separation Hence also check section 9 3 Low purities can also result from using an inappropriate separation program or labeling approach Please also refer to section 6 Check the exchange date of the autoMACS Columns Using the autoMACS Columns for longer than two weeks or for more than 100 separations w
110. en niemals umgehen oder manipulieren Abbildung 1 2 Der Kreis zeigt das Warnsymbol f r Quetsch und Schergefahr 1 Wichtige Informationen 39 40 1 3 4 Laserstrahlung WARNUNG Das Ger t ist zur automatischen Reagenzglasgestell erkennung Klasse 1M mit vier oberflachenemittierenden Lasern mit vertikalem Resonator VSCLs ausgestattet Die Strahlung ist nicht sichtbar Schauen Sie nicht direkt mit optischen Instrumenten z B Objektiven Vergr erungsgl sern und Mikroskopen hinein Das Hineinschauen mit optischen Instrumenten in die VSCL ffnung innerhalb eines Abstandes von 100 mm kann Ihre Augen sch digen Das Ger t ist auch mit einem 2D Code Leser ausgestattet der einen sichtbaren Halbleiterlaser als Target Pointer Zeiger zur Einstellung der Leseposition verwendet sowie leistungsstarken Leuchtdioden LEDs zur Beleuchtung des Lesefeldes und der Fl ssigkeitsbeh ler und des Abfallbeh lters Entfernen Sie nicht die Flaschenk rbe Gem der internationalen Norm IEC 62471 hat dieses Lampen system einen Gefahrenwert der Exposition EHV von 0 91 und fallt in die Freie Gruppe Der Gef hrdungsabstand HD f r die Freie Gruppe betr gt 61 cm Der Gefahrdungsabstand f r die Risikogruppe 1 betr gt 20 cm Schauen Sie weder direkt in die Laser oder LED Strahlung noch in eine durch eine Spiegelfl che reflektierte Laser oder LED Strahlung Dies kann sonst zu Augenverletzungen f hren Richten Sie den Laserstrahl ni
111. ened Check for leakage or air bubbles in tubing from uptake needle to valve 1 Verify that connectors are appropriately fastened Calibrate needle arm Check for buffer supply Otherwise call Technical Support Provide all necessary reagents in Reagent Rack and correct Reagent Rack definition Try reinitialization by touching Retry Otherwise call Technical Support Push back needle to it s proper position in the needle holder check for smooth running of the needle in its support then touch Continue Otherwise calibrate needle arm especially if using a Chill 15 Rack Check for objects hindering the movement of the needle Remove the uptake needle from the needle holder and verify that there are no physical obstructions Press Continue Check that cover of the washing station is properly closed Remove any objects hindering the movement of the needle If needle hits rim of tube or top of rack calibrate needle arm using Calibr_1 program Touch Lift for needle retreat Check that cover of the washing station is properly closed Remove any objects hindering the movement of the needle If needle hits rim of tube or top of rack calibrate needle arm Touch Retry to try again Check that cover of the washing station is properly closed Remove any objects hindering the movement of the needle If needle hits rim of tube or top of rack calibrate needle arm Touch Retry to try again Do not use
112. ern Im Bereich des Rechtecks befinden sich verdeckt montierte Laser zur Erkennung von Gestellen Der sichtbare 2D Barcodeleser ist hier mit einem Kreis gekennzeichnet 1 4 Sichere Ger teinstallation In diesem Abschnitt werden die Anforderungen an einen betriebssicheren Standort f r Ihren autoMACS Pro Separator beschrieben Lesen Sie die Anleitungen in diesem Abschnitt und stellen Sie sicher dass der Ger testandort entsprechend vorbereitet ist bevor Sie das Ger t in Betrieb nehmen Ber cksichtigen Sie bei der Aufstellungsplanung die in diesem Abschnitt beschriebenen Vorkehrungen um Ger test rungen zu vermeiden und die Wahrscheinlichkeit umgebungsbedinget Ger teausf lle zu verringern 1 4 1 Montagezubeh r Stellen Sie den autoMACS Pro Separator niemals auf einen instabilen Labortisch Laborwagen Untersatz Laborstativ oder Laborkonsole Als Folge davon k nnte das Instrument zu Boden fallen dadurch k nnten schwere Verletzungen und oder Sch den am Ger t verursacht werden Benutzen Sie daher nur von Miltenyi Biotec empfohlenes oder direkt mit dem Ger t erh ltliches Labormobiliar Versuchen Sie nicht den autoMACS Pro Separator in eine Einbaukonstruktion zu integrieren oder auf begrenztem Raum z B in einem Laborregal unterzubringen es sei denn es wurde eine spezielle Vorrichtung daf r entwickelt es ist f r ausreichende Bel ftung gesorgt und die Montageanleitung f r das Ger t wurde befolgt 1 Wichtige Inform
113. es Le niveau de tension mesur sur ces connecteurs ne doit pas 1 d passer le niveau de tension dangereux de 30 Veff ainsi qu un pic de 42 4 V ou 60 V DC Seul le cable de capteur de flacon autoMACS Pro peut tre raccord au connecteur tiquet Bottle Sensor Seul un lecteur de code 2D recommand par Miltenyi Biotec peut ue tre branch au connecteur RS232 BCR Les appareils laser externes raccord s au connecteur tiquet RS232 BCR doivent r pondre a la norme IEC 60825 1 Utilisez uniquement des cables de connexion de 3 m tres de longueur maximum 1 5 S curit d utilisation maintenance transport et limination Respectez les consignes suivantes afin de garantir la s curit d utilisation la maintenance le transport et l limination de votre s parateur autoMACS Pro 1 5 1 S curit d utilisation Si l appareil ne fonctionne pas correctement et que les instructions ou les messages sur l cran d affichage conseillent de contacter le service technique la s curit d utilisation de l appareil n est plus garantie D connectez imm diatement le s parateur autoMACS Pro d branchez l appareil et contactez un prestataire de services Miltenyi Biotec agr ou l quipe du service apr s vente de Miltenyi Biotec 1 5 2 Entretien et r paration IMPORTANT N effectuez pas vous m me l entretien du s parateur autoMACS Pro sauf autre consigne sp cifique donn e dans ce mode d emploi ou dans un autre
114. ew Select OK 5 Select OK when asked to switch OFF the instrument 6 Switch OFF the instrument 7 Unscrew the syringe from the dilutor valve housing Figure 7 6 To remove the pump syringe the top screw must be turned counter clockwise 8 Carefully remove the plunger from the syringe 9 Remove salt crusts with distilled or deionized water 7 Maintenance 4 Figure 7 7 Washing the pump syringe with distilled water 10 Gently push the plunger back into the syringe but not all the way Dry the plunger lock screw before proceeding with installation of the syringe 11 Fasten the syringe at the dilutor valve by turning it until a resistance can be felt Unfasten again by one rotation Figure 7 8 The syringe is fastened at the dilutor valve 12 Pull the plunger out of the syringe until it reaches its fitting in the plunger holder In order to fit the plunger must have the same orientation as its fitting Tighten the plunger lock screw 7 Maintenance 151 152 Note If symptoms of wear such as leakage persist contact Technical Support Figure 7 9 The plunger is placed in its fitting in the plunger holder The plunger screw is fastened 13 Tighten the syringe at the diluter valve by turning until a resistance can be felt 14 Switch ON the instrument 15 Prime the autoMACS Pro Separator as described in section 7 2 1 16 Run the program Calibr_2 to calibrate the fluidic volume control of the instrume
115. ewahren Sie dieses Benutzerhandbuch an einem sicheren Ort zug nglich f r jeden Anwender des autoMACS Pro Separators auf Dieses Kapitel beschreibt die Sicherheitsvorschriften und die Anforderungen an den Ger testandort Ihres autoMACS Pro Separator Die folgenden Warnhinweise und Vorsichtsma regeln sollen Ihnen dabei behilflich sein Verletzungen f r sich oder Sch den am Ger t zu vermeiden 1 1 Symbole und Gefahrenstufen Warnhinweis Beispiel CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard Die Gefahrenhinweise informieren den Anwender Uber Gefahren die bei Missachtung der nachstehend beschriebenen Warnungen und Vorsichtsma nahmen bestehen k nnen Das Gefahren symbol links versinnbildlicht die Gefahr Die Gefahrenwarnung oben bezeichnet die Schwere der vorliegenden Gefahr wie vorstehend beschrieben Schwere Art und Ursprung der Gefahr sowie m gliche Folgen Verbote und Vorsichtsma nahmen werden im Anschluss beschrieben 1 Wichtige Informationen 35 36 WARNING CAUTION eo O P Pb PPP gt if Symbole und Gefahrenstufen Im folgenden Glossar werden alle in diesem Benutzerhandbuch oder auf dem autoMACS Pro Separator verwendeten Symbole bildlich dargestellt und erklart WARNUNG bezeichnet eine Gefahrensituation die falls sie nicht vermieden wird zu sschweren Verletzungen oder Tod f hren kann VORSICHT bezeichnet eine Gefahrensituati
116. f diluted reagents at 4 C or 20 C may lead to degradation 6 Cells were not labeled with MicroBeads because the MicroBeads were degraded Check the expiration date Check for sterility of the MicroBeads if the vial has been opened before Positive fraction shows poor purity 1 The concentration of the primary antibody was too high leading to non specific binding of unwanted cells The primary antibody should be titrated carefully For tips and hints on titration refer to the FAQs under Customer Support at www miltenyibiotec com 2 The primary antibody shows non specific binding Add blocking reagents such as BSA or immunoglobulin before labeling the cells 3 The incubation time during labeling was too long Do not exceed the incubation times specified in the respective data sheet 4 Check the primary antibody for its specificity When antiserum is used it is recommended to pre adsorb the antiserum e g on cells which do not express the antigen or to purify it by affinity chromatography ammonium sulfate precipitation ion chromatography etc in order to avoid non specific cross reaction 9 Troubleshooting 183 Note Generally MACS Fluorochrome conjugated Antibodies are used in a 1 11 dilution resuspend up to 107 cells in 100 uL of buffer and add 10 uL of MACS Fluorochrome conjugated Antibodies For details or exceptions refer to the MACS Reagent data sheet If other than MACS Antibodies are used carefully titrate
117. f the instrument Sleep program for regular use Store program for long term storage Shutdown button Decision tree for the optimal separation program Maintenance General considerations Daily maintenance Instrument priming Clean uptake outlet ports Instrument shutdown Periodic maintenance Column exchange Clean the pump syringe Clean the washing station Instrument decontamination Instrument long term storage Planned maintenance Valve exchange Exchange of dilutor valve 111 111 111 112 113 115 115 115 116 118 118 120 122 122 122 122 124 125 126 126 130 131 133 133 134 136 139 139 140 142 142 142 143 144 145 146 146 146 147 148 149 149 149 152 153 155 155 155 159 74 3 744 7 5 7 6 7 7 7 7 1 7 7 2 7 8 8 1 8 2 8 3 9 1 9 1 1 9 1 2 9 1 3 9 1 4 9 1 5 9 1 6 9 1 7 9 1 8 9 1 9 9 2 9 2 1 9 2 2 9 2 3 9 2 4 9 2 5 9 2 6 9 3 9 3 1 9 3 2 9 3 3 9 4 9 4 1 9 4 2 9 5 9 6 10 10 1 10 2 Content Exchange of the peristaltic pump head Exchange of hydrophobic air filter Exchange of the tubing system Exchange of fuses Rescue procedure Rescue procedure A Rescue procedure B Spare parts list Quality control of separations performed with the autoMACS Pro Separator Recovery of cells Purity of isolated cell population Viability of the cells Troubleshooting Hardware problems not indicated by a warning or error screen Column leakage Tubing le
118. g the instrument If there is any sign of leakage e g salt deposits tighten the respective tubing connection Close the front door and proceed with the separation 5 When priming is finished the instrument will display Ready for separation in the Status menu The fluid bottles are now illuminated green Ready for separation RINSE N ix Figure 3 29 The instrument has been primed rinsed 3 3 6 Checking the fluid levels The autoMACS Pro Separator automatically ensures that the uptake port is filled with buffer that fluid bottles carry enough fluid for one separation and that the waste bottles can collect fluid from at least one separation If more than one separation is performed ensure that the bottles contain sufficient fluid for all the separations and that the waste bottle is empty To verify the status of the bottles select the menu Status from the upper navigation bar On the left hand side of the menu four symbols display the solution bottles and their filling status If the fluid bottles are full and the waste bottle is empty the symbols are green If the solution bottles are empty or the waste bottle is full the respective symbols are red The symbol for the storage solution bottle remains gray It is recommended to select the Status menu when the autoMACS Pro Separator is in operation Ready tor separation QRINSE DA 02 00 Figure 3 30 Status of the fluid bottles is green for ready 3 Assembly
119. g a reagent vial using the 2D code reader In this example the vial CD4 MicroBeads human was scanned 3 The vial is automatically recognized by the software The next reagent rack position R2 will be automatically highlighted 6 Cell separation using the autoMACS Pro Separator 127 128 Define reagent rack E Figure 6 9 CD4 MicroBeads human 130 045 101 was identified by the 2D code reader software and assigned to the reagent rack position R1 4 Using the same procedure another reagent vial can be scanned using the 2D code reader The reagent rack position R3 is automatically assigned Having entered the desired reagent s assign MicroBead reagent vials to positions on the MACS Reagent Rack Click the Separation tab to proceed with programming a cell separation refer to section 6 2 5 Manual entry of reagents 1 Select Reagent tab and highlight the position where the vial will be placed on the reagent rack Four positions are available R1 R2 R3 and R4 Detine reagent rack i i Figure 6 10 Position R2 was selected for the reagent vial 2 Select Enter Reagent from the lower navigation bar Enter the reagent specific product order number The order number is located on the product data sheet In the event that the data sheet is misplaced visit www miltenyibiotec com to download a printable PDF of the document 6 Cell separation using the autoMACS Pro Separator Define reagent rack
120. ggio laser o a riflessi da una superficie a specchio Durante l installazione del autoMACS Pro Separator accertarsi che il percorso del raggio laser non si trovi alla stessa altezza degli occhi di chi esegue il lavoro Evitare che acqua grasso polvere o altre sostanze estranee si depositino sulla finestra di lettura del lettore di codici a barre 2D Ci potrebbe comportare errori di lettura Accertarsi di aver arrestato l emissione laser prima di pulire il lettore Un esposizione al raggio laser potrebbe danneggiare i propri occhi Utilizzare un panno morbido e asciutto per rimuovere qualsiasi sostanza dal lettore di codici a barre Non utilizzare alcol o altre sostanze detergenti Il autoMACS Pro Separator classificato come prodotto laser della classe 1M secondo lo standard IEC 60825 1 1993 A1 1997 A2 2001 1 Informazioni importanti ATTENZIONE L utilizzo di comandi o regolazioni o l esecuzione 1 di procedure diverse da quanto qui specificato pud essere causa di esposizione a radiazioni pericolose w N ES Figura 1 3 Posizione dei laser Laser invisibili per il rilevamento dei portacampioni portareagenti sono collocati nell area indicata dal rettangolo Il lettore per codice 2D visibile collocato all interno del cerchio 1 4 Installazione sicura Questa sezione descrive i requisiti che deve soddisfare il luogo in cui viene collocato il autoMACS Pro Separator onde garantirne un installazione sicura e i
121. h an einen autorisierten Miltenyi Biotec Service Provider oder Ihren lokalen Stromversorger Benutzen Sie keine Verl ngerungskabel oder Steckdosenleiste berladen Sie eine Steckdose nicht Die Gesamtlast darf 80 der Zweigstromkreis bemessung nicht berschreiten 1 4 6 Zug nglichkeit der Stromverbindungen Der Hauptstromschalter ebenso wie der Netzstecker f r das Stromkabel sollten leicht zuganglich sein und sich in m glichst unmittelbarer Nahe zum Bediener des Ger tes befinden Sollte es erforderlich sein die Stromzufuhr zu unterbrechen ziehen Sie den Netzstecker 1 4 7 Peripherieger te An die mit COM beschriftete RS 232 Buchse d rfen nur Peripherieger te angeschlossen werden die UL 60950 erf llen Die mit RS232 AUX beschriftete Buchse ist nicht belegt An die mit External CAN CAN1 und CAN2 beschrifteten Buchsen d rfen nur Original autoMACS Pro Ger te angeschlossen werden Die Spannung an diesen Buchsen darf die gef hrlichen Werte von 30 Veff und 42 4 V Spitze bzw 60 V DC nicht Ubersteigen An die mit Bottle Sensor beschriftete Buchse darf nur das autoMACS Pro 1 Wichtige Informationen Flaschensensorkabel angeschlossen werden An die mit RS232 BCR 1 beschriftete Buchse darf nur ein von Miltenyi Biotec empfohlener 2D Barcodeleser angeschlossen werden Externe Laserger te die an die mit RS232 BCR beschriftete Buchse angeschlossen werden mussen IEC 60825 1 erf llen Es d rfen nur Ans
122. he possibility of environmentally caused shutdowns 1 4 1 Mounting accessories Do not place the autoMACS Pro Separator on an unstable table cart stand tripod or bracket As a consequence the instrument might fall down This may cause serious bodily harm and or serious damage to the instrument Use only on a table cart stand tripod or bracket recommended by Miltenyi Biotec or sold with the instrument Do not place the autoMACS Pro Separator within a built in apparatus or a confined space such as a shelf rack unless the apparatus has been specifically designed to accommodate the instrument proper ventilation is provided and the mounting instructions for the instrument have been followed 1 Important information 17 1 4 2 Air circulation The instrument should not be placed next to radiators heat registers stoves or other pieces of equipment including amplifiers that produce heat Allow sufficient air circulation around the autoMACS Pro Separator at least 15 cm on all sides during operation to ensure adequate cooling of the instrument Prevent direct exposure of the instrument to sunlight Slots and openings of the instrument are provided for ventilation and should never be blocked or covered as these ensure reliable operation ofthe autoMACS Pro Separator and protect the instrument from overheating Never push a foreign object through an opening into the instrument 1 4 3 Water and moisture Do not use the instrument in a wet
123. her the guiding of the MACS MiniSampler is connected properly to the connector at the autoMACS Pro Separator labeled External CAN 2 Check whether the bolt below the rack detection protrudes from the instrument If this is the case push it in and turn it clockwise to lock the bayonet mount 3 Check the cable connection between MACS MiniSampler and autoMACS Pro Separator Check for cable damages 4 Check whether the MACS MiniSampler can freely move to both sides and check for any resistance or collision 5 Ifthe problem persists contact Technical Support 9 1 8 Touchscreen remains dark 1 Switch OFF the instrument wait five seconds and switch ON again 2 If the autoMACS Pro Separator still does not initialize check if the power cord is plugged in correctly and if the electric power is switched ON 3 Replace the fuses refer to section 7 6 Spare fuses are included in the autoMACS Pro Separator Starter Kit 4 Ifthe problem persists contact Miltenyi Biotec Technical Support 9 1 9 Disruption of power supply during cell separation In the unexpected event of a power supply failure during the cell separation procedure follow the rescue procedure to recover the cell sample trapped in the autoMACS Pro Separator fluidic system For detailed information refer to section 7 7 1 Switch ON the autoMACS Pro Separator 2 Disconnect the outlet tubing from the waste cap and put the end of the tubing in a 50 mL tube The rescued
124. ibration value 4 recommended or reset it Calibrate washing station 11 35 Figure 11 4 Calibration of the washing station To use already saved settings select Use To reset calibration settings not recommended select Reset 6 Select Height and check the central positioning of the uptake needle by using the buttons Move up and Move down Select Position and adjust by using the Move back and Move fwd buttons Select Done after making necessary adjustments Figure 11 5 Test calibration of the needle arm position in the washing station The position of the uptake needle should appear as shown above 11 Hardware and software installation 11 199 200 7 Select Save to save the new configuration Calibration 1 Step 1 4 Washing station Save new value 4 old 4 Calibrate washing station Figure 11 6 Saving new settings for calibration of the washing station Calibration 1 Calibration of the tube rack step 2 4 8 Select Calibrate to continue with the calibration process 9 Place a Chill Rack with sample tubes onto the MACS MiniSampler It is recommended to use a Chill 15 Rack 10 Select Done The needle arm will move to position A1 of the tube rack 11 Check the central positioning of the uptake needle on the bottom of the tube by using the needle navigation buttons Move back Move fwd Height Ensure the needle is positioned at the bottom center of the tube in row A of the
125. ie arri re de l autoMACS Pro Separator en bas a gauche et sur la bouteille poubelle en haut a droite 1 5 5 Transport Le s parateur autoMACS Pro doit tre transport avec soin dans un emballage sp cifi par Miltenyi Biotec Un dommage interne peut survenir si l appareil est soumis des vibrations excessives ou s il chute Si l appareil doit tre r exp di pour tre remis en tat d contaminez l appareil afin d liminer toute mati re dangereuse avant le transport Si vous avez des questions concernant la propre d contamination ou l exp dition n h sitez pas contacter notre service technique assistance 1 5 6 Elimination de l appareil Si vous souhaitez liminer votre appareil veuillez contacter notre service technique qui vous aidera 1 Informations importantes 33 34 1 Informations importantes Wichtige Informationen 1 Vor Inbetriebnahme bitte lesen Bitte lesen Sie vor Inbetriebnahme sorgfaltig alle in diesem Benutzerhandbuch enthaltenen Informationen Werden die folgenden Anleitungen nicht gelesen und beachtet so k nnte dies zu unsachgem er oder unvorschriftsm iger Anwendung Bedienung oder Wartung des Ger tes fiihren Auf diese Weise k nnten Anwender gef hrdet Ihr Ger t besch digt oder dessen Betrieb beeintr chtigt werden Auch k nnte sich vorzeitiger Verschlei einstellen und somit die Lebensdauer des Ger tes verk rzen Hierdurch k nnten Ihre Garantieanspr che verfallen B
126. illement 1 Informations importantes 27 28 1 3 4 Rayonnement laser AVERTISSEMENT Lappareil est quip pour la d tection automatique de porte prouvettes classe 1M par quatre diodes laser a mission par la surface a cavit verticale VCSEL Le rayonnement est invisible Ne pas regarder directement dans ce rayon au travers d instruments optiques p ex lentilles verres grossissants et microscopes Regarder dans l ouverture du VCSEL une distance inf rieure 100 mm au travers d instruments optiques peut endommager les yeux L appareil est galement quip d un lecteur de code 2D utilisant un laser a semi conducteur visible comme pointeur pour le r glage de la position de lecture ainsi que de diodes lectroluminescentes DEL haute puissance pour l clairage du champ de lecture des bouteilles de r actifs ainsi que de la poubelle Ne pas d monter le support a bouteille Selon la norme internationale IEC 62471 de s curit photobiologique ce systeme de DEL a une valeur de risque d exposition EHV de 0 91 et est au dessus du groupe de risque 0 exempt risque La distance de risque HD pour le groupe de risque 0 est de 61 cm La distance de risque pour le groupe de risque 1 risque faible est de 20 cm Ne diriger le regard ni dans le rayon laser ou DEL direct nile rayon laser ou DEL r fl chi sur une surface de miroir Sinon des l sions oculaires peuvent en r sulter Ne pas diriger intentionnellement le
127. inazione in caso di utilizzo di materiale a rischio biologico Indica il rischio di morte lesioni gravi a danno dell operatore o danni alle attrezzature a causa di materiale biologico potenzialmente pericoloso Indica il rischio di morte o di lesioni gravi a danno dell operatore a causa di un livello pericoloso di tensione Terminale per conduttore di protezione Il simbolo affisso all interno dello strumento Avvertenza per il personale di assistenza tecnica On acceso alimentazione Off spento alimentazione E necessario consultare la documentazione prima di procedere con l installazione e l utilizzo dell apparecchio 1 Informazioni importanti 1 2 Avvertenze e precauzioni 1 Il separatore autoMACS Pro Separator utilizza tecnologie all avanguardia Si tratta di un dispositivo gestito mediante computer per la separazione automatica delle cellule marcate magneticamente le mediante la tecnologia MACS Il campionatore MACS MiniSampler collegato al separatore autoMACS Pro Separator e rappresenta pertanto un componente del dispositivo di separazione delle cellule Il separatore autoMACS Pro Separator e il campionatore MACS MiniSampler sono concepiti in modo da garantire un funzionamento sicuro dopo l installazione se utilizzati da personale addestrato secondo le norme generali di sicurezza e le istruzioni fornite nel presente manuale d uso Le linee guida riportate nella presente sezione illustrano i potenziali rischi associa
128. ing the autolabeling feature 1 Check that the recommended maximal sample processing time of 90 minutes was not exceeded In the event of an elevated room temperature the cooling capacity of the Chill Racks may be compromised Higher labeling temperatures could cause unspecific binding which may lead to lower purities when using positive selection kits Check whether the correct sample volume was programmed Higher sample input volumes will result in an excess concentration of MicroBeads leading to non specific labeling Recovery of magnetically labeled cells is low Refer to the beginning of this section Magnetically labeled cells have not been retained on the column 9 Troubleshooting Viability of magnetically labeled cells is low Dead cells have been co enriched in the positive fraction due to non specific binding Remove dead cells before separation by using the MACS Dead Cell Removal Kit Refer to section 9 2 2 This is prone to occur if the number of dead cells in the starting material exceeds the number of target cells Overall recovery is low Cells are often lost during washing steps Determine the number of cells immediately before magnetic separation to check whether the low recovery is due to this or due to problems during the cell separation 9 3 2 Depletion Too many cells are retained on the column 1 Magnetic labeling of the cells was non specific In order to block non specific binding use MACS FcR Blocking R
129. ion Informations importantes 1 lire avant utilisation Veuillez imp rativement lire toutes les informations fournies dans ce mode d emploi avant d utiliser l appareil Le non respect de ces consignes peut donner lieu une utilisation une manipulation et une maintenance inappropri e ou incorrecte de votre appareil ce qui pourrait mettre en danger les utilisateurs fournir des r sultats non attendus entra ner le dysfonctionnement ou la d t rioration de l appareil son usure pr matur e et r duire la dur e de vie de l appareil ainsi qu annuler votre garantie Conservez ce mode d emploi dans un endroit s r et accessible aux utilisateurs de l unit de autoMACS Pro Separator s parateur autoMACS Pro Ce chapitre d crit les consignes de s curit et les exigences d installation s appliquant votre s parateur autoMACS Pro Les avertissements et les mesures de pr caution suivantes ont pour objectif de vous aider viter toute blessure corporelle ainsi que toute d t rioration de l appareil 11 Symboles et niveaux de danger Mise en place des consignes de s curit Example CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard Les consignes de s curit informent l utilisateur des risques potentiels pouvant survenir si les avertissements et les mesures de pr caution d crits ci dessous ne sont pas respect s Le pictogramme visible
130. ion Kits based on this strategy are available for fast and convenient isolation of specific cell subsets Two subsequent positive selections Multiparameter sorting with MACS MultiSort MicroBeads allows the performance of two sequential positive selections according to two different markers Labeling cells with MACS MultiSort MicroBeads specific for the first marker allows the first positive selection After the separation the cells are incubated with the MultiSort Release Reagent which enzymatically removes the MultiSort MicroBeads from the cells In the next step the target cells are magnetically labeled with MACS MicroBeads directed against the second marker and again subjected to positive selection 6 Cell separation using the autoMACS Pro Separator 117 118 Note When using the program Posselwb the whole blood sample will be diluted to the 3 fold of the starting volume 6 1 3 autoMACS Pro separation wash and maintenance programs 6 1 3 1 Separation programs The autoMACS Pro Separator provides a selection of twelve pre set separation programs The appropriate program is generally chosen depending on the separation strategy the target cell frequency and the level of antigen expression A decision tree on what cell separation strategy to follow is located on the last page of this chapter section 6 3 Positive selection programs Possel Positive selection in standard mode for the isolation of cells with freq
131. ire de services Miltenyi Biotec agr d effectuer tous les contr les de s curit requis par la proc dure de r paration afin de garantir que l appareil est parfaitement op rationnel Utilisez uniquement les options et les mises jour recommand es par Miltenyi Biotec 1 5 3 Nettoyage D branchez le s parateur autoMACS Pro avant le nettoyage N utilisez pas d agents nettoyants liquides ou en a rosol utilisez toujours un chiffon humide 1 5 4 Mati res dangereuses WARNING Risk of loss of life if biohazardous material is used Wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes Operate device in a safety hood Decontaminate device after spilling of biohazardous material Si une mati re nocive pour l organisme est ou a t utilis e l op rateur doit choisir et porter un quipement de protection individuelle conforme aux avertissements et aux pr cautions pour les substances utilis es Portez des gants de protection des v tements de protection et des lunettes de s curit afin d viter tout contact avec la peau et les yeux Prot ger aussi la bouche et le nez des a rosols qui pourraient provenir du syst me Un quipement de s curit d fectueux ou inad quat peut mettre l op rateur en danger Le s parateur autoMACS Pro doit tre manipul dans un couvercle de protection si des mati res dangereuses ou inconnues sont trait es Si une mati re dangereuse a
132. ithin two weeks may affect purity Check whether the autoMACS Pro Separator has been set to Sleep Store correctly Inappropriate storage or allowing the autoMACS Pro Separator fluidic system to run dry will affect the columns and therefore the purity Make sure that the appropriate Running Buffer has been used Perform the Qrinse program If pumps are filled with air during the process refer to section 9 1 4 Perform a test run using PBS as a mock sample and check output volumes For correct volumes refer to table 6 1 in section 6 1 3 1 If volumes are not correct go to section 9 2 1 Run the Safe program refer to section 4 7 If the problem persists contact Miltenyi Biotec Technical Support 9 Troubleshooting When using the autolabeling feature 9 1 Check whether the uptake needle is positioned in the middle of the sample tube during the mixing procedure If not mixing will be inadequate 2 Check whether the reagent volume contents of reagent vial is sufficient for labeling 3 Check that the recommended maximal sample processing time of 90 minutes was not exceeded In the event of an elevated room temperature the cooling capacity of the Chill Racks may be compromised Higher labeling temperatures could cause non specific binding which may lead to lower purities when using positive selection strategies 9 2 4 Low recovery of isolated cells 1 Low recoveries and low purities may be due to similar problems Go to section
133. ives the pump during operation 7 Replace pump head with the spare part 8 Reassemble in reverse order Take care not to pinch the tubing at the bottom left when pushing the bottom cover back into place 7 Maintenance Note The hydrophobic air filters should be exchanged once a year 7 4 4 Exchange of hydrophobic air filter Hydrophobic air filters 0 2 um are attached to the bottle closures to vent the liquid bottles and to prevent release of aerosols To avoid clogging of the filters and to prevent contamination of liquids air filters should be exchanged if they come into direct contact with any liquid i e become wet They also should be exchanged once a year to avoid clogging through dust deposits 1 Hydrophobic air filters are attached to all fluid bottles Figure 7 31 Hydophobic air filter attached to a fluid bottle 2 Unscrew the hydrophobic air filter from the fluid bottle Replace the new filter in a similar manner 7 5 Exchange of the tubing system If there is any leakage in the tubing system the affected tubing part should be exchanged Please note that each tubing has a specific length and should be exchanged exclusively with the corresponding spare part 1 Switch OFF and unplug the instrument 2 Remove the affected tubing by loosening the tube connectors using the black wrench 3 Replace the tubing with the correct part 4 Pull back the connector from the tubing so that the tubing can be easi
134. jos de todas las piezas m viles del separador autoMACS Pro y de los accesorios para evitar aplastamientos lesiones de corte o da os en el dispositivo No tocar las bombas de fluido ni ajustar la tuber a durante la operaci n del dispositivo Desactivar siempre el dispositivo antes de ajustar cualquier parte del sistema de fluidos El procedimiento debe pararse o interrumpirse antes de maniobrar los accesorios p ej el MACS MiniSampler o antes de colocar o retirar los tubos del soporte de tubo colocado en el sampler No omitir ninguna de las medidas o los dispositivos de seguridad Figura 1 2 El c rculo abierto muestra la se al de advertencia para el peligro de aplastamiento o corte 1 Informaci n importante 63 64 1 3 4 Radiaci n l ser ADVERTENCIA El dispositivo est equipado de cuatro unidades l ser de emisi n superficial de cavidad vertical VSCLs para la detecci n automatizada de reactivos clase 1M La radiaci n no es visible No mire directamente con instrumentos pticos por ejemplo gafas lupas y microscopios Puede ser peligroso para el ojo mirar el puerto VSCL con instrumentos pticos a una distancia de 100 mil metros El dispositivo tambi n est equipado de un lector de c digo 2D que utiliza un l ser semiconductor visible como indicador de blanco para ajustar la posici n de lectura y diodos electroluminosos de gran alcance LED para iluminar el rea de la lectura las botellas de fluidos
135. k nnen ACHTUNG Der autoMACS Pro Separator ist ausschlie lich ausgelegt f r den Betrieb in Innenr umen Bitte benutzen Sie das Ger t nicht in ausgewiesenen Gefahrenzonen wie etwa sauerstoffangereicherten Arbeitsumgebungen rtliche Arbeitsschutzbestimmungen Laborrichtlinien Sicherheitsnormen und Unfallverh tungsvorschriften m ssen auf jeden Fall beachtet werden Wenden Sie sich an die rtlichen Beh rden und Ihren Stromversorger f r weitere Informationen zur Stromversorgung Geb udeinstallationen Wartung und Sicherheit f r die Installation dieses Ger tes Wenn Sie Sicherheitsbedenken in Bezug auf die Ger tenutzung haben setzen Sie sich bitte mit Ihrem autorisierten Miltenyi Biotec Service Provider in Verbindung oder kontaktieren den Miltenyi Biotec Customer Support 1 Wichtige Informationen 37 38 1 3 Allgemeine Sicherheitshinweise Um m gliche mit dem Betrieb des autoMACS Pro Separators verbundene Sicherheitsrisiken zu verringern beachten Sie bitte die folgenden allgemeinen Sicherheitshinweise Eine Nichteinhaltung dieser Vorsichtsma nahmen k nnte Feuer Gesundheitssch den und oder Sch den am Ger t verursachen 1 3 1 Gefahr des Stromschlags und der Ausbreitung von Feuer WARNING Hazardous voltages Risk of loss of live severe personal injury Unplug before cleaning Do not use if device is opened or damaged if liquids have been spilled into device if objects device through ventilation slo
136. k the staining reagent for its specificity 9 5 Contamination of tubing system 1 Decontaminate the instrument Refer to section 7 3 4 for details 2 Ifthe problem persists call Technical Support 9 6 Problems indicated by error or warning screens Iferrors or warnings are displayed on the screen of the instrument please refer to the following table Ifthe below measures do not clear the fault call Miltenyi Biotec Technical Support To assist in the troubleshooting process please have the instrument serial number and details of the error message at hand i e error number module number file and error line If this is not possible please go to the instrument log list and view the log list details taking care to note the displayed parameters exactly To discuss the table below please call Technical Support 9 Troubleshooting Error Cause code 5 Hardware module is not initialized After a module malfunction an initialization ofthe module is neccessary This might also be a subsequent error if another error has been displayed shortly before 27 Standard valve initialization failed Valve may be blocked or worn and cannot be turned correctly or valve drive is damaged 28 Motor rotation detection failed Valve may be blocked or worn and cannot be turned correctly Otherwise valve or magnet drive malfunction 29 Valve rotation hindered Valve may be blocked or worn and cannot be turned correctly 257 Needle arm movement
137. l funzionamento corretto Leggere le istruzioni fornite nella presente sezione e accertarsi che il luogo prescelto sia adeguatamente predisposto prima di collegare lo strumento alla presa di alimentazione Nel predisporre l ambiente di installazione e nell organizzare la disposizione delle attrezzature tenere in considerazione le precauzioni descritte nella presente sezione in modo da evitare guasti allo strumento e ridurre la possibilit di spegnimenti causati dall ambiente 1 4 1 Accessori di montaggio Non collocare il autoMACS Pro Separator su un piano un carrello un supporto un treppiede o una staffa poco stabile che potrebbe provocare la caduta dello strumento con la possibile conseguenza di lesioni personali gravi o di gravi danni allo strumento Utilizzare unicamente piani carrelli supporti treppiedi o staffe raccomandati da Miltenyi Biotec o venduti insieme allo strumento Non collocare il autoMACS Pro Separator su sistemi incassati o in spazi angusti come uno scaffale a meno che non siano stati concepiti specificamente per accogliere lo strumento e a condizione che vi sia una ventilazione adeguata e che siano state seguite le istruzioni di montaggio dello strumento 1 Informazioni importanti 53 54 1 4 2 Circolazione dell aria Lo strumento non deve essere collocato in prossimit di radiatori stufe o altri dispositivi inclusi gli amplificatori che producono calore Accertarsi che via sia un adeguata circola
138. l lato sinistro specifica il tipo di rischio Il livello di rischio in alto classifica il tipo di pericolo come indicato in precedenza Il livello il tipo e la fonte del pericolo nonch le possibili conseguenze i divieti e le misure da adottare vengono indicati come segue 1 Informazioni importanti 47 48 WARNING CAUTION o OP gt EEE Al Simboli e livelli di pericolo Di seguito viene presentato un glossario illustrato che descrive i simboli utilizzati nel presente manuale d uso e sul autoMACS Pro Separator AVVERTENZA indica una situazione pericolosa che potrebbe comportare lesioni gravi o morte qualora non venga evitata ATTENZIONE indica una situazione pericolosa che potrebbe comportare lesioni di grado lieve o moderato qualora non venga evitata Attenzione consultare il manuale d uso per ulteriori istruzioni e procedere con cautela Le avvertenze includono il rischio di danni alle attrezzature gravi lesioni personali o morte Pericolo di schiacciamento o di taglio Rischio di schiacciamento o di taglio di parti del corpo a causa di componenti meccanici Radiazione Laser Rischio di danni permanenti agli occhi ed alla pelle Forte campo magnetico Il campo magnetico pu interferire con oggetti magnetizzati e dispositivi elettronici o danneggiare i supporti magnetici per le informazioni Rischio di lesioni personali gravi per soggetti portatori di pacemaker o impianti medici elettronici Rischio di contam
139. le di avvertenza per forte magnete permanente 1 3 3 Pericolo di schiacciamento e di taglio CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard ATTENZIONE Non aprire i coperchi di accesso anteriore mentre il dispositivo in funzionamento Non impedire il movimento del braccio automatizzato e degli accessori hardware durante il funzionamento Tenere le dita ecc lontano da tutte le parti in movimento del dispositivo autoMACS Pro Separator e degli accessori hardware per evitare lesioni da schiacciamento e ferite da taglio o danni al dispositivo Non toccare le pompe dei liquidi e non spostare i tubi mentre il dispositivo in funzione Spegnere sempre il dispositivo prima di regolare eventuali parti del sistema della fluidica Interrompere o sospendere sempre una procedura prima di maneggiare gli accessori hardware ad es il campionatore MACS MiniSampler o prima di caricare rimuovere le provette dal portaprovette collocato sul campionatore Non bypassare alcun dispositivo o misura di sicurezza Figura 1 2 Il cerchio indica il segnale di avvertenza per il pericolo di schiacciamento e di taglio 1 Informazioni importanti 51 52 1 3 4 Radiazioni laser AVVERTENZA L apparecchio dotato di quattro laser a emissione superficiale a cavit verticale VSCL per il rilevamento automatizzato classe 1M La radiazione non visibile Non osservare direttame
140. lease refer also to section 1 3 4 of the autoMACS Pro Separator user manual for associated warning and precautionary information 2D code reader Figure 10 7 Expanded view of the autoMACS Pro Separator 2D code reader barcode reader 192 10 Technical data and specifications 10 2 Technical data and specifications 1 0 of the autoMACS Pro Separator Technical data Model autoMACS Pro Separator Color Blue orange Footprint 605x343 mm WxD Footprint with MiniSampler 605x455 mm WxD Height 392 5 454 mm adjustable touchscreen Weight 25 kg Input voltage 100 240 VAC 50 60 Hz Power consumption 200 VA Fuses 2xT4A 250 Programs 12 preset Sample volume input 0 25 50 mL Sample volume output 0 5 52 mL Column capacity 4x10 cells sample 2x10 magnetically labeled cells sample 15 mL of whole blood RS232 Interface Pins 1 4 6 9 NC labeled COM Pin 2 RXD Pin 3 TXD Pin 5 GND Pin 7 RTS Pin 8 CTS RS232 Interface Pins 1 4 6 7 8 9 NC labeled RS232 AUX Pin 2 RXD Not in use Pin 3 TXD Pin 5 GND RS232 Interface DC Output Pins 4 6 NC labeled RS232 BCR Pin 1 Input Pin 2 RXD Pin 3 TXD Pin 5 GND Pins 7 8 Shorted Pin 9 5 VDC 0 5 A CAN Bus DC Output Pins 1 4 8 NC labeled External CAN Pin 2 CAN L Pins 3 6 GND Pins 5 9 24 VDC 2A Pin 7 CAN H AC Output Pins 1 2 3 4 5 5 VAC 10kQ labeled Bottle Sensor Pins 6 7 8 14 15 GND Pi
141. ly inserted into the appropriate port 5 Insert the connector cautiously and precisely and fasten it by hand Make sure not to overtighten the screw 6 Plugin and switch ON the instrument Run the program Rinse and check for leakage 7 Run the Calibr_2 program to calibrate the volume control of the instrument For details refer to section 11 2 7 Maintenance 163 164 8 If there are any leakage refer to to section 10 1 2 for advice on how to improve the tubing connection 7 6 Exchange of fuses If the instrument fails to start upon switching it ON or if operation suddenly stops and the screen is dark an exchange of the fuses might be required CAUTION Fuse specifications are given in section 11 2 and on the marking plate on the rear of the instrument close to the fuse holder Do not use other fuses than specified 1 Switch OFF the instrument 2 Unplug the main power cord from the power outlet as well as from the instrument The fuse holder is located below the main power connector on the rear panel of the instrument 3 Pull out the fuse holder from the housing and exchange fuses Figure 7 32 Removing the fuse holder 4 Push the fuse holder back into the housing and reconnect the main power cord 7 7 Rescue procedure Should the separation be interrupted before target cells are eluted it is possible to perform a cell rescue procedure to recover the sample If the instrument can be restarted follow procedure A
142. m that will be performed automatically after each initialization to prime the instrument O_led Activating deactivating fluid bottle illumination The autoMACS Pro Separator has a bottle illumination designed to facilitate monitoring the instrument s status The bottle illumination can be switched ON or OFF O_progs Enabling disabling special separation protocols The O_progs are used to enable or disable special separation programs To enable or disable these protocols perform the following steps 1 Select Option User settings and O_progs 2 Select Run A dialog box will report the current status Option Special separation programs are enabled RE 1012 Figure 4 30 O_progs Enabling or disabling special separation protocols Select Disable to disable special separation programs 3 Select OK if the current reported status should not be changed Alternatively select Disable or Enable to change the status Sepcount Displaying the number of performed separations The Sepcount or Sep counter program is used to display the number of separations that have been on the autoMACS Pro Separator To view this statistic perform the following steps 1 Select Option User settings and Sepcount 2 Select Run A dialog box will report the number of separations 4 An overview of the autoMACS Pro Separator user interface have been 0 separations started on this device E E Figure 4 31 Sepcount viewing the
143. manutenzione offerti da Miltenyi Biotec contattare il rappresentante Miltenyi Biotec locale o consultare il sito www miltenyibiotec com support 1 Informazioni importanti 55 56 IMPORTANTE Qualora si rendano necessari la sostituzione o l impiego di pezzi di ricambio accertarsi che il tecnico utilizzi unicamente pezzi di ricambio originali Miltenyi Biotec o ricambi di altri fabbricanti specificati e raccomandati da Miltenyi Biotec L impiego di pezzi di ricambio non autorizzati pu causare il malfunzionamento del dispositivo e compromettere il risultato della separazione cellulare Miltenyi Biotec non fornisce alcuna garanzia n si assume la responsabilit per eventuali guasti o danni derivanti dall impiego di pezzi di ricambio inappropriati Al termine dell intervento di assistenza o di riparazione chiedere al tecnico Miltenyi Biotec autorizzato di eseguire tutti i controlli di sicurezza previsti dalla procedura di riparazione onde assicurarsi che lo strumento funzioni correttamente Utilizzare esclusivamente le funzioni opzionali e gli aggiornamenti raccomandati da Miltenyi Biotec 1 5 3 Pulizia Staccare il autoMACS Pro Separator dalla presa di corrente prima di eseguire la pulizia Non utilizzare detergenti liquidi o aerosol utilizzare sempre un panno umido 1 5 4 Materiale pericoloso WARNING Risk of loss of life if biohazardous material is used Wear protective gloves protective clothing and safety glasses to preve
144. me and dale j t Be Esa 10 45 Figure 3 15 Select the desired field and use the numeric keypad to modify the date and or time 2 Select Ok to return to the Option menu The procedure can be cancelled at anytime by selecting Cancel 3 Assembly and installation 83 3 3 3 Performa test calibration Note If test calibration 1 During test calibration ensure the correct location of the fails perform a calibration 1 needle arm at the following positions Refer to section 11 2 for Washing station needle should be positioned directly above details the center of the rear opening of the washing station figure 3 16 Chill Rack positions A1 B1 C1 needle should be positioned at the bottom center of the tube figure 3 17 X position needle should be positioned in the middle of the x axis of tube B1 Reagent Rack positions 1 4 needle should be located at the lowest point of the vial to ensure full uptake of the substrate figure 3 18 Figure 3 16 Test calibration of the needle arm position in the washing station a Figure 3 18 Test calibration of the needle arm position in the reagent vial 84 3 Assembly and installation 1 Goto the Option menu Under User settings select Calibr_1 3 Click Run and Test running Calibration 1 Selec made lies Figure 13 19 Instructing the instrument to perform a test calibration 2 Place a Chill 15 Rack containing three 15 mL plastic tubes in
145. n no action is required If the symbol is red the columns must be exchanged The level of the green fill on the column symbol indicates the remaining operation life of the autoMACS Columns Touch the column symbol to activate the column status pop up text box A 111 Figure 4 9 Column status By touching the column symbol a pop up box displays a text report of the column status 4 An overview of the autoMACS Pro Separator user interface 4 3 3 Tube rack status A Figure 4 10 Tube rack status graphic a 5 mL Chill 5 Rack was detected If no tube rack is detected no graphic is displayed Rack detection occurs prior to starting the separation process Before cell labeling and or cell separation is performed the instrument will not attempt to detect the rack Touch the position of the rack status graphic in order to view the tube rack pop up text table RE 11 50 Figure 4 11 Tube rack status text box No rack is detected until cell separation is performed 4 3 4 MACS MiniSampler status Figure 4 12 MACS MiniSampler status graphic Left The MiniSampler was successfully installed Right No MiniSampler was detected If the MACS MiniSampler has been detected correctly a rectangular symbol is displayed If it has not been detected a hand symbol will be displayed in the same field 4 An overview of the autoMACS Pro Separator user interface 97 98 4 4 Separation menu The Separation menu schematically represen
146. n from a mirrored surface Take care during installation of the autoMACS Pro Separator that the path of the laser beam is not at the same height as that of the human eye during operation Do not allow water oil dust or other foreign substances to stick to 2D Code Reader aperture window This may cause read errors Be sure to stop the laser emission before cleaning the scanner Otherwise exposure to the laser may cause eye injury Use a soft dry cloth to wipe any substances from the scanner Do not use alcohol or other cleaning substance The autoMACS Pro Separator is classified as a Class 1M laser product per standard IEC 60825 1 1993 A1 1997 A2 2001 CAUTION Use of controls or adjustments or performance of procedures other than those specified herein may result in hazardous radiation exposure 1 Important information Figure 1 3 Position of lasers Invisible rack detection lasers are located within the rectangle area The 2D code reader visible is located within the open circle 1 4 Secure installation This section describes the requirements your site must meet for safe installation and operation of your autoMACS Pro Separator Read the instructions in this section and ensure that your site is properly prepared before you connect the instrument to its power source When planning your site layout and equipment locations keep in mind the precautions described in this section to help avoid instrument failures and reduce t
147. n gelangt sind sollten vor Entsorgung autoklaviert werden Fl ssigabfall sollte autoklaviert oder unter Verwendung eines f r das jeweilige spezifische Pathogen geeigneten Desinfektionsmittels dekontaminiert werden z B 10 Bleichmittel Isopropylalkohol oder 70 Ethanol 1 Wichtige Informationen Die Entsorgung der Verbrauchsmaterialien muss gem lokal geltender Bestimmungen erfolgen autoMACS Pro Separator SN 0563 Order na 130 092 545 Power Supply 100 240 VAL 50 00 Hz 200 VA Fuses 2XT4A 2504 c Abbildung 1 4 Warnhinweise f r biologische Gefahren befinden sich r ckseitig auf dem autoMACS Pro Separator links und oben auf dem autoMACS Pro Separator Fl ssigkeitsbeh lter rechts 1 5 5 Transport Der autoMACS Pro Separator sollte vorsichtig gehandhabt in der von Miltenyi Biotec bereit gestellten Verpackung transportiert werden Im Ger t k nnen innere Sch den auftreten falls es groBer Ersch tterung ausgesetzt oder fallengelassen wird Sollte wegen Reparatur oder Wartungsleistungen ein R cktransport zum Hersteller notwendig werden dekontaminieren Sie das Ger t vor Versand von jeglichen biologischen Gefahrenstoffen Wenn Sie Fragen zur vorschriftsm igen Dekontaminierung oder zum Versand des Ger tes haben wenden Sie sich bitte direkt an unseren Technical Support 1 5 6 Ger teentsorgung Setzen Sie sich bitte direkt mit unserem Technical Support in Verbindung falls Sie Ihr Ger t entsorgen m
148. n live and dead cells The most common method to discriminate between live and dead cells is based on trypan blue staining and analysis by light microscopy Trypan blue crosses the cell membrane of dead cells and stains the cells Live cells are not stained Propidium iodide 5 0 g mL is most often used for flow cytometry and fluorescence microscopy It crosses the permeable cell membrane of dead cells enters the nucleus and interacts with DNA Therefore the nucleus of dead cells is fluorescently stained Other fluorescent dyes for example DAPI 4 6 diamidino 2 phenylindole can be used depending on the properties of the flow cytometer i e its excitation wavelength capabilities particularly in the UV range When working with fixed cells it is recommended to use the Fixation and Dead Cell Discrimination Kit 130 091 163 for both the cell fixation as well as the discrimination of dead cells The viability can be calculated as follows No of live cells Viability 100 x No of total cells live amp dead 170 8 Quality control of separations performed with the autoMACS Pro Separator Troubleshooting 9 In the unlikely event of problems using the autoMACS Pro Separator Note If general hardware the following section aims to address any relevant issues If the errors occur it is outcome of a cell separation procedure is deemed unsatisfactory recommended to run the this may either be due to incorrect function
149. n merito all impiego sicuro dello strumento contattare il rivenditore Miltenyi Biotec autorizzato o chiamare il servizio di assistenza clienti Miltenyi Biotec 1 Informazioni importanti 49 50 1 3 Precauzioni generali Per ridurre i potenziali rischi associati all utilizzo di autoMACS Pro Separator osservare le precauzioni generali di seguito elencate Il mancato rispetto di tali precauzioni pu comportare il rischio di incendi lesioni personali e o danni allo strumento 1 3 1 Pericolo di scossa elettrica e di incendio WARNING Hazardous voltages Risk of loss of live severe personal injury Unplug before cleaning Do not use if device is opened or damaged if liquids have been spilled into device if objects device through ventilation slots AVVERTENZA dispositivi elettrici presentano il rischio di scosse elettriche Per ridurre i rischi di scossa elettrica non aprire alcun coperchio tranne i coperchi di accesso anteriore di autoMACS Pro Separator n altri accessori hardware forniti da Miltenyi Biotec Tutti gli altri coperchi dello strumento e gli accessori hardware possono essere rimossi esclusivamente da personale autorizzato Prestare particolare attenzione quando si manipolano liquidi Pulire immediatamente eventuali schizzi Non lasciare che i liquidi penetrino all interno del dispositivo Staccare il cavo dell alimentazione prima di pulire manualmente lo strumento autoMACS Pro Separator AVVERTENZA
150. n volume 0 2 mL max final labeling volume 2 mL Max number of samples 5 min first incubation volume 0 2 mL max final labeling volume 6 5 mL 3 Max number of samples 3 min first incubation volume 4 mL max final labeling volume 8 mL When working with fewer cells than the necessary minimal volume resuspend cells in the stipulated minimal volume Table 6 3 Dilution volumes for the first labeling step and MACS Chill Rack specifications for autolabeling including minimal and maximal volumes and cell numbers 6 Cell separation using the autoMACS Pro Separator Maximal total cell number 2x10 2x10 2x10 2x10 2x10 6 5x10 6 5x10 6 5x10 6 5x10 6 5x10 6 2 2 Select appropriate tube rack Three different tube racks are available for processing sample volumes between 0 2 mL and 50 mL Unless otherwise specifically indicated in the Cell Separation Reagent data sheet the magnetically labeled cell samples are resuspended at 10 total cells per 500 uL when using manual labeling The cell numbers and volumes for autolabeling are slightly different Refer to table 6 3 for further details Regarding the tube positioning of the Chill Racks please note that Row A of the Chill Rack holds the sample tubes e Row B of the Chill Rack holds the tubes for the non labeled fractions negative fractions e Row C of the Chill Rack holds the tubes for labeled fractions po
151. nce and setup of the autoMACS Pro Separator Each program is briefly discussed below About About informs the user about the software version serial number of the instrument and other hardware information Calibr_1 Performing calibration 1 of the needle arm positioning This program is used for the calibration of the needle arm to the washing station and tube racks Refer to section 11 2 for more information Calibr_2 Performing calibration 2 of the instrument tubing This program is used for the calibration of the instrument tubing Refer to section 11 2 for more information Check_up Performing a system check up The Check_up program allows the user to perform a system check up It is recommended to use the program if hardware errors occur The program starts after highlighting check_up and pressing Run O_bcr 2D code reader setup configuration and initialization The O_ber program allows the user to setup configure and initialize a recently installed autoMACS Pro Separator 2D code reader 4 An overview of the autoMACS Pro Separator user interface 107 108 Note In case of malfunction during cell separation the option O_init must be disabled in order to perform a cell rescue procedure O_init Optional priming of the instrument at startup By default the autoMACS Pro Separator does not perform a wash program after initialization The option O_init allows the user to add an initial rinse progra
152. next column replacement if the Column exchange program Col_ex has been used for column installation Columns do not have a top or bottom and do not require special orientation in the column holder Test position ORI tube Select action RE 11 32 Fig 3 21 To test a position select Test position To skip the test position select Next To test a previous test position select Previous To end the test calibration select Test end Verify sample tube position Test calibration Control position of needle in ORI tube Fig 3 22 After selecting Test position the Move up and Move down buttons can be use to check the exact positioning of the needle 6 After testing the fourth vial position as the last position select Done Select Done once again to leaf the test calibration Select Cancel to leave the calibration program 7 Test calibration is completed 3 3 4 Installation of the autoMACS Columns Remove the column substitutes and install the autoMACS Columns according to the following instructions 1 Open the front door and note the position of the tubing and autoMACS Column slots in the magnet cover column 1 to the left column 2 to the right Column 2 Figure 3 23 Position of the autoMACS Columns 3 Assembly and installation 2 Ensure that the fluid bottles are filled with solutions 3 3 Using the touch screen select Option and Special from the Main submenu 4 Select C
153. nformations importantes pacemakers dispositifs de mesure et de contr le ordinateurs et 1 montres Ces l ments peuvent tre affect s ou endommag s par le champ magn tique Figure 1 1 Symbole d avertissement pour aimant puissant permanent 1 3 3 Risque d crasement et de cisaillement CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard ATTENTION N ouvrez pas les capots frontaux pendant le fonctionnement de l appareil Ne bloquez pas le mouvement du bras automatique ni des pi ces correspondantes pendant le fonctionnement N approchez pas vos doigts etc des pi ces mobiles du s parateur autoMACS Pro et des pi ces correspondantes afin d viter toute blessure ainsi que toute d t rioration de l appareil due l crasement et au cisaillement Ne touchez pas les pompes liquide et ne modifiez pas les tuyaux pendant le fonctionnement de l appareil D connectez toujours l appareil avant de modifier une partie du syst me fluidique Stoppez ou annulez toujours le processus avant de manipuler les appareils suppl mentaires comme le mini chantillonneur MACS ou de charger retirer les prouvettes du r telier prouvettes plac sur l chantillonneur Ne bloquez pas les dispositifs de s curit et respectez les mesures de s curit Figure 1 2 Le symbole d avertissement entour d un cercle blanc indique un risque d crasement et de cisa
154. ng cells that tend to aggregate e g tissue cells it may be helpful to dilute the sample 1 2 2 Remove the sample tube Wipe the uptake port needle with a tissue soaked with 70 ethanol Run the Qrinse program 3 Check for cell clumps in the tubing system If cell clumps are suspected run the Safe program 5 Run the Qrinse program Check tubing system and pump syringe for air inlet 6 Make sure that all tubing are connected properly especially the tubing connected to the columns and to the buffer bottles 7 Verify that the uptake port needle is connected correctly and that no air inlet is possible Tighten if necessary 8 Make sure that both columns are installed correctly 9 Clean pump syringe as described in section 7 3 2 to remove salt deposits Salt crusts might allow air to enter the fluidic system 10 Check whether the hydrophobic air filters connected to the bottles are clogged If filters are clogged replace them with new hydrophobic air filters 11 Run the Check_up program to identify hardware malfunctions 12 If problem persists call Technical Support 9 2 6 Reagent vial runs dry As the liquid levels in the reagent vials are not controlled by the system before uptake of reagent it will not be detected if the reagent volume provided in the vial is sufficient for the current separation process or not Ensure enough reagent is present in all vials for all programmed processes As the residual volume in the vial is
155. nicos Riesgo de contaminaci n si se usan sustancias peligrosas biol gicamente Indica el riesgo de muerte lesiones graves al operario del aparato o da os al equipo debido a sustancias biol gicas potencialmente peligrosas Indica el riesgo de muerte o lesiones graves al operario del aparato debido a un voltaje peligroso Corriente continua El equipo est marcado con el tipo de suministro corriente continua Encendido suministro Apagado suministro La documentaci n del equipo debe ser consultada antes de proceder con la instalaci n y operaci n del sistema 1 Informaci n importante 1 2 Advertencias y precauciones 1 El separador autoMACS Pro emplea tecnologia puntera Se trata de un dispositivo controlado por ordenador desarrollado para la separaci n de c lulas magn ticamente rotuladas con la tecnologia bee MACS El MACS MiniSampler conecta el separador autoMACS Pro y forma de esta manera parte del dispositivo de separaci n de c lula El separador autoMACS Pro y el MiniSampler est n dise ados para funcionar con seguridad una vez instalados y siempre que sean manejados por personal cualificado de conformidad con las normas generales de seguridad y las instrucciones contenidas en este manual del usuario Las directrices del presente cap tulo explican los potenciales riesgos asociados al manejo del instrumento y suministran informaci n importante a fin de reducir dichos riesgos al m nimo Si sigue cuidadosamen
156. ning Buffer It is recommended to use this program between separations of cells with normal frequency Rinse is an extensive rinsing program that uses Washing Solution and Running Buffer It is recommended to use this program between separations of rare cells e g stem cells the separation of cells from different species and is mandatory between whole blood separations Clean is an optional very extensive rinsing program that uses storage solution Washing Solution and Running Buffer It may be used after whole blood and bone marrow applications Sleep is mandatory as the last wash program before overnight storage Upon completion of this program the fluidic system contains 70 ethanol Maintenance programs Safe is designed for decontamination of the fluidic system For details refer to section 7 Upon completion of this program the fluidic system contains Running Buffer The program includes an exchange of autoMACS Columns Store should be applied to prepare the instrument for long term storage Upon completion of this program the fluidic system contains 70 ethanol For details refer to section 7 Col_ex is used for column exchange Upon completion of the Col_ex program the fluidic system contains Running Buffer Program Washing Running Storage Qrinse Rinse Clean Sleep Safe Store Col_ex Solution Buffer 96 mL 96 mL 96 mL 96 mL 96 mL 96 mL 48 mL 48 mL 48 mL 96 mL 96 mL soluti
157. nmediatamente el separador autoMACS Pro desench felo y p ngase en contacto con un proveedor de servicios de Miltenyi Biotec autorizado o con el servicio t cnico de Miltenyi Biotec Est expresamente prohibido utilizar un instrumento estropeado o cuyo cable de corriente est da ado 1 3 2 Campo magn tico intenso WARNING Extremely strong magnetic fields Risk of severe personal injury to persons carrying pacemakers or electronical medical implants stay clear Keep all magnetically sensitive objects away from device ADVERTENCIA El separador autoMACS Pro esta equipado con un iman extremadamente potente Mantenga todos los portadores de informaci n magn tica tarjetas de cr dito cintas magn ticas y medios de almacenamiento y todos los equipos electr nicos aud fonos marcapasos instrumentos de medici n y control ordenadores y relojes a una distancia m nima de 20 cm de la cubierta magn tica El campo magn tico puede alterar o da ar dichos objetos 1 Informaci n importante Figura 1 1 Se al de advertencia para im n permanente potente 1 3 3 Peligro de aplastamiento y corte CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard ATENCI N No abrir las cubiertas de acceso frontal durante el servicio del dispositivo No obstruir el movimiento del brazo automatizado y de los accesorios durante la operaci n Mantener los dedos etc le
158. ns 9 10 11 12 13 Input 10 Technical data and specifications 193 194 CAN Bus Pins 1 4 5 8 9 NC labeled CAN1 or CAN2 Pin 2 CAN L Pins 3 6 GND Pin 7 CAN H Model MACS MiniSampler Footprint without lid 182x148x47 mm WxDxH Footprint with lid 280x153x172 mm WxDxH Weight 1 5 kg Input voltage 24 VDC Current 0 8A Sub D9 interface Pins 1 4 8 NC with shielding Pin 2 CAN L Pins 3 6 GND Pins 5 9 24 VDC 2A Pin 7 CAN H Depending on the type of power plug chosen the depth increases by 27 mm or 62 mm Table 10 1 Technical data The autoMACS Pro Separator is labeled as a protection class instrument and must be plugged into a grounded power outlet refer to section 1 The MACS MiniSampler is labeled as a protection class III instrument and must only be plugged into the connector labeled with External CAN of the autoMACS Pro Separator refer to section 1 2 The main power supply cord and plug of the instrument shall comply with following specifications USA and Canada only UL listed and KAM cord minimum type SJ minimum 18 AWG 3 conductors Rated for a minimum temperature of 60 C Provided with grounding type NEMA 5 15P attachment plug rated 125 Vac 10 A Opposite end terminates in IEC 320 style connector rated 125 Vac 10 A Conditions of operation 15 30 C with 0 85 humidity non condensing at a maximum altitude of 2000 m Supply voltage fluctuations up to 10 of th
159. nt Refer to section 11 2 for details 7 3 3 Clean the washing station The washing station is designed for the automated rinsing of the outlet and uptake ports as well as surface cleaning of the uptake port needle The washing station should be cleaned as necessary to remove spills and salt crusts 1 Switch OFF and unplug the instrument 2 Make sure that the needle arm is in the uppermost position 3 Swivel the front cover to the right side swivel the cover of the washing station to the left side The cover can be removed by lifting it 4 Press the tubing clamp on the right hand side of the washing station and pull to remove the tubing 7 Maintenance Note Do not autoclave the washing station or wash using a dish washer Figure 7 10 Remove the peristaltic pump tube from the washing station 5 Unscrew the thumb screw that attaches the washing station to the instrument y Figure 7 11 Unscrew the thumb screw 6 Pullout the washing station 7 Clean the washing station by soaking it in 10 bleach and 70 ethanol for 15 min each Optionally sonicate it in water Rinse with distilled water 8 Reassemble the unit in reverse order 9 Runa Safe program 7 3 4 Instrument decontamination The Safe program is a disinfectant procedure which uses MACS Bleach Solution for cleaning and decontamination of the autoMACS Pro Separator Depending on the level of use and general instrument maintenance it is recommended to
160. nt contact with skin and eyes Operate device in a safety hood Decontaminate device after spilling of biohazardous material Qualora si utilizzi o sia stato usato del materiale a rischio biologico l operatore deve indossare dispositivi di protezione personale conformi alle avvertenze e alle precauzioni relative alle sostanze impiegate Indossare guanti protettivi indumenti e occhiali di protezione per prevenire il contatto con la pelle e gli occhi Proteggere sempre la bocca ed il naso dagli aereosol che possono provenire dal sistema L impiego di dispositivi di protezione difettosi o inadeguati rappresenta un rischio per l incolumit dell operatore Lo strumento autoMACS Pro Separator va utilizzato con uno schermo protettivo in caso di trattamento di materiali sconosciuti o pericolosi Qualora sia stato usato o versato del materiale pericoloso il sistema deve essere accuratamente decontaminato 1 Informazioni importanti Ispezionare sempre il sistema fluidico il sistema di tubi le bottiglie 1 e le relative chiusure le valvole le colonne il dispositivo di diluizione e gli aghi prima di accendere lo strumento In caso di perdita di liquidi sostituire le parti danneggiate prima di accendere lo strumento Nel caso non sia possibile sostituire le parti danneggiate ei disconnettere lo strumento e non utilizzarlo Il malfunzionamento di parti che contengono materiale potenzialmente infettivo o liquidi venuti in contatto con tali mate
161. nte il laser con strumenti ottici p es lenti lenti di ingrandimento e microscopi Osservando la porta VSCL a una distanza inferiore a 100mm con uno strumento ottico si potrebbero danneggiare i propri occhi L apparecchio dotato anche di un lettore di codici 2D dotato di un laser a semiconduttore visibile come puntatore per regolare la posizione di lettura e di potenti diodi luminosi LED per illuminare la zona di lettura e le bottiglie per i fluidi cos come quella per la bottiglia del liquido di scarto Non rimuovere i cestelli per le bottiglie Rispetto allo standard IEC 62471 la lampada ha un valore di rischio relativo all esposizione EHV pari a 0 91 ed in eccesso rispetto al Gruppo Esente da Rischio La distanza a rischio HD per il Gruppo Esente da Rischio 61 cm La distanza a rischio per il Gruppo a Rischio 1 di 20 cm Non guardare direttamente le radiazioni di diodi laser o LED o le radiazioni di diodi laser o LED riflesse da una superficie a specchio In caso contrario si potrebbero danneggiare i propri occhi Non rivolgere intenzionalmente il raggio laser verso altre persone Non smontare modificare o rimuovere le sorgenti di radiazioni laser o LED installate o le loro staffe di montaggio Le sorgenti di radiazioni laser o LED non cessano automaticamente di emettere radiazioni una volta smontate Le radiazioni di unit smontate possono essere causa di lesioni agli occhi Fare attenzione al percorso del ra
162. number of performed separations In this example no separations have been performed using this instrument 3 Select OK to return to the Option menu Set_time Setting the time and date To set the time and date perform the following steps 1 Select Option User settings and Set_time Select Run 2 Highlight either the time or date fields by touching the display 3 Use the keypad to change the date or time accordingly 4 Select OK to return to the Option menu The procedure can be cancelled at anytime by selecting Cancel Valve_ex Exchanging the instrument valves This program is used for valve exchange Use of this program turns the valve to the exchange position for removal Valves may require periodical exchange Refer to section 7 4 for more details 4 An overview of the autoMACS Pro Separator user interface 109 110 4 An overview of the autoMACS Pro Separator user interface Materials required 3 This chapter describes the consumables and accessories required for the operation of the autoMACS Pro Separator IMPORTANT Please be advised that the autoMACS Pro Separator is specified for use with MACS MicroBeads autoMACS Columns and other genuine Miltenyi Biotec consumables and accessories only Please only use consumables and accessories recommended by Miltenyi Biotec Failure to use recommended consumables and accessories may result in inaccurate results instrument malfunction or damage premature wear and reduced life
163. ny programs beyond this position The autoMACS Pro Separator automatically performs the Sleep program if the device is inactive for more than six hours Ensure enough storage solution is available The fluid bottle illumination can be switched ON OFF 1 Select the Option menu and User settings 2 Highlight O_led and press Run The fluid bottle illumination can now be enabled or disabled 6 2 6 Shut down of the instrument 6 2 6 1 Sleep program for regular use 1 Combine a separation program and the Sleep program for the last position in the programming field 2 Upon completion of the Sleep program switch OFF the autoMACS Pro Separator using the main power switch 6 2 6 2 Store program for long term storage To store the autoMACS Pro Separator for a period longer than two weeks the tubing system should be cleaned and the columns should be replaced with column substitutes The Store program automatically performs the cleaning procedure and prompts the user to install column substitutes 1 Select Option and Special 2 Select Store and press Run The system will be rinsed automatically Rinsing RE 13 35 Figure 6 33 Running the Store program 3 Install the column substitutes refer to section 3 3 4 for details on column exchange 4 Select Done 5 Switch OFF the autoMACS Pro Separator using the main power switch 6 Cell separation using the autoMACS Pro Separator 6 2 6 3 Shutdown button 6 1 Press the sh
164. of an electric shock To reduce the risk of an electric shock do not open any cover other than the front access covers of the autoMACS Pro Separator nor any other accessory hardware supplied by Miltenyi Biotec All other covers of the instrument and accessory hardware are to be removed by authorized personnel only Special care must be taken while handling fluids Clean up spillages immediately Do not allow fluids to enter the interior of the instrument Unplug the power cord before manually cleaning the autoMACS Pro Separator WARNING A potential risk exists if an opened dropped or damaged autoMACS Pro Separator is used if liquids are spilled into the instrument if an object has entered the instrument through the ventilation slots or if an object has been dropped into the instrument If flames or smoke appear immediately switch OFF the autoMACS Pro Separator unplug the instrument from the electrical outlet and contact an authorized Miltenyi Biotec service provider or the Miltenyi Biotec Customer Support team Use of a damaged instrument or an instrument with a damaged power cable is expressly prohibited 1 3 2 Strong magnetic field WARNING Extremely strong magnetic fields Risk of severe personal injury to persons carrying pacemakers or electronical medical implants stay clear Keep all magnetically sensitive objects away from device WARNING The autoMACS Pro Separator is equipped with an extremely powerful magnet Keep
165. of the instrument or to Check_up program Select inappropriate sample preparation Both of these factors are the Options menu from discussed in this section At the end of this section a list of the upper navigation bar numerically encoded errors and warning messages are presented Then select User settings along with user actions for troubleshooting Highlight Check_up and press Run This section addresses problems that are not indicated by a warning or error screen but might occur during the separation or rinsing The program automatically programs Identify the problem and refer to the appropriate section hay analyzes the functionality of moving hardware components A report is 9 1 Hardware problems not indicated dp tame analysis by a warning or error screen of each single component The procedure can be 9 1 1 Column leakage canceled after each step 1 Ifafreshly installed autoMACS Column shows signs of leakage or continued by pressing check if the column is installed properly The column should be OK Following hardware inserted precisely into the column connector and fastened to the components are analyzed point of resistance If this is not the case loosen the column dilutor valve valves 1 4 connector insert the column precisely and tighten the connector peristaltic waste pump again magnet 1 2 needle arm movement along the z axis 2 Run the Qrinse program Select the Separation menu from the and the y axis and MACS u
166. ol_ex column exchange from the Detail panel 5 Select Run to start the Col_ex program RE 130 Figure 3 24 Selecting Col_ex from the Option menu 6 Wait until the instrument prompts you to exchange the autoMACS Columns before proceeding 7 Using both hands take the top and bottom of the column 1 substitute and pull gently but firmly to remove it from its slot Figure 3 25 Removal of the column substitutes from the column slots 3 Assembly and installation 87 88 Note The program will then proceed to wash the columns with autoMACS Running Buffer Check that the columna are securely fastened to the column connectors and that no buffer is leaking 8 Place a wide mouth bottle under the column substitute Hold the column substitute in one hand and gently unscrew the bottom column connector counter clockwise Unscrew the top column connector while holding the column over the bottle to catch any fluids Store the column substitute in the autoMACS Pro Separator Starter Kit box Figure 3 26 Unscrewing the top column connector for column removal 9 10 11 12 13 14 Insert one end of a fresh column into the bottom column connector and gently screw in the column by turning it clockwise until you feel a resistance Point the column towards the top of the instrument and screw in the top column connector Align the column with the top column connector sitting on the guiding of the magnet cover Pre
167. on MACS Bleach Time Solution Not including the time required for column exchange Not including the time required for disconnecting and reconnecting bottle tubing 1 5 min 4 min 7 min 5 min 21 min 8 min 6 min Table 6 2 Liquid usage and time of wash and maintenance programs 6 Cell separation using the autoMACS Pro Separator 6 121 122 6 2 Experimental considerations 6 2 1 Preparation of samples In order to obtain optimal separation results some crucial points must be considered regarding sample preparation 6 2 1 1 Prepare single cell suspensions Cell aggregates may contain mixtures of target and non target cells and therefore can impair the separation results MACS Separation Buffer MACS Running Buffer should be used during sample handling steps to minimize the risk of cell aggregation Resuspend cells carefully after centrifugation Moreover large cell aggregates may interfere with the separation process and may cause pressure variations in the autoMACS Pro fluidic system It is recommended to use Pre Separation Filters 30 um 130 041 407 or Pre Separation Filters 70 um 130 095 823 to remove cell clumps that may clog the column Dead cells and cell debris may bind non specific to MACS MicroBeads antibodies and antibody conjugates To remove dead cells it is recommended to use density gradient centrifugation or the Dead Cell Removal Kit 130 090 101 For specific recommenda
168. on die falls sie nicht vermieden wird zu leichteren oder mittelschweren Verletzungen f hren kann Achtung schlagen Sie im Benutzerhandbuch fir weitere Anweisungen nach und gehen Sie umsichtig vor Warnhinweise deuten auf das Risiko von Besch digungen des Ger tes bzw der Ausr stung die Gefahr von schweren Verletzungen oder Lebensgefahr hin Quetsch und Schergefahr Durch mechanische Gef hrdungen k nnen Br che oder Quetschungen von K rperteilen verursacht werden Laserstrahlung Gefahr von schweren Augen und Hautverletzungen Starkes Magnetfeld Das Magnetfeld kann auf andere magnetisch aufladbare Gegenst nde und elektronische Ger te st rend einwirken oder magnetische Datentr ger besch digen F r die Tr ger eines Herzschrittmachers oder von eletronischen Implantaten besteht die Gefahr schwerer Verletzungen Gefahr der Kontaminierung bei Verwendung von biologischen Gefahrenstoffen Dieses Symbol weist auf Lebensgefahr das Risiko schwerer Verletzungen f r den Bediener des Ger tes oder Sch den am Ger t selbst hin die durch Arbeit mit m glicherweise gef hrlichen biologischen Substanzen entstehen k nnen Weist auf Lebensgefahr oder das Risiko schwerer Verletzungen f r den Benutzer des Ger tes aufgrund gef hrlicher Stromspannung hin Schutzleiteranschluss Dieses Symbol ist innerhalb des Ger ts angebracht und stellt einen Hinweis f r das Servicepersonal dar Stromversorgung AN Stromversorgung AUS Vor
169. on potentially bending surfaces The angle connectors reduce the depth of the instrument to 455 mm including MACS MiniSampler for placement in a location with limited space The operating environment should be stable and vibration free dust free sufficiently ventilated and free from sources of electromagnetic radiation Before operating the autoMACS Pro Separator for the first time carefully read the user manual and contact your local Miltenyi Biotec representative for assistance The following components are included in the delivery of the autoMACS Pro Separator Starter Kit 130 092 545 autoMACS Pro Separator MACS MiniSampler MACS Reagent Rack 4 e 1 MACS Chill 5 Rack for 5 mL tubes e 1 MACS Chill 15 Rack for 15 mL tubes e 1 MACS Chill 50 Rack for 50 mL tubes Software Barcode reader 5x2 autoMACS Columns 2x2 column substitutes autoMACS Pro buffer combination User manual and short instructions One year warranty 3 Assembly and installation 75 76 Note The top compartment holds the autoMACS Pro Separator user manual the short instructions and various bags containing accessories Carefully remove these parts 3 1 Unpacking and installing the autoMACS Pro Separator Read through the following instructions carefully before commencing the installation procedure Before opening the transportation box check for any visible external damage to the box Check also to see if
170. ore the autoMACS Pro Separator for a period longer than two weeks the tubing system should be cleaned and the columns should be replaced with column substitutes The Store program automatically performs the cleaning procedure and prompts the user to install column substitutes 1 Select Option and Special 2 Select Store and press Run The system will be rinsed automatically 7 Maintenance Rinsing A 1 Figure 7 4 Running the Store program 3 Install the column substitutes refer to section 3 3 4 for details on column exchange 4 Select Done 5 Switch OFF the autoMACS Pro Separator using the main power switch 7 3 Periodic maintenance 7 3 1 Column exchange autoMACS Pro Separator Columns must be exchanged every two weeks or after 100 separations whichever comes first Refer to section 3 3 4 for more details 7 3 2 Clean the pump syringe The pump syringe must be cleaned every 1 3 months 1 Goto the Option menu Select User settings and Syrin_ex from the Main and Detail submenu respectively 2 Press Run to start the program 3 Loosen the plunger lock screw at the bottom of the plunger as prompted on the screen Press OK The plunger holder will move to the lowest position 7 Maintenance 149 150 Figure 7 5 Top Unscrewing the plunger lock screw Bottom The plunger holder is in the lowest position 4 Thesceen prompts to switch the instrument OFF replace the syringe and tighten the plunger scr
171. orrect servicing or repair of your autoMACS Pro Separator can cause hazards to users lead to unpredictable results instrument malfunction or damage premature wear and reduced life time of the instrument and may void your warranty Inquire with your local Miltenyi Biotec representative about Miltenyi Biotec s extensive instrument service and support arrangements or refer to www miltenyibiotec com support IMPORTANT When replacement or spare parts are required make sure that the service provider uses only genuine Miltenyi Biotec parts or third party parts specified and recommended by Miltenyi Biotec Using unauthorized replacement or spare parts can cause malfunction of the instrument and impair cell separation results Miltenyi Biotec does not honor any warranty or accept any responsibility for instrument failure or damages resulting from the use of inappropriate replacement or spare parts After completing any service or repair work have your authorized Miltenyi Biotec service provider perform all safety checks required by the repair procedure to ensure that the instrument is in proper operational condition Only use options and upgrades recommended by Miltenyi Biotec 1 Important information 19 20 1 5 3 Cleaning Unplug the autoMACS Pro Separator from the outlet before cleaning Do not use liquid or aerosol cleaning agents always use a damp cloth 1 5 4 Hazardous material WARNING Risk of loss of life if biohazardou
172. pareil qu une ventilation appropri e est garantie et que les instructions de montage relatives l appareil ont t respect es 1 linformations importantes 29 30 1 4 2 Circulation de l air Ne placez pas l appareil a proximit de radiateurs de registres de chaleur de fours ou d autres pi ces d quipement amplificateurs qui produisent de la chaleur Veillez a ce que suffisamment d air puisse circuler autour du s parateur autoMACS Pro au moins 15 cm de chaque c t pendant le fonctionnement afin de garantir le refroidissement ad quat de l appareil vitez d exposer l appareil un ensoleillement direct Les encoches et les fentes de l appareil sont pr vues pour la ventilation et ne doivent jamais tre bloqu es ou recouvertes car elles assurent le fonctionnement fiable du s parateur autoMACS Pro et prot gent l appareil de la surchauffe N introduisez jamais d objet tranger dans l appareil par l une des fentes 1 4 3 Eau et humidit N utilisez pas l appareil dans un endroit humide ou mouill vitez l humidit et la condensation et prot gez la machine des claboussures 1 4 4 Produit reli la terre L appareil est quip d une fiche lectrique trifilaire avec mise la terre dont la troisi me borne est pr vue pour relier l appareil la terre Cette fiche fonctionne uniquement dans une prise de courant avec contact de mise la terre Il s agit d un dispositif de s curit N essayez pas d ins
173. por el proceso de 1 Informaci n importante 67 68 reparaci n para asegurarse de que el instrumento est en buenas condiciones de funcionamiento Utilice exclusivamente las opciones y actualizaciones recomendadas por Miltenyi Biotec 1 5 3 Limpieza Desenchufe el separador autoMACS Pro de la toma de corriente antes de proceder a su limpieza No emplee productos de limpieza l quidos ni aerosoles y use siempre un trapo h medo 1 5 4 Sustancias peligrosas WARNING Risk of loss of life if biohazardous material is used Wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes Operate device in a safety hood Decontaminate device after spilling of biohazardous material Si se utilizan o se han utilizado sustancias biol gicamente peligrosas el operario deber a utilizar el equipo de seguridad que aparece en las se ales de aviso de las sustancias empleadas P ngase guantes ropa y gafas de seguridad para evitar el contacto con la piel y con los ojos Tambi n proteja la boca y la nariz pues los aerosoles podr an escaparse del sistema Un equipo de seguridad defectuoso o inadecuado puede poner en peligro al operario El separador autoMACS Pro deber ser manejado dentro de una campana de seguridad si se procesan sustancias peligrosas o desconocidas Si se han utilizado sustancias peligrosas o stas se han derramado se debe velar por una desinfecci n meticulosa del sistema
174. pper navigation bar and Wash now from the lower navigation MiniSampler Furthermore bar Select Qrinse and press Run Check if the leakage persists If the calibration data is so unscrew the column and check if the luer connectors of the checked columns are damaged If this is the case exchange the leaking column with a new autoMACS Column refer to section 3 3 4 3 Check if the column connector is fastened properly If not use second wrench to counter and tighten another quarter turn 4 Ifthe problem persists contact Miltenyi Biotec Technical Support 9 Troubleshooting 171 Note Do not remove the connector from the tubing Note Please note that each tubing has a specific length and should be exchanged with the corresponding spare part only 172 9 1 2 Tubing leakage 1 Identify the location of the leaky tubing by running the Rinse program Select Wash Now from the lower menu bar in the Separation menu Highlight Rinse and click Run 2 Check whether the tubing is tightened properly If this is not the case tighten the tube connector The connector should be inserted precisely 3 If the problem persists loosen the tube connector and pull back the connector from the tubing 4 Check the ends of the tubing for wear and fissures If necessary replace tubing with the appropriate spare part Insert the tubing into the appropriate port Then cautiously insert and fasten the tube connector 5 Run the Rinse program and che
175. ppropriate positions and reposition up take needle in needle holder 7 7 2 Rescue procedure B If it is not possible to restart the instrument the cells retained on the columns can be recovered 1 Switch OFF the instrument and disconnect from the power supply 2 Prepare two 50 mL tubes in a rack and fill two 5 mL syringes with Running Buffer Separation Buffer 3 Open the front cover and place absorbent tissue underneath the columns 4 Pull out the column from the column holder and replace the top connector with a 5 mL syringe filled with Running Buffer 5 Undo the bottom connector and flush the column into a 50 mL tube Discard column and syringe appropriately 6 Repeat steps 4 to 5 with the second column 7 Centrifuge tubes at 350xg for 10 minutes 7 Maintenance 165 8 Discard the supernatant and perform reseparation on the recovered cells as soon as possible Keep cells on ice until the separation 9 Install new columns or column substitutes in place of the discarded ones 7 8 Spare parts list Pump Seal 130 022 101 Pump Syringe 130 090 339 Square Key 130 090 378 Hydrophobic Air Filter 130 090 385 O Ring for Bottle Closure 130 090 386 Air Filter Connector 130 090 387 Power Cord D 130 090 389 Power Cord USA 130 090 391 Plunger Lock Screw 130 090 644 Column Connector 130 090 676 4 port 4 way Valve 130 090 684 4 port Distribution Valve 130 090 685 Fuse Holder 130 090 834 Column substitutes 2 pieces 130 0
176. r a caerse causando lesiones corporales graves y o da os graves al equipo Utilice exclusivamente una mesa carrito estante tripode o soporte recomendado por Miltenyi Biotec o vendido junto con el instrumento No coloque el separador autoMACS Pro en un mueble empotrado o un espacio reducido como por ejemplo una estanter a a menos que el mismo haya sido dise ado espec ficamente para dar cabida al instrumento que provea una ventilaci n adecuada y que se hayan seguido las instrucciones de montaje del equipo 1 Informaci n importante 65 66 1 4 2 Circulaci n de aire El instrumento no deber a ser colocado cerca de radiadores rejillas de calor estufas o cualquier otra pieza de equipamiento incluidos los amplificadores que produzca calor Permita que circule suficiente aire alrededor del separador autoMACS Pro deje al menos 15 cm de separaci n en todas direcciones mientras est en funcionamiento para garantizar que el instrumento se enfr e adecuadamente Evite la exposici n directa a la luz solar Las ranuras y aberturas del instrumento sirven para que se ventile y no deben ser bloqueadas o cubiertas puesto que contribuyen a un funcionamiento seguro del separador autoMACS Pro evitando que se recaliente No introduzca ning n cuerpo extra o por las aberturas del instrumento 1 4 3 Agua y humedad No use el instrumento en un lugar h medo Evite la elevada humedad o la condensaci n y proteja a la m quina de salpicadu
177. r documentation provided by Miltenyi Biotec it does not cover Product issues not arising out of defects in material or workmanship including but not limited to Product issues resulting from failure to follow installation operating and or maintenance instructions or environmental conditions prescribed in this user manual or other Product documentation misuse abuse neglect mishandling unauthorized or improperly performed maintenance or repairs accident acts of God limitations of technology electrical current fluctuations modification of or to any part of the Product use of accessories spare parts and or consumables other than those recommended by Miltenyi Biotec or normal wear and tear Miltenyi Biotec s product warranty does not cover products sold AS IS or WITH ALL FAULTS or which had its serial number defaced altered or removed or any consumables or parts identified as being supplied by a third party those third party accessories or parts may be covered by a separate warranty from their manufacturer Miltenyi Biotec must be informed immediately if a claim is made under such warranty If a material or manufacturing defect occurs within the warranty period Miltenyi Biotec will take the appropriate steps to restore the full usability of your Product Limitation on damages Miltenyi Biotec shall not be liable for any incidental or consequential damages for breach of any express or implied warranty or condition on this Produc
178. rack can handle If the tube rack does not match the template definition a warning screen will be displayed upon starting the separation 6 2 5 3 Example of application entry of sample separation instructions The purpose of the following example is to demonstrate how sample processing conditions can be easily modified and to clarify the differences between autolabeling and manual labeling Alternatively each sample could be programmed individually Two samples were placed in positions 1 and 2 of a Chill 15 Rack Sample 1 was already manually labeled using CD4 MicroBeads human Sample 2 has not yet been labeled the autoMACS Pro Separator will perform autolabeling using CD4 MicroBeads human For further details on using the Reagent menu please refer to section 4 5 1 Highlight sample positions 1 and 2 on the sample template PEA 02 lt 3 Figure 6 23 Sample positions 1 and 2 were selected 2 Select CD4_MB human under the Labeling submenu By default the software selects the separation program Possel and the wash program Qrinse 6 Cell separation using the autoMACS Pro Separator Note If the Clean program has been enabled it will also appear in the Wash submenu Note The Sleep program can only be selected as the last step in the program sequence PA 035 Figure 6 24 The current display indicates that autolabeling will be performed on both samples 3 Select the Volume submenu to enter the required sample vol
179. range exceeded Figure 11 9 Adjust MACS MiniSampler position P6 13 10 18 Select Save to store new settings Calibration 1 Test current calibration settings step 4 4 19 Select Test to test new configurations The autoMACS Pro Separator will perform a complete test of Calibration 1 settings Calibration 1 Step 4 4 Test Test calibration data Figure 11 10 It is important to test the new calibration settings To navigate back through the calibration steps select Previous 11 Hardware and software installation 11 201 202 20 Place a Chill 15 Rack onto the MACS MiniSampler 21 Select Done 22 Place the MACS Reagent Rack 4 onto the MACS MiniSampler 23 Select Done 24 Inthe next steps you can test the calibration settings of the washing station the tube rack position A1 B1 and C1 the x position and vials 1 4 in the Reagent Rack To test a postion select Test postion To skip a postion select Next Check all approached positions by using the needle navigation buttons Move up and Move down then select Done After testing the last postion the Calibr_1 screen figure 11 3 will reappear Select Cancel to leave the calibration program 25 If any errors or misalignments were noted repeat the entire process Calibration 2 1 Select menu Option 2 Highlight User settings and select Calibr_2 3 Select Run RUN to start Figure 11 11 Performing Calibration 2 by selecting program
180. ras 1 4 4 Producto conectado a tierra El instrumento est equipado con un tipo de enchufe de tres varillas que tiene un tercer contacto para toma de tierra Este enchufe s lo encaja en una toma conectada a tierra Se trata de una medida de seguridad No intente enchufarlo en una toma de electricidad no conectada a tierra Si el enchufe no encaja p ngase en contacto con un electricista para que reemplace la toma de corriente 1 4 5 Fuentes el ctricas S lo se deber a maniobrar el instrumento utilizando la fuente el ctrica indicada en la etiqueta con los valores el ctricos del producto Si tiene alguna pregunta sobre qu tipo de fuente el ctrica utilizar p ngase en contacto con su proveedor de servicios de Miltenyi Biotec autorizado o con su compa a el ctrica No utilice alargadores o regletas No sobrecargue la toma de corriente La carga total del sistema no debe sobrepasar el 80 de la rama del circuito 1 4 6 Accesibilidad Aseg rese de poder acceder f cilmente tanto al interruptor principal como al conector para el cable de corriente el ctrica y de que stos est n situados tan cerca del operario como sea posible Si es necesario desconectar el suministro el ctrico desenchufe el cable de la toma de corriente 1 4 7 Aparatos perif ricos Solamente los dispositivos perif ricos que cumplen con la UL 60950 pueden ser conectados con el conector RS232 rotulado con COM No se utiliza el conector rotulado con R
181. rayon laser sur d autres personnes Ne pas d monter changer ni retirer des sources de rayon laser ou DEL encastr es ni leur support Les sources de rayon laser ou DEL ne cessent pas forc ment d mettre un rayonnement une fois d mont es Le rayonnement d appareils d mont s peut entrainer des l sions oculaires Prendre garde au chemin optique du rayon laser ou de sa r flexion sur une surface de miroir Veiller lors de l installation de l autoMACS Pro Separator a ce que le chemin optique rayon laser ne se trouve pas a hauteur des yeux de personnes pendant le service Eviter que de l eau de l huile de la poussi re ou d autres corps tranger adh rent sur la fen tre de lecture du lecteur de code 2D comme ceci risque d entra ner des erreurs de lecture S assurer avant le nettoyage du scanner qu aucun rayon laser n est plus mis La manipulation du laser risque sinon de provoquer des l sions oculaires Utiliser un chiffon doux pour essuyer des substances sur le scanner Ne pas utiliser d alcool ou d autres produits de nettoyage L autoMACS Pro Separator est d sign comme produit laser de la classe 1M selon la norme CEl 60825 1 1993 A1 1997 A2 2001 1 Informations importantes ATTENTION Lutilisation d organes de commande autres que ceux 1 mentionn s dans ce document ainsi que toute adaptation ou utilisation d autres proc d s que ceux mentionn s dans ce document peuvent d gager des rayonnements dangereux
182. rda especifica el riesgo de que se trate El nivel de peligro que encabeza la advertencia clasifica el peligro de que se trate tal y como se mencion m s arriba El nivel tipo y fuente del peligro as como las posibles consecuencias prohibiciones y remedios se indican tal y como aparece a continuaci n 1 Informaci n importante 59 60 WARNING CAUTION e0o gt gt gt b P p ii S mbolos y niveles de peligro El siguiente recuadro recoge un glosario ilustrado que describe los s mbolos usados en el presente manual de usuario y en el separador autoMACS Pro ADVERTENCIA indica una situaci n de peligro que si no se impide puede producir la muerte o lesiones graves ATENCI N indica una situaci n de peligro que si no se impide puede producir lesiones leves o moderadas Atenci n consulte el manual de usuario para obtener m s informaci n y proceda con precauci n Las advertencias incluyen el riesgo de da ar el equipo las lesiones personales graves o la muerte Peligro de aplastamiento o corte Riesgo de que se aplaste o corte alguna parte del cuerpo debido a peligros mec nicos Radiaci n l ser Riesgo de graves lesiones oculares y de piel Campo magn tico intenso El campo magn tico puede interferir en objetos magnetizables e instrumentos electr nicos o da ar portadores magn ticos de informaci n Riesgo de producir lesiones personales graves a personas que lleven un marcapasos o implantes m dicos electr
183. riali possono rappresentare un pericolo Le colonne le piastre le provette e tutti gli altri materiali di consumo entrati in contatto con campioni a rischio biologico vanno sterilizzati in autoclave prima di essere smaltiti rifiuti liquidi vanno autoclavati o decontaminati utilizzando un disinfettante idoneo per il patogeno specifico ed es candeggina al 10 alcol isopropilico o etanolo al 70 Lo smaltimento dei rifiuti deve avvenire in conformita alle disposizioni locali vigenti en a utoMACS Pro Separator SN 0563 Order na 130 097 545 Power Supply 100 280 VAL 30700 Hz 200 VA Fuser 274250 ui Figura 1 4 simboli di pericolo biologico sono localizzati sulla superficie posteriore dello strumento sinistra e sulla parte superiore della bottiglia del fuido di scarto destra 1 5 5 Trasporto Lo strumento autoMACS Pro Separator va trasportato con cautela in imballaggi specificati da Miltenyi Biotec In caso di cadute o di vibrazioni eccessive lo strumento pu subire dei danni interni Qualora si renda necessario rispedire lo strumento al produttore per interventi di manutenzione decontaminare lo strumento da eventuali materiali pericolosi prima della spedizione In caso di domande circa la decontaminazione o il trasporto adeguati contattare l assistenza tecnica 1 5 6 Smaltimento Contattare il servizio di assistenza tecnica per lo smaltimento dello strumento 1 Informazioni importanti 57
184. rol figure 10 2 and 10 3 Color coded sensor cable Color coded sensor cable Main power switch Figure 10 3 Right side view of the autoMACS Pro Separator Plugs connections and guidings Sockets for the main plug the fluid sensor cables the 2D code reader cable and the MACS MiniSampler are installed at the rear of the instrument figure 10 4 Additional sockets are implemented for further instrument development The main power switch is located at the right hand side of the instrument figure 10 3 Several guidings at the rear and sides of the instrument ease the safe connection of tubing and sensor cables vacant Sensor cable plug 2D code reader plug MACS MiniSampler plug Figure 10 4 Rear view of the autoMACS Pro Separator 190 10 Technical data and specifications MACS MiniSampler Chill Racks and Reagent Rack The MACS MiniSampler figure 10 5 can be loaded with one of three different chill racks that carry cell samples and fraction collection tubes and the MACS Reagent Rack 4 The upper plate of the MACS MiniSampler moves in an x direction and aligns the tube openings with the port of the automated arm The guiding of the MACS MiniSampler is directly attached to the corresponding slot below the washing station When attaching the MACS MiniSampler sensor cable to the corresponding socket at the rear of the instrument the MACS MiniSampler will be automatically detected The type of tube rack carried by the MAC
185. rument is not to be used for an extended period of time the store procedure must be performed refer to 7 2 3 for further details 7 2 1 Instrument priming A rinse program must be performed before performing the first cell separation after the instrument has been switched ON Performing an instrument rinse 1 Click the Separation menu and Wash Now from lower menu bar 2 Select Rinse 3 Click Run O_init Optional priming of the instrument at startup By using the O_init program the instrument can be instructed to perform an automated rinse sequence at startup The Rinse program is used for efficient washing and equilibration of the fluidic system 1 Select Option User settings and O_init Press Run 2 Follow the prompt on the screen to enable or disable the initial wash Option initial Wash is disabled Om CS Figure 7 1 Initial wash program is used to activate or deactivate an automatic instrument prime on instrument startup 7 Maintenance Note To prevent the formation of salt deposits wipe the outlet ports with a tissue soaked with distilled or deionized water before each Sleep program 7 2 2 Clean uptake outlet ports It is recommended to clean uptake and outlets daily This should be performed before running the Sleep program shutdown 1 2 Switch OFF the instrument The ports can be wiped with tissue soaked with 70 ethanol isopropyl alcohol or MACS Bleach Solution followed by distilled or
186. s and bottle closures using detergent 1 hypochlorite or 70 ethanol Rinse thoroughly using deionized water 7 Reconnect all tubing Select OK 8 Clean the entire needle manualley using 70 alcohol swaps Select OK 9 Install fresh columns Select OK 7 Maintenance Note Run the Store program and exchange the autoMACS Columns with substitute columns if you intend to store the instrument for a period longer than two weeks Note It is recommended to exchange valves once a year Note If wear of a valve is suspected use the Check_up program to analyze functionality 7 3 5 Instrument long term storage The program Store should be applied to prepare the instrument for long term storage Upon completion of the Store program the fluidic system contains 70 ethanol To store the autoMACS Pro Separator for a period longer than two weeks the tubing system should be cleaned and the columns should be replaced with column substitutes The Store program automatically performs the cleaning procedure and prompts the user to install column substitutes 1 Select the Option Special and Store Select Run The system will be rinsed automatically RE 150 Figure 7 13 Store program is underway 2 Install the column substitutes as described by the section 3 3 4 3 Select Done 4 Switch OFF the autoMACS Pro Separator using the main power switch 7 4 Planned maintenance 7 4 1 Valve exchange The fluidic system of
187. s have already been assigned to positions on the reagent rack using the Reagent menu Figure 4 16 Labeling submenu Using the MACS Reagent Rack 4 a total of four reagents may be used at any given time Assigning sample volumes The Volume submenu 3 is used to inform the instrument about the available sample volume To enter or modify a volume 1 Ensure that the appropriate sample template position is highlighted 2 Select the Volume submenu 3 Using the numeric keypad enter the sample volume For autolabeling enter the sample volume for the first labeling step as outlined in the corresponding data sheet e g 160 uL 2x10 cells for labeling with CD4 MicroBeads human or 120 uL 4x10 cells for labeling with Monocyte Isolation Kit II human 4 An overview of the autoMACS Pro Separator user interface 99 100 Figure 4 17 Entering the sample volume in microliters uL for Monocyte Isolation Kit Il human 4 Note that the minimal incubation volume after the addition of the first reagents may not be less than 200 uL 5 Select Enter Assigning a cell separation program The Separation submenu 4 is used to instruct the autoMACS Pro Separator which cell separation program should be applied to each sample Several sample positions can be highlighted to assign a separation condition for multiple samples Refer to section 6 1 2 for a detailed explanation of the various cell separation strategies To assign
188. s material is used Wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes Operate device in a safety hood Decontaminate device after spilling of biohazardous material If biohazardous material has been used the operator shall choose and wear personal safety equipment in accordance with warnings and precautions for the used substances Wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes Also protect mouth and nose as aerosols might leak from the system Defective or inadequate safety equipment might endanger the operator The autoMACS Pro Separator shall be operated in a safety hood if hazardous or unknown materials are processed If hazardous material has been used or spilled care must been taken to thoroughly decontaminate the system For details refer to section 7 3 4 Always inspect the fluidic system complete tubing set bottles and their closures valves columns diluter and needles before switching ON the instrument If leakage has been detected replace all damaged parts before switching ON the instrument If damaged parts cannot be replaced unplug and do not use the instrument Failure of parts containing biohazardous material or liquids that have been in contact with such material could cause a hazard Columns tubes and any other consumables that were in contact with biohazardous samples shall be autoclaved prior to dispos
189. s potenciales riesgos asociados al manejo del separador autoMACS Pro por favor observe las siguientes precauciones generales En caso de no seguir estas precauciones puede producirse un incendio o causar da os corporales o al equipo 1 Informaci n importante 61 62 1 3 1 Peligro de descarga el ctrica y propagaci n de fuego WARNING Hazardous voltages Risk of loss of live severe personal injury Unplug before cleaning Do not use if device is opened or damaged if liquids have been spilled into device if objects device through ventilation slots ADVERTENCIA Los aparatos el ctricos pueden producir una descarga el ctrica Para reducir este riesgo no abra ninguna cubierta ni abra tampoco ningun otro accesorio de harware suministrado por Miltenyi Biotec Cualquier cubierta asi como hardware accesorios deber n ser retirados nicamente por personal autorizado Se debe tener especial cuidado cuando se manejen l quidos Limpie inmediatamente los l quidos vertidos Impida que stos accedan al interior del aparato Desenchufe el cable de electricidad antes de proceder a limpiar manualmente el separador autoMACS Pro ADVERTENCIA Existe un riesgo potencial en caso de usar separador autoMACS Pro abierto que se haya ca do o que est averiado si se han derramado l quidos en el equipo si se ha colado alg n objeto por las ranuras de ventilaci n o si ha ca do algo dentro del equipo Si salen llamas o humo apague i
190. s the target cells Deplete Depletion program standard mode a normal cell deposition rate is used The non labeled cells are eluted in row B of the sample rack This is the optimal program to be used in combination with MACS Cell Isolation Kits or to achieve the highest recovery rate of untouched cells DepleteS Depletion program sensitive mode a slow cell deposition rate is used 1 mL min The non labeled cells are eluted in row B of the sample rack This program is optimized for depletion of those cells that weakly express the antigens used for magnetic labeling or to achieve optimal purity of the untouched cell fraction Depl05 Special depletion program the cell deposition rate is 0 5 mL min The non labeled cells are eluted in row B of the sample rack This program is used for strong depletion of those cells that weakly express the antigens used for magnetic labeling or to achieve the highest purity of the untouched cell fraction However choosing Depl05 might result in a reduced recovery of the target cell fraction Depl025 Special depletion program the cell deposition rate is 0 25 mL min The non labeled cells are eluted in row B of the sample rack This program is used for very strong depletion of those cells that weakly express the antigens used for magnetic labeling or to achieve the highest purity of the untouched cell fraction However choosing Depl025 might result in a reduced recovery of the target cell fraction
191. sample is remaining in tube Plunger movement blocked because of column clogging blocked tubing set or any other cross section constriction during output of the positive fraction The positive fraction has not been eluted completely Positive cells are still remaining in the system Resuspensed sample could not be taken up completely no final air intake detected Leakage of air into system behind bubble sensor Portion of resuspended sample is remaining in tube Resuspensed sample could not be taken up completely no final air intake detected Leakage of air into system behind bubble sensor Portion of resuspended cells are remaining in tube Required volume cannot be provided in given Chill Rack Miscalculation of diluter movement Target position is negative 9 Troubleshooting Possible remedies Do not use sample volumes exceeding the maximum whole blood sample volume specified for the used Chill Rack type Split sample into several tubes reprogram separation parameters and restart separation If volumes are in specified range but error is displayed anyway please call Technical Support Check buffer supply Check for leakage of system unintended air intake Start a rinse program Wash only and then retry calibration Otherwise call Technical Support Set time and date to actual values Make sure foam ontop of sample is not higher than 5 mm above liquid level Check for leakage at transition of needle
192. ser interface 93 94 Tab Function Reagent To define the position of MACS Reagents on the MACS Reagent Rack 4 Separation To define an autolabeling and or cell separation strategy for up to six samples In addition cell processing procedures can be saved as templates for regular use Status The instrument status is displayed at a glance Log list The log list details completed actions and errors Option Allows for special procedures such as MACS Columns exchange instrument calibration and service steps Table 4 1 Feature overview of the upper tabbed menu After instrument initialization the screen automatically displays the Status menu which helps to monitor the instrument during installation and operation Color coded symbols indicate the status of the hardware components Further information on a particular component can be obtained by touching the symbol RE lt lt Figure 4 4 The main root menu screen displays instrument Status at startup 4 3 Status menu The autoMACS Pro Separator is a sensor controlled instrument that allows easy monitoring during operation At a glance the instrument status can be determined by viewing the Status menu at any time Sample status Fluid bottle filling status Column status Tube rack status MiniSampler status VO lt lt Figure 4 5 Overview of the instrument status panels 4 An overview of the autoMACS Pro Separator user interface A description of how
193. sitive fractions 1 Select the appropriate tube rack according to table 6 4 2 Recommended Cool down the tube rack for 3 4 hours in a refrigerator 2 8 C or until the coolant becomes solid Do not cool below 0 C as samples may freeze 3 Equip the tube rack with sample tubes and fraction collection tubes Rack type Maximal number Manual labeling Autolabeling Selo ofsamples Maximal Minimal first Maximal final sample volume incubation volume labeling volume Chill 5 24x5 mL 6 2 5 mL 0 2 mL 2 0 mL 5 mL tubes 0 25 mL 1ml Chill 15 15x15mL 5 12 5mL 0 2 mL 6 5 mL 5x5 mL 15 mL tubes 1ml 4 m Chill 50 6x50mL 3 50mL 4m 8 ml 3x15 mL 50 mL tubes 3x5 mL Volumes refer to whole blood samles only Table 6 4 MACS Chill Rack specifications for manual labeling and autolabeling For further details on samples volumes for autolabeling refer to table 6 3 124 6 Cell separation using the autoMACS Pro Separator Note It is assumed that the MACS MiniSampler has been already installed the waste bottle is empty and that all fluid bottles are filled with recommended solutions refer to section 3 for more details Note 70 ethanol does not contain electrolytes Therefore the fluid level cannot be determined and the symbol for the storage solution bottle is always gray Please ensure that the volume of each solution is sufficient for the number of separation and washing programs to be performed Note The autoMACS
194. ss the column into the slot until you feel the guides click Verify that the column is placed in the center of the magnet cover Repeat steps 8 through 11 to install the second column Ensure that the tubes are securely fastened to the columns and that the tubing is neither pinched nor obstructed Press Done Close the front door The unit is now ready to perform cell separations 3 3 5 Priming the autoMACS Pro Separator Priming implicates the initial cleaning and filling of the autoMACS Pro Separator tubing system before cell separations are performed The autoMACS Pro Separator must be primed each time the instrument is switched ON WARNING Read the warnings and precautions section before priming the autoMACS Pro Separator for the first time 3 Assembly and installation WARNING When working with biohazardous samples it is 3 recommended to fill the waste bottle with 100 mL of disinfectant before use e g MACS Bleach Solution 130 093 663 For proper disposal please follow local regulations and carefully read the chapter 1 1 Fill all bottles with the appropriate solutions and empty the waste bottle 2 Switch ON the instrument After initialization is completed the Status menu will be displayed Verify that the touchscreen symbols for all fluid bottles are green If this is not the case check if the fluid sensor cables are connected to the correct fluid bottles The symbol for the ethanol bottle remains gray fl
195. ssary Unexpected air in system during sample uptake The reagent designated for the current labeling process is not assigned a position in the Reagent Rack Needle could not be retreated completely Collision of the needle with the bottom of the tube or any other object has been detected but resetting the collision sensor failed although the needle has been lifted Collision of the needle with an unexpected object Resetting the collision sensor failed as the needle could not be lifted already too close to top Collision of the needle with an unexpected object far above expected tube bottom Collision of the needle with an unexpected object far above expected tube bottom Collision of the needle with an unexpected object far above expected tube bottom Sample volume exceeds maximum volume specified for rack or program type Remaining portion of sample will not be processed The sample volume has been underestimated The needle has been rinsed but might still be contaminated Collision of needle with bottom of tube could not be detected at expected position 9 Troubleshooting Possible remedies Make sure foam on top of sample is not higher than 5 mm above liquid level Verify that sample was filtered before separation and that uptake needle is not clogged Check for leakage at the point where the needle and tubing meet and where the tubing to bubble sensor meet Verify that connectors are appropriately fast
196. suario u otra documentaci n de Miltenyi Biotec especifique lo contrario no revise usted mismo el separador autoMACS Pro Las revisiones y reparaciones deben ser llevadas a cabo por personal cualificado Las revisiones y reparaciones del separador autoMACS Pro incorrectamente realizadas pueden poner en peligro a sus usuarios producir resultados impredecibles derivar en un mal funcionamiento del aparato o que ste sufra da os as como causar un desgaste prematuro y reducir el tiempo de vida del instrumento pudiendo anular su garant a Pregunte a su representante de Miltenyi Biotec por los contratos de servicios de asistencia t cnica de Miltenyi Biotec o consulte www miltenyibiotec com support IMPORTANTE Cuando se necesiten piezas de recambio o de repuesto aseg rese de que el proveedor de servicios utiliza exclusivamente piezas originales de Miltenyi Biotec o de otros fabricantes especificados y recomendados por Miltenyi Biotec El uso de piezas de recambio o de repuesto no autorizadas puede producir un mal funcionamiento del aparato y alterar los resultados de la separaci n celular Miltenyi Biotec no cubrir la garant a ni aceptar ninguna responsabilidad por la aver a de aparatos o por los da os resultantes del uso de piezas de recambio o de repuesto inapropiadas Una vez completado el servicio o la reparaci n haga que su proveedor de servicios autorizado por Miltenyi Biotec realice todos los controles de seguridad requeridos
197. t Some states or jurisdictions do not allow the exclusion or limitation of incidental or consequential damages so the above limitations or exclusions may not apply to you This warranty statement gives you specific legal rights and you may have other rights which may vary from county to country or jurisdiction to jurisdiction 13 Limited warranty 205 206 13 Limited warranty Glossary Air filter Hydrophobic 0 2 um air filter attached to the bottle closure Used to vent the bottle and at the same time prevent contaminants from entering the fluid bottle Air filter connector Luer to thread connector for attaching the air filter to the threaded bottle closure vent APC Allophycocyanin autoMACS Columns Specifically designed autoMACS Columns reusable for two weeks or for 100 separations within these two weeks autoMACS Column 1 First autoMACS Column in which labeled cells are retained during positive selection and depletion programs The autoMACS Column 1 occupies the left slot of the black magnet cover autoMACS Column 2 The second autoMACS Column is used during double selection programs The autoMACS Column 2 occupies the right slot of the black magnet cover autoMACS Pro Separator Automated magnetic cell separator also referred to as device or instrument autoMACS Running Buffer Sterile and ready to use solution for cell separation and washing programs The tubing connector is color coded blue autoMACS Pro
198. t veuillez vous adresser votre organisme local responsable de l approvisionnement en lectricit des travaux de b timent de la maintenance et de la s curit Si vous avez un doute s rieux quant la s curit d utilisation de votre appareil veuillez contacter votre prestataire de services Miltenyi Biotec agr ou appelez le service apr s vente de Miltenyi Biotec 1 Informations importantes 25 26 1 3 Pr cautions d ordre g n ral Afin de r duire les risques potentiels li s l utilisation du s parateur autoMACS Pro veuillez observer les pr cautions g n rales suivantes La non observation de ces pr cautions peut provoquer un incendie des dommages corporels et ou d t riorer l appareil 1 3 1 Risque d lectrocution et de propagation de feu WARNING Hazardous voltages Risk of loss of live severe personal injury Unplug before cleaning Do not use if device is opened or damaged if liquids have been spilled into device if objects device through ventilation slots AVERTISSEMENT Les appareils lectriques pr sentent un risque d lectrocution Afin de r duire le risque d lectrocution n ouvrez aucun cache du s parateur autoMACS Pro ni d autres quipements accessoires fournis par Miltenyi Biotec Tous les caches de l appareil et des quipements accessoires doivent tre d mont s par le personnel agr uniquement Soyez particuli rement prudent pendant la manipulation de fl
199. te las instrucciones se proteger a s mismo y al equipo de posibles peligros y crear un ambiente de trabajo seguro Si este instrumento es manejado de un modo no previsto por su fabricante la seguridad se ver mermada IMPORTANTE Por favor lea y siga todas las instrucciones de uso recogidas en el presente manual de usuario y preste atenci n a todas las advertencias que aparecen en el instrumento Guarde para futuras consultas este manual y todas las dem s instrucciones de seguridad o de funcionamiento recibidas junto con el instrumento en un lugar accesible para todos sus usuarios IMPORTANTE El separador autoMACS Pro est dise ado para ser usado exclusivamente en interior No utilice el equipo en reas clasificadas como peligrosas como ambientes con alta concentraci n de ox geno En todo momento se debe observar la normativa nacional de seguridad en el trabajo las normas del laboratorio y los est ndares de salud y seguridad en el laboratorio y de prevenci n de accidentes P ngase en contacto con la autoridad local competente para el suministro de electricidad construcci n de edificios mantenimiento o seguridad para obtener m s informaci n sobre la instalaci n del equipo Si tiene serias dudas sobre el manejo seguro del equipo p ngase en contacto con su proveedor de servicios de Miltenyi Biotec autorizado O llame al servicio de atenci n al cliente de Miltenyi Biotec 1 3 Precauciones generales Para reducir lo
200. the A autoMACS Pro Separator An overview of the log list table follows Name Definition Name Definition Date Indicates the date Program Name of program the action was for example performed Deplete Cell depletion Possel Positive selection Qrinse Quick rinse performed Type Description of the Protocol If a protocol is associated A action for example with the log its name will Rack Action involving be listed under this rack heading for example Init Initiation of CD4 h CD4 MicroBeads instrument human were used for Wash only Wash only autolabeling and cell performed separation Service Service step performed Special wash Special wash performed Pos Corresponding Status The status of the log is position on sample depicted as follows tube rack EA successfully completed EA action failed Table 4 3 Overview of the log list table parameters To display further details about an individual log 1 Select Log list tab and highlight a log from the log list 2 Select Details A detailed view of the program status is shown 3 Select Ok to return to the log list screen or select Log Details to view a detailed log of performed actions 4 An overview of the autoMACS Pro Separator user interface 105 106 ee 02 13 ii Figure 4 28 Viewing log details A A positive selection using CD4 MicroBeads human was performed on March 26th 2009 B An overview of program actions and corresponding times are displaye
201. the column symbol is green no action is required If the symbol is red the columns must be exchanged The level of the green fill on the column symbol indicates the remaining service life of the autoMACS Columns Rack detection Rack detection only occurs upon starting the separation process MACS MiniSampler detection If the MACS MiniSampler has been detected correctly a rectangular symbol is displayed If it has not been detected a hand symbol will be displayed in the same field 6 2 5 4 2 Status menu during the separation process It is recommended to monitor the instrument s status during cell separation using the Status menu Programs yet to be processed appear in yellow fields Programs currently undergoing autolabeling appear in lilac Programs in progress in which no autolabeling is being performed switch to orange completed programs switch to white The current action is always displayed in the status bar located below the lower navigation bar The status bar also displays the overall progress in minutes min Rising E Figure 6 30 Monitoring the status during cell separation Sample processing at position 1 is completed white color Sample at position 2 has finished the separation and is now performing a Rinse Interrupting cell separation The cell separation process can be paused at any time from all menu screens by selecting Stop 1 From any menu screen select Stop The autoMACS Pro Separator will then immediatel
202. the shock and position indicators if present suggest incorrect transportation of the instrument If there is apparent damage please contact Technical Support for assistance refer to section 12 CAUTION Due care must be taken while lifting the autoMACS Pro Separator Miltenyi Biotec accepts no liability for potential injuries and damages sustained during lifting and or movement of the instrument 1 Open the box and remove the top compartment of the packaging to reveal the instrument and associated packaging Figure 3 1 Top compartment of the autoMACS Pro Separator box 2 Remove boxes containing the MACS MiniSampler and cover and the MACS Chill Racks Remove the foam packaging from both sides of the autoMACS Pro Separator Figure 3 2 Packing format of the autoMACS Pro Separator and accessories 3 Place the instrument onto a stable worktop surface e g laboratory bench Remove the plastic bag surrounding the instrument 3 Assembly and installation Note Two people are required to lift the autoMACS Pro Separator The instrument must be gripped at the base of the orange bottle baskets located at both sides of the instrument Note that the instrument is heavier at the front Ensure the front of the instrument is stabilized while lifting it Figure 3 3 Positioning the autoMACS Pro Separator securely onto a laboratory bench 4 Note the positions of the MACS MiniSampler guiding 2 and its corresponding slot
203. ti al funzionamento dello strumento e forniscono importanti informazioni in materia di sicurezza atte a ridurre al minimo detti rischi Seguendo scrupolosamente le istruzioni possibile proteggere se stessi e le attrezzature dai possibili pericoli e garantire un ambiente di lavoro sicuro Qualora lo strumento venga impiegato in modo non conforme alle istruzioni fornite dal fabbricante la sicurezza potrebbe risultare compromessa IMPORTANTE Leggere e seguire tutte le istruzioni operative contenute nel presente manuale e prestare particolare attenzione alle avvertenze indicate sullo strumento Conservare il presente manuale d uso e tutte le altre istruzioni operative e di sicurezza fornite insieme alla strumento in un luogo accessibile a tutti gli operatori per future consultazioni IMPORTANTE autoMACS Pro Separator sono destinati unicamente all impiego interno Non utilizzare lo strumento in ambienti classificati come luoghi pericolosi quali gli ambienti con forte concentrazione di ossigeno Vanno sempre osservate le istruzioni di sicurezza sul luogo di lavoro le procedure di laboratorio e le norme relative alla salute e alla sicurezza del laboratorio nonch alla prevenzione degli infortuni Per ulteriori informazioni relative all installazione della strumentazione contattare le autorit locali competenti per la fornitura di energia elettrica in materia di edilizia manutenzione o sicurezza Qualora abbiate serie preoccupazioni i
204. tically labeled and depleted During separation the unlabeled target cell type is collected in the flow through i e the negative fraction The mixture of magnetically labeled non target cells is retained within the autoMACS Column and will be eluted at the end of the separation process MACS Cell Isolation Kits for untouched isolation contain a cocktail of titrated antibodies and MACS MicroBeads for indirect magnetic labeling They are the preferred choice if binding of antibodies to the target cells is not desired Sequential sorting A combination of two subsequent separations is applied to isolate Se cell subsets that can be distinguished from other cell types through their expression of two different markers This includes cell types for which a specific marker has not been defined Depletion followed by positive selection This separation strategy is useful if undesired cells and target cells have one marker of interest needed for separation in common In this case the target cells cannot be isolated in a single positive selection step Therefore the undesired cells expressing the common marker are magnetically labeled via antigens distinct from the common marker and depleted Cells in the flow through fraction of the depletion step including the target cells are subsequently labeled with MACS MicroBeads that bind to the common marker Target cells are then isolated by positive selection Sophisticated MACS Cell Isolat
205. tically sensitive objects away from device WARNUNG Der autoMACS Pro Separator enth lt einen extrem starken Magneten Halten Sie mit magnetischen Datentr gern Kreditkarten Magnetb nder Speichermedien und dergleichen und elektronischen Ger ten wie H rger te Herzschrittmacher 1 Wichtige Informationen Mess und Steuerger te PCs Uhren mindestens 20 cm Abstand 1 zur Magnetabdeckung da diese durch das Magnetfeld gest rt und gesch digt werden k nnen Abbildung 1 1 Warnsymbol f r starken Dauermagnet 1 3 3 Quetsch und Schergefahr CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard VORSICHT Frontabdeckungen nicht 6ffnen wenn das Ger t l uft Bewegung des Automatikarms und der zugeh rigen Komponenten im Betrieb nicht behindern Finger usw von allen bewegten Teilen des autoMACS Pro Separator und zugeh rigen Komponenten fernhalten es besteht sonst Gefahr von Quetsch und Scher verletzungen und Sch den am Ger t Fl ssigkeitspumpen nicht ber hren und Leitungen nicht ver ndern wenn das Ger t l uft Ger t immer ausschalten bevor Arbeiten am Fl ssigkeitssystem vorgenommen werden Laufenden Vorgang immer anhalten oder abbrechen bevor Arbeiten an Zusatzger ten wie dem MACS MiniSampler ausgef hrt oder Reagenzgl ser aus dem Reagenzglasgestell im Sampler entnommen oder dort eingesetzt werden Sicherheitsma nahmen und vorrichtung
206. time of the instrument Miltenyi Biotec does not honor any warranty or accept any responsibility for damages resulting from the use of inappropriate consumables or accessories 5 1 Materials required for operation 5 1 1 Buffers and Solutions For daily operation the following solutions are required autoMACS Running Buffer autoMACS Pro Washing Solution and storage solution Solution bottles can be identified by color code and symbols table 3 1 For proper operation of the autoMACS Pro Separator solution bottles must be filled with a minimum volume of 150 200 mL It is recommended to use ready to use MACS Buffers or fresh filter sterilized solutions to prevent potential contamination of the tubing system Please note that autoMACS Running Buffer contains azide as a preservative To obtain an azide free running buffer prepare a 1 20 dilution using the autoMACS Rinsing Buffer and bovine serum albumin BSA MACS BSA Stock Solution e g add 10 mL of MACS BSA Stock Solution to 190 mL of autoMACS Rinsing Buffer The autoMACS Pro Washing Solution is a filter sterilized and Note Do not use denatured ready to use solution to rinse the fluidic system after any ethanol technical ethanol autoMACS Pro cell separation It contains a detergent that as the autoMACS Columns dissolves cell aggregates It was developed for optimal cleaning are not resistant to oxidative of the autoMACS Pro Separator tubing system compounds Use 100 eth
207. tions on the preparation of single cell suspensions please refer to the respective Cell Separation Reagent data sheet 6 2 1 2 Adjusting sample volumes Typically 1x10 cells are resuspended in 80 uL of buffer and labeled with 20 uL of MicroBeads leading to a total labeling volume of 100 uL When working with higher cell numbers scale up all reagent volumes and total volumes accordingly For example for 2x10 total cells use twice the volume of all indicated reagent volumes and total volumes indicated in the respective data sheet When working with fewer than 1x10 cells do NOT scale down the volumes but use the same volumes as indicated In the table below the dilution volumes account for the first step of labeling For manual labeling please refer to the respective Cell Separation Reagent data sheet for ongoing procedures Minimal and maximal volumes and total cell numbers in table 6 3 account for autolabeling samples only Autolabeling protocols are being continually developed and optimized by Miltenyi Biotec For a current list of Cell Separation Reagents that are optimized for cell separations with the autoMACS Pro Separator autolabeling feature please contact Technical Support 6 Cell separation using the autoMACS Pro Separator Cell Separation Reagent Chill 5 Rack Direct MicroBeads human rat non human primate Direct MicroBeads mouse Whole Blood MicroBeads Cell Isolation Kits Cell Isolation Kits
208. to monitor the instrument s status using the A Status menu follows 4 3 1 Status of fluid bottles Figure 4 6 Fluid bottle status symbols Left Fluid bottles are ready Right Fluid bottles need to be replaced The status of fluid bottles is indicated by color coded graphic symbols and by a text table Bottle Symbol Symbol color and user action Running Buffer Green no action required Red refill bottle Gray connect bottle sensor Washing Solution Green no action required Red refill bottle Gray connect bottle sensor Storage solution Gray no liquid detection visually check volume Waste Green no action required Red empty waste Gray connect bottle sensor Table 4 2 Status of fluid bottles displayed in the Status menu In addition to color coded graphic symbols of the fluid bottles a pop up text table also reports fluid bottle status Touch any fluid bottle symbol to activate the pop up textbox To close the pop up table select e Moreover the symbols are also red when the sensor cables are connected to a wrong bottle closure 4 An overview of the autoMACS Pro Separator user interface 95 96 o Figure 4 7 Fluid bottle status By touching any fluid bottle symbol a pop up box displays a text report of the fluid bottle status 4 3 2 Column status Figure 4 8 The column status graphic Left The column is ready Right The column must be changed If the column symbol is gree
209. to tubing and tubing to bubble sensor Touch Cont to process currently uptaken sample volume Touch Cancel to abort Make sure foam on top of sample is not higher than 5 mm above liquid level Check for leakage at transition of needle to tubing and tubing to bubble sensor Touch Cont to discard the remaining positive cells into the waste bottle To output positive fraction again at lower speed exchange positive tube with an empty tube and touch Retry Check for air inlet into system behind bubble sensor Touch Cont to process currently uptaken sample volume Touch Cancel to abort Check for leakage of air into system behind bubble sensor Readjust tubing connectors and verify correct fastening of columns Exchange rack with rack able to provide requested volume e g Chill 50 instead of Chill 15 Touch Retry to try again Touch Cancel to abort Touch Retry to use target position 0 instead of negative value for current diluter move Touch Cancel to abort Please contact the Technical Support in all cases also if the sample has been processed completely after touching Retry Technical data and specifications 10 10 1 Schematics of the autoMACS Pro Separator Integrated computer for control of cell processing All interactions with the computer are performed with a TFT color touchscreen figure 10 1 A memory card is used to run all programs and to log processes
210. toMACS Pro Separator for removal of cells with low antigen expression 6 for untouched isolation with MACS Cell Isolation Kits if purity is highest priority Depl05 Depletion in sensitive mode Il for removal of cells with low antigen expression special program for very sensitive depletion Depl025 Depletion in sensitive mode Ill for removal of cells with low antigen expression special program for very sensitive depletion A_Depl07 Depletion in standard mode via loading of sample in separate 1 mL stages for removal of cells with normal antigen expression if recovery Sa is the highest priority This special program is disabled by default To enable A_Depl07 select Option User settings and O_progs A_Depls7 Depletion in sensitive mode via loading of sample in separate 1 mL stages for removal of cells with low antigen expression if purity is the highest priority This special program is disabled by default To enable A_Depls7 select Option User settings and O_progs During programs Possel and Possel_s the magnetically labeled target cells are retained in the autoMACS Column 1 The unlabeled cells are released as the negative fraction into the negative fraction collection tube i e row B of the tube rack After automated retraction of the magnet the magnetically labeled cells are eluted as the positive fraction into the positive fraction collection tube i e row C of the tube
211. ts WARNUNG Der Umgang mit elektrischen Ger ten birgt das Risiko eines Stromschlags Um diese Gefahr zu minimieren ffnen Sie weder das Geh use des autoMACS Pro Separators noch anderes Zubeh r Alle Abdeckungen sowie das Ger te Zubeh r d rfen nur von geschultem Personal entfernt werden Besondere Vorsicht ist geboten beim Umgang mit Fl ssigkeiten Beseitigen Sie ausgetretene oder versch ttete Fl ssigkeit sofort Es darf unter keinen Umst nden Fl ssigkeit in das Innere des Ger tes eindringen Ziehen Sie den Netzstecker vor manueller Reinigung des autoMACS Pro Separators WARNUNG Eine potentielle Gefahrenquelle liegt auch im Betrieb eines ge ffneten zu Boden gefallenen oder besch digten Ger tes Ebenfalls sollte unbedingt vermieden werden dass Fl ssigkeit in das Ger t gelangt Fremdk rper durch die Bel ftungs ffnungen eindringen oder von au en in das Instrument hinein gelangen Bei Auftreten von Flammen oder Rauchentwicklung schalten Sie das Ger t sofort aus trennen es von der Stromzufuhr und kontaktieren einen autorisierten Miltenyi Biotec Service Provider oder das Miltenyi Biotec Customer Support Team Der Betrieb eines besch digten Ger tes oder eines Ger tes mit schadhaftem Stromkabel ist ausdr cklich verboten 1 3 2 Starkes Magnetfeld WARNING Extremely strong magnetic fields Risk of severe personal injury to persons carrying pacemakers or electronical medical implants stay clear Keep all magne
212. ts a sample rack and allows definition of sample processing strategies for each sample rack position For each sample rack position it is possible to define cell labeling cell separation and washing programs A single wash program can be performed by selecting Wash Now from the lower navigation bar A 0 23 Figure 4 13 The Separation menu An overview of the Separation menu follows Sample rack template Sample labeling options Sample processing volume Separation program Wash procedure 0 3 Figure 4 14 Overview of the submenus in the Status menu Sample rack template The positions in the sample rack programming field 1 correspond to the sample positions in the tube rack Positions 1 6 are used in combination with Chill 5 Rack positions 1 5 with Chill 15 Rack and positions 1 3 with Chill 50 Rack 1 Toselect deselect a sample position touch to highlight the desired position 1 6 2 Select deselect a sample position by touching the display 4 An overview of the autoMACS Pro Separator user interface VE gt Figure 4 15 Sample positions 1 3 and 4 were selected Sample labeling options The Labeling submenu 2 is used to instruct the instrument if autolabeling is to be performed on a sample The default setting is which indicates that NO autolabeling will be performed the type of autolabeling that will be performed A list of product options is only visible if reagent
213. uencies higher than 5 and normal antigen expression Possel_s Positive selection in sensitive mode for the isolation of cells with frequencies higher than 5 and low antigen expression for the isolation of cells with frequencies higher than 5 and normal antigen expression if recovery is the highest priority Posseld Positive selection in standard mode I double column program for the isolation of cells with frequencies lower than 5 and normal antigen expression in a small elution volume Posseld2 Positive selection in standard mode ll double column program for the isolation of cells with frequencies lower than 5 and normal antigen expression if purity is the highest priority Posselds Positive selection in sensitive mode double column program for the isolation of cells with frequencies lower than 5 and low antigen expression Posselwb Special positive selection in special mode double column program e for the isolation of cell subsets from whole blood cell samples are automatically diluted with Running Buffer Depletion programs Deplete Depletion in standard mode e for removal of cells with normal antigen expression if recovery is the highest priority for untouched isolation with MACS Cell Isolation Kits Depletes Depletion in sensitive mode e for removal of cells with normal antigen expression if purity is the highest priority 6 Cell separation using the au
214. uid level can not be checked by system At this point the fluid bottle illumination is yellow E Figure 3 27 The Status menu The fluid level status and column are green for ready A wash or instrument prime has not yet been performed 3 Go to the Separation menu and select Wash Now from the lower menu You now have the option to perform a quick rinse Qrinse or full rinse Rinse Figure 3 28 Select Rinse to prime the autoMACS Pro Separator 4 Select Rinse and Run to start the priming process The progress will be displayed at the bottom of the touchscreen menu 3 Assembly and installation 89 90 Note A cell separation template can already be programmed or reloaded during the priming procedure When selecting Run the user will be asked to confirm that enough running buffer is available thereafter priming will begin The autoMACS Pro Separator will display a warning screen if the buffer supply is low or if the waste bottle is full If no wash program has been performed before the first separation a warning screen will ask to rinse the system You may interrupt any program by selecting Stop The Stop button is located at the bottom left hand corner in all menus When priming the instrument for the first time or when the instrument was not serviced for a long period of time it is recommended to visually inspect the fluidic system for potential leaks Open the front door after primin
215. uides Nettoyez imm diatement si des liquides se sont d vers s Veillez ce que les fluides ne s infiltrent pas l int rieur de l appareil D branchez le c ble lectrique avant de nettoyer manuellement le s parateur autoMACS Pro AVERTISSEMENT Un risque potentiel existe si le s parateur autoMACS Pro utilis est ouvert d form ou endommag si des liquides se d versent dans l appareil si un objet est entr dans l appareil par les fentes de ventilation ou si un objet a chut dans l appareil En cas d apparition de flammes ou de fum e d connectez imm diatement le s parateur autoMACS Pro d branchez l appareil et contactez un prestataire de services Miltenyi Biotec agr ou l quipe de support technique de Miltenyi Biotec Il est formellement interdit d utiliser un appareil endommag ou un appareil dont le cable lectrique est endommag 1 3 2 Champ magn tique puissant WARNING Extremely strong magnetic fields Risk of severe personal injury to persons carrying pacemakers or electronical medical implants stay clear Keep all magnetically sensitive objects away from device AVERTISSEMENT Le s parateur autoMACS Pro est quip d un aimant extr mement puissant Veillez laisser un espace d au moins 20 cm entre le cache de l aimant et les supports information magn tiques cartes de cr dit bandes magn tiques et supports de donn es ainsi que l quipement lectronique appareils auditifs 1 I
216. ume Select Enter For this experiment a minimal starting volume of 160 uL is required for autolabeling for autolabeling with CD4 MicroBeads 40 uL MicroBeads are added to the entire 160 uL sample volume For details please refer to the data sheet BEE 14 Figure 6 25 A volume of 160 uL was entered for both sample positions 4 Optional If a change of the pre set wash program is necessary e g when working with rare cells highlight the wash program to be performed before cell separation In this case the program was not changed and Qrinse stayed selected Figure 6 26 Qrinse was selected as a wash program 5 The sample in position 1 was manually labeled with CD4 MicroBeads and therefore must be setup for a cell separation without autolabeling Deselect sample position 2 by touching the display at this position Only sample position 1 is now highlighted 6 Cell separation using the autoMACS Pro Separator 137 138 N 14 47 Figure 6 27 Only sample position 1 is selected 6 Deselect autolabeling for sample 1 by selecting in the Labeling submenu Change the sample volume as recommended in the data sheet After manual labeling it is recommended to dilute cells to a volume of 500 uL per 10 total cells For further information please refer to the corresponding data sheet RE 14 47 Figure 6 28 The separation program Possel was selected for sample 1 7 View and recheck the sample setup 8 Select Run
217. unsere Website www miltenyibiotec com support WICHTIG Wenn Ersatzteile ben tigt werden stellen Sie sicher dass Ihr Service Provider nur Originalteile der Miltenyi Biotec GmbH oder Teile von Drittanbietern verwendet die von der Miltenyi Biotec GmbH spezifiziert und empfohlen werden Die Verwendung unautorisierter Ersatzteile kann Fehlfunktionen des Ger tes verursachen und die Ergebnisse von Zellseparationen beieintr chtigen Die Miltenyi Biotec GmbH akzeptiert keinerlei Garantieanspr che und haftet nicht f r Fehlfunktionen oder Sch den am Ger t die auf Verwendung ungeeigneter Verschlei oder Ersatzteile zur ckzuf hren sind Nach jedweder erfolgter Wartungs oder Reparaturleistung lassen Sie Ihren autorisierten 1 Wichtige Informationen 43 44 Miltenyi Biotec Service Provider alle notwendigen Sicherheits pr fungen durchf hren um sicherzustellen dass das Ger t sich in vorschriftsm igem Zustand befindet Nutzen Sie nur von Miltenyi Biotec empfohlenes Zusatzger t und von uns empfohlene Upgrades zu Ihrem Ger t 1 5 3 Reinigung Ziehen Sie den Netzstecker vor Reinigung des Ger tes Verwenden Sie keine Fl ssig oder Spr hreinigungsmittel sondern nur ein feuchtes Tuch 1 5 4 Gef hrliches Material WARNING Risk of loss of life if biohazardous material is used Wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes Operate device in a safety hood Decontaminate
218. use only and not for therapeutic or diagnostic use autoMACS Pro Separator User manual Version 4 Original instructions Miltenyi Biotec GmbH Friedrich Ebert Stra e 68 51429 Bergisch Gladbach Germany Phone 49 2204 8306 0 Fax 49 2204 85197 macs miltenyibiotec de www miltenyibiotec com Content EN FR DE ES 2 1 2 2 3 1 3 2 3 3 3 3 1 3 3 2 3 3 3 3 3 4 3 3 5 3 3 6 3 3 7 4 1 4 2 4 3 4 3 1 4 3 2 4 3 3 4 3 4 44 4 5 4 6 4 7 4 7 1 4 7 2 Content Important information Informations importantes Wichtige Informationen Informazioni importanti Informacion importante Introduction MACS Technology the gold standard in cell separation The autoMACS Pro Separator Assembly and installation Unpacking and installing the autoMACS Pro Separator Replacing fluid bottles and the connection of fluid sensor cables Running the autoMACS Pro Separator for the first time performing a test calibration Switch ON the autoMACS Pro Separator Check the date and time Perform a test calibration Installation of the autoMACS Columns Priming the autoMACS Pro Separator Checking the fluid levels Test of barcode scanner An overview of the autoMACS Pro Separator user interface Working with the lower menu bar The main menu screen Status menu Status of fluid bottles Column status Tube rack status MACS MiniSampler status Separation menu Reagent menu Log list menu Option menu Special
219. use wear of the pump seal and thus may lead to leakage The pump syringe should not run dry at any time This can damage the pump seal and thereby may lead to leakage of the pump syringe Note To prevent the formation of salt deposits wipe the outlet ports with a tissue soaked with distilled or deionized water before each Sleep program 9 1 4 Pump syringe is filled with air during operation If there is any air inlet into the pump syringe during operation the correct proceeding of a separation will be impaired 1 5 Check all tubings that are connected to the fluid bottles Make sure that all tubings are fastened properly If a screw thread is damaged order and install new tubing Check if the hydrophobic air filters connected to the fluid bottles are clogged Clogging may cause positive or negative pressure in the fluid bottles which can lead to pressure problems in the fluidic system If filters are clogged replace them with new hydrophobic air filters refer to section 7 4 4 Check if the connections and pump syringe are leaky refer to section 9 1 3 Check if the uptake port needle is connected correctly and no air inlet is possible If not unscrew and check screw threads If they are undamaged reinsert precisely and fasten Then use the wrench to turn an extra quarter turn Do not to overtighten the screw If the problem persists contact Miltenyi Biotec Technical Support 9 1 5 Washing station overflow
220. utdown button on the upper right hand corner of the screen 2 Select Yes from the pop up dialog box ption and select RUN to start Do vou really want to shutdown the device N 150 Figure 6 34 Shutting down the autoMACS Pro Separator 3 The autoMACS Pro Separator will automatically perform a Sleep program 4 Upon completion of the Sleep program you will be prompted to shut down the instrument The fluid bottle illumination is purple at this point Switch OFF the autoMACS Pro Separator by using the main power switch 6 Cell separation using the autoMACS Pro Separator 143 6 3 Decision tree for the optimal separation program Frequency of target cells 4 Depletion Depletion H gt 5 to 10 lt 5 to 10 usingmacse Sing MACS Cell Isolation MicroBeads i Level of Level of Level of Purity is antigen antigen antigen highest expression expression expression priority Normal to i Normal to i L Normal to high oi high ow oN high Recovery Purity is Recovery is highest highest is highest priority priority priority y y tt v o 2 3 E 2 u E 9 3 3 2 2 6 a E ri v 6 2 TD a 2 v v 2 2 D Ro o 2 3 a E E o E a o 2 3 s 3 E E a z gt 5 E S S S 3 Tn 3 Ki f a 5 B 5 N gt Gi o o ha o 3 2 ve 95 DA v WS gt i El El El v5 a 2 MIES w 2 5 a y y v vu 5 5
221. utoMACS Pro Separator is an innovative instrument for automated sensor controlled multisample labeling and separation of various cells types At the touch of a button target cells are magnetically labeled separated and eluted in a fully automated fashion Purify and progress Important information 1 Please read before use Please read all information contained in this user manual before use Failure to read and follow these guidelines could lead to improper or incorrect use handling or care of your instrument and could cause hazards to users unpredictable results instrument malfunction or damage premature wear and reduced life time of the instrument and may void your warranty Keep this user manual in a safe place accessible for anyone using the autoMACS Pro Separator This chapter describes the safety instructions and site requirements for your autoMACS Pro Separator The following warnings and cautions are provided to help you prevent injury to yourself or damage to the instrument 1 1 Symbols and hazard levels Setup of safety notices Example CAUTION Moving parts stay clear Hazard of puncturing or crushing fingers Operate only with attached needle guard The safety notices inform the user about potential risks if warnings and precautions outlined below are not followed The icon on the left side specifies the risk The hazard level at the top classifies the hazard as mentioned above The level type
222. y stop operation and will display a pop up warning screen from which the current program can be continued or canceled 6 Cell separation using the autoMACS Pro Separator Frocass paused Autolabeling a 10 40 Figure 6 31 Select Stop to pause cell separation To continue cell separation select Continue To cancel the entire procedure select Cancel 2 Select Cancel to cancel the procedure Alternatively select Continue to carry on with cell separation Ready for separation GRINSE py Figure 6 32 The process has been cancelled Fluid bottle illumination The autoMACS Pro Separator has a fluid bottle illumination that facilitates monitoring of the instrument s status The table below summarizes the color code of the fluid bottle illumination and the respective user action required Code Status User action Green Ready for separation No action required Blue Instrument operating No action required Yellow Not ready for separation Run wash program Rinse or Orinse before starting a separation Red Error Check screen for error detection Purple Program Sleep Switch OFF is completed autoMACS Pro Separator Blinking Action required Check screen for required action Table 6 6 Various fluid bottle illumination statuses 6 Cell separation using the autoMACS Pro Separator 141 142 Note If Sleep is chosen as a wash program the autoMACS Pro Separator will not allow definition of a
223. zardous voltage Protective conductor terminal Symbol is attached on the inside of the instrument Warning for service personnel ON supply OFF supply Documentation needs to be consulted before proceeding with installation and operation of the system 1 Important information 1 2 Warnings and precautions The autoMACS Pro Separator employs state of the art technology It is a computer controlled instrument for the automated separation of magnetically labeled cells using MACS Technology The MACS MiniSampler connects to the autoMACS Pro Separator and thus represents a part of the cell separation instrument The autoMACS Pro Separator and the MACS MiniSampler are designed to operate safely after installation and when used by trained personnel according to general safety practices and the instructions set forth in this user manual The guidelines in this section explain the potential risks associated with the operation of the instrument and provide important safety information in order to minimize these risks By carefully following the instructions you can protect yourself and the equipment from potential hazards and create a safe work environment If this instrument is used in a manner not specified by the manufacturer protection may be impaired IMPORTANT Please read and follow all operating instructions in this user manual and pay attention to all warnings displayed on the instrument Retain this user manual and any other
224. zione d aria attorno al autoMACS Pro Separator mantenendo uno spazio libero di almeno 15 centimetri su ogni lato durante il funzionamento onde assicurare un adeguato raffreddamento dello strumento Evitare l esposizione diretta ai raggi solari Le fessure e le aperture dello strumento sono necessarie per la ventilazione e non devono essere in alcun caso ostruite o coperte poich garantiscono il funzionamento corretto del autoMACS Pro Separator e proteggono il dispositivo dal surriscaldamento Non inserire mai corpi estranei nelle aperture dello strumento 1 4 3 Acqua e umidit Non utilizzare lo strumento in ambienti bagnati o umidi Evitare ambienti ad elevata umidit o condizioni di condensa e proteggere lo strumento dagli schizzi d acqua 1 4 4 Prodotto con messa terra Lo strumento dotato di un sistema di alimentazione a tre fili ed provvisto di spina con un terzo polo per la messa a terra La spina pu essere inserita unicamente in una presa di corrente con messa a terra per ragioni di sicurezza Non cercare di inserire la spina in una presa sprovvista di messa a terra Qualora non riusciate ad inserire la spina nella presa vi consigliamo di contattare il vostro elettricista di fiducia per sostituire la presa 1 4 5 Fontidi alimentazione Lo strumento deve essere alimentato unicamente dalla fonte indicata sulla targa relativa alle caratteristiche di alimentazione del prodotto In caso di domande sul tipo di alimentazione da

autoMACS® Pro Separator User manual

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autoMACS® Pro Separator User manual