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autoMACS® Pro Separator User manual

1. 4 3 2 2 Assign one labeling option to several samples 1 Highlight several samples using the sample rack template 2 Scroll through the Labeling submenu using the arrows 3 Select the desired Labeling reagent 4 3 2 3 Apply an assigned labeling option to other samples 1 Assign a program to the first sample refer to above 2 Select subsequent samples The same program will be automatically applied to the selected samples 4 3 3 Assigning sample volumes The Volume submenu El is used to enter or modify sample volumes It is man datory to assign a volume for autolabeling For manually labeled samples it is not mandatory to assign a volume However the autoMACS Pro Separator requires this information to calculate and display the total sample processing time Volumes are given in microliter uL 91 USER INTERFACE 92 1 Highlight one or more sample s using the sample rack template 2 Touch the Volume submenu A dialog box will appear 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 the Monocyte Isolation Kit Il human Note The minimal incubation volume after the addition of the first reagents may not be less than 200 uL 4 Press Enter 4 3 4 Assigning a cell separation program The Separation
2. Make sure that the peristaltic waste pump works properly Run the Check_up program Go to Option gt User settings gt Check_up and press 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 3 3 Clean the washing station refer to section 7 2 4 Reassemble the unit and check whether the problem persists If the problem persists replace the pump head refer to section 7 3 3 9 2 6 Outlet port is clogged Wipe the outlet port with a tissue soaked with 70 ethanol or double distilled water Flush the outlet port manually using a syringe filled with 70 ethanol or double distilled water 9 2 7 Contamination of tubing system To prevent contamination run a Sleep program before turning off the instrument Decontaminate the instrument Refer to section 7 2 5 for details If the problem persists call Technical Support 9 2 8 MACS MiniSampler does not move properly Check whether the guiding of the MACS MiniSampler is connected properly to the connector at the autoMACS Pro Separator labeled External CAN 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 Check the cable connection between MACS MiniSampler and autoMACS Pro Separator Check for cable damages Check whether the MACS MiniSampler can freely move to both sid
3. livelli di tensione su questi connettori non devono superare i livelli di tensione ammessi pari a 30 V rms e 42 4V 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 60825 1 Utilizzare unicamente cavi di connessione inferiori ai 3 metri di lunghezza Forte campo magnetico autoMACS Pro Separator provvisto di un magnete estremamente potente Esiste un forte rischio per la salute di portatori di pacemaker drenaggi cerebrali o impianti medicali 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 51 al 3 m 5 IMPORTANTI ISTRUZIONI DI SICUREZZA 5 52 gt gt una distanza minima di 20 cm dal coperchio del magnete Questi oggetti potrebbero essere danneggiati o il loro funzionamento essere compromesso dal campo magnetico Rischi meccanici 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 l
4. 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 6 2 2 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 2 3 Untouched isolation To isolate a particular target cell type in an unlabeled i e untouched form non target cells are magnetically 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 6 2 4 Sequential sorting A combinat
5. CAUTION Le s parateur autoMACS Pro est con u pour une utilisation s re si il est correctement install et mani par un personnel qualifi conform ment aux mesures de s curit g n rales et aux consignes libell es dans ce manuel 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 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 quipement veuillez vous adresser a votre organisme local responsable de l approvisionnement en lectricit des travaux de b timent de la maintenance et de la s curit Des mots cl sont utilis s pour identifier des messages sur des d t riorations mat riels et de s curit Les mots cl suivants sont utilis s dans ce manuel ou WARNING indique une situation potentiellement dangereuse pouvant entra ner la mort o
6. Miltenyi Biotec autoMACS Pro Separator User manual Version 5 Original instructions Copyright 2014 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 GmbH however notwithstanding the foregoing the owners of the gentleMACS Octo Dissociator with Heaters 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 GmbH in the preparation of this user manual However Miltenyi Biotec GmbH 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 gentleMACS and MACS are registered trademarks or trademarks of Miltenyi Biotec GmbH All other trademarks mentioned in this publication 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 use only and n
7. Press Height and check the positioning ofthe uptake needle at the bottom of the tube To do so press Down until the needle hits the bottom and the Down button disappears Go up again by one increment The needle should be centered If the needle tip is not located directly in the middle of the conical tube press Position Use the needle navigation buttons Move back Move fwd to correct the needle position Press Done to continue with calibration Press Save to store the new settings 103 104 5 1 1 5 Calibration of the x axis step 4 5 1 Press Calibrate to proceed with calibration Service tools Calibr_2 Check_up Calibration 1 Step 4 5 x axis Current calibration value from unknown x position 0 Cancel Previous Calibrate Next 11 44 Figure 5 13 Calibration 1 Step 3 5 check needle position above the chill rack 2 If you haven t done so yet place a Chill 15 Rack onto the MACS MiniSampler Place three 5 mL conical centrifugation tubes in positions A1 B1 and C1 Press Done to confirm The needle arm will move to position A1 of the tube rack Select Use to start the calibration at current position recommended Press Reset to use factory settings Press Height and check the positioning of the uptake needle at the bottom of the tube To do so press Down until the needle hits the bottom and the Down button disappears Go up again by one increment The needle should be centered If the
8. co 2 O a E Check configuration and connection of the protective cover the MACS MiniSampler and the front support at the autoMACS Pro Separator then touch Continue Touch Cancel to abort Please close protective cover and touch Continue Touch Cancel to abort Check electrical connection of MACS MiniSampler If detected the MACS MiniSampler symbol would be displayed in the status screen 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 153 154 7030 7031 7032 7033 7035 7036 7038 7039 7048 The instrument has been shut down without using Sleep 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 Uptaken sample has been processed Portion of 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
9. If material like whole blood or tissue is primarily used it is recommended to run the Safe program once a month Store prepares the instrument 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 Upon completion of the Store program the fluidic system contains 70 ethanol 4 5 2 User settings User settings contain programs necessary for maintenance and setup of the autoMACS Pro Separator About System and hardware information This program informs the user about the software version serial number of the instrument and other hardware information Calibr_1 Calibrating the needle arm position This program is used for the calibration of the needle arm to the washing station and tube racks Refer to section 5 for more information Calibr_2 Calibrating the liquid volume control This program is used for the calibration of the liquid volume control Refer to section 5 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 Refer to section 9 1 for details O_bcr 2D code reader setup configuration and initialization The O_ber program allows th
10. Kontaktieren Sie Miltenyi Biotec um Ersatzaufkleber zu erhalten Allgemeine Sicherheitshinweise Falls Ihr Ger t nicht einwandfrei funktioniert und oder Anzeigen auf dem Display Sie dazu auffordern den technischen Kundendienst zu kontaktieren ist die Betriebs sicherheit des Ger tes nicht l nger gew hrleistet Schalten Sie das Ger t sofort aus ziehen Sie den Netzstecker und kontaktieren Sie einen autorisierten Miltenyi Biotec Servicedienstleister oder den Miltenyi Biotec Technical Support Elektrische Gef hrdung und Brandgefahr 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 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 auBen in das Instrument hinein gelangen Bei Auftreten von Flammen oder
11. 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 Product 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 163 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 Australia Miltenyi Biotec Australia Pty Ltd Unit 16A 2 Eden Park Drive Macquarie Park NSW 2113 Australia Phone 61 2 8877 7400 Fax 61 2 9889 5044 macs miltenyibiotec com au Benelux Miltenyi Biotec B V Schipholweg 68 H 2316 XE Leiden The Netherlands macs miltenyibiotec nl Customer service The Netherl
12. 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 If the outcome of a cell separation procedure is deemed unsatisfactory this may either be due to incorrect function of the instrument or to inappropriate processing of the sample This section discusses possible instrument malfunctions that may appear In section 9 3 a list of numerically encoded errors and warning messages are presented along with user actions for troubleshooting If an error with moving hardware components occurs it is recommended to run the Check_up program The program automatically analyzes the functionality of moving hardware components A report is displayed after the analysis of each single component Following hardware components are analyzed dilutor valve valves 1 4 peristaltic waste pump magnet 1 2 needle arm movement along the z axis and the y axis and MACS MiniSampler Furthermore the calibration data is checked 1 Goto Option gt User settings gt Check_up and press Run 2 Aftereachstep press Cancel to quit or press OK to continue Possible hardware problems will be detailed on the screen 145 146 9 2 Hardware issues not indicated by an error screen 9 2 1 Column leakage 1
13. rat and non human primate cells 6 1 2 Indirect magnetic labeling Indirect magnetic labeling is a two step procedure First cells are labeled with a primary antibody directed against a cell surface marker The primary antibody can either be unconjugated biotinylated or fluorochrome conjugated Second 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 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 During depletion the unwanted cells are labeled and depleted from the target cells Furthermore sequential sorting allows the performance of two consecutive separations 6 2 1 Positive selection To isolate a certain cell type the target cells are magnetically labeled During separation the target cells are retained within the column Unlabeled cells flow through and are collected as the negative fraction After automated retraction of
14. rinsed but might still be contaminated Collision of needle with bottom of tube could not be detected at expected position 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 Press 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 Press 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
15. the correct proceeding of a separation will be impaired 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 2 2 Check if the connections and pump syringe are leaky refer to section 7 2 3 e 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 e If the problem persists contact Technical Support 9 2 5 Washing station overflow Verify that the washing station is not clogged with salt deposits Take out the washing station and clean as indicated in section 7 2 4 To prevent the formation of salt deposits wipe the outlet ports with a tissue soaked with distilled or deionized water before each Sleep program TROUBLESHOOTING 147 148 Reassemble washing station and run a Rinse program Go to the Separation menu and press Wash Now Select Rinse and press Run If the problem persists follow the steps below
16. the next step the target cells are magnetically labeled with MACS MicroBeads directed against the second marker and again subjected to positive selection 6 3 Separation programs The autoMACS Pro Separator provides a selection of fourteen 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 6 3 1 Positive selection programs During the positive selection programs Possel and Possels 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 rack During the 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 ar
17. 4 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 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 6 4 1 Wash programs Qrinse Standard short wash program that only uses Running Buffer It is recommended to use this program between separations of cells with normal frequency Rinse 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 An optional very extensive rinsing program It uses storage solution Washing Solution and Running Buffer It may be used after whole blood and bone marrow applications Clean has to be enabled first to appear in the Wash menu Sleep Mandatory as the last wash program before overnight storage Upon completion of this program the fluidic system contains 70 ethanol 6 4 2 Maintenance programs Safe Designed for decontamination of the fluidic system Upon completion of this program the fluidic system contains Running Buff
18. Goto Option gt User settings gt Set_time Press Run 2 Select the time or date fields by touching the respective fields on the display 3 Use the keypad to change the date or time please input hours 0 23 Ft A 0 46 16 10 46 Figure 3 6 Select the desired field and use the numeric keypad to modify the date and or time 4 Press Ok to return to the Option menu The procedure can be cancelled at anytime by pressing Cancel 3 1 1 4 Testing the bar code scanner CAUTION Do not look directly at the light emitted by the 2D code reader Eye injury may result Do not activate the 2D code reader if the washing station cover is open 1 Go to Reagent gt Read Reagent 2 Place a barcode e g the test bar code below in front of the 2D code reader Refer also to section 4 2 The name of the scanned reagent will appear in the display in this case CD4 Microbeads human 130 045 101 ul he Figure 3 7 Test barcode 3 1 1 5 Testthe needle calibration Correct needle calibration is necessary for proper sample and reagent uptake Before using the autoMACS Pro Separator for the first time check whether the uptake and output needle are positioned correctly when moving into in the washing station sample tubes and reagent vials uptake needle only Test needle calibration also if the instrument was moved to a different place Please refer to section 5 for details 3 1 2 Checking fluid levels The closures for the fluid
19. If a freshly installed autoMACS Column shows signs of leakage check if the column is installed properly The column should be inserted precisely into the column connector and fastened to the point of resistance If this is not the case loosen the column connector insert the column precisely and tighten the connector again Run the Qrinse program Go to the Separation menu from the upper navigation bar and press Wash now Select Qrinse and press Run Check if the leakage persists If so unscrew the column and check if the luer connectors of the columns are damaged If this is the case exchange the leaking column with a new autoMACS Column refer to section 7 2 1 Check if the column connector is fastened properly If not use second wrench to counter and tighten another quarter turn If the problem persists contact Technical Support 9 2 2 Tubing leakage 1 Identify the location of the leaky tubing by running the Rinse program Press Wash Now from the lower menu bar in the Separation menu Select Rinse and press Run Check whether the tubing is tightened properly If this is not the case tighten the tube connector The connector should be inserted precisely If the problem persists loosen the tube connector and pull back the connector from the tubing Do not remove the connector from the tubing Check the ends of the tubing for wear and fissures If necessary replace tubing with the appropriate spare part Insert the tubing in
20. 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 while unlabeled cells pass through and are collected as the negative fraction Columns rapidly demagnetize when the column is removed from the separator or in case of the autoMACS Pro Separator the magnet is automatically retracted from the autoMACS Column The labeled cells are eluted as the positive fraction 57 58 Isolated cells can be immediately used for downstream applications including flow cytometric analysis cell culture and molecular analysis Thus with MACS Tech nology both labeled and unlabeled cells can easily be isolated with high purity and recovery 1 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 depending on 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 ce
21. Reagent Rack 4 in the correct order 6 Goto the Separation menu to proceed with programming a cell separation procedure see section 4 3 for details 4 2 2 Manual entry of reagents 1 Goto the Reagent menu and highlight the position where the vial will be placed on the reagent rack 2 Select Enter Reagent from the lower navigation bar Enter the reagent specific product order number Press Ok entr Product no 130 045 kxx 3 4 11 02 Figure 4 11 Reagent information is entered manually 89 USER INTERFACE 90 3 Ifa correct number is inserted the software will recognize the reagent or kit Select the reagent from the list by using the touch screen 4 Press Ok to confirm the identified reagent and its reagent vial position 4 2 3 Deleting reagents 4 2 3 1 Deleting a single reagent from the reagent list 1 Highlight the reagent name that is to be deleted 2 Press Delete Selected to remove the highlighted reagent 4 2 3 2 Deleting the entire reagent list 1 Highlight an unassigned position on the reagent rack If there are no unassigned positions delete an individual reagent as described above 2 Press Delete All 4 3 The Separation menu The Separation menu schematically represents a sample rack For each sample rack position it is possible to define cell labeling cell separation and washing programs Program combinations can be saved as a template for later use by pressing the Save Template butt
22. Risk of crushing and shearing of bodily parts Hazardous optical radiation Hazardous laser radiation Biohazard Risk of contamination if potentially dangerous biological material is used Protective conductor terminal Symbol is attached on the inside of the instrument Information for service personnel On Power supply Off Power supply Safety labels Please notice the hazard points and safety symbols of the autoMACS Pro Separator Risk of crushing and shearing of bodily parts Hazardous optical radiation Risk of crushing and shearing of bodily parts Hazardous laser radiation Figure 1 Hazard sources and the position of the safety symbols on the front of the autoMACS Pro Separator Figure 2 Position of the safety symbols on the back of the autoMACS Pro Separator g m E a IMPORTANT SAFETY INFORMATION av 12 The above mentioned safety labels and safety markings must be kept clean and legible Periodically inspect the safety labels and safety markings and replace them if they are not legible or perceptible from a safe viewing distance Contact Miltenyi Biotec for replacement labels General safety instructions If the instrument is not working properly and or the displayed instructions or messages advise you to contact Technical Support safe operation of the instrument is no longer possible Immediately switch off and unplug the instrument from the power outlet
23. Step 2 5 NEG washing statio Current calibration value from 20140715 NEG position at washing station 55 y Cancel Previous Calibrate Next 11 43 Figure 5 11 Calibration 1 Step 2 5 check NEG needle position in the washing station 2 Chose either current position to start with recommended press Use or reset needle position to factory settings press Reset The needle arm will automatically move towards the washing station The needle should be located directly above the center of the front opening of the washing station see figure 5 5 3 4 5 1 Select Height and check the central positioning of the 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 Select Save to save the new configuration 1 1 4 Calibration of the z axis step 3 5 Select Calibrate to proceed with calibration Special Service tools Check_up Calibration 1 Step 3 5 Rack Current calibration value from unknown Rack position O Cancel Previous Calibrate Next 11 44 Figure 5 12 Calibration 1 Step 3 5 check needle position above the chill rack 2 5 CALIBRATION If you haven t done so yet place a Chill 15 Rack onto the MACS MiniSampler Place three 15 mL conical centrifugation tubes in positions A1 B1 and C1 Confirm be pressing Done The needle arm will move to rack position A1
24. Technical data of the autoMACS Pro Separator CE Technical data 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 Table 10 2 Technical data of the MACS MiniSampler 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 used in accordance with the instructions may cause harmful 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 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 technician for
25. The washing station should be cleaned as necessary to remove spills and salt crusts Do not autoclave the washing station or wash using a dishwasher Note This procedure requires a column exchange 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 Figure 7 7 Remove the peristaltic pump tube from the washing station 5 Unscrew the knurled screw that attaches the washing station to the instrument Figure 7 8 Unscrew the knurled screw MAINTENANCE 131 132 6 Pulloutthe 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 2 5 Instrument disinfection The Safe program is a disinfectant procedure that uses MACS Bleach Solution for cleaning of the autoMACS Pro Separator Depending on the level of use and general instrument maintenance it is recommended to run the Safe program every 3 to 6 months to disinfect the fluidic system If predominantly whole blood bone marrow or tissue samples are separated the Safe procedure can be performed more o
26. 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 t zen und ein sicheres Arbeitsumfeld zu garantieren Bei Verwendung des Ger ts unter Missachtung der Herstelleranweisungen ist die Sicherheit nicht gew hrleistet 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 Gefahrensymbole sollen auf Sicherheits und Sachschadenshinweise aufmerksam machen Folgende Gefahrensymbole werden in diesem Benutzerhandbuch verwendet oder WARNING bezeichnet eine Gefahrensituation die falls sie nicht vermieden wird zum Tode oder schwerwiegenden Verletzungen f hren kann oder CAUTION bezeichnet eine Gefahrensituation die falls sie nicht vermieden wird zu leichteren oder mittelschweren Verletzungen f hren kann Es kann auch verwendet werden um vor unsicherem Gebrauch zu warnen 17 18 PPPRPP ODE gt Gefahrensymbole Die folgenden Symbole werden benutzt um Gefahrensituationen anzuzeigen welche zu Sach und Personenschaden f hren k nnen Warnzeichen Gefahrenrisiko Dieses Benutzerh
27. Unscrewing the tubing connected to the dilutor valve 4 Unscrew the two hexagonal socket screws using the allen key that was delivered with the new or exchange valve Figure 7 17 Unscrewing the dilutor valve s socket srews 5 Pull the dilutor valve out of the coupling 6 Insert the new valve Contact Tech Support for assistance 7 When the dilutor valve is properly inserted mount the hexagonal socket screws 8 Connect the tubing according to the letters on the diluter valve and tubing Figure 7 18 Connect tubing to the dilutor value as indicated by the letters 9 Guide the draining tube towards the washing station as shown below MAINTENANCE 137 138 Figure 7 19 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 Figure 7 21 Dismantle 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 22 Removal of the waste distributor 12 Mount the new draining tube and finger tighten Take care to clean spilled fluids with ethanol or disinfectant Figure 7 23 The draining tube is attached 13 Install the waste distributor back to its former position and reinstall the washing station 14 Install the syringe pump 15 Switch on the autoMACS Pro Separator 16 Ch
28. Yeoksam dong Gangnam gu Seoul South Korea Phone 82 2 555 1988 Fax 82 2 555 8890 macs miltenyibiotec co kr Spain Miltenyi Biotec S L C Luis Bufiuel 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 www miltenyibiotec com 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 or trademarks of Miltenyi Biotec GmbH All other trademarks mentioned in this publication are the property of their respective owners and are used for identification purposes only Copyright 2014 Miltenyi Biotec GmbH All rights reserved 140 002 016 07
29. a causa di differenze legislative Si prega di notare che a seconda del tipo e dell utilizzo delle apparecchiature possono essere necessari requisiti aggiuntivi Per ulteriori informazioni o se si desiderassero smaltire le apparecchiature Miltenyi Biotec in disuso si prega di contattare il Responsabile di Zona o il Supporto Tecnico Miltenyi Biotec 55 al 3 m 5 IMPORTANTI ISTRUZIONI DI SICUREZZA 5 56 MACS Technology has become the standard method in cell separation It is based on MACS MicroBeads highly specific antibodies coupled to superparamagnetic 50 nm particles MACS Columns containing a matrix of ferromagnetic spheres and MACS Separators providing a strong magnetic field Cells in a single cell suspension are paramagnetically labeled with MACS MicroBeads The colloidal suspension of MACS MicroBeads allows easy handling and short incubation times Cells can be separated in less than 30 minutes As minimal amounts of MicroBeads are required for magnetic labeling a large number of epitopes remains available for fluorescent labeling and subsequent cell analysis Labeling with MACS MicroBeads and fluorochrome conjugated antibodies can be performed silmultaneously which further decreases handling times 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 When a
30. 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 vorrichtungen niemals umgehen oder manipulieren Gefahr durch optische Strahlung Das Ger t ist zur automatischen Reagenzglasgestell erkennung Klasse 1M mit vier oberfl chenemittierenden 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 Hinein schauen 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 verwen det 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 f llt in die Freie Gruppe Der Gef hrdungsabsta
31. check needle position in the washing station CALIBRATION 5 1 1 2 Calibrate the washing station Uptake needle step 1 5 1 Chose current position to start with recommended press Use The needle arm will automatically move towards the washing station The uptake needle should be located directly above the center of the rear opening of the washing station see figure 5 3 Separation Status flog list About calor DE Check _up Calibration 1 Step 1 5 Washing station Adiust uptake needle centrically to rear orifice Current offset O Move Move Height Done back Fwd Calibrate washing station 11 34 Figure 5 9 Calibrate the uptake needle position in the washing station 101 102 Press Height 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 Calibr_2 ck_up Calibration 1 Step 1 5 Washing station Adiust uptake needle centrically to rear orifice Current offset O Move Move Position Done up down Calibrate washing station 11 35 Figure 5 10 Calibrate the uptake needle position in the washing station 4 Select Save to save the new configuration 5 1 1 3 Calibrate the washing station NEG needle step 2 5 1 Select Calibrate to proceed with calibration Service tools Calibration 1
32. 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 cells are remaining in tube Required volume cannot be provided in given Chill Rack Miscalculation of diluter movement Target position is negative Table 9 1 Error codes and possible remedies Always use Sleep to shut down the instrument 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 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 leakage of air into system behind bubble sensor Readjust tubing connectors and verify correct fastening of columns Ex
33. 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 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 Upgrades zu Ihrem Ger t Fragen Sie Ihren rtlichen Miltenyi Biotec Vertriebsmitarbeiter nach Miltenyi Biotecs weit reichenden Vereinbarungen zum Ger teservice und Technical Support oder besuchen unsere Website www miltenyibiotec com support 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 gro er 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 Dekontaminie
34. 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 other 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 161 162
35. have a serious detrimental effect on the environment and or human health That is why all equipment must be specifically collected and treated by designated waste facility centres and by qualified WEEE compliance schemes By ensuring that you dispose of your unwanted electrical and electronic equipment according to the applicable WEEE and hazardous waste disposal legislation you are helping to preserve our natural resources and protect human health Miltenyi Biotec is committed to protecting the environment Miltenyi Biotec offers product end of life return programs in many countries and partners with licensed WEEE compliance schemes throughout the world Miltenyi Biotec lets you recycle your end of life Miltenyi Biotec equipment free of charge The terms and availability of this offer vary by geography because of differences in regulatory requirements Please note that depending on the type and use of your equipment additional requirements may apply For more information or if you wish to dispose of your end of life Miltenyi Biotec equipment please contact your local Miltenyi Biotec representative or Miltenyi Biotec Technical Support NWARNING WARNING CAUTION Der autoMACS Pro Separator ist f r einen sicheren Betrieb ausgelegt wenn das Ger t richtig installiert von geschultem Personal bedient und entsprechend der Sicherheitsrichtlinien und Anweisungen in diesem Benutzerhandbuch eingesetzt wird Dieses Kapitel erkl rt die im
36. needle tip is not located directly in the middle of the tube press Position Use the needle navigation buttons Move left Move right to correct the needle position Press Done to continue with calibration Press Save to store new settings 5 1 1 6 Test current calibration settings step 5 5 1 Press Test to check the new configuration The autoMACS Pro Separator will perform a complete test of Calibr_1 settings Reagent Separation Status fLog list Option Calibration 1 Step 5 5 Test Test calibration data Previous Test End 11 38 Figure 5 14 Calibration 1 Step 3 5 check needle position above the chill rack 2 If you haven t done so yet place a Chill 15 Rack onto the MACS MiniSampler Place 5 mL conical centrifugation tubes in positions A1 B1 and C1 Press Done to confirm 3 If you haven t done so yet place the MACS Reagent Rack 4 containing 4 sample vials onto the MACS MiniSampler Press Done to confirm In the 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 press Done After testing the last postion the Calibr_1 screen will reappear Select Cancel to leave the calibration program If any errors or misalignments were noted re
37. rack e g Chill 5 Rack onto the MACS MiniSampler into the right recess ensuring that the rack bar code is facing the autoMACS Pro Separator Figure 2 11 Install the Chill Rack onto the MiniSampler 9 Attach the protective cover of the MACS MiniSampler Keep the protective cover attached and closed during operation Note The protective cover is sensor controlled Running a separation is not possible if the protective cover is open Figure 2 12 Attach the protective cover of the MACS MiniSampler 71 INSTALLATION 72 NWARNING 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 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 ofthe branch circuit rating Make sure that the main switch and the connector of the power cable are easily accessible and located as close to the operator of the instrument as possib
38. rim of tube or top of rack calibrate needle arm Touch Retry to try again Do not use 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 7010 7011 7012 7013 7014 7015 7018 7021 7022 7023 7024 7026 7027 7028 7029 Not certain if liquid surface of sample has been detected correctly Restart of the instrument is required Calibration data not found Calibration data not found Calibration data not found Columns are not installed Calibration data not found A separation program has been started but system had not been rinsed properly Columns are overdue Plunger movement blocked because of column clogging blocked tubing set or any other cross section constriction during output of the negative fraction The negative fraction has not been eluted completely Negative cells are still remaining in the system The number of programmed sample positions exceeds the actual positions of the rack on the MACS MiniSampler The protective cover of the MACS MiniSampler seems to be o
39. segura Apague y desenchufe inmediatamente el instrumento de la toma de corriente y contacte un proveedor de servicios autorizado de Miltenyi Biotec o un servicio t cnico autorizado Incendios y riesgos el ctricos 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 Unicamente por personal autorizado Se debe tener especial cuidado cuando se manejen liquidos Limpie inmediatamente los liquidos vertidos Impida que stos accedan al interior del aparato Desenchufe el cable de electricidad antes de proceder a limpiar manualmente el separador autoMACS Pro Existe un riesgo potencial en caso de usar separador autoMACS Pro abierto que se haya caido o que est averiado si se han derramado liquidos 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 inmediatamente 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 El separador autoMACS Pro est dise ado para ser usado exclusivamente en interior No utilice el equipo e
40. submenu El 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 one separation program to multiple samples Refer to section 6 2 and 6 3 for a detailed explanation of the various cell separation strategies 1 Highlight one or more sample s using the sample rack template 2 Scroll through the Separation submenu using the arrows 3 Select a Separation program 4 3 5 Assigning a wash program The Wash submenu El is used to instruct the autoMACS Pro Separator which wash program should be applied to each sample after separation Four wash programs are available to choose from Qrinse Rinse Sleep and Clean Refer to section 6 4 for a detailed explanation of the respective programs 1 Highlight one or more sample s using the sample rack template 2 Scroll through the Wash submenu using the arrows 3 Select the desired Wash program 4 3 6 Delete program combinations To erase selected program combinations from the programming field first highlight the respective positions and choose Delete selected from the lower navigation bar If positions are not highlighted the button can be used to delete the entire template In this case the button Delete selected will switch to Delete all 4 4 The Log list menu The Log list menu displays a complete log of actions performed by the autoMACS Pro Separator Completed processes are mar
41. the top screw only Do not compress the glass as it might break 8 Carefully remove the plunger from the syringe Place a container underneath to prevent spilling Remove salt crusts with distilled or deionized water 129 130 Figure 7 5 Wash the syringe with distilled water 9 Gently push the plunger halfway into the syringe Dry the plunger lock screw before proceeding with installation of the syringe 10 Fasten the syringe at the dilutor valve by turning the top screw clockwise until a resistance is met Unfasten again by one rotation Grip the syringe at the top knurled screw only Do not compress the glass as it might break 11 Pull the plunger out of the syringe until it reaches its fitting in the plunger holder The plunger must have the same orientation as the fitting see figure 7 2 Tighten the plunger lock screw NY Figure 7 6 Place the plunger in its fitting and tighten the plunger lock screw 12 Tighten the syringe at the diluter valve by turning until a resistance is met 13 Switch on the instrument The plunger holder will move up again 14 Prime the autoMACS Pro Separator as described in section 3 1 3 15 Run the program Calibr_2 to calibrate the fluidic volume control of the instrument Refer to section 5 for details 7 2 4 Cleaning the washing station The washing station is designed for automated rinsing of the outlet and uptake ports as well as surface cleaning of the uptake and outlet port needle
42. 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 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 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 l 80 della potenza del circuito 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 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 originali ai connettori contrassegnati con External CAN CAN1 e CAN2
43. washing program for each sample position The selected programs will be displayed in the programming field Press Run Note The autoMACS Pro Separator ensures that there is enough solution for one separations If more than one separation is to be performed the user should check whether there is enough solution in the bottles Therefore a pop up window will ask the user to check and confirm the filling status before proceeding to separation Press Continue to proceed 6 10 3 Working with templates For convenience it is possible to save reagent and separation templates Reagent templates can be used in combination with separation templates 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 rack can handle If the tube rack does not match the template definition a warning screen will be displayed upon starting the separation 6 10 3 1 Saving a reagent template 1 Inthe Reagent menu assign reagent vials to reagent rack positions 2 Press Save Template 3 Allocate a name to the template 4 Press Ok 6 10 3 2 Saving a separation template 1 Inthe Separation menu configure the sample template 2 Press Save Template 3 Allocate a name to the template 4 Press Ok 6 10 3 3 Loading a template 1 Inthe respecive menu Reagent or Separation menu press the Load Template button To scroll through the list of saved temp
44. 0 1 9 2 10 2 9 3 10 11 12 Valve exchange Dilutor valve exchange Exchange of the peristaltic pump head Exchange of tubing system Exchange of fuses Quality Control Cell recovery Purity of isolated cell population Viability of the cells Troubleshooting Issues with moving hardware Hardware issues not indicated by an error screen Column leakage Tubing leakage Pump syringe leakage Pump syringe is filled with air during operation Washing station overflow Outlet port is clogged Contamination of tubing system MACS MiniSampler does not move properly Touchscreen remains dark Disruption of power supply during cell separation Rescue procedure A Rescue procedure B Hardware problems indicated by an error screen Technical data and specifications Technical support Limited warranty 133 136 140 141 141 143 143 144 144 145 145 146 146 146 147 147 147 148 148 148 148 149 149 149 150 155 159 161 NWARNING WARNING CAUTION The autoMACS Pro Separator is designed for safe use when installed correctly operated by trained personel operated in accordance with general safety practices and the instructions 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 t
45. 0 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 reflection 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 sur
46. 097 037 Chill5 15 50Set Set of three Chill Racks 130 097 038 MACS Reagent Accommodates 4 vials of MACS Reagents 130 094 574 Rack 4 Air Filter Extension tube and hydrophobic air filter for the 130 091 339 Extension Set autoMACS Fluid Bottles autoMACS Pro Metal plate for operating the autoMACS Pro Separator 130 093 246 Laminar Hood in a laminar flow hood Plate autoMACS Pro Space saving angle adaptors for cables connecting to 130 093 245 Angle Connector the autoMACS Pro Separator Set autoMACS Pro Protection foil for long term storage of the autoMACS 130 093 532 Protection Cover Pro Separator Table 2 3 autoMACS Pro Separator accessories 2 4 Unpacking the autoMACS Pro Separator WARNING Read the instructions in the chapter Important safety information and ensure that your site is properly prepared before continuing with installation and assembly If there is any damage do not use the instrument but contact your local Milteny Biotec representative or Miltenyi Biotec Technical Support Incorrect installation or operation of a damaged instrument can lead to the spread of fire explosion the risk of electric shock or biohazards 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 the shock and position indicators if present suggest incorrect transportation of the instrument If there i
47. 1 7 2 2 7 2 3 7 2 4 7 2 5 7 3 Depletion Untouched isolation Sequential sorting Depletion followed by positive selection Two subsequent positive selections Separation programs Positive selection programs Depletion programs Wash and maintenance programs Wash programs Maintenance programs Preparation of single cell suspensions Adjusting sample volumes Selecting the appropriate tube rack Monitoring the separation process Program status Interrupting cell separation Fluid bottle illumination Priming the autoMACS Pro Separator Cell labeling and separation Cell separation using the autolabeling function Cell separation after manual labeling Working with templates Saving a reagent template Saving a separation template Loading a template Deleting a template Shutting down the instrument Maintenance Daily maintenance Instrument priming Automatic instrument priming at startup Clean uptake outlet ports Periodic maintenance Column exchange Exchange of hydrophobic air filters Clean the pump syringe Cleaning the washing station Instrument disinfection Corrective maintenance 109 109 109 109 110 110 110 111 114 114 114 115 116 118 119 119 119 120 120 121 122 122 123 123 123 123 123 124 125 125 125 126 126 127 127 128 128 131 132 133 7 3 1 7 3 2 7 3 3 7 3 4 7 3 5 8 1 8 2 8 3 9 1 9 2 9 2 1 9 2 2 9 2 3 9 2 4 9 2 5 9 2 6 9 2 7 9 2 8 9 2 9 9 2 10 9 2 1
48. 42 4 V Spitze bzw 60 V DC nicht bersteigen An die mit Bottle Sensor beschriftete Buchse darf nur das autoMACS Pro Flaschensensorkabel angeschlossen werden An die mit RS232 BCR 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 m ssen IEC 60825 1 erf llen Es d rfen nur An schlusskabel mit einer L nge von max 3 Metern verwendet werden Starkes Magnetfeld Der autoMACS Pro Separator enth lt einen extrem starken Magneten Es besteht die Gefahr von schweren Verletzungen f r Personen mit Herzschrittmacher Cerebralshunts oder elektronischen medizinischen Implantaten Halten Sie mit magnetischen Datentr gern Kreditkarten Magnetb nder Speichermedien und dergleichen und elektronischen Ger ten wie H rger te Herzschrittmacher Mess und Steuerger te PCs Uhren mindestens 20 cm Abstand zur Magnetabdeckung da diese durch das Magnetfeld gest rt und gesch digt werden k nnen 21 D m EJ m WICHTIGE SICHERHEITSINFORMATIONEN 22 Pb Mechanische Gefahr Frontabdeckungen nicht ffnen wenn das Ger t l uft Bewegung des Auto matikarms 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 Scherverletzungen und Sch den
49. 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 clean washing station verify that uptake needle is not clogged Switch on instrument if error persists press Details Exchange displayed valve if applicable 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 O E Ir n Lu co gt O a E Restart instrument and try a Qrinse to wash out clogged material If error persists run the Safe program Otherwise exchange diluter valve or standard valves depending on where a constriction is suspected Please
50. 97 A2 2001 als Klasse 1M Laserprodukt eingestuft Die Verwendung von anderen als hierin genannten Bedienungselementen sowie die Anpassung oder Durchf hrung von anderen als hier genannten Verfahren kann gef hrliche Strahlung freisetzen Chemische und biologische Gefahren 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 nic
51. C 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 raggio 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 qualsi
52. For rinsing of the autoMACS Pro 6x1 5L 130 092 987 Washing Solution Separator s fluidic system Storage solution 70 v v analytical grade ethanol in distilled water MACS Bleach Used in combination with Safe 1x1L 130 093 663 Solution program Table 2 2 Materials required for operation 63 INSTALLATION 64 2 3 autoMACSPro Separator accessories The autoMACS Pro Separator is a benchtop instrument that fits in laminar flow hoods or biosafety cabinets The operating environment should be stable and vibration free dust free sufficiently ventilated and free from sources of electro magnetic radiation 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 auto MACS Pro Angle Connector Set 130 093 245 The laminar hood plate provides a stable and even surface even 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 Accessory Description Order no Chill 5 Rack Chill Rack for 5 mL tubes 130 092 951 Chill 5 Racks 3x Chill Racks for 5 mL tubes 130 097 041 Chill 15 Rack Chill Rack for 5 and 15 mL tubes 130 092 952 Chill 15 Racks 3x Chill Racks for 5 and 15 mL tubes 130 097 036 Chill 50 Rack Chill Rack for 5 15 and 50 mL tubes 130 092 953 Chill 50 Racks 3x Chill Racks for 5 15 and 50 mL tubes 130
53. 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 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 El uso de controles el ajuste o la realizaci n de procedimientos con excepci n de los aqu especificados puede dar lugar a una peligrosa exposici n de radiaci n Riesgos qu micos y biol
54. MACS Pro Separator Bottle Sensor 2D Code Reader plug External CAN Power socket Figure 1 2 Back view of the autoMACS Pro Separator 59 2 E U 2 Q O ce 60 NWARNING Read the instructions in the chapter Important safety information and ensure that your site is properly prepared before continuing with installation and assembly Incorrect installation can lead to the spread of fire explosion the risk of electric shock or biohazards Before operating the autoMACS Pro Separator for the first time carefully read the user manual Contact your local Miltenyi Biotec representative for assistance 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 61 62 2 1 Components included in delivery The following components are included in the delivery of the autoMACS Pro Separator Starter Kit 130 092 545 Component Position in transportati
55. Pro Separator using the main power switch 81 E SWITCHING ON OR OFF 82 The autoMACS Pro user interface is composed of five main menus Reagent Separation Status Log list and Option They are accessed through the upper tabbed menu bar see figure 4 1 Six buttons at the bottom of the screen allow for easy interaction with the instrument The function of the buttons may vary depending on which main menu is chosen see figure 4 3 Also depending on the menu and the instrument s status not all buttons are available at all times If a button is unavailable its backgroung color changes from white to grey Menu 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 Displays the current instrument s status 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 Menu options 83 84 4 1 The Status menu The autoMACS Pro Separator is a sensor controlled instrument that allows easy monitoring during operation After initialization the screen automatically displays the Status menu which helps to monitor the instrument It displays the current instrument status and can be accessed at any ti
56. RTANT SAFETY INFORMATION av 16 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 Inquire with your local Miltenyi Biotec representative about Miltenyi Biotec s extensive instrument service and support arrangements or refer to www miltenyibiotec com support 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 Waste of Electrical and Electronic Equipment WEEE customer information Please dispose of your end of life Miltenyi Biotec products in accordance with the applicable WEEE and hazardous waste disposal legislation which may differ by country or region Electrical and electronic equipment may contain hazardous substances which may
57. Rauchentwicklung schalten Sie das Ger t sofort aus trennen es von der Stromzufuhr und kontaktieren einen autorisi erten 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 Der autoMACS Pro Separator ist ausschlie lich ausgelegt f r den Betrieb in Innen r umen Bitte benutzen Sie das Ger t nicht in ausgewiesenen Gefahrenzonen wie etwa sauerstoffangereicherten Arbeitsumgebungen 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 Offnungen und Schlitze am Ger t sind zur Bel ftung gedacht und sollten niemals bedeckt oder blockiert werden da sie das Ger t vor Uberhitzung sch tzen und sicheren Betrieb erm glichen F hren Sie niemals Fremdk rper durch Offnungen in das Ger t ein ap Setzen Sie das Instrument niemals in einer feuchten oder nasskalten Umgebung ein Setzen Sie das Ger t nie hoher Feuchtigkeit oder Kondensation aus und schitzen Sie es gegen Spritzwasser Ziehen Sie den Netzstecker vor Reinigung des Ger tes Verwenden Sie kei
58. TCHING ON OR OFF 75 76 5 Tighten the syringe at the diluter valve by turning the top screw until a resistance is met 6 Switch on the instrument The plunger holder will move up again 3 1 1 2 Installing autoMACS Columns 1 Note the position of the tubing and autoMACS Column slots in the magnet cover column 1 to the left column 2 to the right see figure 1 1 2 Ensure that the fluid bottles are filled with the appropriate solutions 3 Goto Option gt Special Refer to section 4 for details on how to work with the user interface 4 Select Col_ex column exchange from the Detail panel 5 Select Run to start the Col_ex program 6 Wait until the instrument prompts you to exchange the autoMACS Columns 7 Using both hands take the top and bottom of column 1 substitute and pull gently but firmly to remove it from its slot Figure 3 4 Column exchange 8 Place a wide mouth bottle under the column substitute to catch any fluids 9 Hold the column substitute in one hand and gently unscrew the bottom column connector counter clockwise Unscrew the top column connector clockwise see figure 3 5 Store the column substitutes for later usage Note Replace columns with column substitutes if the instrument is shut down for a longer period of time See section 3 2 2 for details Top Column Connector autoMACS Column Bottom Column Connector Figure 3 5 Exchange of autoMACS columns 10 Insert one end of a fresh colu
59. a Chill 15 Rack onto the MACS MiniSampler Place 15 mL conical centrifugation tubes in positions A1 B1 and C1 and confirm by pressing Done 3 When prompted place the MACS Reagent Rack 4 containing four sample vials onto the MACS MiniSampler Confirm by pressing Done 4 A pop up window will appear Test position washing station Select Test position The instrument will automatically place the needle in the washing station Ensure that the needle is centered above the rear opening of the washing station To skip this position select Next To end testing select Test end running Pleas pel aS Check_up Test position washing station Select action Test position Verify washing station position 11 48 Figure 5 2 Running a test calibration 98 Figure 5 3 Uptake needle position in the washing station arrow 5 When done with testing the current position press Done The next position NEG needle will be addressed automatically Log listf ERU ait s running Ple Detail Calibr_2 Check _up Test position NEG at washing station Select action Previous Test Next Test position end Verify washing station NEG position 11 48 Figure 5 4 Test position NEG needle in the washing station Figure 5 5 NEG needle in the washing station 99 CALIBRATION 6 Ensure that the NEG needle is centered above the front opening of the washing station 7 Inthe following the needle
60. accendere lo strumento Nel caso non sia possibile sostituire le parti danneggiate disconnettere lo strumento e non utilizzarlo Il malfunzionamento di parti che contengono materiale potenzialmente infettivo o liquidi venuti in contatto con tali materiali 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 conformit alle disposizioni locali vigenti al 3 m 5 IMPORTANTI ISTRUZIONI DI SICUREZZA 5 53 54 Manutenzione transporto e smaltimento sicuri 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 garanz
61. ads 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 recommendations on the preparation of single cell suspensions please refer to the respective Cell Separation Reagent data sheet 115 CELL SEPARATION 116 6 6 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 Cell Separation Strategy Nr of Dil
62. and contact an authorized Miltenyi Biotec service provider or Miltenyi Biotec Technical Support Electricity and fire hazards Electrical devices pose the risk 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 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 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 The instrument should no
63. andbuch muss immer konsultiert werden wenn dieses Warnzeichen benutzt wird um mehr Uber die m glichen Gefahren und entsprechende Handlungsanweisungen zu erfahren Gefahr eines Stromschlags Starkes Magnetfeld Personen mit Herzschrittmacher oder elektronischen medizinischen Implantaten m ssen Abstand halten Quetsch und Schergefahr Gefahr durch optische Strahlung Gefahr durch Laserstrahlung Biologische Gef hrdung Risiko der Kontamination wenn mit m glicherweise gef hrlichen biologischen Substanzen gearbeitet wird Schutzleiteranschluss Dieses Symbol ist innerhalb des Ger ts angebracht und stellt einen Hinweis f r das Servicepersonal dar An Stromversorgung Aus Stromversorgung Sicherheitskennzeichnungen Bitte achten Sie auf die Gefahrenpunkte und die Gefahrensymbole des autoMACS Pro Separator Instruments Quetsch und Schergefahr Gefahr durch optische Strahlung Gefahr durch Laser strahlung Quetsch und Schergefahr Figure 3 Gefahrenquellen und Gefahrensymbole auf der Vorderseite des autoMACS Pro Separators 1 0 0 0 Figure 4 Gefahrensymbole auf der R ckseite des autoMACS Pro Separators D m EJ m WICHTIGE SICHERHEITSINFORMATIONEN 19 20 Alle angegebenen Sicherheitsaufkleber und Markierungen m ssen sauber und lesbar bleiben berpr fen Sie die Aufkleber regelm Rig und ersetzen Sie diese sollten sie aus sicherem Abstand nicht mehr lesbar sein
64. ands 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 Technology amp Trading Shanghai Co Ltd Room 2309 No 319 Xianxia Road Changning District 200051 Shanghai 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 33 156 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 miltenyibiotec com sg South Korea Miltenyi Biotec Korea Petra building 659
65. areil 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 D chets d Equipements Electriques et Electroniques DEEE Information Client Pensez recycler vos produits Miltenyi Biotec en fin de vie en conformit avec la directive DEEE en vigueur dans votre pays Les quipements lectriques et lectroniques peuvent contenir des substances dangereuses qui peuvent avoir un effet n faste sur l environnement et ou la sant humaine C est pourquoi tous les quipements doivent tre sp cifiquement collect s et trait s par les centres d sign s et ce en conformit avec la R glementation DEEE En vous assurant que vous liminez vos quipements lectriques et lectroniques en accord avec la l gislation en vigueur vous contribuez pr server nos ressources naturelles et prot ger la sant humaine La protection de l environnement est au coeur de nos pr occupations Ainsi nous finangons de nombreux programmes de collecte et de recyclage des quipements lectriques que nous mettons sur le march en reversant une partie du prix de vente de nos produits des co organismes agr s travers le monde Miltenyi Biotec vo
66. as 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 por el proceso de reparaci n para asegurarse de que el instrumento est en buenas condiciones de funcionamiento Utilice solo opciones y actualizaciones recomendadas por Miltenyi Biotec Consulte con su representante local de Miltenyi Biotec sobre el amplio servicio del instrumento o dir jase a www miltenyibiotec com support El separador autoMACS Pro debe ser transportado con cuidado en el embalaje 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 Informaci n sobre la Directiva de Residuos de Aparatos El ctricos y Electr nicos Waste of Electrical and Electronic Equipment WEEE Por favor al final de la vida til de su equipo de Miltenyi Biotec disponga del mismo de acuerdo a la ley WEEE aplicable la cual puede ser diferente seg n el pa s o regi n Los equipos el ctricos y electr nicos pueden contener sustancias peligrosas que pueden tener graves efectos perju
67. asi 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 L utilizzo di comandi o regolazioni o l esecuzione di procedure diverse da quanto qui specificato pu essere causa di esposizione a radiazioni pericolose Rischi chimici e biologici 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 Ispezionare sempre il sistema fluidico il sistema di tubi le bottiglie 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
68. assistenza clienti Miltenyi Biotec severamente vietato utilizzare uno strumento danneggiato o provvisto di cavo dell alimentazione danneggiato 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 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 circolazione 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 ap autoMACS Pro Separator e proteggono il dispositivo dal surriscaldamento Non inserire mai corpi estranei nelle aperture dello strumento 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 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 Lo strumento dotato di
69. ation strategy Please be advised that the autoMACS Pro Separator is specified for use with MACS MicroBeads autoMACS Columns and other genuine Miltenyi Biotec consumables and asseccories only Please use only consumables and accessories recommended by Miltenyi Biotec The use of not recommended consumables and accessories may result in inaccurate results instrument malfunction or damage premature wear and reduced lifetime of the instrument Miltenyi Biotec does not honor any warranty or accepts any responsibility for damages resulting from the use of inappropriate consumables or accessories 107 108 6 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 6 1 1 Direct magnetic labeling Direct labeling with MACS MicroBeads is the fastest way of magnetic labeling MACS MicroBeads specifically bind to antigens on the cell surface Only one incubation step is necessary Direct magnetic labeling requires only 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
70. ative port and place into a 50 mL tube 4 Take out the uptake port needle from the needle holder and place it into a 50 mL tube 5 Undo the tubing connector of the waste tube at the waste bottle and place it into a 50 mL tube place a second 50mL tube beside this one 6 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 7 Combine all fractions and centrifuge at 350xg for 10 minutes 8 Discard the supernatant and apply cells to a reseparation as soon as possible Keep cells on ice until the separation 9 Reconnect all tubing at the appropriate positions and reposition up take needle in needle holder 9 2 10 2 Rescue procedure B If it is not possible to restart the instrument the cells retained on the columns can be recovered as follows 1 Switch off the instrument using the main power switch and disconnect the instrument from the power supply TROUBLESHOOTING 149 150 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 c
71. bottles are equipped with a sensor to measure electrolyte conductivity A pop up window will warn the user if the bottles do not carry enough buffer for at least one separation or if the waste bottle is full However the exact filling status cannot be determined Thus if more than one separation is to be performed check the filling status of all bottles visually It is recommended to monitor the bottle status when the autoMACS Pro Separator is in operation To do so go to the Status menu 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 respective symbols are green If the solution bottles are empty or the waste bottle is full the respective symbols are red Note that the symbol for the storage solution bottle always remains gray as its filling status cannot be determined by the instrument 10 45 Figure 3 8 Status of fluid levels E SWITCHING ON OR OFF 79 80 3 1 3 Priming Priming implicates the initial cleaning and filling of the autoMACS Pro Separator fluidic system before cell separations are performed The autoMACS Pro Separator must be primed each time the instrument is switched on If no wash program has been performed before the first separation a pop up window will remind the user to rinse the system 1 Ensure that all bottles are filled with the appropriate s
72. branch au connecteur RS232 BCR Les appareils laser externes raccord s au connecteur tiquet RS232 BCR doivent r pondre la norme IEC 60825 1 Utilisez uniquement des c bles de connexion de 3 m tres de longueur maximum Champ magn tique puissant Le s parateur autoMACS Pro est quip d un aimant extr mement puissant Il y a un risque d accident pour Les personnes qui portent un stimulateur cardiaque une d rivation c r brale ou un implant m dical lectronique 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 pacemakers dispositifs de mesure 41 Z m un EN Q m INFORMATIONS DE S CURIT IMPORTANTES n po 42 gt et de contr le ordinateurs et montres Ces l ments peuvent tre affect s ou endommag s par le champ magn tique Dangers m caniques 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 mod
73. call Technical Support if error is displayed again 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 fastened 151 152 6009 6216 7001 7002 7003 7004 7005 7006 7007 7008 7009 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 ofthe needle with an unexpected object far above expected tube bottom Collision of the needle with an unexpected object far above expected tube bottom Collision ofthe 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
74. change 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 WARNING Read the chapter Important safety information before installation and use The autoMACS Pro Separator is labeled as a protection class instrument and must be plugged into a grounded power outlet The MACS MiniSampler is labeled as a protection class Ill instrument and must only be plugged into the connector labeled with External CAN of the autoMACS Pro Separator 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 the 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
75. croBeads 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 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 Nt N1 2 Therefore cell counting might be associated with large statistical errors 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 100x 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 ce
76. diciales sobre el medio ambiente y o la salud humana Por este motivo todos los equipos deben ser espec ficamente recogidos y tratados por los centros de residuos designados y seg n planes de cumplimiento de la WEEE cualificada Al asegurarse de que se est deshaciendo de su equipo el ctrico y electr nico no deseado de acuerdo con la legislaci n y la WEEE aplicable de disposici n de residuos peligrosos estar ayudando a preservar nuestros recursos naturales y a proteger la salud humana Miltenyi Biotec est comprometido con la protecci n del medio ambiente Miltenyi Biotec ofrece productos que se encuentran en el final de su vida a programas de retorno de muchos pa ses y a socios con esquemas de cumplimiento de licencias WEEE en todo el mundo Miltenyi Biotec le ofrece reciclar su equipo de Miltenyi Biotec en el final de su vida til forma gratuita Los t rminos y la disponibilidad de esta oferta pueden variar seg n la regi n geogr fica debido a las diferencias en los requisitos reglamentarios Tenga en cuenta que dependiendo del tipo y uso del equipo pueden aplicarse requisitos adicionales Para mas informacion o si desea eliminar su equipo de Miltenyi Biotec en el final de su vida util por favor p ngase en contacto con su representante local de Miltenyi Biotec o con el Servicio T cnico de Miltenyi Biotec 35 Z T m un eT m INFORMACI N IMPORTANTE DE SEGURIDAD 36 Z WARNING WARNING
77. e Separation menu In the sample rack template field highlight the desired sample position s Assign a reagent for autolabeling from the Labeling submenu to each position Reagents can be assigned manually or using the 2D code reader Please refer to section 4 2 for details The recommended separation and wash programs will be automatically displayed in the Separation menu Optional The pre selected separation and wash programs can be changed in the corresponding submenus Go to the Volume submenu and enter the sample volume using the numeric keypad Press Enter Press Run Note The autoMACS Pro Separator ensures that there is enough solution for one separation If more than one separation is to be performed the user should check whether there is enough solution in the bottles Therefore a pop up window will ask the user to check and confirm the filling status before proceeding to separation Press Continue to proceed 6 10 2 Cell separation after manual labeling 1 2 Go to Separation menu Highlight the desired position s in the sample rack template field Select from the Labeling submenu for manual labeling 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 Press Enter Select a separation program and a
78. e 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 Rischi di radiazioni ottiche 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 direttamente 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 IE
79. e eluted into the positive fraction collection tube i e row C of the tube rack Possel Positive selection in standard mode Isolation of cells with frequencies higher than 5 and normal antigen expression Possels Positive selection in sensitive mode Isolation of cells with frequencies higher than 5 and low antigen expression 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 Isolation of cells with frequencies lower than 5 and normal antigen expression in a small elution volume Posseld2 Positive selection in standard mode II double column program 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 Isolation of cells with frequencies lower than 5 and low antigen expression Posselwb Special positive selection in special mode double column program Isolation of cell subsets from whole blood cell samples are automatically diluted with Running Buffer Posselbm Special positive selection single column program Isolation of cell subsets from bone marrow cell samples are automatically diluted with Running Buffer Pmalaria Special positive selection Special program for isolation of erythrocytes infected with Plasmodium falciparum 6 3 2 Depletion pr
80. e 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 Use of controls or adjustments or performance of procedures other than those specified herein may result in hazardous radiation exposure Chemical and biological hazards 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 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 Fa
81. e user to setup configure and initialize the autoMACS Pro Separator 2D code reader 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 program to automatically prime the instrument after initialization O_led Activate deactivate 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 95 USER INTERFACE 96 O_progs Enabling disabling special separation protocols The O_progs are used to enable or disable special separation programs 1 Goto Option gt User settings gt O_progs 2 Press Run A dialog box will report the current status 3 Press 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 program is used to display the number of separations that have been performed on the autoMACS Pro Separator 1 Goto Option gt User settings gt Sepcount 2 Press Run A dialog box will report the number of separations 3 Press OK to return to the Option menu Set_time Setting the time and date To set time and date refer to section 3 1 1 Syrin_ex Installing or cleaning the syringe pump This program is u
82. eck for correct function by running the Rinse program 17 Run the program Calibr_2 to calibrate the fluidic volume control of the instrument MAINTENANCE 139 7 3 3 Exchange of the peristaltic pump head 1 Switch off and unplug the instrument 2 Remove the washing station as described in section 7 2 4 steps 1 6 3 Press the tubing clamp on the left hand side of the waste distributor and remove the tubing Figure 7 24 Disconnecting the tubing from the waste distributor 4 If necessary pull out the bottom cover as depicted in figure 7 9 5 Press clamps on both sides of the pump and pull out the pump head Figure 7 25 Detachment of the pump head 6 This will uncover a pin that protrudes from the instrument The pin drives the pump during operation Clean the pin using 70 ethanol but do not pull it out 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 140 7 3 4 Exchange of tubing system If there is any leakage in the tubing system try to tighten all connections first refer to section 9 2 2 for details However if leaking persists an exchange of tubing might be required Please note that each tube has a specific length and should be exchanged with the corresponding spare part only 1 Switch off and unplug the instrument 2 Remove the affected tubing by loosening the tube connector
83. edizione In caso di domande circa la decontaminazione o il trasporto adeguati contattare l assistenza tecnica Informazioni per i clienti riguardo lo smaltimento delle apparecchiature elettriche ed elettroniche secondo le normative WEEE Waste of Electrical and Electronic Equipment Si prega di smaltire i prodotti Miltenyi Biotec in disuso secondo le disposizioni di legge riguardanti lo smaltimento di apparecchiature elettriche ed elettroniche WEEE e dei rifiuti pericolosi che possono differire da paese a paese Le apparecchiature elettriche ed elettroniche possono contenere sostanze pericolose che possono avere un grave effetto dannoso per l ambiente e o la salute umana per questo che tutte le attrezzature devono essere specificamente raccolte e trattati da centri di smaltimento abilitati e qualificati in conformit con la normativa WEEE Il corretto smaltimento delle apparecchiature elettriche ed elettroniche secondo le disposizioni di legge WEEE e dei rifiuti pericolosi possono aiutare a preservare le risorse naturali e a tutelare la salute umana Miltenyi Biotec si impegna a proteggere l ambiente Miltenyi Biotec offre programmi per lo smaltimento dei prodotti dismessi in molti paesi e partner accreditati per lo smaltimento secondo le normative WEEE in tutto il mondo Miltenyi Biotec consente di riciclare le apparecchiature Miltenyi Biotec gratuitamente Le condizioni e la disponibilit di questa offerta variano da zona a zona
84. electrocution 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 fluides 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 cable lectrique avant de nettoyer manuellement le s parateur autoMACS Pro 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 Le s parateur autoMACS Pro est destin a une utilisation int rieure uniquement N utilisez pas l appareil dans des zones class es dangereuses telles que des environnements a teneur lev e en oxyg ne Ne placez pas l appareil a proximit de radiateurs de registres de chaleur de fours
85. 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 pi ces 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 Les colonnes les plaques les tubes et tous les autres consommables qui ont t en contact avec des chantillons nocifs pour l organisme doivent tre trait s a l autoclave avant l limination Les d chets liquides doivent tre trait s a l autoclave ou d contamin s l aide d un d sinfectant adapt l agent pathog ne sp cifique par ex 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 Z m un EN Q m INFORMATIONS DE S CURIT IMPORTANTES n po 43 44 Maintenance transport et limination 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 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 apparei
86. er 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 Col_ex Used for column exchange Upon completion of the Col_ex program the fluidic system contains Running Buffer Washing Running Storage MACS Bleach Running program Solution Buffer solution Solution time Qrinse 48 mL 1 5 min Rinse 96 mL 48 mL 4min Clean 96 mL 48 mL 48 mL 7 min Sleep 96 mL 48 mL 5 min Safe 96 mL 96 mL 40 21 min Store 96 mL 96 mL 8 min Col_ex 96 mL 96 mL 6 min Not including the time required for column exchange Table 6 2 Liquid usage and time of wash and maintenance programs 6 5 Preparation of 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 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 MicroBe
87. ervice 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 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 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 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 Only a 2D code reader recommended by Miltenyi Biotec should be 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 Strong magnetic field The autoMACS Pro Separator is equipped with an extremely powerful magnet There is a risk of severe personal injury for persons carrying pacemakers brain shunts or electronic medical implants Keep any magnetic informatio
88. es and check for any resistance or collision If the problem persists contact Technical Support 9 2 9 Touchscreen remains dark 1 2 Switch off the instrument wait five seconds and switch on again 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 Replace the fuses refer to section 7 3 5 Spare fuses are included in the autoMACS Pro Separator Starter Kit If the problem persists contact Technical Support 9 2 10 Disruption of power supply during cell separation 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 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 Rescue procedure A if the instrument cannot be restarted follow Rescue procedure B 9 2 10 1 Rescue procedure A 1 Restart the instrument by switching it off and on again 2 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 3 Undo the tubing connector at the neg
89. esinfectante 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 Servicio transporte y eliminaci n seguros Salvo que el presente manual de usuario 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 33 Z T m un eT m INFORMACI N IMPORTANTE DE SEGURIDAD 34 Cuando se necesiten piezas de recambio o de repuesto asegurese 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 piez
90. ften without harming the instrument e g every four weeks As the Safe program requires a column exchange the periodic column exchange can be omitted if running the Safe program regularly 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 Neither autoclave the washing station nor wash it in a dishwasher In case of spillage it is recommended to use an appropriate disinfectant e g MACS Bleach Solution isopropyl alcohol or 70 ethanol to decontaminate surfaces with tissue or swabs Switch off and unplug the instrument before disinfection Dispose tissues and swabs appropriately It is recommended to wear protective gloves protective clothing and safety glasses to prevent contact with skin and eyes 1 Goto Option gt Special gt Safe and press Run to start the program Follow the screen prompts 2 Disconnect the tubings from storage solution and buffer bottles Press OK 3 Place the ends of the tubing in a minimum of 15 mL of disinfectant solution Press OK 4 Fill a sample tube with 25 mL of disinfectant solution Press Done 5 Optional Wash fluid bottles and bottle closures using detergent 1 hypochlorite or 70 ethanol Rinse thoroughly using deionized water 6 Reconnect all tubing Press OK 7 Clean the entire needle manually using 70 alcohol Press OK 8 Install fresh columns Press OK 7 3 C
91. h the instrument ON turned correctly or valve drive is again If error persists press the button damaged Details Exchange displayed valve If error is displayed again call Technical Support 28 Motor rotation detection failed Valve may Press the Details button Exchange be blocked or worn and cannot be turned displayed valve if applicable If error is correctly Otherwise valve or magnet drive displayed again or magnet is displayed call malfunction Technical Support 29 257 263 264 769 775 777 6006 Valve rotation hindered Valve may be blocked or worn and cannot be turned correctly Needle arm movement has been hindered Unable to read rack bar code correctly Unable to read rack bar code correctly Diluter plunger could not be initialized Syringe not mounted correctly syringe damaged or diluter valve not positioned correctly Diluter is 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 Plunger movement blocked because of column clogging blocked tubing set or any other cross section constriction 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 Switch off the instrument wait
92. hange is necessary 4 1 4 Tube rack status Tube rack detection occurs prior to the separation process The instrument will not attempt to detect the rack before cell labeling and or cell separation is performed If no graphic is displayed no tube rack was detected E If tube rack recognition fails a dialog box will open and ask for manual selection of the correct tube rack Touch the tube rack status window to open a pop up window for further information Figure 4 6 Tube rack status graphic Chill 5 Rack was detected 4 1 5 MACS MiniSampler status If the MACS MiniSampler has been detected correctly a rectangular symbol is displayed EJ If it has not been detected a hand symbol will be displayed in the same field USER INTERFACE Figure 4 7 MACS MiniSampler status graphic Left The MiniSampler was successfully installed Right No MiniSampler was detected 4 2 The Reagent menu The Reagent menu is used to assign reagent vials to one of the four reagent rack slots R1 R2 R3 and R4 to perform automated magnetic labeling Enter reagents manually or use the 2D code reader The panel on the left side of the screen represents the reagent rack Highlight a rack position by touching the corresponding position on the display Touch it once again to deselect a position CAUTION Do not look directly at the light emitted by the 2D code reader Eye injury may result Do not activate the 2D code reader if the washing station c
93. he 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 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 con structions maintenance or safety for more information regarding the installation of the equipment Signal words are used to identify safety and property damage messages The following signal words are used throughout this user manual or WARNING indicates a potentially hazardous situation which if not avoided could result in death or serious injury or CAUTION indicates a potentially hazardous situation which if not avoided could result in minor or moderate injury It may also be used to alert against unsafe practices 10 OPPPPOPP gt Safety symbols The following symbols are used to highlight conditions that could cause injury to personnel or damage to equipment Safety alert Risk of danger The documentation needs to be consulted in all cases where this safety symbol is used in order to find out the nature of the potential hazard and any actions that have to be taken Risk of electric shock Strong magnetic field Persons carrying pacemakers or electronic medical implants must maintain distance
94. 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 TECHNICAL DATA AND SPECIFICATIONS 157 For other safety considerations refer to the product label or visit www miltenyibiotec com Design and specifications are subject to change without notice 158 For technical support please contact your local Miltenyi Biotec representative or the Technical Support team at Miltenyi Biotec headquarters Miltenyi Biotec GmbH Friedrich Ebert StraBe 68 51429 Bergisch Gladbach Germany Phone 49 2204 8306 830 Fax 49 2204 8306 89 macstec miltenyibiotec de Visit www miltenyibiotec com local to find your nearest Miltenyi Biotec contact 159 160 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
95. ht 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 Gefahrenstoffen 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 Die Entsorgung der Verbrauchsmaterialien muss gem lokal geltender Bestimmungen erfolgen 2 n D m EJ m WICHTIGE SICHERHEITSINFORMATIONEN 23 24 Wartung Transport und Ger teentsorgung 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 ausdricklich 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 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
96. ia 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 Si usino solo opzioni ed aggiornamenti raccomandati da Miltenyi Biotec Si chiedano informazioni al rappresentante locale di Miltenyi Biotec in merito al servizio esteso di strumenti Miltenyi Biotec s e le modalit di supporto o ci si riferisca awww miltenyibiotec com support 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 sp
97. ic symbols 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 If the sensor cables are connected to a wrong bottle closure the symbols are also displayed in red Note that the symbol for the storage solution bottle remains gray as its filling status cannot be determined Touch any fluid bottle symbol to open a pop up window for further information Figure 4 4 Fluid bottle status Left No action required Right Check fluid bottles for possible problems 85 USER INTERFACE 86 Bottle Symbol User action Running buffer Green no action required Red refill bottle Grey connect bottle sensor Washing Solution Green no action required Red refill bottle Grey connect bottle sensor Storage Solution Gray No liquid detection possible check visually Waste Green no action required Red refill bottle Grey connect bottle sensor Table 4 2 Fluid bottle status 4 1 3 Columnstatus If the column symbol El is green no action is required If the symbol is red the columns must be exchanged refer to section 7 2 1 for details on column exchange The level of the green fill on the column symbol indicates the remaining operation life of the autoMACS Columns Touch the column symbol to open a pop up window for further information Figure 4 5 Column status Left No action required Right Column exc
98. ifiez pas les tuyaux pendant le fonc tionnement 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 chan tillonneur Ne bloquez pas les dispositifs de s curit et respectez les mesures de s curit Dangers de radiation optique L appareil est quip pour la d tection automatique de porte prouvettes classe 1M par quatre diodes laser mission par la surface 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 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 bouteille Selon la norme internationale IEC 62471 de s curit photobiologique ce syst me de DEL a une valeur de risque d exposition EHV de 0 91 et est au de
99. ilure 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 disposal 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 70 ethanol is used in Sleep and Store programs The solvent is flammable Therefore keep the instrument away from fire Servicing transport and disposal 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 incorrect 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 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 g m E a IMPO
100. ing White Process is completed Red Process was cancelled Table 6 5 Program status during separation 6 8 2 Interrupting cell separation Cell separation can be paused at any time from all menu screens by pressing the Stop button at the lower left side of the screen The autoMACS Pro Separator will then immediately stop operation and will display a pop up dialog box Press Cancel to quit the current process Press Continue to resume the current process po osp O Pri running P 7 Q Ol Process paused Cancel automatic labeling 31m 11 20 Figure 6 2 Pausing or canceling cell separation CELL SEPARATION 119 120 11 21 Figure 6 3 Cell separation has been canceled 6 8 3 Fluid bottle illumination The autoMACS Pro Separator has a fluid bottle illumination that facilitates monitoring of the instrument s status Blinking indicates that user action is required In this case check the screen for required action The fluid bottle illumination can be switched on or off see section 4 5 2 Color 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 Qrinse before starting a separation Red Error Check screen for error detection Purple Program Sleep is completed Switch off autoMACS Pro Separator Blinking Action required Check screen for required action Table 6 6 Fluid bottle illumi
101. ingungen und die Verf gbarkeit dieses Angebots unterscheiden sich geographisch aufgrund unterschiedlicher regulatorischer Anforderungen Bitte beachten Sie dass je nach Art und Nutzung Ihres Ger tes zus tzliche Anforderungen gelten k nnen F r weitere Informationen oder wenn Sie Ihr Miltenyi Biotec Altger t entsorgen m chten wenden Sie sich bitte an Ihren lokalen Miltenyi Biotec Vertreter oder den Miltenyi Biotec Technical Support D m EJ m WICHTIGE SICHERHEITSINFORMATIONEN 25 26 Z WARNING WARNING CAUTION El separador autoMACS Pro est dise ado para un uso seguro si se instala correctamente y se maneja por personal cualificado de acuerdo con las pr cticas generales de seguridad y las instrucciones de este manual de 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 cuidadosamente 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 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 compete
102. ion of two subsequent separations is applied to isolate 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 6 2 5 Depletion followed by positive selection Use this strategy if both undesired cells and target cells carry the marker that is needed for separation 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 Isolation Kits based on this strategy are available for fast and convenient isolation of specific cell subsets CELL SEPARATION 109 110 6 2 6 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
103. ith the 2D code reader Manual entry of reagents Deleting reagents Deleting a single reagent from the reagent list Deleting the entire reagent list The Separation menu Sample rack template Sample labeling options Assign a labeling option to a single sample Assign one labeling option to several samples Apply an assigned labeling option to other samples Assigning sample volumes Assigning a cell separation program Assigning a wash program Delete program combinations The Log list menu The Option menu Special User settings Calibration Calibrate the needle position Running a test calibration Calibrating the needle position Calibrate the washing station Uptake needle step 1 5 Calibrate the washing station NEG needle step 2 5 Calibration of the z axis step 3 5 Calibration of the x axis step 4 5 Test current calibration settings step 5 5 Calibrating the uptake volume Cell separation Cell labeling strategies Direct magnetic labeling Indirect magnetic labeling Cell separation strategies Positive selection 86 86 87 87 88 89 90 90 90 90 91 91 91 91 91 91 92 92 92 93 94 94 95 97 97 97 101 101 102 103 104 104 106 107 108 108 108 108 108 6 2 2 6 2 3 6 2 4 6 2 5 6 2 6 6 3 6 3 1 6 3 2 6 4 6 4 1 6 4 2 6 5 6 6 6 7 6 8 6 8 1 6 8 2 6 8 3 6 9 6 10 6 10 1 6 10 2 6 10 3 6 10 3 1 6 10 3 2 6 10 3 3 6 10 3 4 6 11 71 7 1 1 7 1 1 1 7 1 2 72 7 2
104. its tighten the tubing connections Close the front door 3 2 Shutting down the instrument 3 2 1 Sleep modus Make sure to run the Sleep program before switching off the instrument If the device is inactive for more than six hours the autoMACS Pro Separator automatically performs the Sleep program To prevent the formation of salt deposits wipe the outlet ports with a tissue soaked with distilled or deionized water before running the Sleep program For convenience you can also choose Sleep instead of a wash program after your last separation See section 6 4 and 6 11 for details 1 Press the shutdown button at the upper right hand corner of the screen see figure 3 9 2 Select Yes from the pop up dialog window 3 Upon completion of the Sleep program you will be prompted to shut down the instrument Switch off the autoMACS Pro Separator using the main power switch 3 2 2 Long term storage If the autoMACS Pro Separator will not be used for more than two weeks clean the fluidic system and replace the columns with column substitutes The Store program automatically performs the cleaning procedure and prompts the user to install column substitutes Column substitutes are installed the same way as regular separation columns 1 Goto Option gt Special 2 Select Store and press Run The system will be rinsed automatically 3 Install the column substitutes as described in section 7 2 1 4 Press Done 5 Switch off the autoMACS
105. ked with a green checkmark If a process is listed in red and is marked by an X the process was either aborted or some problem occurred See section 9 3 for details 11 28 Figure 4 13 The Log list menu 1 Goto Log list and highlight a log from the log list 2 Press Details A detailed view of the program status is shown 3 Press Ok to return to the log list screen or press Log Details to view an even more detailed log of performed actions Press Current step to display the log of the last step performed Figure 4 14 The Log list menu Details 93 USER INTERFACE 94 4 5 The Option menu The Option menu allows maintenance procedures such as exchange of autoMACS Columns or disinfection of the system The menu is divided into two main categories Special and User settings Choose option and select RUN to start Main Detail User settings Figure 4 15 The Option menu 4 5 1 Special Special comprises three commonly used programs column exchange Col_ex instrument decontamination Safe and cleaning of the instrument for long term storage Store Col_ex Column exchange program Applied when installing or exchanging autoMACS Columns Safe disinfectant program The Safe program is to be used with MACS Bleach Solution for cleaning and disinfection of the autoMACS Pro Separator Depending on the level of instrument use it is recommended to run the Safe program at least every 3 to 6 months
106. l son usure pr matur e et r duire la dur e de vie de l appareil ainsi qu annuler votre garantie 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 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 votre prestataire 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 des options et extensions recommand es par Miltenyi Biotec Renseignez vous aupr s de votre repr sentant local Miltenyi Biotec sur les arrangements extensives de support et de maintenance de l appareil ou bien r f rez vous www miltenyibiotec com support 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 app
107. l cells or up to 15 mL of whole blood The autoMACS Running Buffer is a ready to use cell separation buffer that contains azide as a preservative To prepare an azide free running buffer dilute the MACS BSA Stock Solution 1 20 with autoMACS Rinsing Solution The autoMACS Pro Washing Solution is a filter sterilized and ready to use solution to rinse the fluidic system after any cell separation It contains a detergent that dissolves cell aggregates It has been developed for optimal cleaning of the autoMACS Pro Separator tubing system The storage solution consists of 70 analytical grade ethanol and has to be prepared from absolute ethanol p a grade and distilled water Do not use denatured ethanol technical ethanol as the autoMACS Columns are not resistant to oxidative compounds The MACS Bleach Solution is used in combination with the Safe program for disinfection of the fluidic system For special disinfection procedures contact Miltenyi Biotec Technical Support Product Description Content Order no autoMACS Specifically designed columns 5x2 columns 130 021 101 Columns for use with the autoMACS Pro Separator autoMACS Cell separation buffer containing 6x1 5L 130 091 221 Running Buffer azide as preservative autoMACS Rinsing For preparation of preservative 6x1 45 L 130 091 222 Solution free cell separation buffer MACS BSA Stock For preparation of preservative 6x0 75 L 130 091 376 Solution free cell separation buffer autoMACS Pro
108. lates using the navigation arrows 2 Select the desired template s 3 Press Ok 4 Press Run to start the separation 6 10 3 4 Deleting a template 1 Inthe respective menu Reagent or Separation menu press Load Template from the lower navigation bar 2 Scroll through the list of saved templates using the navigation arrows 3 Select the template for deletion 4 Press Delete Template CELL SEPARATION 123 124 6 11 Shutting down the instrument If the instrument is to be shut down after separation the Sleep program can be chosen as a wash program after the last separation instead of Rinse or Qrinse However if Sleep is chosen as a wash program the autoMACS Pro Separator will not allow definition of any programs beyond this position Alternatively press the shutdown button on the upper right hand corner of the screen and confirm Upon completion of the Sleep program the fluid bottles are illuminated purple Switch off the autoMACS Pro Separator using the main power switch Note The autoMACS Pro Separator automatically performs the Sleep program if the device is inactive for more than six hours For long term storage of the instrument refer to section 3 2 2 NWARNING Read the chapter Important safety information Do not service the equipment yourself unless the procedure is described in this manual All other service shall be carried out only by trained experts authorized by Miltenyi Biotec An improperly serviced instr
109. le If it is necessary to disconnect the power supply unplug the cable from the power outlet WARNING Before operating the autoMACS Pro Instrument for the first time carefully read the user manual and contact your local Miltenyi Biotec representative for assistance Always wear protective gloves and eyewear in order to protect against potential biohazard exposure 73 74 When delivered the pump syringe is not installed The autoMACS Pro fluidic system is filled with double distilled water and the column slots contain column substitutes Before operation install the pump syringe first then remove the column substitutes and install the autoMACS Columns Rinse the fluidic system before first usage The autoMACS user interface is composed of five main menus Reagent Separation Status Log list and Option They are accessed through the upper tabbed menu bar see figure 4 1 Six buttons at the bottom of the screen allow for easy interaction with the instrument The function of the buttons may vary depending on which main menu is chosen see figure 4 3 Also depending on the menu and the instrument s status not all buttons are available at all times If a button is unavailable its backgroung color changes from white to grey After initialization the screen automatically displays the Status menu which helps to monitor the instrument during installation and operation For more information about the autoMACS user interface please refe
110. ler refer to section 2 6 6 The autoMACS Pro Separator is now ready for priming Go to the Separation menu and press Wash Now You now have the option to perform a quick rinse Qrinse or full rinse Rinse 7 Select Rinse and press Run to start the priming process The progress will be displayed at the bottom of the touchscreen menu CELL SEPARATION 8 When priming is finished the instrument will display Ready for separation in the Status menu The fluid bottles are now illuminated green 6 10 Cell labeling and separation After completing the wash program the autoMACS Pro Separator is ready for separation The status of the instrument is displayed in the Status menu The fluid bottles are illuminated green Cells can be labeled manually or using the autoMACS Pro Separator autolabeling function Autolabeling and manual labeling may be assigned to independent samples and run in one separation program For detailed information about manual labeling refer to to the respective reagent data sheet For a variety of labeling reagents Miltenyi Biotec has developed autolabeling protocols Please contact Miltenyi Biotec for a list of labeling reagents that are optimized for autolabeling CAUTION Do not look directly at the light emitted by the bar code reader Eye injury may result Do not activate the 2D bar code reader if the washing station cover is open 121 122 6 10 1 Cell separation using the autolabeling function 1 2 Go to th
111. ll subsets by sequential sorting Fourteen 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 two weeks 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 which 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 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 Touchscreen Robotic Ejection needle arm button for Software card Syringe pump 2D code MACS reader Column slots Rack detector Mini A Sampler Main switch Figure 1 1 Front view of the auto
112. lls in neg fraction Overall cell recovery 100 x No of cells in orig sample 143 144 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 e g the MACSQuant Intrument 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 between 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 ug 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
113. local qui remplacera la prise de courant 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 pouvezutiliser 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 Assurez vous que l interrupteur principal et le connecteur du c ble 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 Seuls les appareils p riph riques conformes 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 Le niveau de tension mesur sur ces connecteurs ne doit pas d passer le niveau de tension dangereux de 30 Veff ainsi qu un pic de 42 4 V ou 60 V DC Seul le c ble 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 tre
114. me see figure 4 1 Color coded symbols indicate the status of the hardware components For further information on a particular component touch the symbol A pop up window will open and detail more information To close the pop up window touch the black circle in the upper right corner of the window see figure 4 2 Sample status Fluid bottle filling status Column status Tube rack status MiniSampler status Figure 4 1 The status menu ration Run quieta Fluid container Storage Solution nia Waste OK Running Buffer OK Washing Solution OK 10 53 Figure 4 2 Touch any symbol to display the current status e g Fluid bottle filling status 4 1 1 Sample status The background color of the sample fields Ii indicates the sample status Programs yet to be processed appear in yellow fields Programs currently undergoing autolabeling appear in purple Programs in progress in which no autolabeling is being performed switch to orange completed programs switch to white Refer to section 6 8 for details The current action is displayed in the status bar located below the lower navigation bar The status bar also displays the overall progress in minutes running Ple it 1 6 z Joje ie o dooo ET Executing separation Possel Com 11 12 Figure 4 3 Sample status during separation 4 1 2 Fluid bottle status The status of fluid bottles PA is indicated by color coded graph
115. mn 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 11 Align the column with the top column connector sitting on the guiding of the magnet cover Press the column into the slot until you feel the guides click Verify that the column is placed in the center of the magnet cover 12 Remove column 2 from its slot and repeat steps 8 through 11 13 Ensure that the tubing is neither pinched nor obstructed 14 Press Done The program will then proceed to wash the columns with autoMACS Running Buffer Check that the columns are securely fastened to the column connectors and that no buffer is leaking 15 Close the front door Note The instrument automatically records the date of the column exchange if the program Col_ex has been used for column installation and can display the due date for the next column exchange 16 Prime the autoMACS Pro Separator as described in section 3 1 3 17 Run the program Calibr_2 to calibrate the fluidic volume control of the instrument Refer to section 5 for details 77 E SWITCHING ON OR OFF 78 3 1 1 3 Setthedate and time When switching on the autoMACS Pro Separator for the first time the user is prompted to set the correct time and date Note Set the correct date and time to properly schedule maintenance procedures such as column exchanges 1
116. n pi possibile un uso sicuro dello strumento Si spenga immediatamente lo strumento e lo si scolleghi dalla presa di corrente quindi si contatti il service provider Miltenyi Biotec o Miltenyi Biotec Technical Support Rischi elettrici e pericolo d incendio 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 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
117. n reas clasificadas como peligrosas como ambientes con alta concentraci n de ox geno 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 ap las aberturas del instrumento No use el instrumento en un lugar h medo Evite la elevada humedad o la condensaci n y proteja a la m quina de salpicaduras 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 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 co
118. n 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 g m E a IMPORTANT SAFETY INFORMATION av 14 Pb Mechanical hazards 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 Optical radiation hazards 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 10
119. n gt User settings gt O_progs A_Depls7 Depletion in sensitive mode sample loading in 1 mL batches Removal of cells with low antigen expression if purity is the highest priority This special program is disabled by default To enable select Option gt User settings gt O_progs Volume of Volume of non labeled fraction labeled fraction Program A x RE Loading rate i e negative fraction i e positive fraction Possel 2 mL sample volume 2 mL 4 mL min Possels 2 mL sample volume 2mL 1 mL min Posseld 2 mL sample volume 0 5 mL 4 mL min column 1 1 mL min column 2 Posseld2 2 mL sample volume 2mL 4 mL min columns 1 and 2 Posselds 2 mL sample volume 2mL 1 mL min columns 1 and 2 Posselwb 2 mL 3x sample volume 2 mL 4 mL min predilution columns 1 and 2 Deplete 2 mL sample volume 2mL 4 mL min Depletes 2 mL sample volume 2mL 1mL min Depl05 2 mL sample volume 2mL 0 5 mL min Depl025 2 mL sample volume 2mL 0 25 mL min A_Depl07 3 mL per 1 mL sample volume 2 mL per 1 mL 4 mL min sample volume A_Depls7 3 mL per 1 mL sample volume 2 mL per 1 mL 1 mL min sample volume This program must be used with manual labeling 3 fold predilution of the sample volume Table 6 1 Output volumes and loading rates of separation programs Purity will slightly decrease 2 Recovery will slightly decrease Figure 6 1 Selecting the optimal separation program CELL SEPARATION 113 114 6
120. n un electricista para que reemplace la toma de corriente 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 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 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 RS232 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 el conector RS232 BCR debe conectarse solamente un lector de c digo 2D recome
121. nation status 6 9 Priming the autoMACS Pro Separator Priming implicates the initial rinsing and filling of the autoMACS Pro Separator tubing system before performing cell separations The autoMACS Pro Separator must be primed each time the instrument is switched on Ensure that the MACS MiniSampler has been installed the waste bottle is empty and that all fluid bottles are filled with recommended solutions refer to section 2 and 3 for more details Fill waste bottle with 100 mL disinfectant if necessary At this point the fluid bottles are illuminated yellow If the instrument has not been primed yet the autoMACS Pro Separator will automatically ask to run a Rinse program before starting the separation It is possible to include an initial Rinse program that will be performed automatically upon switching on the instrument The setting of this option O_init is described in section 4 5 2 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 see section 4 1 for more information about the Status menu 3 Ensure that the symbols for the fluid bottles are colored green Note that the symbol for the rinse bottle always remains gray 4 Ensure that the symbol for the columns is colored green 5 Ensure that the MACS MiniSampler is installed correctly For more details concerning correct installation of the MACS MiniSamp
122. nd HD f r die Freie Gruppe betr gt 61 cm Der Gef hrdungsab stand 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 nicht absichtlich auf andere Personen Demontieren wechseln oder entfernen Sie nicht eingebaute Laser oder LED Strahlungsquellen oder deren Halterung Die Laser oder LED Strahlung squellen 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 Spieg eloberfl 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 Offnungsfen ster 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 A1 19
123. ndado 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 Campo magn tico intenso El separador autoMACS Pro est equipado con un im n extremadamente potente Existe un riesgo de lesi n grave para las personas que llevan marcapasos derivaciones cerebrales o implantes m dicos electr nicos 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 31 Z T m un eT m INFORMACI N IMPORTANTE DE SEGURIDAD 32 gt gt Riesgos mec nicos 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 lejos 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 si
124. nductores de protecci n El s mbolo est pegado en el interior del instrumento Esta informaci n es para el personal de servicio Encendido Fuente de energ a Apagado Fuente de energ a Etiqueta de seguridad Por favor tenga en cuenta los puntos de peligro y los simbolos de seguridad del autoMACS Pro Separator Instrument Riesgo de que se aplaste o corte alguna parte del cuerpo Radiaci n ptica peligrosa Radiaci n l ser peligrosa Riesgo de que se aplaste o corte alguna parte del cuerpo Figure 5 Puntos de peligro ylocalizaci n de s mbolos de seguridad que puede encontrar en la parte delantera del autoMACS Pro Separator Be ED EDO eae Figure 6 Localizaci n de los s mbolos de seguridad en la parte transera del autoMACS Pro Separator Instrument Z n m a un eT m INFORMACI N IMPORTANTE DE SEGURIDAD 29 30 Las etiquetas de seguridad arriba mencionadas y las marcas de seguridad deberan mantenerse limpias y legibles Inspeccionar periddicamente las etiquetas y marcas de seguridad y reemplazarlas en caso de que estas no sean legibles o perceptibles a una distancia visual segura Contacte con Miltenyi Biotec para reemplazo de etiquetas Instrucciones generales de seguridad Si el instrumento no funciona correctamente y o las instrucciones o mensajes mostrados le aconsejan contactar con el servicio t cnico no es posible utilizar el instrumento de una manera
125. ne Fl ssig oder Spr hreinigungsmittel sondern nur ein feuchtes Tuch Das Ger t ist ausgestattet mit einem Schutzkontaktstecker und einer dreiadrigen Anschlussleitung Fuhren Sie den Stecker nicht in eine Steckdose ohne Schutzkon takt ein Wenn Ihre Steckdose keinen Schutzkontakt aufweist bitten Sie einen ortsans ssigen Elektriker diese zu ersetzen Das Ger t sollte nur von einer Stromquelle aus betrieben werden die den elektrisch en Angaben auf dem Typschild entsprechen Sollten Sie Fragen zur Art der Stromver sorgung haben wenden Sie sich an einen autorisierten Miltenyi Biotec Service Provider oder Ihren lokalen Stromversorger Benutzen Sie keine Verlangerungskabel oder Steckdosenleiste Uberladen Sie eine Steckdose nicht Die Gesamtlast darf 80 der Zweigstromkreis bemessung nicht Uberschreiten Der Hauptstromschalter ebenso wie der Netzstecker f r das Stromkabel sollten leicht zug nglich sein und sich in m glichst unmittelbarer N he zum Bediener des Ger tes befinden Sollte es erforderlich sein die Stromzufuhr zu unterbrechen ziehen Sie den Netzstecker An die mit COM beschriftete RS 232 Buchse d rfen nur Peripherieger te ang eschlossen 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
126. ng 7 Elevate the touchscreen and insert the memory card into the memory card slot at the right side of the touchscreen INSTALLATION Figure 2 7 Insert the software memory card 2 5 Installing and replacing fluid bottles WARNING When working with biohazardous 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 your local regulations Operating the autoMACS Pro Separator requires Running Buffer Washing Solution and storage solution absolute analytical grade ethanol diluted to 70 with double distilled water It is recommended to operate the instrument with ready to use MACS Buffers table 2 2 The autoMACS Pro Separator is delivered with four empty fluid bottles which connect to the instrument with specifically designed bottle closures To keep buffers sterile each bottle closure is equipped with a hydrophobic air filter The bottle closures consist of a fluid uptake port or in case of 67 68 the waste bottle a fluid outlet port To measure the filling status bottle closures also have a sensor for measuring electrolyte conductivity However the storage solution 70 ethanol does not contain electrolytes Therefore the filling status of the storage solution bottle cannot be measured by the instrument and must be checked visually For your convenience the fluid bottles bottle closures and fluid sen
127. not specified for use in the cold room The emission sound pressure level at the workstation is lt 70 dB A 155 Technical data Model Color Footprint Footprint with MiniSampler Height Weight Input voltage Power consumption Fuses Programs Sample volume input Sample volume output Column capacity RS232 Interface labeled COM RS232 Interface labeled RS232 AUX Not in use RS232 Interface DC Output labeled RS232 BCR CAN Bus DC Output labeled External CAN AC Output labeled Bottle Sensor CAN Bus labeled CAN1 or CAN2 autoMACS Pro Separator Blue orange 605x343 mm WxD 605x455 mm WxD 392 5 454 mm adjustable touchscreen 25 kg 100 240 VAC 50 60 Hz 200 VA 2xT4A 250 12 preset 0 25 50 mL 0 5 52 mL 4x10 cells sample 2x10 magnetically labeled cells sample 15 mL of whole blood Pins 1 4 6 9 NC Pin 2 RXD Pin 3 TXD Pin 5 GND Pin 7 RTS Pin 8 CTS Pins 1 4 6 7 8 9 NC Pin 2 RXD Pin 3 TXD Pin 5 GND Pins 4 6 NC Pin 1 Input Pin 2 RXD Pin 3 TXD Pin 5 GND Pins 7 8 Shorted Pin 9 5 VDC 0 5 A Pins 1 4 8 NC Pin 2 CAN L Pins 3 6 GND Pins 5 9 24 VDC 2A Pin 7 CAN H Pins 1 2 3 4 5 5 VAC 10 kQ Pins 6 7 8 14 15 GND Pins 9 10 11 12 13 Input Pins 1 4 5 8 9 NC Pin 2 CAN L Pins 3 6 GND Pin 7 CAN H Depending on the type of power plug the depth increases by 27 mm or 62 mm Table 10 1
128. nt If installed correctly the autoMACS Pro Separator automatically recognizes the MACS MiniSampler upon initialization Note Do not use the same MACS MiniSampler with several different instruments e g the MACSQuant Instruments 1 Open the front door of the autoMACS Pro Separator 2 Tilt the MACS MiniSampler and slide the guiding into the receiving slot until a resistance is met Lower the rack to a horizontal position until the rack is locked in the position INSTALLATION Figure 2 8 Insert the MACS MiniSampler guiding into the receiving slot 3 Ensure that the MACS MiniSampler is completely inserted Secure the connection by fastening the MiniSampler with the supplied knurled screw Figure 2 9 Secure the MACS MiniSampler 69 70 4 Close the front door 5 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 Fasten the screws of the connecter using the screwdriver 6 Guide 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 Fasten the screws of the connecter using the screwdriver included in the starter kit 7 Secure the MACS Reagent Rack 4 onto the MACS MiniSampler into the left recess The engagement hook has to snap into the undercut Figure 2 10 Secure the MACS Reagent Rack 4 8 Seta tube
129. nt safety information Wichtige Sicherheitsinformationen Informacion importante de seguridad Informations de s curit importantes Importanti istruzioni di sicurezza Introduction MACS Technology the gold standard in cell separation The autoMACS Pro Separator Installation Components included in delivery Materials required for operation autoMACS Pro Separator accessories Unpacking the autoMACS Pro Separator Installing and replacing fluid bottles 09 17 27 37 47 57 57 58 61 62 62 64 64 67 Installing the MACS MiniSampler MACS Reagent Rack 4 and tube rack 69 Switching on or off Switching on the instrument When switching on the instrument for the first time Installing the pump syringe Installing autoMACS Columns Set the date and time Testing the bar code scanner Test the needle calibration Checking fluid levels Priming Shutting down the instrument Sleep modus Long term storage The autoMACS Pro user interface The Status menu Sample status Fluid bottle status 73 73 73 75 76 78 78 79 79 80 81 81 81 83 84 85 85 4 1 3 4 1 4 4 1 5 4 2 4 2 1 4 2 2 4 2 3 4 2 3 1 4 2 3 2 4 3 4 3 1 4 3 2 4 3 2 1 4 3 2 2 4 3 2 3 4 3 3 4 3 4 4 3 5 4 3 6 4 4 4 5 4 5 1 4 5 2 5 1 5 1 1 5 1 1 1 5 1 1 2 5 1 1 3 5 1 1 4 5 1 1 5 5 1 1 6 5 2 6 1 6 1 1 6 1 2 6 2 6 2 1 Column status Tube rack status MACS MiniSampler status The Reagent menu Entry of reagents w
130. nte para el suministro de electricidad construcci n de edificios mantenimiento o seguridad para obtener m s informaci n sobre la instalaci n del equipo Se usan se ales para identificar mensajes de seguridad y de da o de la propiedad Las siguientes se ales se usan a trav s de este manual de usuario o WARNING indica una situaci n potencialmente peligrosa la cual si no se evita puede resultar en muerte o da os graves o CAUTION indica una situaci n potencialmente peligrosa la cual si no se evita puede resultar en un da o menor o moderado Tambi n puede ser usado para alertar contra pr cticas peligrosas 27 28 PPPP OD gt Simbolos de seguridad Los siguientes simbolos se usan para destacar condiciones que podrian causar da os al personal o da os al equipo Alerta de seguridad Riesgo de peligro Es necesario consultar la documentaci n del equipo siempre que este s mbolo de seguridad aparezca para de esta manera poder averiguar la naturaleza del peligro potencial existente y las acciones necesarias a realizar Riesgo de shock el ctrico Campo magn tico intenso Personas con marcapasos o implantes m dicos electr nicos deben mantenerse alejadas Riesgo de que se aplaste o corte alguna parte del cuerpo Radiaci n ptica peligrosa Radiaci n l ser peligrosa Riesgo biol gico Riesgo de contaminaci n si se usa material biol gico potencial mente peligroso Bloques de conexi n para co
131. o 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 Segnali di avviso sono in uso per identificare un uso sicuro e messaggi in caso di danni a cose personali seguenti segnali saranno in uso in questo manuale o WARNING indica una situazione potenzialmente pericolosa che se non evitata potrebbe causare morte o gravi danni o CAUTION indica una situazione potenzialmente pericolosa che se non evitata potrebbe causare danni minori e lesioni moderate Pu essere impiegato per segnalare procedure pericolose 47 48 6 bbbbobb Bd Simboli di sicurezza seguenti simboli sono impiegati per sottolineare condizioni che potrebbero causare lesioni a persone o danni all attrezzatura Avviso di sicurezza Rischio di pericolo Consultare il manuale di istruzioni ogni volta che questo simbolo di sicurezza viene utilizzato per identificare la natura del potenziale pericolo e stabilire quali azioni intraprendere Rischio di scosse elettriche Forte campo magnetico Portatori di pacemaker o dispositive medici elettronici devono mantenere una adeguata distanza di sicurezza Rischio di schiacciamento o di taglio di parti del corpo Radiazione o
132. oBeads bone marrow volume 1 Max number of samples 6 min first incubation 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 117 118 6 7 Selecting the 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 Row A of the Chill Rack holds the sample tubes Ori Row B of the Chill Rack holds the tubes for the non labeled fractions negative fractions Neg Row C of the Chill Rack holds the tubes for labeled fractions positive fractions Pos Optional For your convenience Row D can hold round bottom tubes for further analyses Select the appropriate tube rack according to table 6 4 It i
133. ogicos 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 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 d
134. ograms 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 Deplete Depletion in standard mode Removal of cells with normal antigen expression if recovery is the highest priority Untouched isolation with MACS Cell Isolation Kits Depletes Depletion in sensitive mode Removal of cells with normal antigen expression if purity is the highest priority Removal of cells with low antigen expression Untouched isolation with MACS Cell Isolation Kits if purity is highest priority Depl05 Depletion in sensitive mode Il Removal of cells with low antigen expression special program for very sensitive depletion CELL SEPARATION 111 Depl025 Depletion in sensitive mode III Removal of cells with low antigen expression special program for very sensitive depletion This special program is disabled by default To enable select Option gt User settings gt O_progs A_Depl07 Depletion in standard mode sample loading in 1 mL batches Removal of cells with normal antigen expression if recovery is the highest priority This special program is disabled by default To enable select Optio
135. olumn 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 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 9 3 Hardware issues indicated by an error screen If errors or warnings are displayed on the screen of the instrument please refer to the following table If the below measures do not clear the fault contact 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 contact Technical Support Error Cause Possible remedies code 5 Hardware module is not initialized Aftera Restart instrument Please call Technical module malfunction an initialization of Support if error is displayed again the module is neccessary This might also be a subsequent error if another error has been displayed shortly before 27 Standard valve initialization failed Valve Switch off the instrument wait for 5 may be blocked or worn and cannot be seconds and switc
136. olutions and that the waste bottle is empty 2 Switch on the autoMACS Pro Separator and wait for the instrument to complete initialization 3 After initialization is completed the Status menu will be displayed For more details on the Status menu please refer to section 4 1 4 Verify that the touchscreen symbols for all fluid bottles are green except for the grey symbol for the storage solution bottle 5 Ensure that the symbol for the columns is green If the symbol is red exchange the columns The fill level on the symbol is an indicator for the remaining operation life of the columns 6 Ensure that the MACS MiniSampler is installed correctly For more details concerning correct installation of the MACS MiniSampler refer to section 2 6 7 The autoMACs Pro Separator is now ready for priming Go to the Separation menu and press the Wash Now button at the bottom of the screen Lr Select ia program Basoph_Ik Labeing Baso IK hum a G q Run Cancel Wash Load Now Template Figure 3 9 Select Rinse to prime the autoMACS Pro Separator 10 50 8 Select Rinse and press Run to start the priming process 9 When priming is finished the instrument will display Ready for separation in the Status menu Note When priming the instrument for the first time it is recommended to inspect the fluidic system for potential leaks Open the front door after priming the instrument If there is any sign of leakage e g salt depos
137. on Refer to section 6 10 3 for details on how to use templates It is also possible to perform a single wash program prior to separation by pressing the Wash now button at the bottom of the screen Reagent pa Ch no Che ct RUNE rt 1 Sample Rack template Labeling volume Sample labeling options 2 3 Basoph_IK hum n E Cul AA Sample processing volume y De Wash Load 4 Separation program 10 48 Wash program Figure 4 12 The Separation menu 4 3 1 Sample rack template The positions in the sample rack template field ID 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 See section 6 7 for details on the different tube rack and loading positions Highlight a sample position by touching the corresponding position on the display 4 3 2 Sample labeling options The Labeling submenu EX is used to instruct the instrument if autolabeling is to be performed on a sample and what type of autolabeling will be performed By default no autolabeling will be performed M A list of product options is visible if reagents have already been assigned to positions on the reagent rack using the Reagent menu 4 3 2 1 Assign a labeling option to a single sample 1 Highlight a sample using the sample rack template 2 Scroll through the Labeling submenu using the arrows 3 Select the desired Labeling reagent
138. on box autoMACS Pro Separator Instrument Lower compartment MACS MiniSampler Lower compartment MACS Reagent Rack 4 Lower compartment 1 MACS Chill 5 Rack Lower compartment 1 MACS Chill 15 Rack Lower compartment 1 MACS Chill 50 Rack Lower compartment 2D code reader Built in Memory card with software Upper compartment 5x2 autoMACS Columns Upper compartment 2x2 column substitutes Built in User manual Upper compartment Short instructions Upper compartment Pump syringe Upper compartment Power cord Upper compartment Fluid bottle sensor cables Upper compartment autoMACS Pro Buffer Combination Separate box One year warranty Table 2 1 Components included in delivery 2 2 Materials required for operation For daily operation the following solutions are required autoMACS Running Buffer autoMACS Pro Washing Solutions and storage solution Additionally MACS Bleach Solution may be required for disinfection and decontamination Fluid bottles can be identified by color code and symbols table 2 4 For proper operation of the autoMACS Pro Separator fluid 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 autoMACS Columns can be used for maximal 14 days or 100 separations whichever comes first Their capacity reaches up to 2x10 magnetically labeled cells from up to 4x10 nucleated tota
139. orrective maintenance Four valves regulate the fluidic system of the autoMACS Pro Separator A fifth valve referred to as dilutor valve is connected to the pump syringe Service plans for preventive and corrective maintenance are available at www miltenyibiotec com However if leakage occurs the user can exchange valves and tubing 7 3 1 Valve exchange 1 Goto Option gt User settings gt Valve_ex 2 Press Run to start the program Select valves that have to be exchanged Valves will automatically be turned to exchange position Note In case the valve exchange program Valve_ex cannot be performed e g valve does not turn switch off instrument and turn the valve bracket manually to the position corresponding to the groove in the valve drive 3 Switch off and unplug the autoMACS Pro Separator Open the front door For exchange of the lower valve remove the bottom cover by pulling firmly MAINTENANCE Figure 7 9 The bottom cover must be removed for access to the lower valve 4 Unscrew the valve cover screw that releases the tubing with the supplied wrench 133 Figure 7 10 Unscrew the valve cover screw 5 Detach all tubing and valve port locks or blind screws from the respective valve Figure 7 11 Unscrew the valve cover screw and detach tubing and valve port locks or blind screws 6 Loosen the two valve screws using the screwdriver contained in the autoMACS Pro Separator Starter Kit and pull out the val
140. ot for therapeutic or diagnostic use autoMACS Pro Separator User manual Version 5 EN 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 Read the user manual before using the instrument Before using the instrument read chapter Important safety information and all other information contained in this user manual including all safety and operating instructions Pay special attention to warnings displayed on the instrument Failure to read and follow these guidelines could lead to improper or incorrect usage and result in damage to the instrument Improper usage could also cause severe personal injury death unpredictable results instrument malfunction and premature wear of components shortening the lifetime of the instrument Such actions may void your warranty Keep this user manual and any other safety and operating instructions provided with the instrument in a safe place that is accessible to all users for reference If you are concerned about the safe use of the instrument please contact your authorized Miltenyi Biotec service provider or call Miltenyi Biotec Technical Support Table of contents EN DE ES FR IT 1 1 1 2 2 1 2 2 2 3 24 2 5 2 6 3 1 3 1 1 3 1 1 1 3 1 1 2 3 1 1 3 3 1 1 4 3 1 1 5 3 1 2 3 1 3 3 2 3 2 1 3 2 2 4 1 4 1 1 4 1 2 Importa
141. 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 gt autoMACS Pro et prot gent l appareil de la surchauffe N introduisez jamais d objet tranger dans l appareil par l une des fentes N utilisez pas l appareil dans un endroit humide ou mouill vitez l humidit et la condensation et prot gez la machine des claboussures 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 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 a la terre Il s agit d un dispositif de s curit N essayez pas d ins 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
142. over is open 87 88 11 01 Figure 4 8 The Reagent menu 4 2 1 Entry of reagents with the 2D code reader 1 Gotothe Reagent menu Highlight the reagent rack position where the vial will be placed in the reagent rack 2 Press the Read Reagent button to activate the 2D code reader and present a reagent vial in front of the 2D code reader Ensure that the 2D code is facing the blinking light The optimal reading distance is 0 5 2 5 cm from the code reader tilt the vial as depicted Figure 4 9 Scanning a reagent using the 2D code reader 3 The reagent is automatically recognized and assigned to the currently highlighted reagent rack position The next reagent rack position will be highlighted automatically Note If the 2D code reader cannot identify the reagent please enter the reagent information manually see below Separation Status Log list Option U Define reagent rack Enter Load Save Delete Read Reagent Template Template Selected f Reagent 11 03 Figure 4 10 Successfully scanned reagents appear on the screen 4 Using the same procedure up to four reagent vials can be scanned and assigned using the 2D code reader Combinations of four reagents can be saved as a template for later use by pressing the Save Template button Refer to section 6 10 3 for details on how to use templates 5 After all desired reagents are entered open the MicroBead reagent vials and place them on the MACS
143. peat the entire process CALIBRATION 105 106 5 2 Calibrating the uptake volume 1 Goto Option gt User settings gt Calibr_2 Press Run to start the program 2 Press Calibrate The calibration is performed automatically Calibration 1 1 Tubing Actual calibration values of unknown 0 0 0 Cancel Calibrate 11 52 Figure 5 15 Calibration 2 Step 1 1 3 Press Save to finish Calibr_2 Che ck up Calibration 1 2 Tubing Needle tube 2 old 0 Bubble sensor tube 31 old 0 Plunger backlash 11 old 0 Ignore 11 54 Figure 5 16 Calibration 2 Step 1 2 NWARNING Read the instructions in the chapter Important safety information before operation of the instrument When working with biohazardous 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 your local regulations 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 instrument s autolabeling feature The pre set separation programs of the autoMACS Pro Separator have been developed for optimal separation results independent from the applied labeling and separ
144. pened by the moving needle arm hitting the cover The protective cover of the MACS MiniSampler needs to be closed Bar code on Chill Rack could not be read MACS MiniSampler is not connected properly Unable to read rack bar code correctly Press Ignore to continue without liquid detection 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 Press 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 Please restart instrument Please calibrate needle arm axes Run program Calibr_1 Please calibrate needle arm axes Run program Calibr_1 Please calibrate tubing Run program Calibr_2 Please install columns Please calibrate MACS MiniSampler using program Calibr_1 Please rinse system by selecting Wash or abort with Cancel Please install new columns Touch Cancel to abort and then install columns touch Continue to ignore and use old columns not recommended Touch Cont to discard the remaining negative cells into the waste bottle To output negative fraction again at lower speed exchange negative tube with an empty tube and touch Retry Exchange rack with rack holding more samples and touch OK or touch Cancel to abort and reprogram O E Ir n Lu
145. position can be tested for tube rack position A1 ORI B1 NEG and C1 POS as well as Reagent Rack positions 1 4 To check the exact needle position use the Move up and Move down buttons to raise or lower the uptake needle respectively Ensure that the uptake needle is positioned directly above the center of the tube Lower the needle all the way to the bottom of the tube to check that the needle is centered For the Reagent Rack the needle should be located at the lowest point of the vial to ensure full reagent uptake Select action Previous Test Next position Verify sample tube position 11 49 Figure 5 6 Test position ORI tube Figure 5 7 Test position Uptake needle position in the Chill Rack 8 After testing the last position press Done Press Done once again to leave test calibration and press Cancel to close the calibration program 100 5 1 1 1 Calibrating the needle position If the needles are not positioned correctly in the washing station and or the sample tubes run a Calibr_1 Open the washing station cover and remove the MiniSampler cover for better visibility 1 Goto Option gt User settings gt Calibr_1 2 Press Run and then Calibrate The first of five 1 5 calibration steps will begin namely the needle position in the washing station Current calibration value from unknown Position of washing station 0 Cancel Calibrate 11 33 Figure 5 8 Calibration 1 Step 1 5
146. r to section 4 1 Note the position of the power socket on the rear panel of the autoMACS Pro Separator see figure 1 2 2 Ensure that the main power switch is in position O off 3 Connect the power cord 4 Switch on the autoMACS Pro Separator As the instrument still contains the column substitutes at this point a pop up window will inform the user that the columns are not installed Select Ignore Warning 7016 Columns are not installed Start column exchange program Ignore Run 10 39 Figure 3 1 A warning screen informs the user that no columns are installed 3 1 1 1 Installing the pump syringe 1 Open the front door of the autoMACS Pro Separator 2 Pull out the plunger of the pump syringe and humidify it with distilled water Push it halfway into the glass pipe Figure 3 2 Humidify the plunger 3 Remove the blind screw and the protective tape Fasten the syringe at the dilutor valve by turning the top screw clockwise until a resistance is met see section 7 2 3 for a detailed description on how to clean and install the pump syringe Unfasten the top screw again by one rotation ny Dilutor valve Blind screw Figure 3 3 Fasten the syringe at the dilutor valve 4 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 the fitting see figure 7 2 7 6 Tighten the plunger lock screw E SWI
147. rateur autoMACS Pro Risque d crasement et de cisaillement des membres A Radiation optique dangereuse Radiation Risque laser d crasement et dangereuse de cisaillement des membres corporels Figure 7 Les zones de risque et symboles de s curit sur la face avant du s parateur autoMACS Pro 2 0 6 6 Figure 8 Les symboles de s curit sur la face arri re du s parateur autoMACS Pro Z EL m pe un EN Q m INFORMATIONS DE S CURIT IMPORTANTES 39 40 Les tiquettes et marquage de s curit doivent tre maintenus propres et lisibles Il convient de les inspecter p riodiquement et de les remplacer si elles ne sont pas lisibles ou perceptibles a une distance d observation garantissant la s curit du manipulateur Contactez Miltenyi Biotec pour le remplacement des tiquettes de s curit Consignes de s curit g n rales Si l appareil ne fonctionne pas correctement et ou les instructions ou messages affich s vous avisent de contacter le support technique l utilisation s re de l appareil n est plus longtemps garantie teignez imm diatement l appareil et d branchez le de la prise lectrique puis contactez un prestataire de services Miltenyi Biotec agr ou l quipe de support technique de Miltenyi Biotec Dangers lectriques et incendie Les appareils lectriques pr sentent un risque d lectrocution Afin de r duire le risque d
148. rung oder zum Versand des Ger tes haben wenden Sie sich bitte direkt an unseren Technical Support Kundeninformation zur Entsorgung von Elektro und Elektronik Altger ten Waste of Electrical and Electronic Equipment WEEE Bitte entsorgen Sie Ihre Altger te von Miltenyi Biotec unter Einhaltung der jeweils geltenden Vorschriften f r die Erfassung und Behandlung von Elektro und Elektronik Altger ten und die Entsorgung von Gefahrstoffen Diese k nnen von Land zu Land sowie regional variieren Elektrische und elektronische Ger te k nnen Gefahrstoffe enthalten welche die Umwelt erheblich belasten und oder die Gesundheit gef hrden Deshalb m ssen Altger te speziell gesammelt und durch ausgewiesene Entsorgungsbetriebe im Rahmen der hierf r vorgesehenen Entsorgungssysteme fachgerecht behandelt werden Indem Sie sicherstellen dass Ihr Altger t gem den geltenden Vorschriften zur Behandlung von Elektro und Elektronik Altger ten sowie von Gefahrstoffen entsorgt wird tragen Sie dazu bei unsere nat rlichen Ressourcen zu schonen und die menschliche Gesundheit zu sch tzen Miltenyi Biotec setzt sich f r den Schutz der Umwelt ein Miltenyi Biotec bietet in zahlreichen L ndern eigene R cknahmeprogramme f r Altger te an und arbeitet weltweit mit lizensierten Partnern zusammen die an bestehende Recycling und Entsorgungssysteme angeschlossen sind Miltenyi Biotec erm glicht Ihnen ein kostenloses Recycling Ihres Altger tes Die Bed
149. s apparent damage please contact Technical Support for assistance refer to section 11 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 The instrument is heavier at the front Stabilize the front of the instrument while lifting it 1 Open the box and remove the upper compartment of the packaging to reveal the instrument INSTALLATION Figure 2 1 Upper compartment of the packaging 2 Remove boxes containing the MACS MiniSampler its protective cover and the MACS Chill Racks from the lower compartment Figure 2 2 Lower compartment of the packaging 3 Remove the foam packaging from both sides of the autoMACS Pro Separator 65 66 Figure 2 3 Remove the foam packaging from both sides of the autoMACS Pro 4 Grip the instrument at the base of the orange bottle holders and lift it out of the box Stabilize the front of the instrument while lifting it Place the instrument onto a stable worktop surface and remove the plastic bag surrounding it Figure 2 4 Two people are required to lift the autoMACS Pro 5 Carefully remove the uptake needle from the foam packaging located under the touchscreen Figure 2 5 Remove the uptake needle from the foam packaging arrow 6 Place the uptake needle into its guiding at the needle arm Figure 2 6 Place the uptake needle into its guidi
150. s recommended to 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 Tube racks will stay cool for 2 2 5 hours Equip the tube rack with sample tubes and fraction collection tubes Racktype Slots Maximal Manual Autolabeling and number of labeling symbol samples Maximal Minimal first Maximal final sample incubation labeling volume volume volume Chill 5 24x5 mL 6 2 5 mL 0 2 mL 2 0 mL EE 5 mL tubes 0 25 mL 1mlL Chill 15 15x15 mL DI 12 5mL 0 2 mL 6 5 mL 5x5 mL 15 mL tubes 1mL 4 mlL Chill 50 6x50 mL 3 50 mL 4 mlL 8 mL 333 3x15 mL 50 mL tubes Volumes refer to whole blood samples only Table 6 4 MACS Chill Rack specifications for manual labeling and autolabeling 6 8 Monitoring the separation process It is recommended to monitor the instruments status before and during cell separation Select the Status menu tab to check the program status Further fluid bottle illumination also indicates the instrument s status 6 8 1 Program status The background color of the sample fields indicates the sample status see table 6 5 and section 4 3 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 Color Program status Yellow Yet to be processed Purple Currently undergoing autolabeling Orange No autolabeling manual label
151. s 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 easily 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 Plug in 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 5 8 If there is still any leakage refer to to section 9 2 7 3 5 Exchange of fuses If the instrument fails to start upon switching it on or if operation suddenly stops and the screen turns dark an exchange of the fuses might be required CAUTION Fuse specifications are given 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 MAINTENANCE 141 Figure 7 26 Removing the fuse holder 4 Push the fuse holder back into the housing and reconnect the main power cord 142 To evaluate any MACS Cell Separation the separated cells can be analyzed with regard to purity recovery and viability Using MACS Mi
152. sed during instrument maintenance Refer to section 7 2 3 for details 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 Refer to section 7 3 1 for details NWARNING Read the instructions in the chapter Important safety information before operation of the instrument The autoMACS Pro Separator is calibrated using two programs Calibr_1 and Calibr_2 Calibr_1 is used to calibrate the settings of the needle arm x y and z axis Calibrating the needle arm is recommended after instrument or software installation or after moving the instrument to a different location Ifthe MiniSampler or one of the needles was exchanged needle calibration should also be checked Calibr_2 automatically calibrates the liquid volume control This is crucial for correct measurement and processing of the sample volumes Run the Rinse program before running Calibr_2 to fill the fluidic system with buffer Always run Calibr_2 after the pump syringe any of the valves or the tubing was exchanged Check proper needle calibration by running a test calibration Open the washing station cover and remove the MiniSampler cover for better visibility 97 1 Goto Option gt User settings gt Calibr_1 Press Run and then Test Calibration 1 Select mode Cancel Test Calibrate 11 31 Figure 5 1 Calibration 1 modes 2 When prompted place
153. sor cables are color coded Bottle Symbol Running Buffer blue Washing Solution green Storage Solution black Waste Red Table 2 4 Color coding of fluid bottles 1 Install one fluid bottle at a time Please note the corresponding color coding table 2 4 2 Take out an empty bottle and unscrew bottle closures counter clockwise but do not remove the bottle closures from the bottles Do not disconnect the color coded tubing 3 Place a fresh bottle into its appropriate holder open it and fasten the bottle closure to the new bottle Note the color coding and connect each sensor cable to the respective bottle closure Note To avoid contamination and spillover do not open a fresh bottle before it is placed in its holder Before removing a full waste bottle from the bottle holder fasten a standard lid onto the bottle to avoid spillover 4 Attach the sensor cable plug to the socket labeled Bottle Sensor at the back of the autoMACS Pro Separator and fasten securely 5 Attach the sensor cables to the cable guide at the back of the autoMACS Pro Separator 6 Connect the hydrophobic air filters 0 2 um to the appropriate connectors on the bottle closures Note Avoid any contact of the hydrophobic air filter with fluids as this may cause clogging of the filter 2 6 Installing the MACS MiniSampler MACS Reagent Rack 4 and tube rack The MACS MiniSampler guiding and its corresponding slot located at the front of the instrume
154. ssus 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 ni le 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 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 entra ner 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 ce que le chemin optique rayon laser ne se trouve pas 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 a
155. stema 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 Riesgos de radiaci n ptica 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 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
156. t 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 of the autoMACS Pro Separator and protect the instrument from overheating Never push a foreign object through an opening into the instrument Do not use the instrument in a wet or damp location Avoid high humidity or condensation and protect the machine against water splashes Unplug the autoMACS Pro Separator from the outlet before cleaning Do not use liquid or gt aerosol cleaning agents always use a damp cloth 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 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 s
157. ters and to prevent contamination of liquids air filters should be exchanged if they come into direct contact with any liquid Note Be sure to use hydrophobic air filters These are not the same as filters used for cell culture 7 2 3 Clean the pump syringe Depending on the level of usage the pump syringe must 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 cause 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 Dilutor valve Top screw Plunger Plunger holder Plunger lock screw Figure 7 2 Dilutor valve with pump syringe 1 Goto Option gt User settings gt Syrin_ex 2 Press Run to start the program 3 When prompted loosen the plunger lock screw at the bottom of the plunger Figure 7 3 Unscrewing the plunger lock screw 4 Press OK The plunger holder will move to the lowest position MAINTENANCE Figure 7 4 The plunger holder is in the lowest position 5 Adialog window will prompt the user to switch the instrument off replace the syringe and tighten the plunger screw Press OK to confirm 6 Switch off the instrument using the main switch 7 Unscrew the syringe from the dilutor valve housing by turning the knurled top screw counter clockwise Grip the syringe at
158. to the appropriate port Then cautiously insert and fasten the tube connector Run the Rinse program and check 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 Please note that each tube has a specific length and should be exchanged with the corresponding spare part only If the problem persists contact Technical Support 9 2 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 and retry If the problem persists follow the guidelines below 1 Run the Sleep program Press the shutdown button select Yes to shutdown the instrument 2 Switch off the instrument 3 Wash the pump syringe as described in section 7 2 3 4 Switch on the instrument and run the Qrinse program to ensure that the problem is solved Go to the Separation menu and press Wash Now Select Qrinse and press Run 5 If the leakage persists installing either a new Pump Syringe or a new Pump Seal might be required Contact Tech Support for assistance For details on the installation refer to section 7 2 3 9 2 4 Pump syringe is filled with air during operation If there is any air inlet into the pump syringe during operation
159. ttica pericolosa Radiazione laser pericolosa Rischio biologico Rischio di contaminazione se in uso materiale biologio poten zialmente pericoloso Morsetto di protezione Questo simbolo applicato internamente allo strumento Si tratta di informazioni per il personale di servizio On Alimentazione Off Alimentazione Etichetta di sicurezza Prendere visione dei punti critici e dei simboli di sicurezza del autoMACS Pro Separator Instrument Rischio di schiacciamento o di taglio di parti del corpo A Radiazione ottica pericolosa Radiazione Rischio di laser schiacciamento pericolosa o di taglio di parti del corpo Figure 9 Punti critici e posizioni dei simboli di sicurezza sulla parte frontale del autoMACS Pro Separator n 0 0 a 6 Figure 10 Posizione dei simboli di sicurezza sulla parte posteriore del autoMACS Pro Separator al 3 m 5 IMPORTANTI ISTRUZIONI DI SICUREZZA 5 49 50 Le etichette di sicurezza e i marchi di sicurezza devono essere mantenuti puliti e leggibili Ispezionare periodicamente le etichette di sicurezza e i marchi di sicurezza e sostituirli se illegibili o percettibili da una distanza di sicurezza Contattare Miltenyi Biotec per delle etichette sostitutive Instruzioni generali di sicurezza Se lo strumento non funziona correttamente e o le istruzioni visualizzate dei messaggi di avviso suggeriscono di contattare l assistenza tecnica no
160. ttom of column 1 substitute and pull gently but firmly to remove it from its slot Place a wide mouth bottle under the column 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 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 MAINTENANCE Align the column with the top column connector sitting on the guiding of the magnet cover Press the column into the slot until the guides click Verify that the column is placed in the center of the magnet cover Remove column 2 from its slot and repeat steps 8 through 11 to install the second column Ensure that the tubing is neither pinched nor obstructed Press Done The program will then proceed to wash the columns with autoMACS Running Buffer Check that the columns are securely fastened to the column connectors and that no buffer is leaking Close the front door 127 128 7 2 2 Exchange of hydrophobic air filters Hydrophobic air filters 0 2 um are attached to the bottle closures to vent the liquid bottles and to prevent release of aerosols They should be exchanged once a year to avoid clogging through dust deposits To avoid clogging of the fil
161. u des blessures graves si elle n est pas vit e ou CAUTION indique une situation potentiellement dangereuse pouvant causer des blessures mineures ou mod r es si elle n est pas vit e Il peut galement servir d alerte contre une utilisation risqu e 37 38 PPPP PP gt Symboles de s curit Les symboles suivants sont utilis s pour souligner des conditions qui pourraient entrainer des blessures du personnel ou une d t rioration du dispositif Alerte de s curit Risque de danger La documentation doit tre consult dans les cas o un symbole d alerte de s curit est mentionn afin de connaitre la nature du danger potentiel et les mesures prendre Risque d un choc lectrique Champ magn tique puissant Les personnes qui portent un stimulateur cardiaque ou un implant medical electronique doivent maintenir une distance de s curit Risque d crasement et de cisaillement des membres corporels Radiation optique dangereuse Radiation laser dangereuse Danger biologique Risque de contamination si des mati res biologiques potentiellement dangereuses sont utilis es Borne pour conducteur de protection Ce symbole est fix a l int rieure de l appareil Cette information est destin e au personnel de service On Alimentation lectrique Off Alimentation lectrique tiquette de s curit Il est important d identifier les zones de risque et symboles de s curit sur le s pa
162. ument may lead to severe personal injury or death by exposure to mechanical hazards hazardous optical radiation and the risk of electric shock Unplug the power cord before cleaning the instrument Always wear protective gloves and eyewear in order to protect against potential biohazard exposure Dispose of contaminated tissues and swabs according to their biological security level Three service contracts are available for the autoMACS Pro Separator Full Service Premium Full Service and Planned Maintenance Please visit www miltenyibiotec com support for further information and assistance In addition it is recommended to perform the following maintenance procedures to maintain the instrument s high performance The autoMACS Pro Separator must be primed each time the instrument is switched on The Rinse program is used for efficient washing and equilibration of the fluidic system The instrument automatically prompts the user to perform a rinse before performing a cell separation 1 Gotothe Separation menu and press Wash Now 2 Select Rinse and press Run 125 126 7 1 1 1 Automatic instrument priming at startup By using the O_init program the instrument can be instructed to perform an automated rinse sequence at startup 1 Select Option gt User settings gt O_init Press Run 2 Follow the prompt on the screen to enable or disable the initial wash 7 1 2 Clean uptake outlet ports It is recommended to clean uptake and o
163. us permet de recycler gratuitement vos quipements Miltenyi Biotec en fin de vie Les conditions et la disponibilit de cette offre varient selon les pays et les diff rentes exigences r glementaires Selon le type et l utilisation de votre quipement des exigences suppl mentaires peuvent s appliquer Pour plus d informations ou si vous souhaitez recycler votre quipement Miltenyi Biotec en fin de vie contactez votre interlocuteur Miltenyi Biotec ou notre support technique 45 2 m un EN Q m INFORMATIONS DE S CURIT IMPORTANTES n po 46 NWARNING WARNING CAUTION Il separatore autoMACS Pro Separator stato realizzato per garantire un uso sicuro se installato correttamente impiegato da personale specializzato e secondo la nor mativa generale di sicurezza e le istruzioni d impiego contenute in questo manuale Le linee guida riportate nella presente sezione illustrano i potenziali rischi associati 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 Vanno sempre osservate le istruzioni di sicurezza sul luogo di lavor
164. ution Autolabeling Reagent reagents volume Minimal Minimal Maximal Maximal volume total cell volume total number cell number Chill 5 Rack1 Direct MicroBeads Positive 1 10 cells per 160 uL 2x107 1600 pL 2x10 human rat non selection or 80 uL human primate depletion Direct MicroBeads Positive 1 10 cellsper 180 pL 2x107 1800 uL 2x10 mouse selection or 90 uL depletion Whole Blood Whole blood or 1 Original 0 25 mL 1mL MicroBeads bone marrow volume Cell Isolation Kits Untouched 2 10 cells per 160 uL 4x10 800 uL 2x10 isolation 40 uL Cell Isolation Kits Untouched 3 107 cells per 120 uL 4x10 600 uL 2x108 isolation 30 uL MicroBead Kits Positive 2 10 cells per 120 uL 2x107 1200 uL 2x10 selection or 60 uL depletion Chill 15 Rack2 Direct MicroBeads Positive 1 10 cells per 160 uL 2x107 5200 uL 6 5x10 human rat non selection or 80 uL human primate depletion di Direct MicroBeads Positive 1 10 cells per 180 uL 2x107 5850 uL 6 5x10 mouse selection or 90 uL y depletion Whole Blood Whole blood or 1 Original 1mL 4mL MicroBeads bone marrow volume Cell Isolation Kits Untouched 2 107 cells per 160 uL 4x10 2600uL 6 5x10 isolation 40 uL Cell Isolation Kits Untouched 3 10 cells per 120 uL 4x10 1950 uL 6 5x10 isolation 30 uL MicroBead Kits Positive 2 10 cells per 120 pL 2x107 3900uL 6 5x10 selection or 60 uL depletion Chill 50 Rack3 Whole Blood Wholebloodor 1 Original 4mL 8mL Micr
165. utlet ports daily This should be performed before running the Sleep program shutdown 1 Switch off the instrument 2 The ports can be wiped with tissue soaked with 70 ethanol isopropyl alcohol or MACS Bleach Solution followed by distilled or deionized water 3 Remove the needle guard that is attached underneath the outlet port holder Push up the ring holding the the finger guard in place Remove the finger guard by pulling gently Clean the finger guard as described for the ports Figure 7 1 Removing the needle guard for cleaning 4 Wipe the outlet port for the negative fraction as indicated above Flush the port using a syringe if desired 5 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 6 Push the needle guard back into position 7 Switch on the instrument 8 Run the Sleep program 7 2 Periodic maintenance 7 2 1 Column exchange Exchange autoMACS Pro Separator Columns every other week or after 100 separations whichever comes first Refer also to section 3 1 1 2 1 2 10 11 12 13 14 15 Open the front door of the autoMACS Pro Ensure that the fluid bottles are filled with solutions Got to Option gt Special gt Col_ex Press Run to start the Col_ex program Wait until the instrument prompts you to exchange the autoMACS Columns before proceeding Using both hands take the top and bo
166. utoMACS Pro Separator est d sign comme produit laser de la classe 1M selon la norme CEI 60825 1 1993 A1 1997 A2 2001 L utilisation d organes de commande autres que ceux 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 Dangers chimiques et biologiques 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 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
167. ve 134 Figure 7 12 Loosen the valve screws Figure 7 13 Detach the valve 7 Check if the groove in the valve drive is positioned horizontally and in the lower half of the axis Groove Bracket Hole for Adjust adjustment ment pin pin Figure 7 14 Left Valve plate driving section Right Rear view of the autoMACS Pro Separator valve 8 Make sure that the bracket of the new valve is positioned horizontally MAINTENANCE 135 136 9 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 15 Inserting the new valve 10 Gently rotate the valve The two adjustment pins will slide into their corresponding holes in the valve plate 11 Make sure that the valve is fully inserted into the driving station Fasten valve screws using the screwdriver 12 Connect the tubing with the installed new valve and fasten tubing finger tight 13 Plug in and switch on the autoMACS Pro Separator 14 Perform a Rinse program and check the valves visually for leakage and air inlet 15 Carefully refasten tubing connectors If leakage persists call Technical Support 16 Take care not to pinch the tubings at the bottom left of the instrument when closing the bottom cover 7 3 2 Dilutor valve exchange 1 Switch off and unplug the instrument 2 Remove the pump syringe as described in section 7 2 3 Figure 7 16

autoMACS® Pro Separator User manual

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autoMACS&reg; Pro Separator User manual

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autoMACS® Pro Separator User manual