Hotlink to autoMACS Pro Separtor QuickStart
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1. program 5 4 9 Push column into the magnet housing with the top column connector sitting on the guide in the column slot 5 4 10 Repeat installation for the second autoMACS Column After exchange of separation columns select Done The autoMACS Pro Separator system will be automatically primed with Running Buffer and is then ready for cell separation Short instructions Miltenyi Biotec Germany Austria Switzerland Miltenyi Biotec GmbH Friedrich Ebert Stra e 68 51429 Bergisch Gladbach Germany Phone 49 2204 8306 0 Fax 49 2204 85197 macs miltenyibiotec de USA Canada Miltenyi Biotec Inc 2303 Lindbergh Street Auburn CA 95602 USA Phone 800 FOR MACS Phone 1 530 888 8871 Fax 1 530 888 8925 macs miltenyibiotec com www miltenyibiotec com Australia Miltenyi Biotec Australia Pty Ltd Unit 16A 2 Eden Park Drive North Ryde NSW 2113 Australia Phone 61 2 8877 7400 Fax 61 2 9889 5044 macs miltenyibiotec com au Benelux Miltenyi Biotec B V Postbus 85183 3508 AD Utrecht The Netherlands macs miltenyibiotec nl Customer service Netherlands Phone 0800 4020120 Fax 0800 4020100 Customer service Belgium Phone 0800 94016 Fax 0800 99626 Customer service Luxembourg Phone 800 24971 Fax 800 24984 China Miltenyi Biotec Trading Shanghai Co Ltd Shanghai Office Fareast International Plaza A Rm 2301 No 319 Xianxia Rd Shanghai 200051 P R China Phone 86 21 62351005 Fax2. Miltenyi Biotec autoMACS Pro Separator Short instructions Version 02 Copyright 2010 Miltenyi Biotec All rights reserved No part of this user manual may be reproduced stored in a retrieval system transmitted published or distributed in any form or by any means electronically mechanically by photocopying microfilming recording or otherwise without the prior written consent of Miltenyi Biotec however notwithstanding the foregoing the owners of the autoMACS Pro Separator may make copies solely for purposes of training personnel in the use and servicing of the unit within their business or organization Maximal care has been taken by Miltenyi Biotec in the preparation of this user manual However Miltenyi Biotec shall not be liable for any technical or editorial errors or omissions contained herein or for incidental or consequential damages in connection with the furnishing performance or use of this document The information in this document is provided as is without warranty of any kind and is subject to change without notice autoMACS MACS and the MACS logo are registered trademarks or trademarks of Miltenyi Biotec GmbH or its affiliates in Germany the United States and or other countries Ficoll and Ficoll Paque are trademarks of GE Healthcare companies BD and BD Falcon are trademarks of Becton Dickinson and Company All other trademarks mentioned in this document are the property of their respective owne
3. Sample processing has not yet started Rinsing 2 Sample autolabeling 2 Sample processing is is underway completed 208 O 3 Incubation of cells Progress has been with labeling stopped or reagents cancelled 4 Sample is being processed e g sample uptake 2 10 3 Check bottle illuminations even from across the laboratory The autoMACS Pro Separator has a bottle illumination that facilitates monitoring of the instrument s status Code Status User action Green Ready for separation No action required Blue Instrument operating No action required Yellow Not ready for Run wash program separation Rinse or Qrinse before starting a separation Red Error Check screen for error detection Purple Program Sleep is Switch off autoMACS completed Pro Separator Blinking Action required Check screen for required action Short instructions 15 Note To store the autoMACS Pro Separator for a period longer than two weeks run the Store program Note For daily usage the instrument should not be switched off but placed into SLEEP mode See section 4 2 11 Optional Switch the instrument off for long term storage 1 11 1 Select Option and Special 1 11 2 Select Store and press Run 1 11 3 Replace the columns with column substitutes refer to section 3 3 4 of the User Manual 1 11 4 Select Done 1 11 5 Switch off the autoMACS Pro Separator using the main power
4. 1 Select Reagent menu and highlight the position where the vial will be placed on the reagent rack Four positions are available R1 R2 R3 and R4 es 17 5 2 6 2 Select Enter Reagent from the lower navigation bar Enter the reagent specific product order number The order number is located on the product data sheet In the event that the data sheet is misplaced visit www miltenyibiotec com to download a printable PDF of the document 2 6 3 Ifa correct product number is entered the software will immediately recognize and list the reagent or kit components To confirm your choice touch the listed reagent The reagent will be assigned to a rack position and the next available reagent rack position will be automatically highlighted Repeat the procedure for any remaining reagents or kit components Finally select OK to complete reagent entry The NK Cell Isolation Kit mouse was entered Short instructions 2 7 Place the sample rack onto the MACS MiniSampler and click on the Separation menu 1 Sample rack template 2 Sample labeling options 3 Sample processing volume 4 Separation program ss i 5 Wash procedure 2 8 Define the sample rack template for cell separation 2 8 1 Highlight the desired position s on the sample separation template and assign an autolabeling protocol from the Labeling menu The recommended cell separation and wash program will be automatically d
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6. status 3 Column status 4 Tube rack status E o 57 MiniSampler status 1 4 1 Confirm that the fluid container status is ready Left Fluid containers are shown in green indicating ready Right Fluid containers are shown in red and need replaced Short instructions Status of fluid containers displayed in the Status menu Container Symbol color and user action Green no action required Red refill container Gray connect bottle sensor Running Buffer Green no action required Red refill container Gray connect bottle sensor Washing Solution Gray no liquid detection visually check volume Storage solution Green no action required Red empty waste or wrong sensor cable is connected Gray connect bottle sensor Waste 1 4 2 Confirm that the column status is ready Status of the columns Green no action required Red exchange column Grey when no column has been installed 1 4 3 Confirm that the MiniSampler has been detected and is correctly installed Symbol Features a MACS MiniSampler No action required detected No sampler Connect MACS MiniSampler detected 1 5 The instrument is now ready to perform an experiment Short instructions Rack type amp symbol Chill 5 Chill 15 Chill 50 2 Performing autolabeling and cell separation 2 1 Choose an appropriate Chill Rack and ensure is it pre cooled to 4 C 2 1 1 Racks are automatically detected by the auto
7. switch Short instructions 3 Performing manual labeling and cell separation 3 1 Ensure that the appropriate Chill Rack is pre cooled to 4 C 3 1 1 Choose an appropriate Chill Rack Racks are automatically detected by the autoMACS Pro Separator Rack types and their features are tabulated below 3 1 2 Dilute single cell suspension according to the recommendations in the respective product data sheet Visit www miltenyibiotec com protocols to download product data sheets and protocols Racktype amp Slots Maximum Maximum Maximum number symbol numberof sample of total cells per tube samples volume Chill 5 24x5 mL 6 2 5 mL 5 0x108 5 mL tubes Chill 15 15x15 mL 5 12 5 mL 2 5x10 5x5 mL 15 mL tubes Chill 50 6x50 mL 3 50 mL 4 0x10 3x15 mL 50 mL 3x5 mL tubes Short instructions 17 3 3 Place the sample tubes into row A of the pre cooled Chill Rack Place empty tubes in the corresponding rows B and C these are for the negative and positive cell fractions respectively For example a Chill 5 Rack has six available positions A1 A6 Pr me EN Row A is for the original sample fraction i e Ori Row B is for the negative untouched fraction i e Neg Row C is for the positive enriched fraction i e Pos Reagent Rack positions are also shown 3 4 Place the sample rack onto the MACS MiniSampler and click on the Separation menu 1 Sample rack template 2 Sam
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9. 8 2 Select Store and press Run 3 8 3 Replace the columns with column substitutes refer to section 3 3 4 of the User Manual 3 8 4 Select Done 3 8 5 Switch off the autoMACS Pro Separator using the main power switch Short instructions 4 Setting the autoMACS Pro Separator in SLEEP mode 4 1 Bi Press the shutdown symbol upper right hand corner of the display 4 2 Alternatively select Sleep program as the last washing step Short instructions 24 5 Maintenance of the autoMACS Pro Separator 5 1 Rinsing programs Program Description Recommended usage Qrinse Standard short rinse of Between separation columns and separations of tubing system with frequent cells Running Buffer gt 5 Rinse Rinse of separation Between and columns and tubing before separations system with Washing of rare cells Solution and Running lt 5 Buffer 5 2 Daily maintenance programs Program Description Recommended usage Rinse Rinse of separation Prior to first columns and tubing separation system with Washing Solution and Running Buffer Sleep Rinse with Washing Before switching Solution followed by OFF the autoMACS filling with storage Pro Separator solution Short instructions Duration 1 5 min 4 min Duration 4 min 5 min 5 3 Periodic maintenance Program Description Column Replacement of exchange separation columns Safe Decontamination procedure with MACS Bl
10. I07 Removal of cells with normal to high antigen expression and results in a better target cell yield This special program is disabled by default To enable select Option User settings and O_progs A_Depls7 Removeal of cells with low antigen expression and results in a better target cell purity This special program is disabled by default To enable select Option User settings and O_progs Short instructions 19 20 3 5 3 Optional It is not mandatory to assign a volume for cell separation with manual labeling However the autoMACS Pro Separator requires this information to calculate and display the total sample processing time To assign a corresponding sample volume click on Volume and insert the sample volume Select Enter 3 5 4 Using the Wash menu assign a wash step between cell separation steps It is possible to change the rinse mode between samples or to program the instrument to go into sleep mode after finishing the experiment Qrinse standard short wash program that only uses Running Buffer Rinse Extensive rinsing program that uses Washing Solution and Running Buffer Sleep itis mandatory to use Sleep as the last wash program before overnight storage Upon completion of the Sleep program the fluidic system contains 70 ethanol Short instructions 3 6 Select Run to start the cell separation experiment and click OK to con
11. MACS Pro Separator Rack types and their features are tabulated below Slots Maximum Minimum Maximum Maximum number of number of first final cells per tube samples incubation labeling volume volume 24x5 mL 6 0 2 mL 2 0 mL 2 0 4x10 5 mL depends on cell labeling tubes concentration and column capacity 0 25 mL 1 mL Whole blood only 15x15 mL 5 0 2 mL 6 5 mL 6 5x108 1 3x10 5x5 mL 15 mL depends on cell labeling tubes concentration and column capacity 1 mL 4mL Whole blood only 6x50 mL 3 4mL 8 mL Whole blood only 3x15 mL 50 mL 3x5 mL tubes Short instructions NB renin ne acacia 2 2 Dilute single cell suspension volume required for the according to recommendations in EES ra eI StS the following table MACS Product Strategy Reagents Cell Minimal Minimal concentra volume absolute tion cell number Direct MicroBeads Positive selection 1 107 cells per 160 uL 2 0x107 human or depletion 80 uL rat non human primate Direct MicroBeads Positive selection 1 107 cells per 180 uL 2 0x107 mouse or depletion 90 uL Whole Blood Whole blood or 1 Original 0 25 mL MicroBeads bone marrow volume 8 mL Cell Isolation Kits Untouched 2 107 cells per 160 uL 4 0x107 selection 40 uL Cell Isolation Kits Untouched 3 107 cells per 120 uL 4 0107 selection 30 uL MicroBead Kits Positive selection 2 107 cells per 120 uL 2 0107 or depletion 60 uL When working with fewer cells than the necessary minimal volume resuspend cell
12. each Solution Pump Cleaning of pump syringe syringe refer to user manual Store Rinse with Washing 5 4 Column exchange Solution followed by storage solution replacement of columns and substitutes Recommended Duration usage Every two weeks 6 min OR after 100 separations whichever comes first Every 3 6 months 21 min Every 1 3 months Before storing the instrument for a period longer than two weeks Replace autoMACS Columns every two weeks or after 100 separations whichever comes first Select menu Option 5 4 1 Select Special and Col_ ex Press Run Figure 5 1 Col_ex program When prompted exchange the column ee ey he ni A zg 5 4 4 Open front door and note the positions of the columns column 1 left column 2 right Exchange one column at a time 3 4 5 Remove column from slot unscrew top column connector followed by the bottom column connector as shown in figure 5 2 Short instructions 25 26 5 4 6 Dispose of the expired column 5 4 7 Point the bottom of the fresh column towards the autoMACS Pro Separator 5 4 8 Insert bottom column connector Screw in the column by turning it clockwise Repeat the procedure for the top column connector 1 Top Column Connector 2 autoMACS Pro Separation Column 3 Bottom Column Connector Select comm action ee 14 42 Figure 5 2 Left Exchange of the column Right Starting the Col_ex
13. firm that enough buffer is available for the experiment z Enim Fam a ee LT onl nt las hex UG EEE 7 Standard positive cell separation Possel will be performed on sample positions 1 and 2 Clicking Run will start the experiment As the last wash step is Sleep a full rinse will be performed before the machine goes into sleep mode 3 7 Monitor the cell separation experiment 3 7 1 Use the Status menu display to view the overall instrument status Refer to section 1 4 for more details 3 7 2 Check bottle illuminations even from across the laboratory The autoMACS Pro Separator has a bottle illumination that facilitates monitoring of the instrument s status Code Status User action Green Ready for separation No action required Blue Instrument operating No action required Yellow Not ready for Run wash program separation Rinse or Qrinse before starting a separation Red Error Check screen for error detection Purple Program Sleep is Switch off autoMACS completed Pro Separator Blinking Action required Check screen for required action Short instructions 21 22 Note To store the autoMACS Pro Separator for a period longer than two weeks run the Store program Note For daily usage the instrument should not be switched off but placed into SLEEP mode See section 4 3 8 Optional Switch the instrument off for long term storage 3 8 1 Select Option and Special 3
14. isplayed To change these settings highlight the desired cell separation and wash options using the Separation and Wash menus respectively Sener eGo 03 55 2 8 2 To assign a corresponding sample volume click on Volume and insert the sample volume Select Enter Refer to step 2 2 above for information about using the correct volume Short instructions 13 Tip It is possible to change the rinse mode between samples or to instruct the instrument to go into sleep mode after finishing the last sample Qrinse Rinse oO Sleep Refer to section 3 5 4 for a more comprehensive explanation 2 9 Select Run to start the cell separation experiment and click OK to confirm that enough buffer is available for the experiment GE 132 CD4 positive cell separation will be performed on sample positions 1 and 2 Clicking Run will start the experiment The last wash step is Sleep thus a full rinse will be performed before the machine goes into sleep mode 2 10 Monitor the cell separation experiment 2 10 1 View the status of the fluid containers refer to step 1 4 1 2 10 2 Use the Status menu display to view the overall instrument status Sample processing statuses are shown as color coded graphics The table below shows an example of an autolabeling experiment performed using CD4 MicroBeads human Short instructions Graphic Definition Graphic Definition Status Waiting
15. ple labeling options 3 Sample processing volume 4 Separation program mus 5 Wash procedure Short instructions 3 5 Define the sample rack template for cell separation 3 5 1 Select the desired position s in the sample separation template field Tip Select multiple sample positions to program them simultaneously 3 5 2 Assign a corresponding cell separation program using the Separation menu Positive selection programs Possel lsolation of cells with normal antigen expression and frequencies higher than 5 select if purity is the highest priority Possel_s lsolation of cells with low antigen expression and frequencies higher than 5 select if yield is the highest priority Posseld For isolation of rare cells in low elution volume Posselds For isolation of rare cells with low antigen expression Posseld2 For isolation of rare cells if purity is the highest priority Posselwb For isolation of cell subsets from whole blood Cell samples are automatically diluted with Running Buffer Depletion programs Deplete For removal of cells with normal to high antigen expression and results in better target cell yield Depletes Removal of cells with low antigen expression and results in better target cell purity Depl0O5 Removal of cells with low antigen expression and results in stringent depletion of cells DepI025 Removal of cells with low antigen expression and results in stringent depletion of cells A_Dep
16. rs and are used for identification purposes only Unless otherwise specifically indicated Miltenyi Biotec products and services are for research use only and not for therapeutic or diagnostic use autoMACS Pro Separator Short instructions Manufacturer Miltenyi Biotec GmbH Friedrich Ebert Stra e 68 D 51429 Bergisch Gladbach Germany Technical Support Phone 49 2204 8306 830 Fax 49 2204 85197 macstec miltenyibiotec de www miltenyibiotec com Version 02 Short instructions 1 Setting up and priming the autoMACS Pro Separator 1 1 Check that all bottles are filled with the appropriate solutions Empty the waste bottle 1 2 Check that the MACS MiniSampler Code Reader and Fluid Sensor cables are correctly attached to the back of the instrument Sensor cable plug Code reader plug MACS MiniSampler plug Power socket Green sensor cable for Washing Solution Blue sensor cable for Running Buffer Bottle closure Short instructions Note To prime the instrument select Wash Now from the Separation menu 1 3 Switch on the autoMACs Pro Separator the instrument will automatically initialize After priming the instrument will automatically display the Status menu on off switch Figure 1 1 Location of the on off switch 1 4 Confirm that the instrument status is ready by viewing the Status menu Overview of the Status menu Eg Sample status 2 Fluid container filling
17. s in the stipulated minimal volume 2 3 Place the sample tubes into row A of a pre cooled Chill Rack Load empty tubes into the corresponding positions along rows B and C a al u B Neg Row A is for the original sample fraction i e Ori Row B is for the negative untouched fraction i e Neg Row C is for the positive enriched fraction i e Pos Reagent Rack positions are also shown R1 R4 Short instructions 2 4 Insert the MACS Reagent Rack 4 onto the MACS MiniSampler u The MACS Reagent Rack snaps into position as illustrated above 2 5 Scan reagent vials 2 5 1 On the Reagent menu select Read Reagent and present a reagent vial in front of the 2D code reader Ensure the 2D code is facing the blinking code reader light Short instructions The optimal reading distance is 0 5 2 5 cm from the code reader cover tilt the vial as shown above 2 5 2 After successfully scanning a reagent vial the software will automatically highlight the next available reagent rack position To view details about the scanned reagent vial highlight the appropriate rack position This is R1 in the example below as 245 2 5 3 Insert the reagent vial into the correct rack position 2 6 Optional Manual entry of reagents This is only recommended if the reagent cannot be identified by the barcode reader Please turn over Short instructions 11 2 6
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