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Automacs pro talk @ Garvan Flow

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Garvan Flow Cytometry Instrument talk. Given by Dr. Katharina Winnemöller, Research and Clinical Sales Manager (NSW, ACT) at the Garvan Instituteof Medical Research on the 15th October 2010.

AutoMacs Pro magnetic Cell Seapartion - "the faster way to ultra-pure, viable cells"

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Automacs pro talk @ Garvan Flow

  1. 1. MACS cell separation - the faster way to ultrapure, viable cells + Dr. Katharina Winnemoeller katharinaw@miltenyibiotec.com.au
  2. 2. About us  Germany's largest independent, privately owned biotech company 1300 employees in 19 countries  Cell biologists, biochemists, biophysicists, clinicians, and engineers all acting as a community 20% of staff in R&D, engineering & clinical development  Manufacturing on site minimal reliance on external suppliers
  3. 3. About us  Founded in Germany in 1989  World market leader in magnetic cell separation (MACS) Cologne (Bergisch Gladbach) corporate headquarters  18 000+ publications with products from Miltenyi Biotec
  4. 4. Worldwide sales and support
  5. 5. Technology and services  Sample preparation  Cell Separation  Cell Culture  Cell Analysis  Molecular Analysis  Small animal imaging  Contract MFG  High throughput automation
  6. 6. MACS MicroBead Technology Flexibility comes as standard
  7. 7. We are translational  Miltenyi provides solution from bench to bedside
  8. 8. Cell Separation
  9. 9. MACS® MicroBeads – Key benefits  MACS® MicroBeads are 50nm superparamagnetic particles  Flexible cell sorting strategies:  positive selection – depletion – multiparameter sorting (no stripping!)  Non-toxic, biodegradable:  labeled cells go straightly to downstream experiments such as flow cytometry, cell culture,...  For virtually any cell type: more than 350 separation reagents
  10. 10. MACS® Technology Proven worldwide – for more than 20 years Minimal labeling required due to column
  11. 11. Cell Separation Portfolio 390 cell separation Stem Cells Clinical reagents 93 55 Immunology Neuro 287 62 cytokine prod. cells Whole blood 17 10 various (MHC,…) fluorochromes 17 11 B cells 46 T cells NK cells 93 Granul. 13 Monos/ Mph 15 18 CSC 5 Tumor cells DC 31 Neural cells 21 6 ES/iPS cells 17 HSC 18
  12. 12. Next generation MicroBeads:  CD4+CD25+ T reg Isolation Kit, human  CD4+CD25+CD127dim/– Regulatory T Cell Isolation Kits  CD4+ T Cell Isolation Kit II, human, mouse  CD8+ T Cell Isolation Kit II, human, mouse  Pan T Cell Isolation Kit II, human, mouse  Plasmacytoid Dendritic Cell Isolation Kit II, human  New Pan Monocyte Isolation Kit, human  NK Cell Isolation Kit II, mouse  Naive Pan T Cell Isolation Kit, human (optional TCR γ/δ T cell depletion)
  13. 13. Whole Blood MicroBead Technology Convenience standardized Positive cell isolation directly from peripheral blood Automated with the autoMACS Pro Complete walkaway automation Higher recovery No RBC lysis No density centrifugation
  14. 14. A peek into the future (mouse cells): New Kits Pan-DC Enrichment Kit, mouse Pan-DC Isolation Kit, mouse PDC Isolation Kit II, mouse Classical DC Isolation Kit, mouse Monocyte Isolation Kit (BM), mouse Monocyte Isolation Kit (WB), mouse
  15. 15. Available now: fast protocols (mouse cells):
  16. 16. autoMACS Pro Separator – Key features
  17. 17. Automated multisample cell sorting Designed and manufactured in Germany Up to 6 samples Option for automated labeling Sample cooling racks  Volumes: 0.25-50 mL (for 5, 15, 50 mL sample tubes)  Samples stay at 4 degrees  Choice of 12 separation programs
  18. 18. Re-usable columns  For 14 days, or up to 100 separations  High capacity – cell sorting within a few minutes  105 up to 4 x 109 total nucleated cells  up to 2 x 108 magnetically labeled cells  up to 15 mL of whole blood
  19. 19. Numbers….  2000 autoMACS (Pro) worldwide, 47 in Australia  Up to 10 Mio cells per second
  20. 20. Programs and running time
  21. 21. autoMACS Pro programs Positive selection Possel positive selection in standard mode for isolation of cells with normal antigen expression and frequencies higher than 5% 4 mL/min Possel_s positive selection in sensitive mode for isolation of cells with low antigen expression and frequencies higher than 5%; 1 mL/min Posseld positive selection in standard mode I, double-column program for isolation of rare cells in low elution volume 4 mL/min Posselds positive selection in sensitive mode, double-column program for isolation of rare cells with low antigen expression 1 mL/min Posseld2 positive selection in standard mode II, double-column program for isolation of rare cells 4 mL/min Posselwb special positive selection in double-column program for isolation of cell subsets from whole blood 4 mL/min
  22. 22. autoMACS Pro programs Depletion Deplete Depletion in standard mode for removal of cells with normal to high antigen expression and results in better target cell yield. 4 mL/min Depletes depletion in sensitive mode I for removal of cells with low antigen expression and results in better target cell purity. 1 mL/min Depl05 depletion in sensitive mode II for removal of cells with low antigen expression and results in stringent depletion of cells 0.5 mL/min) Depl025 depletion in sensitive mode III for removal of cells with low antigen expression and results in stringent depletion of cells 0.25 mL/min) A_depl7 special depletion in standard mode for removal of cells with normal antigen expression; used for large sample sizes where magnetically labeled cells exceed autoMACS Column binding capacity (2x108). depends A_deplS7 special depletion in sensitive mode for removal of cells with low antigen expression; used for large sample sizes 4 mL/min
  23. 23. Results, Examples From abundant to rare cells
  24. 24. CD4 MicroBeads, human autoMACS™ program Start percentage CD4 CD4 Purity % CD4 cells in negative fraction POSSEL 39,3% 97,9% 16,7% 2.5 min POSSEL_S 39,3% 94,7% 3,2% 5 min POSSELD 39,3% 99,6% 17,3% 7 min POSSELDS 39,3% 97,2% 2,21% 8.5 min CD4-Cy5 stained sample Results analysed by flow cytometry By Courtesy of A. Sheffold, A. Thiel - Berlin, Germany time
  25. 25. Whole Blood CD14 MicroBeads, human Anticoagulated whole blood; Purity: > 98% program Posselwb /Rinse
  26. 26. CD34 MicroBead Kit, human Original sample Human cord blood, Purity > 95% Program POSSELD / Rinse Count 7-AAD 0.37% CD34+ cells Count 7-AAD 95.38% CD34+ cells CD34-PE CD34-PE CD34+ cells autoMACS Separator, by Courtesy of Dr. Haberstock, Dr. Allam, Hautklinik Bonn, Germany
  27. 27. Maintenance and service
  28. 28.  Daily Maintenance  Priming: Program Rinse (automated before first separation)  Shut down: Program Sleep for overnight storage (fill with Ethanol)  Periodic Maintenance  Exchange Columns: Every 2 weeks or after 100 separations, whatever comes first  Clean pump syringe: Every 1-2 month  Long term storage; (substitute columns)  Service – Service engineer and spare parts located in Sydney
  29. 29. autoMACS Pro - the best way for automated magnetic cell separation Close to 400 Products for cell separation Most flexibility  positive or negative selection  true Whole Blood separation, PBMC, other sources Prettiest instrument!
  30. 30. autoMACS Pro demo – experience the difference From November 1st you can try out the autoMACS Pro by yourself Until November 15th I will be around regularly to help you with your separation Let me know what you would like to separate  please let me know by tomorrow morning I will order the respective samples
  31. 31. Thank you for you attention

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