automacs pro talk @ garvan flow
DESCRIPTION
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"TRANSCRIPT
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 siteminimal reliance on external suppliers
About us
Cologne (Bergisch Gladbach)corporate headquarters
Founded in Germany in 1989
World market leader in magnetic cell separation (MACS)
18 000+ publications with products from Miltenyi Biotec
About us
Worldwide sales and support
Sample preparation Cell Separation Cell Culture Cell Analysis Molecular Analysis Small animal imaging Contract MFG High throughput automation
Technology and services
MACS MicroBead TechnologyFlexibility comes as standard
Miltenyi provides solution from bench to bedside
We are translational
Cell Separation
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
MACS® Technology
Proven worldwide – for more than 20 years
Minimal labeling
required due to column
Cell Separation Portfolio
390 cell
separation
reagents
Clinical
55
Immunology
287
Neuro
62
Stem Cells
93
T cells
93
B cells
46
DC
31
NK cells
18HSC
18
Neural cells
21
ES/iPS cells
17
Monos/Mph
15
Tumor cells
6Granul.
13
various (MHC,…)
17 CSC
5
fluorochromes
11
cytokine prod. cells
17
Whole blood
10
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)
Whole Blood MicroBead TechnologyConvenience standardized
Positive cell isolation directly from peripheral blood
Automated with the autoMACS Pro
Complete walkaway automation
Higher recovery
No RBC lysis
No density centrifugation
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
A peek into the future (mouse cells):
Available now: fast protocols (mouse cells):
autoMACS Pro Separator – Key features
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
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
Numbers….
2000 autoMACS (Pro) worldwide, 47 in Australia
Up to 10 Mio cells per second
Programs and running time
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 isolationof cell subsets from whole blood
4 mL/min
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 Columnbinding 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
Results, Examples From abundant to rare cells
CD4 MicroBeads, human
CD4-Cy5 stained sampleResults analysed by flow cytometryBy Courtesy of A. Sheffold, A. Thiel - Berlin, Germany
POSSEL
POSSEL_S
POSSELD
POSSELDS
39,3%
39,3%
39,3%
39,3%
97,9%
94,7%
99,6%
97,2%
16,7%
3,2%
17,3%
2,21%
2.5 min
5 min
7 min
8.5 min
autoMACS™ program
Start percentage CD4
CD4 Purity
% CD4 cells in negative fraction time
Whole Blood CD14 MicroBeads, human
Anticoagulated whole blood; Purity: > 98%program Posselwb /Rinse
CD34 MicroBead Kit, human
Human cord blood,Purity > 95%
Program POSSELD / Rinse
7-A
AD
7-A
AD
Co
un
tC
ou
nt
CD34-PECD34-PE
autoMACS Separator, by Courtesy of Dr. Haberstock, Dr. Allam, Hautklinik Bonn, Germany
0.37%CD34+ cells
95.38%CD34+cells
Original sample
CD34+ cells
Maintenance and service
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
Close to 400 Products for cell separation
Most flexibility
positive or negative selection
true Whole Blood separation, PBMC, other sources
Prettiest instrument!
autoMACS Pro - the best way for automated magnetic cell separation
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
autoMACS Pro demo – experience the difference
Thank you for you attention