Transcript Flow Cytometry Core

Einstein Flow Cytometry Core Facility
2013
CFAR Retreat, January 15, 2013
Steven A. Porcelli, MD
Scientific Director
416 Forchheimer, x3228
[email protected]
FACS Core Laboratory 2004 - 2013
Main location: 3rd Floor Chanin Building
Recent renovations funded by $850,000
ARRA Award Supplement to AECC.
Completed May 2012
Also:
PRICE CENTER (1st Floor) – FACS Core
Satellite Laboratory (incorporated 2009)
FACS Core Staff - 2013
Jinghang Zhang, MD
Lydia Tesfa, PhD
Operations Director, 2009Asst. Operations Director, 2006-2009
Asst. Operations Director, 2009-
Olisambu Uche, PhD
Grace Wang, B.S.
Asst. Operations Director, 2011-
Facility Technician, 2012-
Analyzers 2013
Partec CyCube 6 (Chanin 309)
FACSCalibur DxP10 (Chanin 309)
FACSCanto II (Price 159A)
LSR II (Chanin 308A and 307A)
Analyzers 2013
Cytometer
Location
Lasers
Detectors
Software
BD FACSCalibur
Chanin 307A
2 lasers
4 colors
Cellquest
CyFlow Cube 6
Chanin 309
2 lasers
4 colors
Partec
BD FACSCanto II
Price 159A
2 lasers
6 colors
Diva
BD FACSCalibur
DxP10
Chanin 309
4 lasers
10 colors
FloJo Collector
BD LSRII
Chanin 308A
4 lasers
12 colors
Diva
BD LSRII-y
Chanin 307A
5 lasers
14 colors
Diva
Analyzers – 2013
The Compucyte iCys Laser Scanning Cytometer (LSC)
Similar to a flow cytometer except without the flow component.
Cells are analyzed while attached to slides or culture vessels by moving
them through the laser with an automated nanostep microscope stage.
Although slower than flow cytometry, this device gathers both microscope
images and quantitative fluorescence signals on each cell.
Quantification of phagocytosis and cell cycle phase in macrophages by LSC
Coelho C & Casadevall A et al. Infect. Immun. 2012;80:1467-1478
Cell Sorters - 2013
Two instrument platforms:
MoFlo (Beckman-Coulter)
Fastest (triggering rate >70,000 events/second)
Most flexible
Best for delicate cells and sorting rare events
FACSAria II (BD Biosciences)
Fast, but not as fast as MoFlo
Good configuration, but less flexible
Simpler operation
Best for applications involving biocontainment
Biohazard Sorting 2013 (human cells including HIV+)
APPROACH TO DELIVERY OF SERVICES
Strong emphasis on training facility users to do their own work using
Facility resources.
For most analytical FACS procedures:
We train you to use the instruments and provide guidance
as needed.
There are exceptions from non-laboratory based researchers;
e.g, CD4 counts by MultiSet, priced very low ($29/sample)
For cell sorting:
We usually do it for you
(i.e., you prepare the samples and bring them to us to be
sorted according to your specifications)
There is an option for self operation of FACSAria for non-biohazard
related sorting
Costs for services – Charge back rates (mostly unchanged since 2007)
20% off
these rates
for CFAR
20% discounts on all services also applied to CFAR and Diabetes Center members