Einstein Flow Cytometry Core Facility 2013 CFAR Retreat

Einstein Flow Cytometry Core Facility 2013 CFAR Retreat, January 15, 2013 Steven A. Porcelli, MD Scientific Director 416 Forchheimer, x 3228 steven. porcelli@einstein. yu. edu

FACS Core Laboratory 2004 - 2013 Main location: 3 rd Floor Chanin Building Recent renovations funded by $850, 000 ARRA Award Supplement to AECC. Completed May 2012 Also: PRICE CENTER (1 st Floor) – FACS Core Satellite Laboratory (incorporated 2009)

FACS Core Staff - 2013 Jinghang Zhang, MD Operations Director, 2009 Asst. Operations Director, 2006 -2009 Lydia Tesfa, Ph. D Asst. Operations Director, 2009 - Olisambu Uche, Ph. D Grace Wang, B. S. Asst. Operations Director, 2011 - Facility Technician, 2012 -

Analyzers 2013 Partec Cy. Cube 6 (Chanin 309) FACSCalibur Dx. P 10 (Chanin 309) FACSCanto II (Price 159 A) LSR II (Chanin 308 A and 307 A)

Analyzers 2013 Lasers Detectors Software BD FACSCalibur Chanin 307 A 2 lasers 4 colors Cellquest Cy. Flow Cube 6 Chanin 309 2 lasers 4 colors Partec BD FACSCanto II Price 159 A 2 lasers 6 colors Diva BD FACSCalibur Dx. P 10 Chanin 309 4 lasers 10 colors Flo. Jo Collector BD LSRII Chanin 308 A 4 lasers 12 colors Diva BD LSRII-y Chanin 307 A 5 lasers 14 colors Diva ITY / COST Location COMPLEX Cytometer

Analyzers – 2013 The Compucyte i. Cys 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: Mo. Flo (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 Mo. Flo 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, CD 4 counts by Multi. Set , 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