Einstein Flow Cytometry Core Facility 2013 CFAR Retreat, January 15, 2013 Steven A. Porcelli, MD Scientific Director 416 Forchheimer, x3228

FACS Core Laboratory 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)

Jinghang Zhang, MD Operations Director, Asst. Operations Director, Lydia Tesfa, PhD Asst. Operations Director, Olisambu Uche, PhD Asst. Operations Director, Grace Wang, B.S. Facility Technician, FACS Core Staff

Partec CyCube 6 (Chanin 309) FACSCanto II (Price 159A) FACSCalibur DxP10 (Chanin 309) LSR II (Chanin 308A and 307A) Analyzers 2013

CytometerLocationLasersDetectorsSoftware BD FACSCaliburChanin 307A2 lasers4 colorsCellquest CyFlow Cube 6Chanin 3092 lasers4 colorsPartec BD FACSCanto IIPrice 159A2 lasers6 colorsDiva BD FACSCalibur DxP10 Chanin 3094 lasers10 colorsFloJo Collector BD LSRIIChanin 308A4 lasers12 colorsDiva BD LSRII-yChanin 307A5 lasers14 colorsDiva COMPLEXITY / COST

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:

Cell Sorters 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+)

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 APPROACH TO DELIVERY OF SERVICES Strong emphasis on training facility users to do their own work using Facility resources.

Costs for services – Charge back rates (mostly unchanged since 2007) 20% discounts on all services also applied to CFAR and Diabetes Center members 20% off these rates for CFAR