BD FACSAria Cell-Sorting System

The BD FACSAria is a 14-parameter plus forward scatter PMT, custom-made high-speed digital bench-top cell sorter equipped with four excitation lasers (20 mW Blue, adjustable 0-150 mW Green, 25 MW Red and 60 mW UV). The instrument is able to perform two- and four-way aseptic sorting for a variety of tube sizes as well as sorting into multi-well plates. The flow cell is able to accommodate 70μm, 100μm and 130μm-sized nozzles to sort a variety of cell sizes. The instrument is also equipped with an aerosol management option to control the risk of biohazard exposure by creating a negative laminar-flow pressure in the sort chamber. We routinely use the FACSAria to analyze surface and intracellular marker expression (eg. phosphorylation events in signaling pathways), fluorescent protein expression (eg. GFP), morphological features based on Forward and Side Scatter characteristics, apoptosis and cell cycle/ploidy in megakaryocytes and other blood cell types as well as bacterial samples. Other applications of flow cytometry include, but are not limited to, immunophenotyping, detection of rare events, absolute cell counting, analysis of cellular biochemistry (eg. intracellular calcium levels, intracellular pH) and functional studies (eg. phagocytosis of fluorescence-labeled bacteria). Fluorescence-activated cell sorting (FACS) can be performed based on these parameters. Our FACSAria is equipped with FACSDiva software, which is used for acquisition and is a user-friendly platform for data analysis.

Design of the collection optics of the BD FACSAria custom-made for the Papoutsakis lab

Flow cytometry resources available online (aka Pani's favorite websites)
http://www.bdbiosciences.com/immunocytometry
(a description of the BD FACSAria Cell-Sorting system)

http://www.cyto.purdue.edu/hmarchiv/Current/index.htm
(Purdue cytometry mailing list archive: an outstanding online resource for finding protocols, asking the flow cytometry experts worldwide, geared towards the beginner as well as the more advanced student of cytology)

http://www.isac-net.org/
(website of the International Society for Analytical Cytology - ISAC)

http://probes.com/resources/education/
(introduction to flow cytometry tutorials from Molecular Probes website)

http://probes.com/resources/spectraviewer/
(fluorescence spectraviewer from Molecular Probes website)

http://probes.invitrogen.com/products/flowcytometry/
(Free online access to Practical Flow Cytometry by Howard M. Shapiro, one of the best flow cytometry reference textbooks, provided through the Molecular Probes website)

http://www.drmr.com/compensation/index.html
(An excellent informal guide to compensation written by Mario Roederer)

Note: For Further details contact Pani Apostolidis, Stephan Lindsey and Bryan Tracy .