FLOW CYTOMETRY

Flow cytometry is primarily used to analyze particles (usually cells but also vesicles and viruses) for the levels of surface proteins.  After fixing and permeabilizing the particles, internal protein-, DNA-, and RNA-levels, can be assessed.  Selected physiological parameters are measurable by flow cytometry, such as Ca2+ flux, oxidative state, protein phosphorylation, mitochondrial activity, and pH.  Aggregation of particles and the formation of conjugates can be assessed. Conventional flow cytometers provide a single aggregate measurement from each particle, but do not provide an image of the cell.  The Amnis ImageStream is an imaging flow cytometer that as the name connotes also provides a detailed (upto 60x magnification) image of the cell.  LJIDI has a 4-color Accuri flow cytometer and an Amnis ImageStreamx Mk II for flow cytometric analysis.  Our flow cytometers are analyzers only and not capable of sorting cells for subsequent analysis.  The Scripps Research Institute (TSRI) core facility has flow cytometers (LSRII, Compucyte Laser Scanning Cytometer, FACSCalibur HTS) and 3 high speed cell sorters (Aria).  The La Jolla Institute for Allergy and Immunology (LIAI) core facility also has flow cytometers (LSRII,LSR Fortessa, and Canto-II) and cell sorters (Aria).  The Aria sorts at 25,000 particles/second.

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