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Flow Cytometry

 

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BD Accuri C6 Flow Cytometer

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  • Flow cytometry (FCM) is a technique for counting and examining (notably the fluorescence) 0.5 - 150 μm particles by suspending them in a stream of fluid and directing a particular wavelength of light at them. This technology can evaluate particles at a rate of 10,000 per second. Detectors in line with the light beam pick up Forward Scatter (FSC) light, detectors perpendicular to it detect Side Scatter (SSC) and a fluorescent detector all collect the scattered light and fluorescence. Ultimately, FSC correlates with the cell volume and SSC with the inner complexity of the particle. The fluorescence will be specific for the particular fluorophore you are using.
  • FCM is routinely utilized to count the number of live cells within a population expressing a particular cell surface protein of interest. Additionally with permeabilized cells intracellular fluorescence can be detected. Flow cytometry has been successfully used in the following applications:
      • Apoptosis
      • Calcium Flux
      • Cell counting
      • GFP Transfection
      • Immunophenotyping
      • Microbiology
      • Cell cycle and DNA Analysis
      • Stem cell markers
  • Optimal use of Accuri C6 Flow Cytometer requires a single-cell suspension & aggregated samples require filtering. This instrument does not support Fluorescence-activated Cell Sorting (FACS). Experiments utilizing smaller particles (e.g. prokaryotic) typically require more optimization than larger (e.g. eukaryotic) ones. The minimum volume required is 50μl. The technical specifications of this instrument include a 488 and 640 solid state laser.