The CABIMER Flow Cytometry and Cell Sorting Facility provides instrumentation and assistance for multi-parameter flow cytometry analysis and sorting.

Flow cytometry is a powerful tool that measures functional and structural characteristics of heterogeneous mixtures of cells and particles in suspension. The properties measured include relative size, relative granularity or internal complexity, and relative fluorescence intensity. These characteristics are determined using an optical-to-electronic coupling system that records how the cell or particle scatters incident laser light and emits fluorescence.  

Cell sorting allows the physical isolation of distinct populations of cells of interest for further downstream applications including cell culture, RNA or protein analysis and single cell cloning.

Frequent applications

-Detection of cell surface and intracellular antigens by

 immunofluorescence (immunophenotyping).

-Cell viability assays.

-Cell toxicity by changes of membrane potential.

-Cell cycle analysis (Propidium Iodide, 7-AAD, BrdU...).

-Analysis of apoptosis and necrosis.

-Gene reporter assays (fluorescent proteins).

-Reactive oxygen species (ROS) analysis (superóxide, peroxide and

 nitric oxide).

-Changes in intracellular pH and Ca2+ fluxes with specific    

 fluorochromes.

-Cellular proliferation measured by the dilution of the fluorescent dye   

 CFSE with each round of division.

-Cell sorting.

Equipment

- 2 BD FACSCalibur analytical cytometers: 488 and 635 nm lasers.

- BD FACSAria high-speed cell sorter: 407, 488 and 633nm lasers.  

  Multiwell plate deposition system (ACDU). Temperature control system for  

  sample and collection tubes.