Flow Cytometry

Flow cytometry is a biophysical tool that rapidly counts and sorts cells and particles in solution. Each particle goes through an electronic optical system which detects the light scattered by it when passing through the laser light beam. The individual light signal scattered when the light source is interfered with is picked up by detectors and processed through a computer program. One of the most important analytical characteristics of flow cytometers is their ability to measure multiple cellular parameters (size, shape, complexity, and fluorescence) and to detect and identify cellular components by using fluorescent markers. These qualitative, multi-parametric measurements make it possible to define the properties of cell populations and even the subpopulations composing them.

The analysis of samples at IMO’s Flow Cytometry Laboratory mainly consists in the characterization of autofluorescent organism populations and subpopulations, such as cyanobacteria and protists capable of photosynthesis, as well as of non-fluorescent particles such as viruses, bacteria and eukaryotic cells from both natural environments and cell lines that can be stained with fluorescent pigments and subsequently analyzed.

Our Flow Cytometry Laboratory is properly equipped and run by specialized technical and professional personnel with extensive experience in the analysis of both culture and environmental samples collected in freshwater and saline solutions such as those found in rivers, lakes, and the sea. We have a FACSCalibur flow cytometer with a blue emission laser (λ = 488 nm), allowing us to identify not only natural but also induced fluorescence (for bacterial analysis using DNA stains) using 5 detection channels, two for the optical parameters "Side Scatter" and "Forward Angle Light Scattering", and three fluorescence detectors at specific wavelengths (580, 610 and 670 nm). Additionally, we have a high-performance experimental cytometer, the InFlux®, consisting of 5 excitation lasers (λ = 355-457-488-532-638 nm) and 16 detectors that can be arranged according to particular needs, producing higher resolution results.

Along with the analysis of each sample, the lab delivers a complete results report in two sections: first, a general report containing information about the equipment, the number of samples and general observations, as well as two annexes called Fluorescence and Cells, including parameters, graphs (cytograms) and descriptive information about each sample observation; and a second section with values for those parameters in each analysis. The type of analysis depends on the parameters defined by the requester.


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