Flow cytometry technology allows simultaneous multiparametric analysis of thousands of cells per second, enabling trained users to rapidly analyze complex cell populations based on phenotypic and functional features. High-speed assisted cell sorting services provide researchers with physical separation of identified cell populations, for any downstream characterizations.
Flow cytometry is a laser-based technology in which cells in suspension are illuminated one-by-one and at high speed by different excitation sources (i.e. violet, blue, yellow-green or red lasers). Cell populations are identified based on physical and fluorescence parameters :
- Scattered light collected at low angle (Forward Scatter or FSC) allows to roughly determined cell size.
- Scattered light collected at 90° (Side Scatter or SSC) allows to roughly determined internal complexity or granularity.
- Autofluorescence and emitted fluorescence of cells stained either by fluorescent compounds or by antibodies coupled to fluorochromes are collected at 90° and allow precise determination of cell populations and functional features.
The scattered light and fluorescences are collected, separated and directed to detectors by various set of filters. The detectors or photon-to-electron multipliers (PMT) have a dual role i) to transform photons into electrons ii) to amplify electronic signals. Finally, electronic signals are digitalized in order to be interpreted by the computer and visualized by users. Raw data are analyzed with a dedicated software called FlowJo.