The Complete Flow Cytometry Guide

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Flow cytometry is a laser-based technique widely used to analyze cell surface and intracellular molecule expression. Flow cytometry tests measure the fluorescence intensity emitted by antibodies labeled with fluorescent dyes that are specific to proteins on or inside cells or ligands that bind to certain cell-associated molecules.

The staining process involves creating a single-cell suspension from either tissue samples or cell culture. These cells are then incubated in tubes or microtiter plates with unlabeled or fluorophore-labeled antibodies and subsequently analyzed using a flow cytometer.

Multicolor flow cytometry expands on this by allowing the simultaneous analysis of several parameters on thousands of cells or particles within seconds. Using fluorescent markers, this technique helps define and identify various cell types of interest in heterogeneous populations, evaluate the purity of subpopulations, and assess cell shape and size.

This guide covers the fundamentals of flow cytometry, aiming to help you better understand how it works, when it is most valuable, and how to conduct an experiment.