Flow cytometry sample preparation refers to the process of obtaining, treating, and labeling cells or particles from a sample so that they can be analyzed using flow cytometry. This includes steps like obtaining a single-cell suspension, staining cells with appropriate fluorochrome-conjugated antibodies, and ensuring that the cells or particles are in an optimal condition for flow cytometric analysis.
Flow cytometry is a sophisticated laser-based technique employed to evaluate the expression of cell surface and intracellular molecules. Central to both research and diagnostics, it enables the simultaneous multi-parameter analysis of individual cells, aids in characterizing varying cell types in mixed populations, and can measure cell size and volume. Flow cytometry assays determine the purity of isolated cell subgroups and even sort diverse cell populations, a process termed fluorescence-activated cell sorting (FACS). This is achieved by detecting fluorescence intensity emanating from cells labeled with fluorescent antibodies specific to cell proteins or ligands, like propidium iodide linked to DNA.
Depending on its configuration and the intended application, a flow cytometer can have various components and capabilities including:
The sample preparation for flow cytometry involves creating a single-cell suspension, typically from cell cultures or tissue samples, and incubating these cells with either unlabeled or fluorophore-tagged antibodies. Here are some common methods of FC Sample Preparation: