Fluorescence Activated Cell Sorting (FACS) Methods, Techniques & Protocols




Definition: Fluorescence-activated cell-sorting (FACS) is a specialised type of flow cytometry. It provides a method for sorting a heterogenous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell.

General Methods & Techniques

Antigen-Antibody Specific Applications

Tissue-Cell Specific Applications

Tumor, Disease & Diagnostic Applications

Questions and Answers about Cell Sorting - Q1: When should I use fluorescence activated cell sorting over bulk separation methods like panning or magnetic bead separations? A: When very high purity (95%-100%) of the target population is required.

FACS staining protocols - Single colour standard method (indirect detection); Dual colour method using local biotin or PE conjugate (after indirect detection of first antibody)

Setting up 2 or 3 Colour FACS Analysis - These detailed instructions are for the FACScans with FACSMates, but the principles also apply to setting up 3 (or even 4) colour analysis on the FACSort.

Four Colour Analysis on the FACSort - The FACSort has recently been upgraded to allow four colour analysis. This is possible through the addition of a second (red) diode laser and a new detector to allow the use of allophycocyanin (APC) conjugates in addition to the usual 3 colours.

FACS STAINING OF PBLs for monitoring CD4 and CD8 depletion - This method is useful for monitoring depletion in vivo using rat IgG2b monoclonal antibodies against CD4 and CD8.