Flow Cytometry (FC) Methods, Techniques & Protocols




Definition: analysis of cells or subcellular components by detecting fluorescence or light-scatter of sample fractions passing in narrow-stream droplets through a laser beam.

General Methods & Techniques

Antigen-Antibody Specific Applications

Tissue-Cell Specific Applications

DNA-RNA Detection

Apoptosis Detection

Virological Applications

Tumor, Disease & Diagnostic Applications

Flow Cytometry Protocols from UCLA - Welcome to the Flow Cytometry Core Laboratory at UCLA. The Core Laboratory provides instrumentation and technical and professional assistance for performing laser-based flow cytometric analysis and sorting. 

Salk Flow Cytometry Protocols - Paraformaldehyde preparation, Fluorescein labeling, Phycoerythrin labeling, FCM of leukocytes in whole blood, Yeast cell cycle

Advanced Methodologies in Flow Cytometry - It has always been a dream of mine to write an introduction, so I thought it worthwhile to organize a course of Flow Cytometry and so publish the relative book. Thus a dream come true.

Flow Cytometry Assay to Measure Internalization of GPCR's - The following protocol is optimized for internalization of FLAG-tagged beta 2-adrenergic receptor (B2AR) stably transfected into HEK293 cells.

Immunodetection of cyclin D1 and D2/D3 using 488/630 nm dual laser flow cytometry - This protocol is for use with the D cyclins and employs 488 nm argon laser excitation of propidium iodide and 630 nm NeNe or diode laser excitation of the fluorochrome Cy5 to detect cell cycle-specific cyclin D expression.

Propidium Iodide (PI) Staining of Dead Cells for Flow Cytometry  - Propidium iodide (PI) intercalates into double-stranded nucleic acids. It is excluded by viable cells but can penetrate cell membranes of dying or dead cells.

Measuring Apoptosis and Necrosis by Dual Laser Flow Cytometry - Apoptotic cells, due to a change in membrane permeability, show an increased up-take of the vital dye HO342 compared to live cells. PI or 7-AAD is added to discriminate late apoptotic or necrotic cells which have lost membrane integrity from early apoptotic cells which still have intact membranes by dye exclusion.

A Sensitive Method for Detection of Apoptosis by Single Laser Flow Cytometry - Apoptotic cells, probably due to a change in membrane permeability, take up some 7-AAD and become 7-AADdim compared to live cells which remain 7-AAD-. Late apoptotic or necrotic cells which have lost membrane integrity appear 7-AADbright.

Flow Cytometry Protocols - The following protocols are designed to give the flow cytometry core facility users at Princeton an idea of the types of protocols that are currently being used in the facility. A more complete and comprehensive collection of protocols can be found in "CURRENT PROTOCOLS IN CYTOMETRY" published by Wiley-Liss. An updated copy of this manual can be found in the facility.

Flow Cytometry Protocols -  This page contains protocols provided by the Telford Lab or by users of our facility.   Protocols are provided both as HTML pages and in Acrobat PDF file format; you will need Acrobat Reader to view the PDF files.

Flow Cytometry Protocols from NIEHS - Annexin V Protocols, Calcium Flux AnalysisFASC Analysis of Cell Cycle using BrdU and PI, CaspaTag Caspase Activity Protocol, DNA Analysis by Flow Cytometry, Membrane Potential Analysis by Flow Cytometry, PARP FITC, Sodium and Potassium Analysis by Flow Cytometry, Immunofluorescent Staining of Rat Thymocytes.