Analysis of DNA Content by Flow Cytometry (CAT#: STEM-CBT-0062-WXH)

Introduction

The measurement of DNA content of cells is one of the earliest and major application in flow cytometry and still is one that is extensively used today. DNA content analysis by flow cytometry can provide valuable information about the cell cycle status of individual cells.<br />DNA content measurement demands high precision because small variations in DNA content can have considerable biological significance.<br />Precise measurement of DNA content becomes possible thanks to the availability of nucleic acid specific fluorescent dyes and the availability of powerful flow cytometers capable of measuring with great precision fluorescence coming from inside cell nucleus.

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Principle Applications Procedure Materials Notes Related Products

Principle

To measure the DNA content, the cells have to be stained with a fluorescent dye that binds to DNA in a manner that reflects accurately the amount of DNA present. Fortunately, there are several suitable compounds available. These dyes are only weakly fluorescent in aqueous solution but fluoresce strongly when bound to DNA due to the hydrophobic nature of their environment.

Applications

• Study cell cycle.
• Monitor cell growth.
• Obtain information about their ploidy.
• Detect perturbations of cellular proliferation due to physical, chemical or biological assaults.
• Clinical applications: tumours diagnosis, characterize solid tumors, chemotherapy monitoring.

Procedure

1.Collect cells
2.Wash cells
3.Add staining solution and incubate
4.Examine by flow cytometry

Materials

DAPI(propidium iodide,ethidium bromide), PI (propidium iodide), DRAQ5, Hoechst 33342, BrdU, PIPES

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