0:00
Overview
0:56
Principle Behind Cell Cycle Analysis
3:33
Generalized Protocol for BrdU and PI Staining
5:43
Flow Cytometric Analysis
7:26
Applications
9:07
Summary
Cell cycle refers to the set of events through which a cell grows, replicates its genome, and ultimately divides into two daughter cells through the process of mitosis. Because the amount of DNA in a cell shows characteristic changes throughout the cycle, techniques known as cell cycle analysis can be used to separate a population of cells according to the different phases of cell cycle they are in, based on their varying DNA content.
This video will cover the principles behind cell cycle analysis via DNA-staining. We will review a generalized protocol for performing this staining using bromodeoxyuridine (BrdU, a thymidine analog that is incorporated into newly synthesized DNA strands) and propidium iodide (PI, a DNA dye that stains all DNA), followed by analysis of the stained cells with flow cytometry. During flow cytometry, a single cell suspension of fluorescently labeled cells is passed through an instrument with a laser beam and the fluorescence of each cell is read. We will then discuss how to interpret data from flow cytometric scatter plots, and finally, look at a few applications of this technique.
Cell cycle refers to the set of cellular and biochemical processes occurring within a cell leading to its division. A cell passes through different stages of the cycle, and the time taken to complete this process is almost constant for a cell type. Any deviation from this duration is indicative of abnormal cell division. Therefore, cell cycle analysis becomes a very useful tool for biologists interested in studying mechanisms of cell division, which will ultimately help in treating patients with diseases like cancer, in wh
Explore More Videos
ABOUT JoVE
Copyright © 2024 MyJoVE Corporation. All rights reserved