How to measure cell proliferation?

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Accepted Answer

Cell proliferation can be determined by measuring such parameters as newly synthesized DNA, total nucleic acid content or cell division (Table 1).
The simplest method would be to monitor cell division using amine-reactive cell tracking indicators, such as CytoTell&reg; dyes. These cell-permeable indicators covalently bind to cytoplasmic proteins. Due to this covalent coupling reaction, CytoTell&reg; dyes cannot be transferred to adjacent cells and are well-retained in cells for several generations (up to 9 generation can be visualized). As cells divide, CytoTell&reg; dyes are distributed equally between daughters cells, and each new generation of cells is marked by a fluorescence intensity half that of its parents. Cells labeled with CytoTell&reg; dyes may be fixed and permeabilized using standard formaldehyde-containing fixatives and saponin-based permeabilization buffers for further intracellular analysis.

Table 1. List of Reagents and Assays for Monitoring Cell Proliferation.

Parameter
Principle
Reagent/Assay
Instrument

Monitor Cell Division

Uses cell-permeable dyes that bind to cytoplasmic proteins. As cells divide, dye is transferred to daughter cells. Each new generation of cells is marked by  fluorescence intensity half its parents.

CytoTell&reg; dyes

Fluorescence microplate reader, fluorescence microscope or flow cytometer

CytoTrace&trade; dyes

ReadiUse&trade; CFSE

Monitor Newly Synthesized DNA

Incorporates modified nucleotides into newly synthesized DNA during the S-phase of the cell cycle. Once incorporated, these nucleoside analogs serve as cell cycle and proliferation markers that can be detected using labeled probes to identify cells that are actively proliferating.

Bucculite&trade; dT Incorporation Cell Proliferation Fluorescence Imaging Kits

Fluorescence microscope

BrdU (requires an enzyme or fluorophore labeled anti-BrdU antibody)

Can be adapted for colorimetric, fluorimetric or chemiluminescent instruments.

Monitor Total Nucleic Acid Content

Uses cell-permeable nucleic acid stains that exhibit emission signals proportional to DNA mass. Flow cytometric analysis of stained populations is then used to generate a DNA histogram to reveal the percentage of cells in each phase of the cell cycle.

Cell Meter&trade; Fluorimetric Live Cell Cycle Assays

Flow cytometer

Cell Meter&trade; Fluorimetric Fixed Cell Cycle Assays

Nuclear Violet&trade; LCS 1

Hoechst 33258, Hoechst 33342 and DAPI

Parameter	Principle	Reagent/Assay	Instrument
Monitor Cell Division	Uses cell-permeable dyes that bind to cytoplasmic proteins. As cells divide, dye is transferred to daughter cells. Each new generation of cells is marked by fluorescence intensity half its parents.	CytoTell® dyes	Fluorescence microplate reader, fluorescence microscope or flow cytometer
		CytoTrace™ dyes
		ReadiUse™ CFSE
Monitor Newly Synthesized DNA	Incorporates modified nucleotides into newly synthesized DNA during the S-phase of the cell cycle. Once incorporated, these nucleoside analogs serve as cell cycle and proliferation markers that can be detected using labeled probes to identify cells that are actively proliferating.	Bucculite™ dT Incorporation Cell Proliferation Fluorescence Imaging Kits	Fluorescence microscope
		BrdU (requires an enzyme or fluorophore labeled anti-BrdU antibody)	Can be adapted for colorimetric, fluorimetric or chemiluminescent instruments.
Monitor Total Nucleic Acid Content	Uses cell-permeable nucleic acid stains that exhibit emission signals proportional to DNA mass. Flow cytometric analysis of stained populations is then used to generate a DNA histogram to reveal the percentage of cells in each phase of the cell cycle.	Cell Meter™ Fluorimetric Live Cell Cycle Assays	Flow cytometer
		Cell Meter™ Fluorimetric Fixed Cell Cycle Assays
		Nuclear Violet™ LCS 1
		Hoechst 33258, Hoechst 33342 and DAPI

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