Assessment of cell proliferation by 5-bromodeoxyuridine (BrdU) labeling for multicolor flow cytometry.

Kristina Rothaeusler, Nicole Baumgarth

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Cell proliferation assays are used for a large variety of applications in the life sciences. This unit describes a flow-cytometry-based method that uses BrdU labeling in conjunction with multiple fluorescently labeled cell surface markers, allowing extensive phenotypic characterization of dividing cells in addition to assessment of their frequency. BrdU labeling has the advantage of constituting a nonradioactive technique that, when combined with additional fluorescent-based reagents, avoids time-consuming and often costly cell isolation procedures. Moreover, because BrdU is stably integrated into the DNA, it can be measured in a cell for many months. The method presented in this unit is based on paraformaldehyde fixation and reversible saponin-based cell membrane permeabilization, which maintains cell integrity as well as fluorescent labeling with a large number of different fluorochromes, allowing 10- to 12-color flow cytometric analysis of proliferating cells.

Original languageEnglish (US)
JournalCurrent protocols in cytometry / editorial board, J. Paul Robinson, managing editor ... [et al.]
VolumeChapter 7
StatePublished - Apr 2007

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Flow cytometry
Cell proliferation
Bromodeoxyuridine
Labeling
Flow Cytometry
Cell Proliferation
Saponins
Cell membranes
Fluorescent Dyes
Assays
Biological Science Disciplines
Cell Separation
Color
DNA
Cell Membrane

ASJC Scopus subject areas

  • Medicine(all)

Cite this

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abstract = "Cell proliferation assays are used for a large variety of applications in the life sciences. This unit describes a flow-cytometry-based method that uses BrdU labeling in conjunction with multiple fluorescently labeled cell surface markers, allowing extensive phenotypic characterization of dividing cells in addition to assessment of their frequency. BrdU labeling has the advantage of constituting a nonradioactive technique that, when combined with additional fluorescent-based reagents, avoids time-consuming and often costly cell isolation procedures. Moreover, because BrdU is stably integrated into the DNA, it can be measured in a cell for many months. The method presented in this unit is based on paraformaldehyde fixation and reversible saponin-based cell membrane permeabilization, which maintains cell integrity as well as fluorescent labeling with a large number of different fluorochromes, allowing 10- to 12-color flow cytometric analysis of proliferating cells.",
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