Flow cytometric analysis of bromodeoxyuridine substituted cells stained with 33258 Hoechst

S. A. Latt, Y. S. George, Joe Gray

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

This paper describes a flow-cytometric application of the quenching of fluorescence from 33258 Hoechst stained Chinese hamster ovary-line cells due to the incorporation of 5-bromodeoxyuridine (BrdU) into the cellular deoxyribonucleic acid. Cells were grown for 24 hr in medium containing BrdU in concentrations ranging from 1 x 10-8 to 1 x 10-4 M. For each concentration we measured the average fluorescence as determined by flow cytometry, the extent of BrdU substitution and the effect of the BrdU on cell growth. We determined that a BrdU concentration of 1 x 10-5 M resulted in sufficient substitution to quench the fluorescence from 33258 Hoechst by a factor of 4, allowing discrimination between cycling and noncycling cells. The extent of BrdU substitution after growth for 24 hr in this concentration of BrdU was 64%. These data indicate the feasibility of detecting deoxyribonucleic acid synthesis in whole cells using the 33258 Hoechst-BrdU methodology.

Original languageEnglish (US)
Pages (from-to)927-934
Number of pages8
JournalJournal of Histochemistry and Cytochemistry
Volume25
Issue number7
StatePublished - 1977
Externally publishedYes

Fingerprint

Bisbenzimidazole
Bromodeoxyuridine
Fluorescence
DNA
Growth
Cricetulus
Ovary
Flow Cytometry

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

Flow cytometric analysis of bromodeoxyuridine substituted cells stained with 33258 Hoechst. / Latt, S. A.; George, Y. S.; Gray, Joe.

In: Journal of Histochemistry and Cytochemistry, Vol. 25, No. 7, 1977, p. 927-934.

Research output: Contribution to journalArticle

@article{c721a08e132a4791858b78064c048eff,
title = "Flow cytometric analysis of bromodeoxyuridine substituted cells stained with 33258 Hoechst",
abstract = "This paper describes a flow-cytometric application of the quenching of fluorescence from 33258 Hoechst stained Chinese hamster ovary-line cells due to the incorporation of 5-bromodeoxyuridine (BrdU) into the cellular deoxyribonucleic acid. Cells were grown for 24 hr in medium containing BrdU in concentrations ranging from 1 x 10-8 to 1 x 10-4 M. For each concentration we measured the average fluorescence as determined by flow cytometry, the extent of BrdU substitution and the effect of the BrdU on cell growth. We determined that a BrdU concentration of 1 x 10-5 M resulted in sufficient substitution to quench the fluorescence from 33258 Hoechst by a factor of 4, allowing discrimination between cycling and noncycling cells. The extent of BrdU substitution after growth for 24 hr in this concentration of BrdU was 64{\%}. These data indicate the feasibility of detecting deoxyribonucleic acid synthesis in whole cells using the 33258 Hoechst-BrdU methodology.",
author = "Latt, {S. A.} and George, {Y. S.} and Joe Gray",
year = "1977",
language = "English (US)",
volume = "25",
pages = "927--934",
journal = "Journal of Histochemistry and Cytochemistry",
issn = "0022-1554",
publisher = "Histochemical Society Inc.",
number = "7",

}

TY - JOUR

T1 - Flow cytometric analysis of bromodeoxyuridine substituted cells stained with 33258 Hoechst

AU - Latt, S. A.

AU - George, Y. S.

AU - Gray, Joe

PY - 1977

Y1 - 1977

N2 - This paper describes a flow-cytometric application of the quenching of fluorescence from 33258 Hoechst stained Chinese hamster ovary-line cells due to the incorporation of 5-bromodeoxyuridine (BrdU) into the cellular deoxyribonucleic acid. Cells were grown for 24 hr in medium containing BrdU in concentrations ranging from 1 x 10-8 to 1 x 10-4 M. For each concentration we measured the average fluorescence as determined by flow cytometry, the extent of BrdU substitution and the effect of the BrdU on cell growth. We determined that a BrdU concentration of 1 x 10-5 M resulted in sufficient substitution to quench the fluorescence from 33258 Hoechst by a factor of 4, allowing discrimination between cycling and noncycling cells. The extent of BrdU substitution after growth for 24 hr in this concentration of BrdU was 64%. These data indicate the feasibility of detecting deoxyribonucleic acid synthesis in whole cells using the 33258 Hoechst-BrdU methodology.

AB - This paper describes a flow-cytometric application of the quenching of fluorescence from 33258 Hoechst stained Chinese hamster ovary-line cells due to the incorporation of 5-bromodeoxyuridine (BrdU) into the cellular deoxyribonucleic acid. Cells were grown for 24 hr in medium containing BrdU in concentrations ranging from 1 x 10-8 to 1 x 10-4 M. For each concentration we measured the average fluorescence as determined by flow cytometry, the extent of BrdU substitution and the effect of the BrdU on cell growth. We determined that a BrdU concentration of 1 x 10-5 M resulted in sufficient substitution to quench the fluorescence from 33258 Hoechst by a factor of 4, allowing discrimination between cycling and noncycling cells. The extent of BrdU substitution after growth for 24 hr in this concentration of BrdU was 64%. These data indicate the feasibility of detecting deoxyribonucleic acid synthesis in whole cells using the 33258 Hoechst-BrdU methodology.

UR - http://www.scopus.com/inward/record.url?scp=0017711979&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0017711979&partnerID=8YFLogxK

M3 - Article

C2 - 70460

AN - SCOPUS:0017711979

VL - 25

SP - 927

EP - 934

JO - Journal of Histochemistry and Cytochemistry

JF - Journal of Histochemistry and Cytochemistry

SN - 0022-1554

IS - 7

ER -