High-resolution measurement of breaks in prematurely condensed chromosomes by differential staining

Michael Cornforth, Joel S. Bedford

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

A relatively simple method has been developed to improve the resolution for measuring breaks produced in interphase chromosomes by X rays or other agents following the induction of premature chromosome condensation (PCC). Mitotic HeLa cells, which induce PCC when fused with interphase cells, were obtained from cultures grown for several generations in 5-bromodeoxyuridine (BrdU). These were fused to cells from low-passage confluent cultures of normal human fibroblasts and subsequently stained by a modified fluorescence-plus-Giemsa (FPG) technique. Following this protocol the prematurely condensed chromosomes stain intensely, whereas the mitotic chromosomes of the inducer cell(s), which are intermingled with them, stain very lightly. With this technique the interphase chromosomes and their fragments can be identified unequivocally, making scoring much easier and more accurate. The frequency of breaks produced in G1 phase AG1522 human fibroblasts immediately following X-ray doses of 58 and 117 rad was 3.68 and 7.38 per cell, respectively. Use of this technique should allow the detection of damage from ionizing radiation at doses lower than 10 rad.

Original languageEnglish (US)
Pages (from-to)315-318
Number of pages4
JournalChromosoma
Volume88
Issue number4
DOIs
StatePublished - Oct 1983
Externally publishedYes

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Chromosomes
Staining and Labeling
Interphase
Coloring Agents
Fibroblasts
X-Rays
G1 Phase
Bromodeoxyuridine
Helper-Inducer T-Lymphocytes
Ionizing Radiation
HeLa Cells
Fluorescence

ASJC Scopus subject areas

  • Genetics

Cite this

High-resolution measurement of breaks in prematurely condensed chromosomes by differential staining. / Cornforth, Michael; Bedford, Joel S.

In: Chromosoma, Vol. 88, No. 4, 10.1983, p. 315-318.

Research output: Contribution to journalArticle

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