Stabilization of chromosomes by DNA intercalators for flow karyotyping and identification by banding of isolated chromosomes

J. A. Aten, C. H C M Buys, A. Y. van der Veen, J. R. Mesa, L. C. Yu, Joe Gray, J. Osinga, J. Stap

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A number of structurally unrelated DNA intercalators have been studied as stabilizers of mitotic chromosomes during isolation from rodent and human metaphase cells. Seven out of the nine intercalators tested were found to be useful as chromosome stabilizing agents. Chromosome suspensions prepared in this way could be preserved for long periods of time. After isolation the chromosomal DNA was longer than 150 kb. With intercalated chromosomes high resolution flow karyotypes could be obtained as jllustrated for the non-fluorescent intercalators 9-methylene-(1,3-dimethyl-2,4-dionepyrimidine-5-yl)-phenanthridiniumchloride and 4′-aminomethyl-4,5′, 8-trimethylpsoralen combined with DAPI and 33258 Hoechst for fluorescent staining and for the fluorescent intercalator propidium iodide used as a stabilizer and as a fluorochrome. Passage of the intercalated chromosomes through the laser beam had no measureble effect on the length of the chromosomal DNA subsequently isolated. After flow analysis and collection on slides human chromosomes could easily be banded by Giemsa staining methods with the same resolution as obtained in conventional metaphase spreads. This allowed a ready indentification of about 80 percent of all chromosomes in the unfractionated suspension collected after passage through the laser beam.

Original languageEnglish (US)
Pages (from-to)359-366
Number of pages8
JournalHistochemistry
Volume87
Issue number4
DOIs
StatePublished - Jul 1987
Externally publishedYes

Fingerprint

Chromosome Banding
Intercalating Agents
Karyotyping
chromosome banding
karyotyping
Chromosomes
chromosomes
DNA
Metaphase
metaphase
Suspensions
Lasers
Trioxsalen
lasers
Staining and Labeling
Bisbenzimidazole
Propidium
Excipients
Human Chromosomes
Karyotype

ASJC Scopus subject areas

  • Anatomy

Cite this

Stabilization of chromosomes by DNA intercalators for flow karyotyping and identification by banding of isolated chromosomes. / Aten, J. A.; Buys, C. H C M; van der Veen, A. Y.; Mesa, J. R.; Yu, L. C.; Gray, Joe; Osinga, J.; Stap, J.

In: Histochemistry, Vol. 87, No. 4, 07.1987, p. 359-366.

Research output: Contribution to journalArticle

Aten, J. A. ; Buys, C. H C M ; van der Veen, A. Y. ; Mesa, J. R. ; Yu, L. C. ; Gray, Joe ; Osinga, J. ; Stap, J. / Stabilization of chromosomes by DNA intercalators for flow karyotyping and identification by banding of isolated chromosomes. In: Histochemistry. 1987 ; Vol. 87, No. 4. pp. 359-366.
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