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Methods for improving the yield and quality of metaphase preparations for FISH probing of human lymphocyte chromosomes
Author(s) -
McFee A.F.,
Sayer A.M.,
Salomaa S.I.,
Lindholm C.,
Littlefield L.G.
Publication year - 1997
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/(sici)1098-2280(1997)29:1<98::aid-em13>3.0.co;2-c
Subject(s) - metaphase , mutagen , fish <actinopterygii> , biology , fluorescence in situ hybridization , mitosis , fluorescence microscope , lymphocyte , in vitro , centrifugation , microbiology and biotechnology , chromosome , fluorescence , genetics , biochemistry , dna , optics , gene , physics , fishery
Procedures are described for the in vitro culture of human lymphocytes, which have been concentrated by density gradient centrifugation, and for a modified slide‐making technique for the fixed cells. The method yields improved percentages of mitotic cells which are largely synchronized at harvest. Controlled placement of fixed cells on slides produces well‐spread metaphase preparations with little background material to interfere with fluorescence in situ hybridization (FISH) probe procedures. The FISH reagents and microscope scanning time required are minimized by concentrating cells in a defined area of the slide. Environ. Mol. Mutagen. 29:98–104, 1997 © 1997 Wiley‐Liss, Inc.