Format

Send to

Choose Destination
Cytometry. 2001 Feb 1;43(2):101-9.

Improvements in cytogenetic slide preparation: controlled chromosome spreading, chemical aging and gradual denaturing.

Author information

1
Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510, USA. henegariu@yahoo.com

Abstract

BACKGROUND:

Metaphase spreading is an essential technique for clinical and molecular cytogenetics. Results of classical banding techniques as well as complex fluorescent in situ hybridization (FISH) applications, such as comparative genomic hybridization (CGH) or multiplex FISH (M-FISH), are greatly influenced by the quality of chromosome spreading and pretreatment of the slide prior to hybridization. Materials and Methods Using hot steam and a metal plate with a temperature gradient across its surface, a reproducible protocol for slide preparation, aging, and hybridization was developed.

RESULTS:

This protocol yields good chromosome spreads from even the most difficult cell suspensions and is unaffected by the environmental conditions. Chromosome spreads were suitable for both banding and FISH techniques common to the cytogenetic laboratory. Chemical aging is a rapid slide pretreatment procedure for FISH applications, which allows freshly prepared cytogenetic slides to be used for in situ hybridization within 30 min, thus increasing analytical throughput and reducing benchwork. Furthermore, the gradually denaturing process described allows the use of fresh biologic material with optimal FISH results while protecting chromosomal integrity during denaturing.

CONCLUSION:

The slide preparation and slide pretreatment protocols can be performed in any laboratory, do not require specialized equipment, and provide robust results.

PMID:
11169574
[Indexed for MEDLINE]

Supplemental Content

Loading ...
Support Center