The Optimal Conditions of Chromosomal Analysis in Peripheral Blood.
- Author:
Hee Gyung SON
1
;
Young Il LEE
;
Byung Joon JUNG
;
Sang Hoon LEE
;
Soon Ja KANG
Author Information
1. Department of Obstetrics and Gynecology, College of Medicine, Chung-Ang University, Korea.
- Publication Type:Original Article
- Keywords:
Chromosomal analysis;
Optimal conditions
- MeSH:
Demecolcine;
Ethidium;
Humans;
Mitotic Index
- From:Korean Journal of Obstetrics and Gynecology
2001;44(1):5-16
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
OBJECTIVE: To find out the optimal conditions of human chromosomal analysis protocol in peripheral blood sample. METHODS: The experiments were made with the variations of phytohaemagglutinin, colcemid, ethidium bromide concentration and the variations of hypotonic solution exposure time. RESULTS: In the experiment on the optimal phytohaemagglutinin concentration, the highest mitotic index in the overall collected cells was obtained in phytohaemagglutinin concentration 15microL/ml. In the experiment on the concentration of mitotic arrestant colcemid, the proper chromosomal state that is meta phase stage and doesn't have many chromosomal crossings or tangles was obtained in colcemid concentration 0.05microg/ml. In the experiment on the optimal exposure time of hypotonic solution(0.075M KCl) treatment, the most suitable intervals between chromosomes were subtained in 20 minutes. In the experiment on the optimal concentration of ethidium bromide to obtain minute chromosomal bands, the best result was when ethidium bromide concentration 5microg/ml or 7.5microg/ml was addition to colcemid concentration 0.02microg/ml. CONCLUSION: The combination of phytohaemagglutinin 15microL/ml, colcemid 0.05microg/ml, hypotonic solution exposure time for 20 minutes is important to the collection of appropriate chromosome state in human chromosomal analysis using peripheral blood. In the case that needs to obtain minute bands, the elongated chromosomes are obtained when ethidium bromide 5microg/ml or 7.5microg/ml in addition to colcemid concentration 0.02microg/ml with the same conditions of phytohaemagglutinin and hypotonic solution.
