Exploration of an improved technique for the preparation of chromosomal specimens from peripheral blood lymphocytes
10.13491/j.issn.1004-714X.2025.05.012
- VernacularTitle:外周血淋巴细胞染色体标本制备改良技术探索
- Author:
Donglin CHEN
1
;
Lina WU
2
;
Xin LIU
3
;
Hongliang GAO
3
;
Yuyang MA
3
;
Caihong DUO
3
;
Jingguang FAN
2
Author Information
1. School of Public Health, Guangdong Pharmaceutical University, Guangzhou 510240, China.
2. School of Public Health, Guangdong Pharmaceutical University, Guangzhou 510240, China;National Center for Occupational Safety and Health, National Health Commission of the People’s Republic of China, Beijing 102308, China.
3. National Center for Occupational Safety and Health, National Health Commission of the People’s Republic of China, Beijing 102308, China.
- Publication Type:OriginalArticles
- Keywords:
Anhydrous ethanol;
Methanol;
Chromosome specimen preparation;
Dispersion index;
Overlapping ratio
- From:
Chinese Journal of Radiological Health
2025;34(5):695-701
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the application value of anhydrous ethanol as an alternative to methanol in the preparation of chromosomal specimens from peripheral blood lymphocytes, and to establish a set of quantitative analytical methods for objectively evaluating the effectiveness of specimen preparation. Methods Residual blood samples from routine laboratory slide preparation were used for lymphocyte culture. The standard slide preparation method was employed. The fixative in the control group was methanol and glacial acetic acid (3∶1). Four experimental groups were set up based on the ratio of anhydrous ethanol to glacial acetic acid in the fixative (volume ratios of 3∶1, 5∶1, 7∶1, and 9∶1 for experimental groups 1, 2, 3, and 4, respectively). A chromosomal analysis was conducted using an automated chromosome scanning/image analysis system to evaluate the morphology and dispersion of metaphase chromosomes in both control and experimental groups. Comparisons were made between the control and experimental groups regarding the dic + r aberration rate, ace aberration rate, chromosomal aberration rate, chromosome dispersion index, chromosome overlapping ratio, and dispersion index/overlapping ratio. Results Microscopic evaluation revealed that the preparation quality of experimental groups 1 and 2 was comparable to the control group. No statistically significant differences were observed in dic + r aberration rate between each of the experimental groups and the control (P > 0.05). All experimental groups except group 4 showed no significant differences in ace aberration rate and chromosome aberration rate compared with the control group (P > 0.05). Experimental groups 1 and 2 showed no significant differences in chromosome dispersion index, overlapping ratio, and dispersion index/overlapping ratio compared with the control group (P > 0.05). Conclusion A mixture of anhydrous ethanol and glacial acetic acid at a 5∶1 ratio is recommended for use as a fixative in the preparation of chromosomal specimens from peripheral blood lymphocytes. A quantitative index system for assessing the quality of chromosomal specimens was established, enabling objective evaluation of slide preparation effectiveness.
