[Improvement of the silver-stain and it's test and verification in comet assay].
In order to find out a better silver-stain method so that to replace EB-stain in comet assay, the selected silver-stain methods were tested via determining fixing, coloring, displaying, fading, and testing the pigmentation temperature. When it was optimized, it was used in testing the DNA damage by two types of compounds, namely potassium dichromate (K2Cr2O7) and 1,2-dichloroethane (DCE) to two kind of cells, namely spleen cells of mouse and human outer lymphocyte, and, the sensitivity was compared with that of EB-stain. The results showed that the modified silver-stain was the first successful non-fluorescent dye method in comet assay that can dye clearly, permanently, with a light background and good contrast, and it was easy to manipulate and it was cheap. The stain can detect the DNA damage by compounds to cells and has the same positive threshold like that in EB-stain. It can even dye the small fragments that EB-stain cannot. It is suggested that in comet assay, the silver-stain can substitute the EB-stain completely.
Zhao
Rong
R
Department of Environmental Health, Huaxi College of Public Health, Sichuan University, Chengdu 610041, China.
Heng
Zhengchang
Z
D020552:Comet Assay / Q000379:methods; D004249:DNA Damage; D005025:Ethylene Dichlorides / Q000633:toxicity; D008214:Lymphocytes / Q000166:cytology; D011192:Potassium Dichromate / Q000633:toxicity; D012680:Sensitivity and Specificity; D016622:Silver Staining; D013154:Spleen / Q000166:cytology
D005025:Ethylene Dichlorides; C024565:ethylene dichloride; D011192:Potassium Dichromate