Application and comparison of two kinds of staining methods for measurement of nuclear DNA contents

Objective To find out the suitable hydrolysis time for two staining methods of Feulgen and to provide guidline on research and clinic.Methods Smears of liver cell suspension were obtained from five healthy rats.All the hepatocytes are,respectively,hydrolyzed in different time at room temperature and 60℃,and then stained with modified Feulgen stain and quick one.DNA content and ploid of intact hepatocyte nuclei was analyzed by TIGER cell image analysis system.Results(1) the nuclear DNA contents are almost identical from same DNA content ploidy of different rats under the same temperature and time of hydrolysis,and the CV of IOD from three types of DNA content ploidy was less than 10 percent.(2) The IOD ratio among diploid,tetraploid and octaploid was close to 2 and 4.(3) The DNA contents of hepatocyte nuclei from same DNA content ploidy of different rats were distinct under the same temperature and different hydrolysis time: hydrolysis at 60℃,IOD(5~7 min)IOD(9~15 min)IOD(1 min,20 min);hydrolysis at room temperature,IOD(50 min)IOD(20~30 min,70~90 min)IOD(5~10 min,100 min).Conclusion nuclei DNA can't be stained better for extra long or short hydrolysis time,the better hydrolysis time of quick Feulgen stain and modified one are,respectively,5~7 min and 20~90 min.