三种Feulgen染色方法的比较

Comparison among three procedures of Feulgen staining

目的探索一种快速的Feulgen染色方法,并尝试其在细胞核DNA含量分析中的应用。方法选取十只健康小白鼠的肝细胞涂片,每只小白鼠的肝细胞涂片分成三组,采用快速Feulgen染色法、改良Feulgen染色法、传统Feulgen染色法对这三组分别染色,用TIGER细胞图像分析仪测量肝细胞涂片内单个完整肝细胞核的DNA含量,比较各组染色效果和肝细胞核DNA含量的测量结果。结果应用快速Feulgen染色法,获得了满意的效果,细胞核染色深,结构清晰,背景着色淡,而所需染色时间仅为15分钟;同一种方法染色的不同小鼠肝细胞涂片,相同DNA含量倍体肝细胞核的DNA含量均值的变异系数(CV)<10%,CV(快速)<CV(改良)<CV(传统);不同方法染色的同一小鼠肝细胞涂片,同一倍体的肝细胞核DNA含量存在差异,IOD(快速)>IOD(改良)>IOD(传统);各组2、4、8倍体肝细胞核DNA含量间的比值均接近2或4;结论此种快速Feulgen染色方法优于其他两种方法,可应用于图像分析系统对细胞核DNA含量与倍体的测量和分析。

Objective To develop a quick Feulgen staining and try its application in cell nuclei DNA content measurement. Methods Smears of liver cell suspension were obtained from ten healthy mice. The specimens were divided into three groups, stained with quick Feulgen staining, modified one and traditional one, respectively. DNA content of intact hepatocyte nuclei was analyzed by TIGER cell image analysis system. The results were compared among different groups. Results It took only 15 minutes to finish the quick Feulgen staining. Results with the quick Feulgen staining was satisfactory, that structure of cell nuclei was clear and were stained well, the CV for DNA content of hepatocyte nuclei with same DNA content ploidy was less than 10 percent among different mice hepatocyte nuclei stained using the same staining procedure, i.e. CV（quick） 〈 CV（modified） 〈 CV（traditional） , the nuclear DNA content of hepatocytes with the same DNA ploidy from the same mice stained by different staining procedures was different, i.e. AIOD（quick） 〉 AIOD〈（modified〉 AIOD〈（traditional） , the AIOD ratio among 2c, 4c and 8c in each group was close to 2 and 4. Conclusion The quick Feulgen staining is better then the others. It is applicable in image analysis system to measure and analyze the cell nuclei DNA content and its ploidy.