Application Of Image Analysis System Used To Measure DNA Content And Ploidy Of Nuclei

Purposes:Part One: To find out current situation of the image analysis system used to measure thecontent of chemical components in China. Part Two: To find out the pattern of DNA ploidy changes of hepatocytes of developing mice and thesome methodological problem.Part Three; To estimate in situ heterogeneity of DNA stemline ploidy of non-small cell lung carcinomas (NSLCs) with image analysis system and to analyze the relations of them with different pathological type, clinical stage.Methods.-Part One: Every two smears of normal mice liver was send to eight laboratories. One smear had been stained with Feulgen and another smear are required stain with Feulgen in each laboratory. DNA content of hepatocyte nuclei on the two slides was measured by each laboratory and the results were returned to us. Histograms of the results from the eight laboratories were compared each other.Part Two: Nuclear DNA contents on the smear of mice liver in different stages were measured with TIGER image analyzer to calculate the percentage of hepatocyte nuclei with various DNA content ploidy.Part Three: 41 archival cases of non-small cell lung carcinomas (NSLCs) were taken, 4um and Sum consecutive tissue section were separately cut on each tumor tissue. Three different carcinomas net or glandular cavity in each tumor tissue section were sampled respectively. TIGER image analyzer measured DNA content in a unit of volume of single intact cell nuclei sampled in each carcinomas net or glandular cavity. The lymophacyte nuclei on the same section were estimated as intra-control for that case, so that TIGER image analyzer can value the DNA stemline ploidy of each carcinomas net or glandular cavity and then DNA stemline ploidy heterogeneity of every case can be analyzed .3Result:Part One: (l)The terms used to express DNA content are lack of united criteria among differentimage analysis systems in China; (2)The accuracy of DNA content of hepatocyte nuclei measured with image analysissystems is largely different;(3) Feulgen staining conditions are different among the eight laboratories. Fortunately, different staining protocols didn't influent obviously the histograms of DNA content that was measured by the same ICM.Part Two: (DThe DNA contents from hepatocyte nuclei of the same DNA content ploidy arealmost identical; (2)The IOD ratio among 2c, 4c, 8c and 16c was close to 2 and 4, and the CV of IODfrom four types of DNA content ploidy is less than 10 percent; (3)The most of hepatocyte nuclei after birth 2-20d mice is 2c (at least 90 percent); the proportion of the 4c nuclei of thirty-day-old mice exceed 2c nuclei; the hepatocyte nuclei of 35d group contain four types of DNA content ploidy, a descending order of frequencies in various DNA content ploidy can be illustrated in the following manner: 4O8O2O16C? the 32c nuclei appear in 60d mice.Part Three: (DDNA stemline ploidy in 123 samples of NSCLs clustered mainly in 0.78-3.18. 20 samples (16.3%) were DNA stemline periteraploidy, and only 32 (26.0%) were DNA stemline diploid;(2)Heterogeneity of DNA stemline ploidy was found in various pathological type of NSCLs, and the percent of heterogeneity was 73.2% (30/41). (3)There was significant difference of the percent of DNA stemline ploidy and heterogeneity of NSCLs in various clinical stage, but it was no statistically difference of the percent of DNA stemline ploidy and heterogeneity of NSCLs in various pathological type.Conclusions:Part One: (l)In view of performance of national image analysis systems is different largely, it is time to standardize the technical characteristics and evaluating methods ofimage analysis systems that will be used to measure the optical density and lightintensity of cells and tissue; (2) The producer of image analysis systems must grasp the technical andengineering characteristics of the ICM instrumentation. (3)User must align and calibrate strictly the image analysis system to satisfy therequirement of the photometric and mophometric pa...