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采用流式细胞术和图像细胞术对新鲜实体瘤进行DNA定量分析:使用罗氏病理工作站图像分析仪的比较研究

Quantitative DNA analysis of fresh solid tumors by flow and image cytometric methods: a comparison using the Roche Pathology Workstation Image Analyzer.

作者信息

Ellison D A, Maygarden S J, Novotny D B

机构信息

Department of Pathology, University of North Carolina, Chapel Hill, USA.

出版信息

Mod Pathol. 1995 Apr;8(3):275-81.

PMID:7617654
Abstract

The clinical utility of DNA ploidy and cell cycle parameters as prognostic indicators has been demonstrated for selected malignant tumors. Previous quantitative DNA analysis studies have used various tumor sample preparation methods and analyzers. We undertook a pilot study to compare the results of DNA analysis of fresh solid tumors by flow cytometry with the new Roche Pathology Workstation Image Analyzer. Flow cytometric DNA analysis was done on cell suspensions of fine needle aspirates from fresh tumor specimens and analyzed for ploidy and cell cycle statistics with a Becton-Dickinson FACScan Analyzer, using a rectangular model. Small aliquots from these same aspirates were prepared as direct cytologic smears and Feulgen stained for DNA analysis with the Roche Image Analyzer. Additional smears were stained with Diff-Quik for morphologic correlation with DNA histograms. The study group consisted of 40 malignant neoplasms. There was a high correlation between the flow and image DNA indices (R = 0.93, slope = 1.0036, P < 0.001) but a weaker relationship between the flow and image estimated S-phase fractions (R = 0.57, slope = 0.5401, P < 0.01). DNA ploidy categorization for the two methods was concordant in 30 (75%) cases, discordant in seven (17.5%) cases, and equivocal in three (7.5%) cases. In our experience, quantitative DNA analysis of fresh tumor aspirates by flow and image cytometric methods yielded comparable and/or complementary results, with each method having certain advantages and disadvantages. Proposed reasons for false and true discordances and an approach for evaluation are discussed.

摘要

DNA倍体和细胞周期参数作为预后指标的临床实用性已在特定恶性肿瘤中得到证实。以往的定量DNA分析研究使用了各种肿瘤样本制备方法和分析仪。我们进行了一项初步研究,以比较通过流式细胞术对新鲜实体瘤进行DNA分析的结果与新型罗氏病理工作站图像分析仪的结果。对来自新鲜肿瘤标本的细针穿刺吸取物的细胞悬液进行流式细胞术DNA分析,并用Becton-Dickinson FACScan分析仪使用矩形模型分析倍体和细胞周期统计数据。从这些相同的吸取物中取出少量样本制成直接细胞学涂片,并用福尔根染色,以使用罗氏图像分析仪进行DNA分析。另外的涂片用Diff-Quik染色,以便与DNA直方图进行形态学对照。研究组由40例恶性肿瘤组成。流式和图像DNA指数之间存在高度相关性(R = 0.93,斜率 = 1.0036,P < 0.001),但流式和图像估计的S期分数之间的关系较弱(R = 0.57,斜率 = 0.5401,P < 0.01)。两种方法的DNA倍体分类在(75%)30例中一致,在(17.5%)7例中不一致,在(7.5%)3例中不明确。根据我们的经验,通过流式和图像细胞术方法对新鲜肿瘤吸取物进行定量DNA分析可产生可比和/或互补的结果,每种方法都有一定的优缺点。讨论了假不一致和真不一致的可能原因以及一种评估方法。

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