Kong J, Ringer D P
Oklahoma Medical Research Foundation, Noble Center for Biomedical Research, Oklahoma City 73104, USA.
Cytometry. 1995 May 1;20(1):86-93. doi: 10.1002/cyto.990200113.
Scanning laser cytometric analysis of fluorochrome-labeled cells was used to survey and quantitate the distribution of proliferating cells in formalin-fixed, paraffin-embedded tissue sections from hyperplastic and neoplastic rat livers. The technique used fluorescent immunochemical staining of proliferating cell nuclear antigen (PCNA) as a marker for proliferating cells and propidium iodide as a fluorescent nuclear counterstain. Use of an antigen retrieval treatment improved detection of PCNA and treatment of tissue sections with crystal violet improved the sensitivity of the method by quenching background autofluorescence. PCNA evaluation of cell proliferation in regenerating rat liver 0-48 h post-partial hepatectomy showed that 3-43% of cells stained positively for PCNA, a pattern closely correlating with previously reported rates of maximum DNA synthesis. The PCNA staining patterns observed among cells in neoplastic nodules were more focal in distribution and indicated that from 5 to 25% of the cell nuclei per nodular region stained positively for PCNA. This use of image analysis for the rapid identification of proliferating cell areas in fixed, paraffin-embedded tissue active in neoplastic growth will expedite in situ cytochemical and molecular studies attempting to identify key differences between hyperplastic and neoplastic growth.
利用扫描激光细胞仪对荧光染料标记的细胞进行分析，以检测和定量来自增生性和肿瘤性大鼠肝脏的福尔马林固定、石蜡包埋组织切片中增殖细胞的分布。该技术使用增殖细胞核抗原（PCNA）的荧光免疫化学染色作为增殖细胞的标志物，碘化丙啶作为荧光核复染剂。采用抗原修复处理可改善PCNA的检测，用结晶紫处理组织切片可通过淬灭背景自发荧光提高该方法的灵敏度。对部分肝切除术后0 - 48小时的再生大鼠肝脏细胞增殖进行PCNA评估，结果显示3 - 43%的细胞PCNA染色呈阳性，这一模式与先前报道的最大DNA合成率密切相关。在肿瘤结节细胞中观察到的PCNA染色模式在分布上更具局灶性，表明每个结节区域有5%至25%的细胞核PCNA染色呈阳性。这种利用图像分析快速识别固定石蜡包埋组织中参与肿瘤生长的增殖细胞区域的方法，将加快原位细胞化学和分子研究，以试图确定增生性生长和肿瘤性生长之间的关键差异。
