Nagashima R
Department of Neurosurgery, Saitama Medical School, Japan.
No To Shinkei. 1990 Apr;42(4):352-9.
Using DAPI-DNA cytofluorometry, the author analyzed nuclear DNA content of formalin fixed, paraffin embedded, glioma material obtained from 14 glioma cases at surgery. Sections of 10 microns were deparaffinized. Following simultaneous DAPI (4,6-diamidino-2-phenylindole dihydroporphyrin chloride)/HP (hematoporphyrin) staining, DAPI binds DNA and DNA-DAPI complexes emit blue fluorescence when exited by ultraviolet (UV) light. Through Zeiss fluorescence microscope, the author measured nuclear fluorescence intensity with histological verification of glioma cells. A DNA histogram was obtained with fluorescence intensity recorded on the abscissa and number of cells plotted on the ordinate. Samples of 20 normal non-neoplastic astrocytes taken from apparently normal brain tissue included in the histological slide were used as diploid (2 C) control. Based on DNA content, tumor cells were classified into 4 groups: N-group composed of cells with 2 C DNA content (normoploid), S-group with less than 2 C (hypoploid), L-group more than 4 C (hypertetraploid), I-group between 2 C and 4 C (intermediate ploidy). Intermediate ploidy was significantly higher and normoploid was significantly lower in glioblastoma compared with those of benign astrocytoma. Thus, DNA content and histological malignancy were well correlated. Due to limitation of measuring diaphragm of turret in the microscope, some extra large cell could not be included in it and was excluded from the measurement.(ABSTRACT TRUNCATED AT 250 WORDS)
作者使用DAPI-DNA细胞荧光测定法,分析了14例神经胶质瘤患者手术时获取的福尔马林固定、石蜡包埋的神经胶质瘤组织的核DNA含量。将10微米厚的切片进行脱蜡处理。在同时进行DAPI(4,6-二脒基-2-苯基吲哚二盐酸盐)/HP(血卟啉)染色后,DAPI与DNA结合,DNA-DAPI复合物在紫外线激发下发出蓝色荧光。通过蔡司荧光显微镜,作者测量了核荧光强度,并对神经胶质瘤细胞进行了组织学验证。获得了一个DNA直方图,横坐标记录荧光强度,纵坐标绘制细胞数量。从组织学切片中包含的明显正常脑组织中获取的20个正常非肿瘤性星形胶质细胞样本用作二倍体(2C)对照。根据DNA含量,肿瘤细胞分为4组:N组由DNA含量为2C的细胞组成(正常倍体),S组DNA含量小于2C(亚二倍体),L组大于4C(超四倍体),I组在2C和4C之间(中间倍体)。与良性星形细胞瘤相比,胶质母细胞瘤的中间倍体显著更高,正常倍体显著更低。因此,DNA含量与组织学恶性程度密切相关。由于显微镜转台测量光阑的限制,一些超大细胞无法包含在内,被排除在测量之外。(摘要截短于250字)