DAPI-DNA cytofluorometric study of glioma cells--application of DAPI-DNA cytofluorometry to paraffin embedded archival glioma tissue for nuclear DNA content analysis

• Published in: No To Shinkei Vol. 42; no. 4; p. 352
• Main Author: Nagashima, R
• Format: Journal Article
• Language: English; Japanese
• Published: Japan 01.04.1990
• Subjects: Adult; Aged; Brain Neoplasms - analysis; Brain Neoplasms - genetics; Brain Neoplasms - pathology; Cell Nucleus - analysis; Child, Preschool; Cytophotometry; DNA, Neoplasm - analysis; Female; Fluorescent Dyes; Formaldehyde; Glioma - analysis; Glioma - genetics; Glioma - pathology; Hematoporphyrins; Humans; Indoles; Male; Middle Aged; Paraffin; Ploidies; Staining and Labeling - methods
• ISSN: 0006-8969

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)