COMBINED PROTEIN AND DNA MEASUREMENTS BY THE NINHYDRIN-SCHIFF AND FEULGEN TECHNIQUES

被引:14
作者
FUKUDA, M
NAKANISHI, K
BOHM, N
KIMURA, J
HARADA, K
FUJITA, S
机构
[1] UNIV FREIBURG,INST PATHOL,D-7800 FREIBURG,FED REP GER
[2] KYOTO PREFECTURAL UNIV MED,DEPT OBSTET & GYNECOL,KYOTO 602,JAPAN
关键词
D O I
10.1007/BF00508010
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Feulgen nuclear staining with pararosanilin-SO2 was combined with the ninhydrin-Schiff technique. The aldehyde groups converted from primary amino groups are stained with an acriflavine-Schiff reaction. This results in a red nuclear fluorescence and a bright yellow cytoplasmic and nuclear fluorescence. The combined fluorescence staining facilitates cytofluorometric determination of total protein and DNA in the same cell. The ninhydrin-Schiff reaction is affected by the fixation procedure and the duration of the ninhydrin reaction. Investigations with a model system showed that proportionality beween the fluorescence intensity of acriflavine and the amount of protein stained by the procedure was obtained after fixation with a fixation mixture suggested by Böhm et al. (1968) and a reaction with ninhydrin at 37° C for 10 h. The ninhydrin-Schiff reaction has no effect on the fluorescence intensity of cells previously treated with pararosanilin-Feulgen staining and it is not affected itself by this previous procedure. Testing this double fluorescence staining on cytology specimens taken from patients with gastric carcinoma and uterine cervial carcinoma, cancer cells were shown to have markedly increased protein and DNA contents compared with those of normal cells. © 1979 Springer-Verlag.
引用
收藏
页码:35 / 45
页数:11
相关论文
共 29 条
[1]   A SELECTIVE STAINING METHOD FOR THE BASIC PROTEINS OF CELL NUCLEI [J].
ALFERT, M ;
GESCHWIND, II .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1953, 39 (10) :991-999
[2]   A MICROPHOTOMETRIC STUDY OF THE SYNTHESES OF DESOXYRIBONUCLEIC ACID AND NUCLEAR HISTONE [J].
BLOCH, DP ;
GODMAN, GC .
JOURNAL OF BIOPHYSICAL AND BIOCHEMICAL CYTOLOGY, 1955, 1 (01) :17-+
[3]  
BOHM N, 1968, HISTOCHEMISTRY, V16, P100
[4]   PROPORTIONALITY ERRORS DURING FEULGEN HYDROLYSIS [J].
BOHM, N ;
SPRENGER, E ;
SCHLUTER, G ;
SANDRITTER, W .
HISTOCHEMIE, 1968, 15 (03) :194-+
[5]  
BOHM N, 1970, ACTA HISTOCHEM, V35, P324
[6]  
BOHM N, 1972, TECHNIQUES BIOCHEMIC, V1, P90
[7]   NEW TYPE OF 2-COLOR FLUORESCENCE STAINING FOR CYTOLOGY SPECIMENS [J].
CORNELISSE, CJ ;
PLOEM, JS .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1976, 24 (01) :72-81
[8]   RAPID, SIMULTANEOUS MEASUREMENT OF DNA, PROTEIN, AND CELL VOLUME IN SINGLE CELLS FROM LARGE MAMMALIAN-CELL POPULATIONS [J].
CRISSMAN, HA ;
STEINKAMP, JA .
JOURNAL OF CELL BIOLOGY, 1973, 59 (03) :766-771
[9]   SYNTHETIC ACTIVITIES DURING SPERMATOGENESIS IN LOCUST [J].
DAS, NK ;
SIEGEL, EP ;
ALFERT, M .
JOURNAL OF CELL BIOLOGY, 1965, 25 (2P1) :387-&
[10]  
FRIEDMAN HP, 1950, AM J OBSTET GYNECOL, V59, P852