Platinum porphyrins as phosphorescent label for time-resolved microscopy

被引：55

作者：

deHaas, RR ^{[1
]}

vanGijlswijk, RPM ^{[1
]}

vanderTol, EB ^{[1
]}

Zijlmans, HJMAA ^{[1
]}

BakkerSchut, T ^{[1
]}

Bonnet, J ^{[1
]}

Verwoerd, NP ^{[1
]}

Tanke, HJ ^{[1
]}

机构：

[1] UNIV AMSTERDAM,DEPT ORGAN CHEM,AMSTERDAM,NETHERLANDS

来源：

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY | 1997年 / 45卷 / 09期

关键词：

metalloporphyrin conjugates; time-resolved luminescence microscopy; FISH; autofluorescence; bleaching; tyramide amplification system; phosphorescence;

D O I：

10.1177/002215549704500911

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

We investigated phosphorescent metalloporphyrins as potential labels for time-resolved microscopy. On the basis of spectroscopic analysis of their physicochemical properties (quantum yield, molar absorption coefficient, decay times) the best candidates were selected. Next, we synthesized antibody and avidin metalloporphyrin conjugates. The optimal F/P ratio with respect to quantum yield, decay time, and retention of biological activity of these immunoreagents was determined. The reagents were then evaluated by in situ hybridization and immunocytochemical procedures for demonstration of hapten-labeled DNA probes, membrane antigens (CD type), and 28S rRNA. All stained samples exhibited bright phosphorescence that could be selectively detected using time-resolved microscopy, especially when glucose/glucose oxidase was added to the embedding medium to deplete oxygen. Applications of time-resolved detection of phosphorescent porphyrins in strongly autofluorescent material (histological sections) are discussed.

引用

页码：1279 / 1292

页数：14

共 33 条

[1] BIOTIN AMPLIFICATION OF BIOTIN AND HORSERADISH-PEROXIDASE SIGNALS IN HISTOCHEMICAL STAINS [J].