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Titolo:
DEMONSTRATION OF DESMOSOMAL ANTIGENS BY ELECTRON-MICROSCOPY USING CRYOFIXED AND CRYOSUBSTITUTED SKIN WITH SILVER-ENHANCED GOLD PROBE
Autore:
SHIMIZU H; MASUNAGA T; ISHIKO A; HASHIMOTO T; GARROD DR; SHIDA H; NISHIKAWA T;
Indirizzi:
KEIO UNIV,SCH MED,DEPT DERMATOL,SHINJUKU KU,35 SHINANOMACHI TOKYO 160JAPAN UNIV MANCHESTER,DEPT CELL & STRUCT BIOL MANCHESTER ENGLAND UNIV YAMANASHI,SCH MED,DEPT BIOL YAMANASHI JAPAN
Titolo Testata:
The Journal of histochemistry and cytochemistry
fascicolo: 5, volume: 42, anno: 1994,
pagine: 687 - 692
SICI:
0022-1554(1994)42:5<687:DODABE>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
CELL-ADHESION MOLECULES; PEMPHIGUS-FOLIACEUS; GLYCOPROTEIN-3 DESMOCOLLINS; IMMUNOELECTRON MICROSCOPY; BIOCHEMICAL-IDENTIFICATION; EPIDERMAL DESMOSOMES; MONOCLONAL-ANTIBODY; EPITHELIAL-CELLS; PLAQUE PROTEIN; DESMOPLAKIN-II;
Keywords:
DESMOSOMAL ANTIGENS; IMMUNOELECTRON MICROSCOPY; SKIN CRYOFIXATION; CRYOSUBSTITUTION; SILVER ENHANCEMENT;
Tipo documento:
Note
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
46
Recensione:
Indirizzi per estratti:
Citazione:
H. Shimizu et al., "DEMONSTRATION OF DESMOSOMAL ANTIGENS BY ELECTRON-MICROSCOPY USING CRYOFIXED AND CRYOSUBSTITUTED SKIN WITH SILVER-ENHANCED GOLD PROBE", The Journal of histochemistry and cytochemistry, 42(5), 1994, pp. 687-692

Abstract

In a previous post-embedding immunogold electron microscopic (EM) studies, localization of various desmosomal antigens was possible at highbut not at low magnification. We developed a method for simultaneous demonstration of epidermal desmosomal antigens at both low- and high-power EM magnifications by a method based on cryofixation and acetone cryosubstitution and the use of a 1-nm gold probe with silver enhancement. Ultra-thin sections of Lowicryl K11M were incubated with primary antibodies against desmoplakin, desmocollin, or desmoglein, followed by1-nm gold-conjugated secondary antibody. Silver enhancement for 12 min provided the ideal labeling size for low-power visualization, whereas silver enhancement for 4-6 min was ideal for high-power EM observation. Each desmosome immunolabeled with the gold probe was dearly demonstrated, even at very low-power magnification. The level of background labeling could be determined easily and the area of interest for high-power observation selected accurately. The fine ultrastructural appearance of desmosomal molecules was precisely demonstrated on high-power observation. This system should be useful for the immunocytochemical study of a variety of desmosomal antigens as well as other molecules ofinterest.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 24/11/20 alle ore 10:40:53