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Immunodetection of RNA on ultra-thin sections incubated with polyadenylate nucleotidyl transferase.
Thiry, Marc
1993In Journal of Histochemistry and Cytochemistry, 41 (5), p. 657-65
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Keywords :
Animals; Carcinoma, Ehrlich Tumor/chemistry; Immunohistochemistry; Mice; Mice, Inbred C57BL; Microscopy, Immunoelectron; Polynucleotide Adenylyltransferase; RNA/analysis; RNA, Neoplasm/analysis
Abstract :
[en] A new method is described for locating RNA on ultra-thin sections. Sections of aldehyde-fixed, plastic-embedded cells were incubated in a medium containing polyadenylate nucleotidyl transferase (PnT) and biotinylated ATP. The labeled nucleotides bound to RNA at the surface of the ultra-thin sections were than visualized by an indirect immunogold labeling technique. The resulting labeling pattern was dependent on the presence of divalent cations in the PnT medium. The method revealed with great precision the specific RNA-containing structures within Ehrlich tumor cells. The method is applicable to Epon sections. However, the labeling intensity varies according to the fixation used. Best results were obtained on acetylated cell sections. The method can be combined with EDTA regressive staining. The in situ PnT method provides a very useful tool for pinpointing the precise location of RNA within biological material at the ultrastructural level.
Disciplines :
Anatomy (cytology, histology, embryology...) & physiology
Author, co-author :
Thiry, Marc  ;  Université de Liège - ULiège > Département des sciences de la vie > Biologie cellulaire
Language :
English
Title :
Immunodetection of RNA on ultra-thin sections incubated with polyadenylate nucleotidyl transferase.
Publication date :
1993
Journal title :
Journal of Histochemistry and Cytochemistry
ISSN :
0022-1554
eISSN :
1551-5044
Publisher :
Histochemical Society, New York, United States - New York
Volume :
41
Issue :
5
Pages :
657-65
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 25 November 2009

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