Journal of Histochemistry and Cytochemistry Priciples for Free Access to Science
  Search:   
    >> Advanced Search

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Büki, A.
Right arrow Articles by Povlishock, J. T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Büki, A.
Right arrow Articles by Povlishock, J. T.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
Journal of Histochemistry and Cytochemistry, Vol. 48, 153-162, January 2000, Copyright © 2000, The Histochemical Society, Inc.

TECHNICAL NOTE

Novel Application of Tyramide Signal Amplification (TSA): Ultrastructural Visualization of Double-labeled Immunofluorescent Axonal Profiles

András Bükia,b, Susan A. Walkera, James R. Stonea, and John T. Povlishocka
a Department of Anatomy, Medical College of Virginia, Virginia Commonwealth University, Richmond, Virginia
b Department of Neurosurgery, University Medical School of Pécs, Pécs, Hungary

Correspondence to: John T. Povlishock, Dept. of Anatomy, Medical College of Virginia, Virginia Commonwealth U., Richmond, VA 23298-0709.

Fluorescent immunocytochemistry (FICC) allows multiple labeling approaches when enzyme-based techniques are difficult to combine, such as in double-labeling experiments targeting small-caliber axonal segments. Nevertheless, the conversion of FICC to a product visible at the electron microscopic (EM) level requires labor-intensive procedures, thus justifying the development of more user-friendly conversion methods. This study was initiated to simplify the conversion of FICC to EM by employing the unique properties of tyramide signal amplification (TSA), which allowed the simultaneous targeting of a fluorescent tag and biotin label to the same antigenic site. Briefly, one of two antigenic sites typically co-localized in damaged axonal segments was visualized by the application of a fluorescent secondary antibody, with the other tagged via a biotinylated antibody. Next, an ABC kit was used, followed by the simultaneous application of fluorophore–tyramide and biotin–tyramide. After temporary mounting for fluorescent digital photomicroscopy, sections were incubated in ABC and reacted with diaminobenzidine before EM analysis. Double-labeling fluorescent immunocytochemistry with TSA clearly delineated damaged axonal segments. In addition, these same axonal segments yielded high-quality EM images with discrete electron-dense reaction products, thereby providing a simple and reproducible means for following fluorescent analysis with EM. (J Histochem Cytochem 48:153–161, 2000)

Key Words: tyramide signal amplification, axonal injury, immunofluorescence, double labeling, trauma, calpain


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Am. J. Physiol. Gastrointest. Liver Physiol.Home page
Z. K. Krowicki, M. A. Burmeister, H.-R. Berthoud, R. T. Scullion, K. Fuchs, and P. J. Hornby
Orexins in rat dorsal motor nucleus of the vagus potently stimulate gastric motor function
Am J Physiol Gastrointest Liver Physiol, August 1, 2002; 283(2): G465 - G472.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurosci.Home page
A. Buki, D. O. Okonkwo, K. K. W. Wang, and J. T. Povlishock
Cytochrome c Release and Caspase Activation in Traumatic Axonal Injury
J. Neurosci., April 15, 2000; 20(8): 2825 - 2834.
[Abstract] [Full Text] [PDF]



Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 2000