Journal of Histochemistry and Cytochemistry
  Search:   
    >> Advanced Search

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

JHC exPRESS: First Published December 23, 2007. doi:10.1369/jhc.2007.950170
Journal of Histochemistry and Cytochemistry
Copyright © 2007 van der Loos

A more recent version of this article appeared on April 1, 2008.
This Article
Right arrow exPRESS PDF
Right arrow All Versions of this Article:
jhc.2007.950170v1
56/4/313    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
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 van der Loos, C. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van der Loos, C. M.
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   Add to Twitter  
What's this?

Articles

Multiple Immunoenzyme Staining: Methods and Visualizations for the Observation With Spectral Imaging

Chris M. van der Loos 1*

1 Department of Pathology, Academic Medical Center, Amsterdam, The Netherlands

* To whom correspondence should be addressed. E-mail: c.m.vanderloos{at}amc.uva.nl.

Submitted on November 19, 2007
Accepted on 26 November 2007


  Abstract
Several staining concepts and color combinations exist to perform successful double immunoenzyme staining on human tissue specimens. Most of these concepts are based on differences between both primary antibodies: animal species, mouse Ig isotype or IgG subclasses, conjugates or concentrations. Traditionally, double immunoenzyme staining has used chromogens selected to provide maximum color contrast when observed with the unaided eye. Unfortunately, visually good color combinations always include at least one diffuse chromogen, due to the paucity of appropriate chromogen colors. This situation is drastically changed with the use of spectral imaging, where multicolor microscopy can be unmixed in individual images based on their spectral characteristics. Spectral unmixing can be performed even up to quadruple immunoenzyme staining. This work contains practical suggestions for immunoenzyme double staining procedures for some frequently encountered primary antibody combinations: rabbit-mouse, goat-mouse, mouse-mouse and rabbit-rabbit. The suggested protocols are all suitable for a classical red-brown color combination plus blue nuclear counterstain that is composed of peroxidase activity (DAB), alkaline phosphatase activity (Liquid Permanent Red) and hematoxylin, respectively. Although the red and brown chromogens do not contrast very well visually, they both show a crisp localization and can be perfectly unmixed by spectral imaging.

Key Words: immuno double staining, immuno quadruple staining, chromogens, spectral imaging, unmixing


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   Add to Twitter Twitter    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
A. S. Flozak, A. P. Lam, S. Russell, M. Jain, O. N. Peled, K. A. Sheppard, R. Beri, G. M. Mutlu, G. R. S. Budinger, and C. J. Gottardi
{beta}-Catenin/T-cell Factor Signaling Is Activated during Lung Injury and Promotes the Survival and Migration of Alveolar Epithelial Cells
J. Biol. Chem., January 29, 2010; 285(5): 3157 - 3167.
[Abstract] [Full Text] [PDF]

Home page
 JNMHome page
L. Zhou and W. S. El-Deiry
Multispectral Fluorescence Imaging
J. Nucl. Med., October 1, 2009; 50(10): 1563 - 1566.
[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 © 2007

 
Purchase HCS Short Course Manual on HCS site