| Complete chromogen separation and analysis in double immunohistochemical stains using Photoshop-based image analysis. | |
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MedLine Citation:
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PMID: 9857219 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Simultaneous detection of two different antigens on paraffin-embedded and frozen tissues can be accomplished by double immunohistochemistry. However, many double chromogen systems suffer from signal overlap, precluding definite signal quantification. To separate and quantitatively analyze the different chromogens, we imported images into a Macintosh computer using a CCD camera attached to a diagnostic microscope and used Photoshop software for the recognition, selection, and separation of colors. We show here that Photoshop-based image analysis allows complete separation of chromogens not only on the basis of their RGB spectral characteristics, but also on the basis of information concerning saturation, hue, and luminosity intrinsic to the digitized images. We demonstrate that Photoshop-based image analysis provides superior results compared to color separation using bandpass filters. Quantification of the individual chromogens is then provided by Photoshop using the Histogram command, which supplies information on the luminosity (corresponding to gray levels of black-and-white images) and on the number of pixels as a measure of spatial distribution. (J Histochem Cytochem 47:119-125, 1999) |
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Authors:
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H A Lehr; C M van der Loos; P Teeling; A M Gown |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society Volume: 47 ISSN: 0022-1554 ISO Abbreviation: J. Histochem. Cytochem. Publication Date: 1999 Jan |
Date Detail:
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Created Date: 1999-02-09 Completed Date: 1999-02-09 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9815334 Medline TA: J Histochem Cytochem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 119-26 Citation Subset: IM |
Affiliation:
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Institute of Pathology, University of Mainz, Germany. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Actins
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analysis Antigens, CD / analysis Antigens, Differentiation, Myelomonocytic / analysis Breast Neoplasms / chemistry Carotid Arteries / chemistry Chromogenic Compounds / analysis* Epithelium / chemistry Humans Image Processing, Computer-Assisted* Immunohistochemistry / methods* Keratins / analysis Software Stromal Cells / chemistry Vimentin / analysis |
| Chemical | |
Reg. No./Substance:
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0/Actins; 0/Antigens, CD; 0/Antigens, Differentiation, Myelomonocytic; 0/CD68 antigen, human; 0/Chromogenic Compounds; 0/Vimentin; 68238-35-7/Keratins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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