Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy
Picrosirius red (PSR) staining is a commonly used histological technique to visualize collagen in paraffin-embedded tissue sections. PSR stained collagen appears red in light microscopy. However it is largely unknown that PSR stained collagen also shows a red fluorescence, whereas live cells have a...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-01-01
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| Series: | MethodsX |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016115000102 |
| _version_ | 1818177291111366656 |
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| author | Benjamin Vogel Hanna Siebert Ulrich Hofmann Stefan Frantz |
| author_facet | Benjamin Vogel Hanna Siebert Ulrich Hofmann Stefan Frantz |
| author_sort | Benjamin Vogel |
| collection | DOAJ |
| description | Picrosirius red (PSR) staining is a commonly used histological technique to visualize collagen in paraffin-embedded tissue sections. PSR stained collagen appears red in light microscopy. However it is largely unknown that PSR stained collagen also shows a red fluorescence, whereas live cells have a distinct green autofluorescence. Both emission patterns can be detected using standard filter sets as found in conventional fluorescence microscopes. Here we used digital image addition and subtraction to determine the relative area of the pure collagen and live cell content in heart tissue in a semi-automated process using standard software. This procedure, which considers empty spaces (holes) within the section, can be easily adapted to quantify the collagen and live cell areas in healthy or fibrotic tissues as aorta, lung, kidney or liver by semi-automated planimetry exemplified herein for infarcted heart tissue obtained from the mouse myocardial infarction model.
• Use of conventional PSR stained paraffin-embedded tissue sections for fluorescence analysis.
• PSR and autofluorescence images are used to calculate area of collagen and area of live cells in the tissue; empty spaces (holes) in tissue are considered.
• High throughput analysis of collagen and live cell content in tissue for statistical purposes. |
| first_indexed | 2024-12-11T20:29:45Z |
| format | Article |
| id | doaj.art-231fd160ebc14b589444010a2afabdd9 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-11T20:29:45Z |
| publishDate | 2015-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-231fd160ebc14b589444010a2afabdd92022-12-22T00:51:52ZengElsevierMethodsX2215-01612015-01-012C12413410.1016/j.mex.2015.02.007Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopyBenjamin Vogel0Hanna Siebert1Ulrich Hofmann2Stefan Frantz3Comprehensive Heart Failure Center (CHCF), Universitätsklinikum Würzburg, GermanyComprehensive Heart Failure Center (CHCF), Universitätsklinikum Würzburg, GermanyComprehensive Heart Failure Center (CHCF), Universitätsklinikum Würzburg, GermanyComprehensive Heart Failure Center (CHCF), Universitätsklinikum Würzburg, GermanyPicrosirius red (PSR) staining is a commonly used histological technique to visualize collagen in paraffin-embedded tissue sections. PSR stained collagen appears red in light microscopy. However it is largely unknown that PSR stained collagen also shows a red fluorescence, whereas live cells have a distinct green autofluorescence. Both emission patterns can be detected using standard filter sets as found in conventional fluorescence microscopes. Here we used digital image addition and subtraction to determine the relative area of the pure collagen and live cell content in heart tissue in a semi-automated process using standard software. This procedure, which considers empty spaces (holes) within the section, can be easily adapted to quantify the collagen and live cell areas in healthy or fibrotic tissues as aorta, lung, kidney or liver by semi-automated planimetry exemplified herein for infarcted heart tissue obtained from the mouse myocardial infarction model. • Use of conventional PSR stained paraffin-embedded tissue sections for fluorescence analysis. • PSR and autofluorescence images are used to calculate area of collagen and area of live cells in the tissue; empty spaces (holes) in tissue are considered. • High throughput analysis of collagen and live cell content in tissue for statistical purposes.http://www.sciencedirect.com/science/article/pii/S2215016115000102Collagen determination in tissue sections by PSR fluorescence |
| spellingShingle | Benjamin Vogel Hanna Siebert Ulrich Hofmann Stefan Frantz Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy MethodsX Collagen determination in tissue sections by PSR fluorescence |
| title | Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy |
| title_full | Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy |
| title_fullStr | Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy |
| title_full_unstemmed | Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy |
| title_short | Determination of collagen content within picrosirius red stained paraffin-embedded tissue sections using fluorescence microscopy |
| title_sort | determination of collagen content within picrosirius red stained paraffin embedded tissue sections using fluorescence microscopy |
| topic | Collagen determination in tissue sections by PSR fluorescence |
| url | http://www.sciencedirect.com/science/article/pii/S2215016115000102 |
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