2018
DOI: 10.1590/abd1806-4841.20187544
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Evaluation of dermal collagen stained with picrosirius red and examined under polarized light microscopy

Abstract: The special picrosirius red staining highlights the natural birefringence of collagen fibers when exposed to polarized light. The results from birefringence allow to evaluate the organization of the collagen fibers in the tissues. The authors intend to elucidate all steps to obtain and capture images of histological sections stained with picrosirius red and evaluated under polarized light microscopy, as well as possible artefacts that may occur.

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Cited by 47 publications
(36 citation statements)
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“…The stained sections were analyzed using an Olympus IX73 inverted microscope (Olympus Corporation; magnification, ×200 or ×400) with a linear polarizer. To avoid missing any details, the filter was tilted to an angle between 0 and 90 until the birefringence became evident and the background became completely black; the focus was then corrected once more ( 25 ). The halogen lamp intensity and exposure time were constant within each image.…”
Section: Methodsmentioning
confidence: 99%
“…The stained sections were analyzed using an Olympus IX73 inverted microscope (Olympus Corporation; magnification, ×200 or ×400) with a linear polarizer. To avoid missing any details, the filter was tilted to an angle between 0 and 90 until the birefringence became evident and the background became completely black; the focus was then corrected once more ( 25 ). The halogen lamp intensity and exposure time were constant within each image.…”
Section: Methodsmentioning
confidence: 99%
“…Relatively inexpensive, the technique relies on the birefringent properties of collagen molecules. While the PSR stain alone does not selectively bind the collagen network, it becomes more specific than the other common collagen stains when combined with polarized light detection [ 9 , 11 ]. Under circularly polarized PSR microscopy, thin bundles of collagen type III appear green-yellow, whereas large bundles of tightly packed, well-organized and mature collagen type I molecules appear yellow-orange [ 9 ].…”
Section: Discussionmentioning
confidence: 99%
“…Under circularly polarized PSR microscopy, thin bundles of collagen type III appear green-yellow, whereas large bundles of tightly packed, well-organized and mature collagen type I molecules appear yellow-orange [ 9 ]. Use of circularly polarized PSR microscopy revealed bundles of yellow-orange collagen I [ 9 , 11 ] along the intrahepatic duct walls, indicating the presence of mature collagen type I, which provides both structural support and tensile strength [ 9 ]. By contrast, there was no accumulation of collagen I or collagen III along the renal cyst walls.…”
Section: Discussionmentioning
confidence: 99%
“…Adjacent sections were stained with Alcian blue and Picrosirius red and subsequently imaged under polarized light to visualize collagen fiber organization 26 . Birefringence of collagen fibers was then determined based on the intensity of individual pixels captured using polarized light microscopy 27 . To confirm effectiveness of the HFD, livers were harvested, fixed in 4% paraformaldehyde in PBS for 16 h at 4 °C, washed 3 times with PBS, and cryoprotected in 30% sucrose for 16 h at 4 °C.…”
Section: Methodsmentioning
confidence: 99%