Picrosirius Red Staining: Revisiting Its Application to the Qualitative and Quantitative Assessment of Collagen Type I and Type III in Tendon

2021 ◽  
Vol 69 (10) ◽  
pp. 633-643
Author(s):  
Consuelo M. López De Padilla ◽  
Michael J. Coenen ◽  
Alejandro Tovar ◽  
Rodolfo E. De la Vega ◽  
Christopher H. Evans ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Quantitative Assessment ◽  
Collagen Type ◽  
Polarized Light ◽  
Polarized Light Microscopy ◽  
Type I ◽  
Collagen Types ◽  
Qualitative And Quantitative ◽  
Type Iii ◽  
Picrosirius Red

Collagen has a major role in the structural organization of tendons. Picrosirius red (PSR) staining viewed under polarized light microscopy is the standard method to evaluate the organization of collagen fibers in tissues. It is also used to distinguish between type I and type III collagen in tissue sections. However, accurate analysis and interpretation of PSR images are challenging because of technical factors and historical misconceptions. The aim of this study was to clarify whether collagen types I and III can be distinguished by PSR staining in rat Achilles tendons, using double immunohistochemistry as the positive control. Our findings showed that PSR staining viewed with polarized light microscopy was suitable for qualitative and quantitative assessment of total collagen but was not able to distinguish collagen types. We found it critical to use a polarizing microscope equipped with a rotating stage; tendon section orientation at 45° with respect to crossed polarizers was optimal for the qualitative and quantitative assessment of collagen organization. Immunohistochemistry was superior to PSR staining for detection of collagen type III. We also compared formalin and Bouin solution as fixatives. Both produced similar birefringence, but formalin-fixed tendons provided higher quality histological detail with both hematoxylin–eosin and immunostaining:

Polarized Light Microscopy for Analyzing Tissue Mechanics During In Vitro Acupuncture

2008 ◽  
Author(s):  
Lowell Taylor Edgar ◽  
Margaret Julias ◽  
David I. Shreiber ◽  
Helen M. Buettner
Keyword(s):  
Connective Tissue ◽  
Light Microscopy ◽  
Type I Collagen ◽  
Polarized Light ◽  
Tissue Mechanics ◽  
Polarized Light Microscopy ◽  
Tissue Response ◽  
Type I ◽  
Collagen Gels

Acupuncture is a traditional therapy originating in China almost 2000 years ago. Acupuncture has slowly been growing in popularity in the West, and clinical evidence has shown the potential for acupuncture as a low-cost ‘alternative’ therapy for an assortment of ailments [1]. The practice of acupuncture involves inserting fine needles into the skin followed by needle manipulation, usually by rotation. Recent studies by Langevin et al demonstrate that this rotation causes the subcutaneous connective tissue to couple to and wind around the needle [2–4], which suggests that mechanotransduction in the connective tissue might play a role in the therapeutic mechanisms that underlay acupuncture [2, 3]. To begin to decompose and quantify this complex mechanism at the tissue level in a controlled setting, we have simulated acupuncture in type I collagen gels in vitro, and have developed algorithms to quantify the tissue response following imaging with polarized light microscopy (PLM).

scholarly journals Evaluation of dermal collagen stained with picrosirius red and examined under polarized light microscopy

2018 ◽  
Vol 93 (3) ◽  
pp. 415-418 ◽  
Author(s):  
Polyana Galvão Bernardes Coelho ◽  
Maria Verônica de Souza ◽  
Lissandro Gonçalves Conceição ◽  
Marlene Isabel Vargas Viloria ◽  
Sirley Adriana Ortiz Bedoya
Keyword(s):  
Light Microscopy ◽  
Polarized Light ◽  
Polarized Light Microscopy ◽  
Dermal Collagen ◽  
Picrosirius Red

scholarly journals Diagnosis of Collagen Type III Glomerulopathy Using Picrosirius Red and PASH/Masson’s Trichrome Stain

2020 ◽  
Vol 57 (5) ◽  
pp. 675-680
Author(s):  
Katelin L. Davis ◽  
Anne L. Burnum ◽  
Jessica A. Beck ◽  
Shannon G. M. Kirejczyk ◽  
Margaret A. Miller ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Collagen Type ◽  
Type Iii Collagen ◽  
Fibrillar Collagen ◽  
Collagen Type Iii ◽  
Type Iii ◽  
Glomerular Capillary ◽  
Collagen Type Iii Glomerulopathy ◽  
Picrosirius Red ◽  
Capillary Walls

Canine collagen type III glomerulopathy (Col3GP) is a rare juvenile nephropathy in which irregular type III collagen fibrils and fibronectin accumulate in glomerular capillary walls and the mesangium. Necropsy findings were reviewed from 5 puppies diagnosed with Col3GP at 6 to 18 weeks of age. Histologically, with hematoxylin and eosin stain, the glomerular capillary walls and mesangium were diffusely and globally expanded by homogeneous pale eosinophilic material. Ultrastructurally, the subendothelial zone and mesangium were expanded by fibronectin and cross-banded collagen type III fibrils, diagnostic of Col3GP. Two additional stains were employed to identify the material within glomeruli as fibrillar collagen using light microscopy. In all 5 cases, the material was red with picrosirius red and birefringent under polarized light, and was blue with periodic acid-Schiff/hematoxylin/trichrome (PASH/TRI), thereby identifying it as fibrillar collagen. Based on these unique staining characteristics with picrosirius red and PASH/TRI, Col3GP may be reliably diagnosed with light microscopy alone.

scholarly journals A Polarized Light Microscopy Study in a Case of Morphea

2020 ◽  
Vol 26 (1) ◽  
pp. 1-4
Author(s):  
Mocanu Liliana ◽  
Deacu Sorin ◽  
Roman Polimaru ◽  
Aschie Mariana
Keyword(s):  
Light Microscopy ◽  
Polarized Light ◽  
Microscopy Study ◽  
Polarized Light Microscopy ◽  
Type I ◽  
Elastic Tissue ◽  
Collagen Fibres ◽  
Reticular Dermis ◽  
Plaque Type ◽  
Sirius Red

Abstract We report a case of plaque type of scleroderma with specific clinical features and conventional histopathology, with sclerosis and hipocellularity of fibroblasts and preservation of elastic tissue. We describe polarized light microscopy findings, on conventional stained slides and on picro sirius red stained slides. We appreciate that picro sirius red stain allows a better characterization of collagen fibres composition in papillary and reticular dermis, that is severely disturbed in morphea, with an inverse distribution of collagen fibres type I and III comparative with normal dermis.

scholarly journals Quantitative assessment of cell contractility using polarized light microscopy

2018 ◽  
Vol 11 (11) ◽  
pp. e201800008 ◽  
Author(s):  
Wenjun Wang ◽  
Joseph P. Miller ◽  
Susan C. Pannullo ◽  
Cynthia A. Reinhart-King ◽  
Francois Bordeleau
Keyword(s):  
Light Microscopy ◽  
Quantitative Assessment ◽  
Polarized Light ◽  
Polarized Light Microscopy ◽  
Cell Contractility

scholarly journals Quantitative and qualitative analysis of collagen types in the fascia transversalis of inguinal hernia patients

2007 ◽  
Vol 44 (3) ◽  
pp. 230-234 ◽  
Author(s):  
Alberto Luiz Monteiro Meyer ◽  
Eduardo Berger ◽  
Orlando Monteiro Jr. ◽  
Paulino Alberto Alonso ◽  
João Norberto Stavale ◽  
...  
Keyword(s):  
Inguinal Hernia ◽  
Collagen Type ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Type Iii ◽  
Collagen Types ◽  
Type Iii ◽  
Fascia Transversalis ◽  
Significant Difference ◽  
Percent Area

BACKGROUND: Inguinal hernia is the second most common surgical case in our field. The anatomical factors alone are not enough to explain the inguinal hernia. Studies show changes in the proportion and quantity of collagen fibers in the developing of inguinal hernia. The greater production of collagen type III compared to the type I could justify the thinning of the fascia transversalis and its weakness. AIM: To determine the quantitative and qualitative changes of collagen in the fascia transversalis in inguinal hernia patients and compare them to findings from corpses without inguinal hernia. METHOD: Prospective case-control study based on the biopsy of fascia transversalis of 27 patients and 24 corpses. The technique used was hematoxylin-eosin and picrosirius colorimetry. RESULTS: The medium percent area of collagen (types I + III) and collagen type I, in both groups, show no statistic difference. The quantity of collagen type III was greater in the patients. Patients classified with Nyhus IIIa presented greater quantity of collagen type III. CONCLUSION: There is no significant difference in the quantity of collagen in the fascia transversalis of patients compared to the controls. An increase in the quantity of collagen type III was found in patients with inguinal hernia and a greater quantity in those patients classified with Nyhus IIIa.

Ferroelastic domains and phase transitions in organic–inorganic hybrid perovskite CH3NH3PbBr3

2021 ◽  
Author(s):  
Maryam Bari ◽  
Alexei A. Bokov ◽  
Zuo-Guang Ye
Keyword(s):  
Phase Transitions ◽  
Electric Field ◽  
Light Microscopy ◽  
Domain Structure ◽  
Polarized Light ◽  
External Stress ◽  
Polarized Light Microscopy ◽  
Inorganic Hybrid ◽  
Hybrid Perovskite ◽  
Ferroelastic Domains

Polarized light microscopy reveals twin domains and symmetry of the phases in CH3NH3PbBr3 crystal; domain structure remains unresponsive to electric field but changes under external stress, confirming ferroelasticity while ruling out ferroelectricity.

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