Quantitative Collagen Imaging in Tissues

In immunohistochemical/immunocytochemical staining, the positivity was evaluated by image analysis performed using the Leica Q500 MC Image Analysis System (Leica, Cambridge, UK) as previously described [18 (link)]. For picrosirius red staining, a total of 10 fields were randomly chosen per mouse and images were viewed with brightfield illumination as well as with polarization contrast illumination at 40x. Picrosirius red is a birefringent molecule that binds to collagens. The complex fibrillar collagen/sirius red can be detected under polarized light, and the collagen bundles are red, yellow, and green. Collagen expression was quantified under brightfield illumination.

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Milanesi S., Garibaldi S., Saio M., Ghigliotti G., Picciotto D., Ameri P., Garibotto G., Barisione C, & Verzola D. (2019). Indoxyl Sulfate Induces Renal Fibroblast Activation through a Targetable Heat Shock Protein 90-Dependent Pathway. Oxidative Medicine and Cellular Longevity, 2019, 2050183.

Publication 2019

Leica Q500 MC Image Analysis System

Birefringent Collagen Fibrillar collagen Illumination Mouse

Corresponding Organization : Ospedale Policlinico San Martino

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Variable analysis

independent variables

Picrosirius red staining

dependent variables

Collagen expression (quantified under brightfield illumination)

control variables

Image analysis performed using the Leica Q500 MC Image Analysis System (Leica, Cambridge, UK)
A total of 10 fields randomly chosen per mouse
Images viewed with brightfield illumination as well as with polarization contrast illumination at 40x

controls

No positive or negative controls were explicitly mentioned in the input protocol.

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