Kidney Fibrosis Quantification with Sirius Red

Paraformaldehyde-fixed paraffin-embedded kidneys were sectioned at 4.5 μm thickness and stained with 0.1% sirius red solution (Electron Microscopy Sciences, Hatfield, PA, United States; Ng et al., 2017 (link)). At least 10 non-overlapping bright field images at 20× magnification were analyzed for each animal using the Carl Zeiss Axio A1 microscope with an AxioCam MRc5 CCD camera (Oberkochen, Germany). Staining was quantified using ImageJ software (NIH, Bethesda, MD; Schneider et al., 2012 (link)) and presented as percentage of stained area within the analyzed image.

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Ng H.H., Soula M., Rivas B., Wilson K.J., Marugan J.J, & Agoulnik A.I. (2021). Anti-apoptotic and Matrix Remodeling Actions of a Small Molecule Agonist of the Human Relaxin Receptor, ML290 in Mice With Unilateral Ureteral Obstruction. Frontiers in Physiology, 12, 650769.

Publication 2021

Axio A1 microscope AxioCam MRc5 CCD camera

Animal Electron microscopy Kidneys Microscope Paraffin Paraformaldehyde

Corresponding Organization : Florida International University

Other organizations : National Center for Advancing Translational Sciences, National Institutes of Health

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Percentage of stained area within the analyzed image

control variables

Paraformaldehyde-fixed paraffin-embedded kidneys
Tissue sectioning at 4.5 μm thickness
Staining with 0.1% sirius red solution
Bright field imaging at 20× magnification using Carl Zeiss Axio A1 microscope with AxioCam MRc5 CCD camera
Image quantification using ImageJ software

controls

No positive or negative controls explicitly mentioned

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