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Elastin autofluorescence and collagen staining

KS Kavita Shirsath
AJ Apeksha Joshi
AV Aliasgar Vohra
RD Ranjitsinh Devkar
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Elastin autofluorescence was recorded from the H × E stained sections using FLoid imaging station (Thermo Fisher Scientific, USA)51. Elastin breaks, defined as discontinuity of an elastic fiber with boundaries at both sides clearly visible, were counted from the images. For collagen staining, paraffin-embedded sections were de-paraffinized with xylene and rehydrated by immersion in a graded series of ethanol. The sections were then stained with 0.1% direct red 80 (Sigma Aldrich, USA) in saturated aqueous solution of picric acid for 1 h at room temperature (RT) and observed under Leica DMRB microscope (Leica Microsystems, Germany). Images were captured and quantitative analysis of the images for collagen and elastin content was done using ImageJ software (NIH, Bethesda, USA)52,53. Aortic stiffness was calculated as ratio of collagen to elastin.

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