Novaultra sirius red stain kit

Formalin-fixed, paraffin embedded liver tissue form study groups were cut in 5μm thick sections. Sections were deparaffinized using standard protocol and stained with picro-sirius red. Picro-sirius red staining of liver sections was carried out by using Nova ultra sirius red stain kit following manufacturer’s protocol (IHC world, Woodstock, MD) and observed under the light microscope using 20× objectives. Stained liver sections were examined for stages of fibrosis using the criteria of the NIH Non Alcoholic Steatohepatitis Clinical Research Network (NIH NASH CRN) scoring system. Stages of fibrosis were determined as 1A: mild, 1C: Portal, 2: Periportal fibrosis and 3: Bridging fibrosis.

The adoptive-transfer model of colitis was induced by intraperitoneal injection of 500,000 CD4⁺CD45RB^hi naive T cells isolated from L-Tg mice to Rag1^−/− mice; Rag^−/−Dr3^−/− or Rag^−/−Dr3^∆Col1a251. Mice used were age-matched, male littermates co-housed under specific pathogen-free conditions in the Animal Facility at Cedars-Sinai Medical Center (CSMC).
DAI was calculated as in previously published studies^{19 (link)}. DAI score was determined twice a week for the adoptive-transfer model. Macroscopic evidence of inflammation was scored blinded to the mice genotype using the established classification^{20 (link),51 (link)}. Mucosal area of collagen deposition was identified by Sirius red staining using the NovaUltra Sirius Red Stain Kit according to manufacturer’s protocol (IHC World, Woodstock, MD). Stained gut sections were quantitated for the relative degree of fibrosis using ImageJ software (imagej.nih.gov), as previously described^{21 (link)}.