The mice liver tissues were fixed in Mildform (Fujifilm Wako Chemicals, Osaka, Japan), processed for paraffin embedding, and sectioned into 3 µm-thick sections. Each section was deparaffinized, rehydrated, and processed for hematoxylin–eosin and Sirius red-Fast Green staining. Sections were incubated in 0.04% Fast Green (Fujifilm Wako Chemicals), saturated in picric acid for 5 min, washed with distilled water, and incubated in 0.1% Sirius Red solution (Fujifilm Wako Chemicals) for 10 min. The histological features after each staining were determined using a Leica DMi8 microscope and LAS X version 3.3 (Leica Microsystems, Inc., Wetzlar, Germany). The steatosis, inflammation, and fibrosis degrees were assessed by an experienced pathologist blinded to the experimental procedures according to previously reported criteria33 (link),34 (link).
Sirius red solution
Manufactured by Fujifilm
Sirius Red solution is a laboratory reagent used in histological staining techniques. It is a solution of the dye Sirius Red, which is used to stain collagen fibers in tissue samples. The core function of Sirius Red solution is to provide a specific and reliable staining method for the identification and visualization of collagen in various types of tissues.
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2 protocols using sirius red solution
1
Histological Assessment of Liver Pathology
2
Sirius Red Staining of Paraffin Sections
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Glass slides with paraffin-embedded sections attached were rinsed with running water for 5 min following deparaffinization and hydrophilic treatment. Sirius red staining solution was created by combining 100 ml of saturated picric acid solution with 3 ml of 1% (w/v) Sirius red solution (Fujifilm Wako). The glass slides were rinsed under running water for 5 min after being stained with Sirius red staining solution for 10 min to remove the color. The slides were then dehydrated using ethanol, permeated with xylene, and sealed with Marinol (Muto Chemical) and cover glass (Matsunami Glass).
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