Cryosections (6 μm) were air-dried and incubated in phosphate buffer 0.05 M pH 7.3 containing 20 mg of diaminobenzidine, 140 mg of cytochrome C, 3 g of saccharose, and 4 ml of catalase (Sigma–Aldrich, St. Louis, MO, United States). Slides were then rinsed in distilled water, dehydrated in three alcohol baths (95°, 100°, and 100°), fixed in Xylene before being mounted with Eukitt® Quick-hardening medium (Sigma–Aldrich, St. Louis, MO, United States). Measurements of COx intensity were performed by converting the image to gray scale to determine optical density. Images were acquired in the same exposure conditions using Zeiss Primo Vert microscope with a 10× objective. The mean relative optical density per pixel was determined by subtracting the optical density of the background using ImageJ software. For each subject, a single value was obtained by averaging measurements from approximately 100 muscle fibers, which were chosen at random.
Eukitt quick hardening medium
Manufactured by Merck Group
Sourced in Sao Tome and Principe
Eukitt® Quick-hardening medium is a laboratory product designed for the rapid mounting and preservation of microscope slides. It is a solvent-based mounting medium that allows for the quick hardening of specimens, facilitating the preparation of permanent microscope slides.
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Lab products found in correlation
2 protocols using eukitt quick hardening medium
1
Cytochrome C Oxidase Quantification
2
Quantitative Cardiac Fibrosis Analysis
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Dissected heart tissue was fixed with 4 % PFA for 24 h at 4 °C, processed in the automated tissue processor (Leica TP1020) with successive alcohol/xylene incubations and embedded in paraffin. Tissues embedded in paraffin were sectioned into 5 μm sections (Leica RM 2125RTS) and mounted onto microscope slides (SuperFrost Plus, WVR cat. 631-0108). Masson's Trichrome (MT) kit (Sigma cat. HT15-1KT) was used to stain heart sections, according to manufacturer's instructions, and the sections were mounted with Eukitt quick-hardening medium (Sigma-Aldrich, cat 03989). The slides were scanned, and images were acquired with Hamamatsu NanoZoomer Scanner S360 (UCL-IQPath histology service) and the amount of fibrosis in sections was quantified with ImageJ software using colour-threshold method and expressed as percentage of total heart tissue area.
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