Kidney, lung, and intestine specimens were separated and fixed in Bouin’s fixative, then dehydrated, embedded in paraffin. The fixed samples were then sectioned (thickness, 4 μm) for hematoxylin and eosin (H&E), Periodic Acid-Schiff stain (PAS), Perls’ Prussian Blue, and immunohistochemistry (IHC) staining. The images were obtained and saved as digital images using Olympus VS200 Virtual Slide System (VS200, Olympus, Tokyo, Japan). Tubular injury was scored using a 5-point scale based on the percentage of brush border loss, tubular dilatation, cast deposition, and necrosis [57 (link)]. The scoring criteria are as follows: 0 point, no involvement; 1 point, < 10% involvement; 2 points, 10–25% involvement; 3 points, 25–50% involvement; 4 points, 50–75% involvement, and 5 points, > 75% involvement. Five random fields (200× magnification) per kidney section were used for quantification. The lung injury and intestine injury were assessed using their scoring criteria as previously described [58 (link), 59 (link)]. Histological analysis was independently graded by two pathologists.
Vs200 virtual slide system
Manufactured by Olympus
Sourced in Japan
The VS200 Virtual Slide System is a digital slide scanning and viewing platform designed for microscopy applications. It captures high-resolution digital images of microscope slides and allows for the viewing and analysis of these digital slides on a computer. The system includes a motorized stage, high-resolution camera, and software for image acquisition, viewing, and management.
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Lab products found in correlation
2 protocols using vs200 virtual slide system
1
Histological Analysis of Organ Injury
2
Tissue Preparation for Microscopic Analysis
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After the behavioral or fiber photometry experiments, rats were anesthetized with 3% pentobarbital sodium (40 mg/kg, ip) and transcardially perfused with physiological saline followed by cold 4% PFA (prepared in 0.1 M phosphate buffer, pH 7.4). The brains were removed from the skull and stored in 4% PFA at 4°C for 24 hours and then transferred to a 30% sucrose solution at 4°C for 48 hours. Coronal sections (30 μm thick) were cut on a freezing microtome (CM3050 S, Leica) and collected in cold PBS (0.01 M, pH 7.4). Slices were washed with PBST (once, 10 min), incubated for 10 min with DAPI (1:2000, D9542, Sigma-Aldrich), and then subjected to three more wash steps of 10 min each in PBST, followed by mounting and coverslipping on microscope slides. The extent of virus expression and the placement of the optical fiber were imaged using an Olympus BX53F fluorescence microscope (Japan) or an Olympus VS200 virtual slide system (Japan). We chose the section of maximum virus expression density as the injection center location. The criterion was consistent during the experiments.
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