After euthanasia, the lungs were inflated with 1mL of 4% paraformaldehyde (PFA) and fixed in 4% PFA. All tissues were routinely processed, embedded in paraffin wax, and sectioned (4μm) according to the revised guide for organ sampling and trimming in rats and mice by the European Experimental and Toxicologic Pathology group [42 (link)]. Cotton rat lung histology slides were routinely stained with hematoxylin and eosin (H&E). Light microscopic evaluation was performed by a veterinary anatomic pathologist (MM) (model CX41, Olympus, B and B Microscopes Limited, Pittsburgh, PA, USA). A semi-quantitative scoring system, adapted from previous studies [43 (link)], was established and applied (S2 Table ). The sum of the mean scores were calculated to produce a total histologic inflammatory score. Immunohistochemistry was performed on the formalin fixed paraffin embedded slides using goat anti-RSV polyclonal antiserum (Virostat, cat# 0601, Westbrook, Maine, USA) at a dilution of 1:800. A semi-quantitative immunohistochemical scoring system was devised (S1 Table ) to compare the expression of the RSV G protein based on percentage of cells staining as well as strength of stain. The sum of the mean scores was calculated to produce a total immunohistochemical score.
Model cx41
Manufactured by Olympus
Sourced in United States, Japan
The Olympus CX41 is a compact and versatile microscope designed for routine laboratory work. It features a binocular observation tube, a range of objective lenses, and a built-in illumination system. The CX41 is capable of bright-field observation and is suitable for a variety of applications in educational and research settings.
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Lab products found in correlation
2 protocols using model cx41
1
Histological Evaluation of Cotton Rat Lungs
2
TUNEL Assay for Apoptotic Cell Detection
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According to the manufacturer's instructions, rehydrated and deparaffinized sections were stained with a commercial kit (Apoptag, S7101, Chemicon, Calif, USA). A CX41 bright-field microscope was used to examine the TUNEL-stained slices (Olympus, Model: CX41, Tokyo, Japan). The TUNEL scores were measured independently by 2 researchers blinded to the experimental information. The positive reactive cell number in all TUNEL-stained slices was accounted and was analyzed from apical to basal switch. The TUNEL reactive cells were accounted in randomly selected fields per case to calculate the average apoptotic cell number. In each case, 100 cells were determined to be TUNEL positive/negative, and the percentage of TUNEL reactive cell number was assigned. All histopathological specimens were assessed by 2 expert pathologists. In evaluation, cells in regions with necrosis, those placed in borders, or poor morphology were excluded.
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