For the immunohistochemical analysis, paraffin-embedded mouse kidney tissue sections (5 μm thick) were employed. The sections were placed on glass slides, de-paraffinized and hydrated with xylene and graded alcohol. The sections were pre-incubated in a boiled sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH = 6.0) for antigen retrieval. Each specific primary antibody against α-SMA, nephrin or HIF-1α was incubated with the tissue sections overnight. For the measurement of α-SMA expression, the tissue section was stained with FITC-conjugated anti mouse IgG. In addition, nephrin and HIF-1α were visualized with 3,3′-diaminobenzidine (DAB) to produce a brown staining, being counterstained with hematoxylin. The stained tissue sections were examined using an optical Axioimager microscope system and five images were taken for each section (Zeiss, Oberkochen, Germany).
Axioimager microscope system
Manufactured by Zeiss
Sourced in Germany
The Axioimager microscope system is a high-performance microscope designed for advanced imaging and analysis applications. It features a modular design that allows for customization to meet specific research or industrial needs. The core function of the Axioimager is to provide detailed, high-resolution imaging capabilities for a wide range of sample types and applications.
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4 protocols using axioimager microscope system
1
Immunohistochemical Analysis of Mouse Kidney
2
Histopathological Analysis of H. pylori Infection
3
Histological Analysis of Renal Tissues
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For the histological analyses of glomerular tissues and renal tubule tissues, tissue specimens were collected at the end of the experiments and fixed in 10% buffered formaldehyde. The paraffin-embedded tissue specimens were sectioned at 5 μm thickness, deparaffinized, and stained with PAS stain to assess interstitial fibrosis. The stained tissue sections were observed using an optical Axioimager microscope system (Zeiss), and 5 images were taken for each section.
4
Quantifying Lung Pathology: Eosinophils and Mucus Cells
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Lung tissues were fixed in 10% neutral buffered formalin for 24 h before being transferred to 70% ethanol. Lungs were paraffin-embedded and sections stained for eosinophil quantification using Congo Red, or for mucus-secreting cells (MSCs) using an Alcian Blue (AB)-PAS staining. The number of MSCs and eosinophils were determined by morphological criteria around major airways under the light microscopy at ×400 original magnification and quantified as described previously46 (link). Scoring for histopathology (inflammatory infiltrates) was performed according to a set of morphological criteria, as previously described47 (link)–49 (link). All scoring was performed blinded. Histological photos were taken using the Zeiss AXIO imager microscope system.
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