Kidneys were immersed in 10% formalin and embedded in paraffin. Sections (5 μm thick) were processed for the histopathology study. The histopathology was investigated using hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) staining. In addition, small pieces of renal cortex were fixed in 2% glutaraldehyde, postfixed in 1% osmium tetroxide, and embedded in Araldite M (Sigma-Aldrich). Ultra-thin sections were counterstained with uranyl acetate and lead citrate and examined using a transmission electron microscope. Electron micrographs were used to determine the glomerular basement membrane (GBM) thickness and podocyte foot process width (FPw) as previously described [17 (link), 18 (link)].
Araldite m
Manufactured by Merck Group
Sourced in United States
Araldite M is a two-component epoxy resin system designed for bonding, sealing, and encapsulating applications. It is characterized by its high mechanical strength, chemical resistance, and thermal stability.
Automatically generated - may contain errors
12 protocols using araldite m
1
Histological and Ultrastructural Kidney Analysis
2
Ultrastructural Examination of Renal Cortex
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Small pieces of renal cortex tissues was obtained and fixed in 2.5% glutaraldehyde. Then, they were washed with PBS (0.01 M), and post-fixed with 1% osmium tetroxide. After gradient dehydration of acetone, the tissues were embedded in Araldite M (Sigma Aldrich). Ultrathin sections were made using an ultramicrotome (Leica, Germany), and stained with uranyl acetate and lead citrate. The sections were examined through a transmission electron microscope (H-7700, Hitachi, Japan).
3
Ultrastructural Renal Cortex Analysis
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Small pieces of the renal cortex tissue block were fixed in 2.5% glutaraldehyde and washed with PBS (0.01 M). Then, they were postfixed with 1% osmium tetroxide, dehydrated with an acetone gradient, and embedded in Araldite M (Sigma Aldrich). Ultrathin sections were counterstained with uranyl acetate and lead citrate and examined with a transmission electron microscope (H-7700, Hitachi, Japan). The thickness of the GBM was measured with Image-Pro Plus 6.0 image analysis software.
4
TEM Sample Preparation Protocol
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For TEM, the cells in different groups were prefixed with 2.5% glutaraldehyde in 0.1 m PBS for 24 h at 4 °C. Then, they were washed with PBS, and post-fixed with 1% osmium tetroxide. After gradient dehydration of acetone, they were embedded in Araldite M (Sigma Aldrich). Ultrathin sections (1 µm) were subsequently cut with an ultramicrotome, mounted on nickel grids, and stained with uranyl acetate and lead citrate. At last, the samples were sent to the electron microscope room at Wenzhou Medical University for subsequent processing and testing using a transmission electron microscope (H-600A-2; Hitachi, Tokyo, Japan).
5
Microscopic Analysis of MIN6-CB Cells
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Bright field and fluorescence images of MIN6-CB cells were obtained using a fluorescence microscope (BZ-9000; Keyence, Osaka, Japan). For transmission electron microscopy, the cultured cells were fixed in 2.5% glutaraldehyde at 4 °C for 12 h. The specimens were dehydrated after post-fixation with 1% OsO4 and embedded in Araldite M (Sigma-Aldrich, St. Louis, MO). Semi-thin sections (1 μm) were prepared and treated with 1% toluidine blue. Ultrathin sections were analyzed using the JEOL 1010 transmission electron microscope (JEOL Ltd., Tokyo, Japan).
6
Renal Cortex Ultrastructural Analysis
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Small pieces of renal cortex were fixed in Trump's fixative (EMS, number 11750) and embedded in Araldite M (Sigma-Aldrich, number 10951). Ultrathin sections were counterstained with uranyl acetate and lead citrate and examined with a transmission electron microscope.
7
Transmission Electron Microscopy of Renal Cortex
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Sections of renal cortex were fixed in Trump’s solution (EMS, 11750) and embedded in Araldite M (Sigma Aldrich, 10951). Ultrathin sections were counter-stained with uranyl acetate and lead citrate and examined with a transmission electron microscope (JEM-1011, JEOL, Peabody, MA).
8
Transmission Electron Microscopy Sample Preparation
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Mouse tissues were fixed in glutaraldehyde, washed with PBS, and subsequently fixed in osmium tetroxide. After dehydration using an acetone gradient, the tissues were embedded in Araldite M (Sigma-Aldrich, St. Louis, MO, USA). Ultrathin sections were obtained using an ultramicrotome (Leica, Wetzlar, Germany) and then stained with uranyl acetate and lead citrate. Finally, the sections were examined using a transmission electron microscope (H-7700, Hitachi, Tokyo, Japan) for further analysis.
9
Renal Tissue Ultrastructural Analysis
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Two percent glutaraldehyde was used to fix renal tissue samples. Then, they were washed in PBS (0.01 M) and postfixed with 1% osmium tetroxide. After gradient dehydration in acetone, the tissue samples were embedded in Araldite M (Sigma Aldrich, St. Louis, MO, USA). Ultrathin sections were cut using an ultramicrotome (Leica, Wetzlar, Germany) and stained with uranyl acetate and lead citrate. The sections were examined with a transmission electron microscope (H-7700, Hitachi, Japan).
10
Transmission Electron Microscopy of Renal Tissue
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Small pieces of renal cortex tissues were obtained and fixed in 2.5% glutaraldehyde. Then, they were washed with PBS and post-fixed with 1% osmium tetroxide. After gradient dehydration of acetone, the tissues were embedded in Araldite M (Sigma-Aldrich). Ultrathin sections were made using an ultramicrotome (Leica, Germany), and stained with uranyl acetate and lead citrate. The sections were examined through a transmission electron microscope (H-7700, Hitachi, Japan).
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