The 1 × 106 monocytes were seeded on the surface of each well, cultured for 1 and 3 days, respectively, and the macrophages were rinsed with PBS and fixed with 2.5% glutaraldehyde (Sangon Biotech) aqueous solution for 20 min. After rinsing for three times, they were dehydrated by 30, 50, 70, 90 and 100% gradient ethanol (Sangon Biotech) solution, each concentration was 10 min ×2 times, then placed in a freeze dryer, lyophilized 1 h and slowly sealed back to temperature. Then gold (platinum) was sprayed on the surface of the sample for 5 min using an ion sputtering apparatus to improve surface conductivity. After the sample preparation was completed, using a FESEM (SU8000) to observe the population morphology at a low magnification, and the individual morphology at a high magnification.
Gradient ethanol
Manufactured by Sangon
Gradient ethanol is a laboratory equipment designed to generate a linear gradient of ethanol concentration. It is used to create a controlled and gradual change in ethanol concentration, which is often required in various experimental and analytical applications.
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Lab products found in correlation
Glutaraldehyde, by Sangon (1 mentions)
Bovine serum albumin (bsa), by Beyotime (1 mentions)
Hematoxylin, by Merck Group (1 mentions)
Diaminobenzidine dab, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Inverted fluorescence microscope, by Nikon (1 mentions)
H e staining kit, by Sangon (1 mentions)
Xylene, by Sangon (1 mentions)
2 protocols using gradient ethanol
1
Morphological Analysis of Cultured Macrophages
2
Comprehensive Histological Staining Techniques
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The HE staining kit (Sangon, Shanghai, China) and Cell Apoptosis Detection Kit (Sangon) were adopted to conduct HE and TUNEL staining by strictly following the kits' instructions. For the completion of IF and IHC staining, the sections were deparaffinated with xylene (Sangon) and rehydrated with gradient ethanol (Sangon). Then, the sections were subjected to antigen retrieval with the microwave method, and endogenous peroxidase was removed with H 2 O 2 (Sangon). After blocking with bovine serum albumin (BSA) (Beyotime), the sections were incubated with primary antibodies at 4°C overnight. Subsequently, the sections were incubated with secondary antibodies. Then, for IHC staining, diaminobenzidine (DAB) (Sigma) and hematoxylin (Sigma) were used to stain the secondary antibody and nuclei; for IF staining, the nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI) (Sigma). Finally, the sections were observed under an inverted fluorescence microscope (Nikon, Tokyo, Japan). The antibodies are listed in Table 1.
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