Native adipose tissue and apoptotic adipose tissue were harvested, fixed in 4% neutral paraformaldehyde (Biosharp, USA), dehydrated in gradient ethanol, transparent in xylene, and finally embedded in paraffin and cut into 5–6 µm thick sections. The morphology was detected by H&E staining (Solarbio, China) according to the manufacturer’s protocol. The apoptosis of adipose tissue was evaluated by TdT-mediated dUTP nick-end labeling (TUNEL) assay, which was usually used for apoptosis detection.17 (link) According to the manufacturer’s instruction of the TUNEL Apoptosis Assay Kit (Beyotime, China), sections were incubated with TUNEL reagent at 37°C for 60 min. Then, DAPI (1:1000, Solarbio, C0050) was used to further mark the nuclei at room temperature for 5 min. Images were captured by confocal microscopy (Olympus FV1000, Japan).
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Adipose Tissue Apoptosis Evaluation
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Other organizations : ShenZhen People’s Hospital, Sichuan University
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Variable analysis
independent variables
- Native adipose tissue
- Apoptotic adipose tissue
- Morphology of adipose tissue
- Apoptosis of adipose tissue
- 4% neutral paraformaldehyde (Biosharp, USA) for fixation
- Gradient ethanol for dehydration
- Xylene for transparency
- Paraffin embedding
- 5-6 µm thick sections
- H&E staining (Solarbio, China)
- TUNEL Apoptosis Assay Kit (Beyotime, China)
- DAPI (1:1000, Solarbio, C0050) for nuclei staining
- Confocal microscopy (Olympus FV1000, Japan) for imaging
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