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Detection of apoptosis in retinal tissue by TUNEL assay

YZ Yihuan Zeng
GM Guangmeng Mo
XW Xiaoyv Wang
YY Yan Yang
YD Yan Dong
RZ Ruiying Zhong
NT Ni Tian
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Retinal cell apoptosis was identified using the transferase-mediated dUTP nick end labelling (TUNEL) method. The TUNEL Staining Kit provided by Elabscience (China) was employed following manufacturer’s directions. Paraffin sections of retina were dewaxed with xylene, rehydrated with gradient alcohol, cleaned with PBS, and treated with ProteinaseK working solution for 20 min. After washing with PBS, an equilibration working buffer of 100 µl TdT was added for 20 min. The liquid was aspirated, and TdT enzyme working solution (50 µl) was added to each slide. Incubation followed in a dark wet box at 37 ℃ for 1 h. After washing, DAPI was incorporated and cells were stained for 5 min. Subsequently, pieces were sealed and re-washed. The staining results were observed using a fluorescence microscope with an excitation wavelength of 450–500 nm and a detection wavelength of 515–565 nm (green fluorescence).

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