One head of an adult fish was subjected to cryosectioning for each genotype in a single experiment. Frozen ocular sections (10 μm thick) were prepared as described in the previous section. The cryosections were subjected to TUNEL staining by using the Click‐iT TUNEL Alexa Fluor 594 Kit (Invitrogen) according to the manufacturer’s protocol. The stained sections were coverslipped with VECTASHIELD Mounting Medium with DAPI (Vector Laboratories) and observed with a microscope (Axioplan2; Carl Zeiss).
Click it tunel alexa fluor 594 kit
Manufactured by Thermo Fisher Scientific
The Click-iT TUNEL Alexa Fluor 594 Kit is a labeling kit used to detect DNA fragmentation, a hallmark of apoptosis. The kit utilizes a modified TUNEL (Terminal deoxynucleotidyl transferase dUTP Nick End Labeling) method, wherein a fluorescent Alexa Fluor 594 dye is incorporated into DNA strand breaks. This allows for the visualization and quantification of apoptotic cells.
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2 protocols using click it tunel alexa fluor 594 kit
1
Cryosectioning and TUNEL Staining of Fish Eyes
2
Cryosectioning and TUNEL Staining of Fish Ocular Tissues
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One head of an adult fish was subjected to cryosectioning for each genotype in a single experiment. Frozen ocular sections (10 μm thick) were prepared as described in the previous section for in situ hybridization. The cryosections were subjected to TUNEL staining by using the Click-iT TUNEL Alexa Fluor 594 Kit (Invitrogen) according to the manufacturer’s protocol. The stained sections were coverslipped with PermaFluor Mountant (Thermo Fisher Scientific) after incubation with DAPI (1 μg/ml) for nuclear staining, and observed with a confocal laser scanning microscope (FV3000, Olympus).
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