4.4.3. TUNEL Assay

Cerebral organoid tissue slices derived from the control and TTX-exposed groups underwent investigation for cell apoptosis utilizing the One Step TUNEL Apoptosis Assay Kit. TUNEL labeling was conducted in conjunction with immunofluorescence staining to discern the various brain cell types experiencing apoptosis post TTX exposure. Primary antibodies targeting neuronal nuclear antigen (NeuN) or GFAP were employed, along with corresponding secondary antibodies (Alexa Fluor 488-conjugated mouse or rabbit IgG, Thermo Fisher Scientific). Cell nuclei were identified using DAPI, and imaging was carried out utilizing a Leica STELLARIS 5 confocal microscope.