Biocytin Staining and Visualization

Biocytin filling and staining were performed as described previously [24 (link)]. To fully infuse the entire cell with the dye, electrophysiological recording lasted for more than 15 min. After electrophysiological recording, brain slices were fixed in neutrally buffered 4% Paraformaldehyde (PFA) solution at 4 °C overnight, washed three times in phosphate buffer saline (PBS) and then, stained with streptavidin-Cy3 (1:100 of 1 mg/ml, S6402, Sigma) for 3 h at room temperature and mounted on glass coverslips for visualization of the biocytin-filled neurons.

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Zhu F., Shi Q., Jiang Y.H., Zhang Y.Q, & Zhao H. (2024). Impaired synaptic function and hyperexcitability of the pyramidal neurons in the prefrontal cortex of autism-associated Shank3 mutant dogs. Molecular Autism, 15, 9.

Publication 2024

Corresponding Organization :

Other organizations : Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Yale University

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Variable analysis

independent variables

Duration of electrophysiological recording (more than 15 min)

dependent variables

Degree of infusion of the dye (biocytin) into the entire cell

control variables

Biocytin filling and staining procedure (as described previously [24])
Fixation of brain slices in neutrally buffered 4% Paraformaldehyde (PFA) solution at 4 °C overnight
Washing of brain slices three times in phosphate buffer saline (PBS)
Staining of brain slices with streptavidin-Cy3 (1:100 of 1 mg/ml, S6402, Sigma) for 3 h at room temperature
Mounting of brain slices on glass coverslips for visualization of the biocytin-filled neurons

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