Preparation of High-Viability Single-Cell Suspensions from African Turquoise Killifish Brain Tissue.

Cold Spring Harb Protoc

Neuroplasticity and Neuroproteomics Research Group, Animal Physiology and Neurobiology Section, Department of Biology, KU Leuven, 3000 Leuven, Belgium.

Published: February 2024

Studying the brain at the single-cell level has become increasingly popular in recent years. This, however, remains challenging, especially in emerging model organisms. To carry out single-cell sequencing, the preparation of a high-viability single-cell suspension is critical. In this protocol, we describe how to prepare a high-viability single-cell suspension starting from brain tissue of the African turquoise killifish (). The protocol consists of dissection, enzymatic and mechanical dissociation of the brain tissue, and debris removal. The protocol described here has been successfully used for 10× Genomics single-cell sequencing of the telencephalon of adult killifish, which requires a cell viability of at least 70%. In addition to single-cell sequencing experiments, the single-cell suspension generated can be used for other applications, including cell culture and flow cytometry.

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http://dx.doi.org/10.1101/pdb.prot107829DOI Listing

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