Protocol for Dissection and Dissociation of Zebrafish Telencephalon for Single-Cell Sequencing
Mehmet İlyas Coşacak, Prabesh Bhattarai, Çağhan Kızıl
Abstract
Single-cell sequencing (sc-Seq) is a powerful tool to investigate the molecular signatures of cell types in a complex mixture of cells. A critical step in sc-Seq is preparing a single-cell suspension with a high number of viable cells. Here, we show how to dissect zebrafish telencephalon and how to dissociate it into a single-cell suspension. This is followed by flow cytometry-based sorting to enrich for neural progenitor stem cells. Our technique typically yields 70,000 live cells from one zebrafish telencephalon. For complete details on the use and execution of this protocol, please refer to Cosacak et al. (2019).
Topics & Concepts
CerebrumZebrafishCell sortingBiologyCellFlow cytometryComputational biologyCell biologyNeuroscienceMolecular biologyGeneGeneticsCentral nervous systemSingle-cell and spatial transcriptomicsCell Image Analysis TechniquesZebrafish Biomedical Research Applications