Protocol for Dissection and Dissociation of Zebrafish Telencephalon for Single-Cell Sequencing
| Autor: | Caghan Kizil, Prabesh Bhattarai, Mehmet Ilyas Cosacak |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Telencephalon
Cell type Biology General Biochemistry Genetics and Molecular Biology Article Flow cytometry methods [Dissection] medicine Animals Humans lcsh:Science (General) Zebrafish Progenitor methods [Single-Cell Analysis] General Immunology and Microbiology medicine.diagnostic_test Cerebrum General Neuroscience Dissection surgery [Zebrafish] biology.organism_classification cytology [Telencephalon] Cell biology medicine.anatomical_structure Single cell sequencing Stem cell Single-Cell Analysis ddc:600 lcsh:Q1-390 |
| Zdroj: | STAR Protocols STAR Protocols 1(1), 100042 (2020). doi:10.1016/j.xpro.2020.100042 STAR Protocols, Vol 1, Iss 1, Pp 100042-(2020) |
| ISSN: | 2666-1667 |
| DOI: | 10.1016/j.xpro.2020.100042 |
| Popis: | SUMMARY 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). Graphical Abstract Highlights • We describe the dissection of adult zebrafish telencephalon. • We detail a protocol for dissociation of telencephalon into single-cell suspension. • We describe a procedure for flow cytometry-based sorting to enrich NSPCs. • Our technique typically yields 70,000 live cells from one zebrafish telencephalon. 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|