Brain Cell Isolation from Zebrafish Larvae: A Technique to Obtain Intact Neurons, Macrophages, and Microglia from Zebrafish Brain Tissue
Published: April 30, 2023
Abstract
Source: Mazzolini, J. et al. Isolation and RNA Extraction of Neurons, Macrophages and Microglia from Larval Zebrafish Brains. J. Vis. Exp. (2018)
This video describes a protocol to isolate neurons, macrophages, and microglia from larval zebrafish brains by mechanically disrupting the brain tissue. The purified cell populations can further be used for genomic analysis.
Protocol
All procedures involving human participants have been performed in compliance with the institutional, national and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Homogenization NOTE: All steps are performed 4 °C. Add 1.5 mL Tricaine (15 mM) to 90 mm Petri dishes containing 50 larvae in 50 mL of E3 embryo medium treated with 200 µM PTU to termin…
Açıklamalar
The authors have nothing to disclose.
Materials
| 1-phenyl 2-thiourea (PTU) | Sigma | P7629 | |
| Hepes | Gibco | 15630-056 | |
| D-Glucose | Sigma | G8644-100ML | |
| HBSS 1X | Gibco | 14170-088 | |
| Percoll | GE Healthcare | 17-0891-02 | |
| HBSS 10X | Gibco | 14180-046 | |
| DPBS 1X | Gibco | 14190-094 | |
| Tricaine (MS222) | Sigma | A5040 | |
| Sterilin standard 90 mm petri dishes | ThermoFisher | 101VIRR | |
| Surgical micro-scissors | Fine Science Tools | 15000-00 | |
| 3 mL Pasteur plastic bulk pipette | SLS | PIP4206 | |
| Glass homogenizer | Wheaton | 357538 | |
| Sterilin standard 55 mm petri dishes | ThermoFisher | P55V | |
| Percoll | GE Healthcare | 17-0891-02 | |
| 40 µm cell strainer | Falcon | 352340 | |
| 50 ml polypropylen conical tube | Falcon | 352070 | |
| Centrifuge | Eppendorf | 5804 R | |
| Normal goat serum (NGS) | Cell Signalling | 5425S | |
| 35 μm cell strainer cap | BD | 352235 |
Etiketler
Bu Makaleden Alıntı Yapın
Brain Cell Isolation from Zebrafish Larvae: A Technique to Obtain Intact Neurons, Macrophages, and Microglia from Zebrafish Brain Tissue. J. Vis. Exp. (Pending Publication), e20637, doi: (2023).