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

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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).

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