Biolistic transfection and expression analysis of acute cortical slices.

ABSTRACT:

Background

Biolistic gene gun transfection has been used to transfect organotypic cultures (OTCs) or dissociated cultures in vitro. Here, we modified this technique to allow successful transfection of acute brain slices, followed by measurement of neuronal activity within a few hours.

New method

We established biolistic transfection of murine acute cortical slices to measure calcium signals. Acute slices are mounted on plasma/thrombin coagulate and transfected with a calcium sensor. Imaging can be performed within 4 h post transfection without affecting cell viability.

Results

Four hours after GCaMP6s transfection, acute slices display remarkable fluorescent protein expression level allowing to study spontaneous activity and receptor pharmacology. While optimal gas pr

SUBMITTER: Hamad MIK 

PROVIDER: S-EPMC7125026 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature