Optical Recording of Neuronal Activity in Brain Slices Stained with a Voltage-Sensitive Dye

In This Article

Abstract

Source: Tominaga, Y., et al. Wide-field Single-photon Optical Recording in Brain Slices Using Voltage-sensitive Dye. J. Vis. Exp. (2019).

This video demonstrates a procedure for observing changes in neuronal activity in a brain slice stained with a voltage-sensitive dye. The dye reflects changes in the neuron's membrane potential by altering its fluorescence intensity. Through the application of electrical stimuli and advanced imaging techniques, this process enables visual tracking of real-time changes in neuronal membrane potential based on VSD fluorescence.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Daily Preparation of Experimental Apparatus

Turn on the amplifier, computer, and camera system, and check that the software is running.
Place artificial cerebrospinal fluid (ACSF) in a 50 mL tube and bubble with carbogen.
Use a peristaltic pump to circulate the ACSF. Adjust the flow rate to approximately 1 mL/min.
Adjust the height of the suction pipette so that the liquid level inside the experiment chamber is always constant.
NOTE: ....

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Results

High-speed imaging system with optical components; diagram shows photonic excitation setup.
Figure 1. Recording system for optical signals from slice preparations. (A) A photograph of the microscope used to image the slices in the current manuscript. The optics consist of an objective lens (5x NA0.60), a mirror box for dichroic filter .......