Selective Single-Neuron Laser Ablation in a Zebrafish Larva Using a Confocal Microscope

Samenvatting

Source: Morsch, M., et. al., Triggering Cell Stress and Death Using Conventional UV Laser Confocal Microscopy. J. Vis. Exp. (2017)

In this video, we explore the technique of high-intensity UV laser ablation for precisely targeting and studying individual neurons in transgenic zebrafish. We demonstrate how to prepare, immobilize, and visualize neuronal responses, highlighting the cellular dynamics of injury and recovery.

Protocol

1. Prepare solutions for zebrafish embedding in agarose.

Resultaten

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Agarose low melting	Fisher-Scientific		Dissolve in egg water with tricaine (0.02%) at 0.8-1.5%
Tricaine	Argent Labs	MS-222	0.02%
Harvard standard wall Borosilicate with filament Capillary Glass	World Precision Instruments, Inc.	GC100F-10 (short) GC100F-15 (long)	Pull to a resistance of 2 -7 MΩ
Egg water			0.6 g Instant Ocean sea salt, 4ml of 1mg/ml methylene blue in 10 l deionized water
Heat block	Select BioProducts	Digital block heater (SBD110_)
Microscope	Leica	SP5
35mm glass bottom dish	MatTek Corporation	P35G-0-20-C