Two-Photon Microscopy for Imaging Neuronal Morphology in a Mouse Pup

Abstract

Source: Mizuno, H., et al. In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice. J. Vis. Exp. (2018)

This video showcases two-photon microscopy being used to image neurons in the brain of an anesthetized transgenic mouse pup. The two-photon laser allows for deeper imaging with high spatial resolution and minimal light scattering, enabling the detailed capture of neuronal morphology, including dendritic projections.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

Results

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
pK031. TRE-Cre	Authors	-	Available from RIKEN BRC and Addgene
pK029. CAG-loxP-STOP-loxP-RFP-ires-tTA-WPRE	Authors	-	Available from RIKEN BRC and Addgene
pK273. CAG-loxP-STOP-loxP-CyRFP-ires-tTA-WPRE	Authors	-	Available from authors
Isoflurane	Wako	099-06571
410 Anaesthesia Unit (isoflurane gas machine)	Univentor	8323101
Vetbond (tissue adhesive)	3M	084-1469SB
MµltiFlex Round (loading tip)	Sorenson	13810
Gelfoam (gelatin sponge)	Pfizer	09-0353-01
Agarose	Sigma	A9793	Low melting point
Round-shaped coverslip	Matsunami	-	Custom made
Unifast 2 (dental cement)	GC	-
Titanium bar	Authors	-	Custom made (see Figure 1G)
Rimadyl (carprofen)	Zoetis	-	Injectable
2-photon microscope	Zeiss	LSM7MP
Titanium-sapphire laser	Spertra-Physics	Mai-Tai eHPDS
Titanium plate	Authors	-	Custom made (see Figure 2A)
IMARIS, FilamentTracer, MeasurementPro	BITPLANE
Goniometer stage	Thorlabs	GN2/M