Imaging Fixture Supported Mouse Liver Staging for Multiphoton Microscopy Based Vascular Visualization: A Stabilization Device-assisted Imaging Technique to Obtain Multi-dimensional Images of Blood Vessels in Mouse Liver

Abstract

Source: Rongrong, W. et al. Multiphoton Microscopic Observation of Vessels in Mouse Liver Tissue. J. Vis. Exp. (2021)

This video describes an imaging procedure to observe and study intravascular dynamics of a mouse liver tissue using multiphoton microscopy following stabilization of the organ. Multiphoton microscopy enables the imaging of biological tissue at high spatial resolution in vivo.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Mouse Preparation Anesthetize the mouse Prepare sodium pentobarbital (50 mg/kg) in a syringe. Grab the mouse (8-week-old male C57BL/6) with the left hand so that its belly is facing up and its head is lower than its tail. Disinfect the abdominal skin with 75% alcohol. Holding the syring…

Representative Results

Figure 1: Body organ imaging frame. (a) Heating pad: It maintains the mouse body temperature. (b) Bracket: It can provide a base to place the mouse and the adjustable suction cup position with knobs. (c) Suction cup: It is fixed by the bracket. The negative pressure provided by the negative pressure pump causes the liver to a…

Materials

1 mL syringe x 2	Hunan Pinan Medical Devices Technology	YA0551
5 W heating pad	BiolinkOptics Technology	BL336
75% absolute ethanol	Guangdong Guanghua Sci-Tech	1.17113.023
Absorbent cotton ball	Healthy Sanitation Kingdom
Mouse surgical instrument	RWD Life Science	SP0001-G	Including scissors and tweezers
Multiphoton microscopy	Olympus	FV1200MPE
Organ imaging fixture	BiolinkOptics Technology	BL336	Including suction cup, hose, negative pressure pump and bracket
Rhodamine B isothiocyanate-Dextran	Sigma	R9379
Sodium pentobarbital	Sigma	P3761-25G
Shaving machine	Lei Wa	RE-3201