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.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Mouse Preparation
2. Fixing the Mouse Liver with the Body Organ Imaging Frame
NOTE: The commercial organ imaging frame has not been released yet.
3. Adjusting the Multiphoton Laser Scanning Microscope
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 adsorb to the transparent lens in the center of the suction cup. (d) Flexible pipe: It connects the suction cup with the pressure pump. The pressure pump provides negative pressure to the suction cup. (e) Centrifuge tube: The centrifuge tube in the middle of the flexible pipe can hold the liquid, to prevent these liquids from being sucked into the vacuum pump. (f) Negative pressure pump: It is connected to the suction cup by the flexible pipe. The negative pressure pump can provide negative pressure to the suction cup to make the liver cling to the lens.
Figure 2: A close-up image of the liver attached to transparent lens. The transparent lens is in the center of the suction cup. The pump provides negative pressure to the suction cup to make it suck the liver and the liver is attached to the transparent lens. Because the suction-faced liver is a bit away from the mouse, the effect of breathing and the heartbeat on artifacts is reduced. The transparent lens can provide a stable view for the multiphoton microscope.
The authors have nothing to disclose.
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 |