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

JoVE Encyclopedia of Experiments

Biologie

0 weergaven • 4:09 min • July 8th, 2025

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

Multiphoton microscopy enables the capture of high-resolution, three-dimensional images of the fluorescently-labeled vasculature within the living tissue. Begin with an anesthetized mouse in the supine position.

Carefully inject a suspension of fluorophore conjugated-dextran into its tail vein. The injected molecules flow through the circulation and reach organs like the liver. The fluorescent dye labels the vascular serum, aiding blood vessels' visualization, while the attached high-molecular-weight dextran prevents their leakage from the vasculature.

Prep the mouse's abdominal area. Transfer it onto the base of the organ imaging frame equipped with an imaging lens-containing suction ring connected to a vacuum pump. Incise along its lower sternal border to expose the liver.

Guiding the suction ring to the liver, apply vacuum or negative pressure which facilitates the adsorption of the liver to the lens in the suction ring. This step reduces the motion artifacts during imaging. Transfer the mouse-containing imaging frame onto the multiphoton laser scanning microscope stage.

Cover the lens with saline which acts as immersion medium for high-resolution imaging. During imaging, the focused long-wavelength lower-energy photons allow deeper tissue penetration and excite fluorophores in the blood vessels at the focal point, causing the emission and detection of fluorescent signals.

A series of scans from the selected optical sections at different depths recreates a complete three-dimensional image of the blood vessels within the tissue.

After confirming a lack of righting reflex in an anesthetized 8-week-old male C57BL/6 mouse, wipe the mouse's tail with 75% alcohol and use a 1-microliter syringe equipped with a 30-gauge needle to inject 10 milligrams per milliliter of freshly prepared Rhodamine B isothiocyanate-dextran in 100 microliters of Rhodamine B isothiocyanate-dextran into the caudal tail vein.

When all of the solution has been delivered, use a cotton swab to apply pressure to the puncture site and use gauze wet with sterile water to soak the abdominal fur. Use a razor to shave the abdomen, making strokes in the direction of the fur, and place the mouse onto a 37 degrees Celsius, alcohol-disinfected heating pad.

To fix the mouse liver with a body organ imaging frame, first, place a clean 5-millimeter diameter suction cup in a fixed position and wipe the heating pad and suction cup with 75% alcohol. Connect the suction cup to the vacuum pump hose and turn on the pump.

On a 75% alcohol-sterilized table, use surgical scissors to cut 2 centimeters of skin from the lower sternal border of the mouse and expose the liver. Place the mouse and the heating pad onto the holder base of the body frame and adjust the organ imaging fixture so that the suction cup can hold the liver. Then, use a negative pressure of 30 to 35 kilopascals for suction so that the liver attaches to the cup.

To set up the multiphoton laser scanning microscope, turn on the microscope and select the 60x objective. Fix the frame and the mouse under the objective and add a drop of normal saline to the cup large enough to cover the lens. Adjust the objective so the lens just touches the normal saline and turn on the laser software.

09:34

Visualiseren van angiogenese door Multiphoton microscopie In Vivo in genetisch gemodificeerde 3D-PLGA/nHAp steiger voor Calvarial kritische bot Defect reparatie

Gerelateerde video's

0 Bekeken

06:53

Een custom multiphoton microscopie platform voor live beeldvorming van muis hoornvlies en conjunctiva

Gerelateerde video's

0 Bekeken

06:29

Real-time intravitale multifotonenmicroscopie om gerichte echografie en microbubbelbehandelingen te visualiseren om de permeabiliteit van de bloed-hersenbarrière te vergroten

Gerelateerde video's

0 Bekeken

10:48

Intravitale Microscopie Imaging van de lever volgende Leishmania Infectie: een beoordeling van de lever Hemodynamica

Gerelateerde video's

0 Bekeken

11:44

Long Term intravitale Multiphoton Microscopie Beeldvorming van immuuncellen in gezonde en zieke lever behulp CXCR6.Gfp Reporter Muizen

Gerelateerde video's

0 Bekeken

07:19

Langdurig High-Resolution intravitale microscopie in de long met een vacuüm gestabiliseerd Imaging Window

Gerelateerde video's

0 Bekeken

07:06

Twee-foton Intravital Imaging van leukocyten tijdens de immuunrespons bij Lipopolysaccharide-behandelde muis lever

Gerelateerde video's

0 Bekeken

06:33

Multifotonen microscopische observatie van vaten in leverweefsel van muizen

Gerelateerde video's

0 Bekeken

07:50

Multifoton intravitale beeldvorming voor het monitoren van leukocytenrekrutering tijdens arteriogenese in een Murine Hindlimb-model

Gerelateerde video's

0 Bekeken

05:27

In vivo beeldvorming van leversferoïden geënt in de voorste oogkamer van het muizenoog

Gerelateerde video's

0 Bekeken

Laatst bijgewerkt: 15 augustus 2026