This study demonstrates a method for imaging immune responses in the eye using optical coherence tomography (OCT) in a mouse model. The approach involves inducing uveitis through the injection of heat-killed Mycobacterium tuberculosis and assessing inflammation via OCT imaging.
Take an anesthetized mouse pre-immunized with heat-killed Mycobacterium tuberculosis, or Mtb, which induces an immune response, and generates Mtb-specific T cells.
Administer eye drops for pupil dilation to enhance light penetration for improved eye imaging. Apply a gel to prevent corneal dehydration.
Wrap the mouse in gauze to maintain body warmth and place it in the optical coherence tomography or OCT system.
Image the eyes to obtain cross-sectional images of the anterior chamber, including the cornea and lens, and the posterior chamber, including the retina.
Examine to ensure the absence of inflammation.
Inject heat-killed Mtb into the vitreous cavity. The immunogen spreads across the eye and triggers an immune response, inducing the infiltration of preformed Mtb-specific T cells and other immune cells, resulting in inflammation termed uveitis.
OCT of the anterior chamber reveals infiltrated immune cells and corneal edema, while OCT of the posterior chamber indicates vitreous inflammation.
Score OCT images based on inflammation severity to quantify uveitis progression.
After anesthetizing the mouse, dilate the pupil with one drop of 2.5% phenylephrine, avoiding excess droplets entering the nose or mouth. After two to three minutes, dab off the excess liquid, and cover the eye with 0.3% hypromellose gel. Next, wrap the mouse in a layer of surgical gauze to maintain body warmth, then place the mouse on the animal cassette, and position the head with the bite bar.
To acquire the optical coherence tomography, or OCT images, turn on the OCT imaging system, and open the imaging software. For posterior chamber imaging, bring the OCT close to the eye's surface, taking care not to bring the surface of the lens in contact with the eye.
Once the eye is correctly positioned, stop the Fast Scan, select the Volume Scan protocol, and activate the scan with the Aim option. For posterior segment images, adjust until the optic nerve is centered in the horizontal B-Scan alignment image and the retina is aligned with the vertical alignment axis. For anterior segment images, adjust the position to center the apex of the cornea in both the horizontal B-Scan alignment image and vertical B-Scan alignment image. Finally, click on Snapshot to capture the volume scan image. Then click on Save.
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