Intracameral Injection of Trypan Blue in Rat Eyes to Assess Delivery into the Anterior Chamber

Begin by loading five microliters of trypan blue into a sterile 10 microliter glass Hamilton syringe, equipped with a 34 gauge blunt needle. Then insert the loaded syringe needle through the self-sealing corneal tunnel into the anterior chamber. Inject the solution in the eye and hold the needle in place for two to three seconds until all fluid clears.

Pull out the needle gently. Examine under slit microscopy. Evaluate the depth of the anterior chamber to exclude shallowing, and verify the presence of trypan blue in the anterior chamber.

Using this protocol, the corneas of Sprague-Dawley rats were injected with trypan blue. Immediate slit lamp examination post-injection revealed that the anterior chamber was visibly stained with trypan blue, confirming the successful delivery of the substance to the target area.