A Hypertonic Saline Injection Technique for Inducing Glaucoma in Rats

JoVE Encyclopedia of Experiments

Neurowetenschap

0 weergaven • 3:15 min • July 8th, 2025

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

Take an anesthetized rat and apply an anesthetic to its eye to numb reflexes.

Trim whiskers and disinfect the area around the eye to prevent contamination.

Under a microscope, clamp the lower eyelid to bulge the eye, exposing the episcleral vein.

This vein connects to the anterior chamber via Schlemm's canal and trabecular meshwork for the outflow of aqueous humor.

Insert a microneedle with hypertonic saline into the exposed vein.

Inject the saline, displacing blood. Perform a second injection opposite the first for a uniform effect across the eye.

The saline enters the trabecular meshwork.

The high salt concentration causes osmotic stress, leading to cell death and tissue scarring. This scarring blocks aqueous humor outflow through the meshwork, increasing intraocular pressure.

The elevated pressure damages the retinal ganglion cells, which are neurons carrying visual information to the brain. The damage leads to glaucoma, a condition that causes vision loss.

In this procedure, after anesthetizing the animal with KAX cocktail, check the reflexes by giving pinches to its feet or tail to ensure the animal is unconscious. Then, trim the whiskers with scissors. Subsequently, saturate a cotton tip applicator with betadine solution and swab the area around the experimental eye.

Under the microscope, use a hemostat to clamp the bottom eyelid in order to bulge the eye, expose the episcleral vein, and restrict the eye movement. Next, carefully pierce the episcleral vein with the micro-needle at an angle between 10 and 20 degrees to the vein. A puncture into the vein is successful when the blood is observed to enter the tip of the micro-needle.

Slowly inject 50 microliters of saline into the vein. The veins will blanch white as the salt circulates through the vasculature, but some regions may maintain a blood-red appearance.

The single most critical step in this procedure occurs during the injection of the two molar saline. It is imperative that blanching in the vasculature occurs to ensure a successful outcome. Maintaining a steady needle in a 20-degree angle are crucial for this procedure.

After that, perform a second injection into the vein to ensure thorough damage of the retinal ganglion cell layer. Within minutes, one should see a distinct cloudy appearance through the iris of the eye as the salt circulates through the vascular system.

07:00

Een Model van glaucoom veroorzaakt door Circumlimbal hechtdraad bij ratten en muizen

Gerelateerde video's

0 Bekeken

07:11

Beoordeling van early stage open-angle glaucoom bij patiënten door geïsoleerde-check visuele opgeroepen potentieel

Gerelateerde video's

0 Bekeken

10:10

Volledige cirkel Cauterisatie van limbale vasculaire plexus voor chirurgisch geïnduceerd glaucoom bij knaagdieren

Gerelateerde video's

0 Bekeken

11:02

Methoden voor experimentele manipulaties na Optic Nerve doorsnijding in het zoogdier CZS

Gerelateerde video's

0 Bekeken

03:04

Experimental Glaucoma Induction via Iridocorneal Angle Obstruction In a Mouse Model

Gerelateerde video's

0 Bekeken

03:04

Induction of Elevated Intraocular Pressure in a Rat Model

Gerelateerde video's

0 Bekeken

06:35

Experimentele Glaucoom geïnduceerd door oculaire injectie van Magnetic microsferen

Gerelateerde video's

0 Bekeken

08:30

Glaucoom-inducerende Procedure in een In Vivo Rat Model en Whole-mount Retina Voorbereiding

Gerelateerde video's

0 Bekeken

09:03

Een omkeerbare silicium olie-geïnduceerde oculaire hypertensie model in muizen

Gerelateerde video's

0 Bekeken

06:19

Intracamerale injectie bij ratten met een laag risico op bijwerkingen

Gerelateerde video's

0 Bekeken

Laatst bijgewerkt: 15 augustus 2026