Immunohistochemical and Calcium Imaging Methods in Wholemount Rat Retina

The overall goal of this procedure is to selectively label retinal ganglion cells in the whole mount preparation via an optic nerve stump injection of a calcium indicator dye. This is accomplished by first injecting a very high affinity calcium indicator dye, such as flow four into the optic nerve stump. Next, the retina is isolated from the eye cup and divided into quadrants to mount onto a glass slide In a recording chamber set up to mimic physiological conditions, the contributions of voltage gated calcium channels to the calcium signal.

In retinal ganglion cells can be measured in depolarized cells using channel blockers. Ultimately, the results can provide semi-quantitative measurements of the contribution of the voltage educated calcium channels to a signal such as a high potassium evoked calcium signal. The main advantage of this technique over existing methods, such as electroporation and bulk loading, is the ability to selectively label retinal ganglion cells and their axons in a whole mount preparation Store, dissected rad eyes and hibernate a aims medium or similar solution.