Abstract
Purpose :
The ex vivo retina preparation allows exceptional access to single-cell physiology, but it is limited in studies of many retinal diseases because the metabolic environment is replaced by an artificial solution, and there is no blood flow. We are developing a new preparation of the intact rodent eye perfused with artificial blood to maintain functional light responses for up to several days outside the body.
Methods :
First, we isolate the eye from a rodent with the optic nerve and ophthalmic artery intact. Then we cannulate the artery and perfuse it with oxygenated artificial blood at 30 µL/min using a microfluidic pump. We will measure the overall health of the outer retina throughout the experiment using the flash electroretinogram (ERG) while monitoring oxygen pressure, glucose, temperature, and pH with inline sensors in our fluid path. Once we establish a reliable system that maintains a healthy ERG for several hours, we will build a projection system for spatially patterned light stimulation and a record retinal ganglion cell (RGC) spikes via dense electrical probes inserted into the optic nerve. A schematic of the preparation is provided in Figure 1.
Results :
We have established successful perfusion of the eye by imaging the vasculature with a fluorophore. By the time of the meeting, we hope to present both ERG data and RGC spike recordings.
Conclusions :
Our endeavors to devise a new preparation for capturing light-induced RGC spikes from the intact rodent eye hold the potential to provide direct insight into the functional output of the retina. This approach ensures precise experimental manipulation of disease-relevant variables, including blood pressure, glucose, and oxygen levels.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.