Organotypic Culture of Adult Rabbit Retina

Hi, my name is, And today I'll show you the incubation methods of a adult rabbit retina and the subsequent gene.Great. Washing several signs. I think it also dries out the retina.

Yeah, that's right. That's right, that's right.Yeah. So you mean all absolve water from the cell so that dries out.

Yeah, I think, yeah. So tissue, that's fine. Do you Oh, it's okay.

It's okay. So now gang sell side up very front. This, let's first introduce ourselves.

Yeah, I'm Tiana. I'm Amari. You've already met.Yeah.

So what can I do with this method? Well, you can culture adult webinars. And since you can also gene gun it and express for instance GFP, it's very useful to see the morphology of sickle cells very clearly.

So you can do anatomy on these cells or also you can label cells with DS red or with GFP to find them for later recording, amp recording for instance. It is also possible to express, for instance, synaptic marker proteins like PSD 95 GFP, and also other proteins which you might be interested in studying in the retina, like ion channels. It could also be used to express inhibitory RNAs.Yeah.

So what are the limitations of this na, Well, I think probably the most important is the time. You cannot keep the retina for longer than six days. I would say we don't trust the retina after six days anymore.

This is because of, this is mized spread Probably, yeah. And now if you just want to express GFP, which comes on after 24 or 48 hours, that's no problem at all. But in some cases, for instance, if you wanna express inhibitory RNAs, now these effects may take four days or even more to develop.

And you have to keep these timelines in frame. In, in, in. You have to, to, to make sure that you will see your effect in a day in, in a time which is shorter than six days because you can't reliably keep it longer.

So this may or may not be possible with your, with the experiment you are, you are planning to do. But this certainly is the biggest limitation. It is medium term, let's say.

So some, some RN ai work within four days, but the others work more.Right. Maybe it's takes five or six days or more. In that case, it's not suitable for this kind of a wrong may.

Yeah, may not be possible. I should say perhaps that during the time we use for incubation, that is up to six days. We do not see any signs of re reorganization of the retina.

Really the retinas do look normal morphologically.Okay. So six days morphology is good, but some visual carry, it looks unreliable.Physiologically.Yes. And also you start to see swelling of den, something like that.

So no, what I meant with the retina looks normal is the layer structure of the retina is still intact. And in principle, some of the cells are still life responsive. It might be that the photoreceptors die or degenerate after six days too.

So that might explain why we can't reliably record life responses anymore. But by and large, mostly the retina looks normal.Okay. Any other points Or, okay.Okay.