Hello, all!
I'm inducing/measuring LTP in in-vitro mouse brain slices. My stimulating electrode is supposed to be in CA3 and my recording electrode is supposed to be in CA1 of the hippocampus. I was having trouble finding the Schaffer collaterals using the usual top lighting from a dissecting microscope, so I've set up a couple different ways to get the backlighting I prefer on the slice (involving such common items as a flashlight on a cell phone and a closet light consisting of a bar of LEDs). I haven't come up with anything ideal yet. I was wondering, what does everyone else use for lighting?
Thanks in advance!
As I anderstand you wish to see Schaffer collaterals. You can't see them
All you need to see is cell layers and stratum radiatum, good anouth to be able to place electrodes. First place stimulation electrode in strat. rad. clouser to cell layer, were SC more dense, and start to place recording electrode somewhere in the middle of strat. rad., where dendrits is, (this is for field potential recording).
Than try to get EPSP (field potential) by increasing stimulation intensity. You may need to replace recording electrode several times to get good shaped potential.
I use a fiber optic ring light attached to a light source. The link below is an example of something similar to what you'll need. It attaches to the underside of your scope.
https://www.edmundoptics.com/f/Fiber-Optic-Ring-Light-Guides/11577/
Thank you Ivan Goussakov and Melinda Peters ! Melinda, are you able to see the Schaffer collaterals with lighting coming from above the slice? I had difficulty with the dissecting scope's original lighting (I think it is LED) shining from above, I had issues with glare and seeing where exactly to place the electrodes.
Hi, sorry for hijacking your channel, but I'm hoping Ivan Goussakov Melinda Peters Jacci Newel can help me.
I am the founder of a nonprofit The Reagent Project, that redistributes donated scientific equipment and reagents to scientists in need. I currently have a university in London offering to donate reagents relating to neuroscience research in mice. The items are too many to list but things relating to electrophysiology/calcium imaging/histology/RTPCR/behavior.
Do you know of any scientists particularly in universities struggling for funding to whom these items might be useful.
Thanks.
Jacci Newel ,sorry for the delay! I do not get notifications when someone mentions my name. I can see it best when I am first placing my slices (they are slightly submerged at that point). Sometimes there is glare, but honestly you get used to understanding where everything is after a while. Good luck!
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