Why no fixatives?

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Why no fixatives?

Post  Brad on Tue Aug 16, 2011 3:21 pm

Greetings,

I am a new user of MEAs, and would like to correlate synaptic architecture with network-level activity. One straightforward way to do this is to use immunocytochemistry to visualize synaptic proteins. However, this usually requires fixation with formaldehyde. The manual states that this should not be done.

My question is, why? What deleterious effects do fixatives have on the MEA?

Thanks,

Brad.

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Re: Why no fixatives?

Post  Frank MCS on Tue Aug 16, 2011 6:54 pm

Dear Brad,
the problem is not so much that the fixative damages the array, but that it is very hard to remove the fixed tissue from the array. That would lead to a insulation layer of fixed cell debris on the array. If you have a safe method to reverse your fixation procedure, you might try that, maybe on an old array.

Best regards Frank MCS
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could proteases remove fixed cell debris?

Post  Brad on Wed Aug 17, 2011 2:25 am

For example, could trypsin remove it?

I suppose this is an empirical question that I can answer, but if you or anyone else can say, I'd appreciate it. That would save me the use of an old array.

cheers,

Brad.

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Re: Why no fixatives?

Post  ThomasMCS on Wed Aug 17, 2011 6:30 pm

We had good success with the use of Terg-a-zyme (that is mostly Trypsin based). However with fixed cell debris you should allow the enzymes about 2-3 days to do their work...for normal cell culture coatings overnight is sufficient.

Thomas

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