Immunostaining on and Cleaning of MEAs

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Immunostaining on and Cleaning of MEAs

Post  portmann.johnny on Mon May 11, 2015 2:54 pm

Hi there,

We have two of the standard MEAs purchased from MCS, the 60MEA100/10-ITO and the 60MEA200/30-ITO. We initially coated the surface of the culture chamber with gelatin before plating our cells. I was wondering how we would have to clean the MEA to ensure the coating is removed entirely (or almost entirely). I noticed that the MEA manual states that normal dishwashing detergents may be used to clean the MEAs. Could this be used to remove most of the gelatin? The manual also mentions two cleaning protocols, a 0.5mM EDTA/Collagenase I protocol and a Terg-A-Zyme protocol. How effective are these at removing gelatin, other protein-based coatings, or other contaminants in general from the MEA surface?

We were also hoping to fix our cells on the MEA surface and perform some immunostaining experiments. I've read that fixing with paraformaldehyde would make the removal of the cells and coating a little bit more difficult. Could it still be done using the Terg-A-Zyme or Collagenase I protocols mentioned in the MEA Manual? Would you recommend against immunostaining?

Thanks a lot for the help,

-Johnny

portmann.johnny

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Re: Immunostaining on and Cleaning of MEAs

Post  Frank MCS on Tue May 12, 2015 12:36 am

Dear Johnny,
the Terg-A-Zyme protocol is used successfully in hundrets of labs, so I assume it's quite efficient. Gelatine specifially dissolves better at temperatures above 40C. We strongly advise against fixation on the MEAs, as fixed tissue is very hard to remove. Fixed tissue debris will form an isolation layer between active cells and the electrodes.

Best regards Frank MCS
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