Sample burned while ETC ?

Shashank20

Hi I was running an ETC of brain 40V/37oC in the designed chamber and the buffer was sampled by Lauda alpha RA8 water circulator. Unfortunately when the chamber was opened after 3 days to check the clearing, the brain sample was lost. Could it be that the brain burned & melted away..if it has come in contact with electrodes ? has anyone had similar experience ? Thank-you

Fabio

Hi,
I never used 40V normally max 20 V. Your sample can melt at higher temperatures. Also contact with electrodes can leave burn marks, try using some mesh to isolate it.

Shashank20

Thanks @ Fabio for you input

lijun

Hi
Check if you brain is in the water filter or other place.

joelrosiene

I have found that inadequate circulation of buffer through the chamber can result in an increase in temperature far above your settings. Perhaps try to get a read of the true chamber temperature, rather than the temperature of your circulator/heater settings?

Of course, increasing the buffer flow causes issues regarding turbulence in the chamber, and the potential for the tissue sample to be sucked up and destroyed by the filter. I had to get clever with bits of perforated plastic to get around this.

...Of course you could always just run it cooler and slower.