recording temperature

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zang
zang's picture
recording temperature

I have question on patching at physiological temperature (35-37C)?
Should I do patch at room temerature first, and then the heating system should be on? I mean, the following 1-3 steps should be applied to each cell? Or can I keep the heating system on always (from the beginning the day until the end)?

1: Achieve whole cell configuration at room temperature
2: Bring the temeprature to the physiological level
3: Start to do experiment

Thanks.

Moonshoon
Moonshoon's picture
Patching at physiological

Patching at physiological temperature is usually hard, that's why people prefer to patch at room temperature (even though it means to have a variable recording temperature). So, the easiest way would be to turn on the heating system after sealing the cell. However, it will take some time before you get the desired temperature (you have a whole bottle of aCSF to heat) and you will have to calibrate the time needed to reach 35C

Moonshoon
Moonshoon's picture
I found this topic in the
The FFM
The FFM's picture
 Thanks for finding that old

 Thanks for finding that old discussion Mooshoon.

The standard technique is to obtain the seal at room temp and then turn on your heated perfusion system to up the chamber temperature to 35 degrees.

You do not necessarily need to heat the whole bottle of aSCF/extracellular solution all at once.

You can get an in-line perfusoin system that passes the perfusion lined through a heating jacket to heat them as you perfuse them into the bath. e.g http://www.autom8.com/thermoclamp.html

You will also need a chamber heater to ensure the temperature is maintained constant in the bath.

Finally an objective heater is recommended to compensate for the heat dissipation through the objective lens

you will have to cycle the system for each cell as it will be hard to patch the next cell if everything is at 35 degrees to start with

JennaMo
JennaMo's picture
I have a different question

I have a different question about recording at physiological temp. I hope you don't mind me tagging my question onto here- I thought you all might be able to help.........

I record from brain slices and I recently switched to recording at physiological temperatures. I have noticed that when I have my slice in the heated bath I get a lot of debris in the bath which looks like dead cells. Could the physiological temp be increasing the metabolic rate of my neurons so much that they are dying quickly? This effect is definitely temperature dependent and the 'debris' is coming from the slice and not my solution. It is causing me problems by blocking my pipettes.

Thanks,

Jenna.