I'm currently patching pancreatic beta-cells using a HEKA amplifier to measure change in cell capacitance due to exocytosis events. I keep on having big oscillations appening every few seconds, which stop me from observing my increase in cell capacitance. Can this oscillations be due to a poor control of chamber temperature from my heating system? I patch at 32°C and temperature oscillates from 30 to 33°C.
Thanks in advance
Dear Vincenzo,
Yes changes in temperature will change the capacitance readings. There are many channels and transporters that are dependent on the temperature, and the cell volume also changes with the temperature. I have previously studied this in the retinal ganglion cells, but the principles applies to pretty much all cells. Please see below publications for details of temperature effects on capacitance measurements.
best wishes, Refik
Conference Paper Characterization of temperature-sensitive leak currents in r...
Data Kanjhan07-IBRO poster-Characterization of temperature-sensit...
If you are near to phase change of the lipids, the temperature will change the capacitance of the lipids (cells). you should perhaps try to work at 35°C
Hi Vincenzo,
As the colleagues already mentioned a change in temperature will certainly affect the cell capacitance. However, your temperature is fluctuating between 30 and 33°C which is not so dramatic, and you mention the occurrence of "big oscillations" - I am not a specialist on this but can a delta of 3°C enough to cause a dramatic change in cell capacitance?
Edit: Attached is an abstract where people measured the effect of temperature on the cell capacitance in cardiomyocytes and have an increase of 0.17 pF/°C - if you assume that a similar effect occurs in beta cells then it should not have much influence on your recordings.
We published a paper in 1996 showing the temperature dependence of exocytosis in pancreatic beta-cells. Exocytosis can not be measured in RT. The largest responses are obtained at 37 degrees, but then the membrane becomes to fragile. We therefore perform most of our capacitance measurements at 31-33 dgeress C.
Please see
Cooling inhibits exocytosis in single mouse pancreatic B-cells by suppression of granule mobilization.Renström E, Eliasson L, Bokvist K, Rorsman P.J Physiol. 1996 Jul 1;494 ( Pt 1):41-52.PMID:8814605. Free PMC Article
for more information.
Ocillations could be due to dispturbance from the surrounding - check grounding properly. Oscillations could also be due to that your flow of solutions is unstable and the surface of your extracellution in the dish is fluxtuating.
And good luck with measurements!
/Lena
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