Which Pipette to use?

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Rahul254
Rahul254's picture
Which Pipette to use?

Hi all,
I am new to this community of scientists(even though I am not the one), I read some of the conversations and found them helpful so Thank you all...
I have a problem in patching L6 skeletal muscle cells, I am trying to make giga seal, but I couldn't form a gigaseal on these cells. I tried different types of pipettes and few other tricks, like applying -ve and +ve voltage, but still I can not seal these cells. Is there any one here, who has experience with these cells before? And if anyone can tell me which specific pipettes are good to seal L6 cells then it would be very helpful...

Fraser Moss
Fraser Moss's picture
I have not worked with these

I have not worked with these cells but you should try contacting the lab who the wrote the paper below to try getting more specific answers to your question.

Skeletal muscle and small-conductance calcium-activated potassium channels. by

Pribnow D, Johnson-Pais T, Bond CT, Keen J, Johnson RA, Janowsky A, Silvia C, Thayer M, Maylie J, Adelman JP.

Department of Cell and Developmental Biology, Oregon Health Sciences University, Portland, USA.

Abstract from Pubmed
Skeletal muscle becomes hyperexcitable following denervation and when cultured in the absence of nerve cells. In these circumstances, the bee venom peptide toxin apamin, a blocker of small-conductance calcium-activated potassium (SK) channels, dramatically reduces the hyperexcitability. In this report, we show that SK3 channels are expressed in denervated skeletal muscle and in L6 cells. Action potentials evoked from normal innervated rat skeletal muscle did not exhibit an afterhyperpolarization, indicating a lack of SK channel activity; very low levels of apamin binding sites, SK3 protein, or SK3 mRNA were present. However, denervation resulted in apamin-sensitive afterhyperpolarizations and increased apamin binding sites, SK3 protein, and SK3 mRNA. Cultured rat L6 myoblasts and differentiated L6 myotubes contained similar levels of SK3 mRNA, although apamin-sensitive SK currents and apamin binding sites were detected only following myotube differentiation. Therefore, different molecular mechanisms govern SK3 expression levels in denervated muscle compared with muscle cells differentiated in culture.