Cardiomyocyte patch clamping

Anatomy & Physiology Electrophysiology
1

Hi,

I am working on adult rat cardiomyocytes. When trying to bring the patch pipette near the cardiomyocytes to form giga ohm seal they start contracting. Any suggestions?

2

Dear Navneet

You can use two different approaches,

  1. use a Calcium free solution or one with low Calcium conc.
  2. Application of a Mild Muscle Relaxant like BDM (2,3-Butanedione monoxime) or EGTA can inhibit contraction by interfering with actin-myosin cross-bridge cycling. Add BDM (5-10 mM) or EGTA (1-5 mM) to your extracellular solution. However, take caution as these agents can affect membrane properties, so use the lowest effective concentration.
3 Likes
3

Thank you Biographix. Can you suggest If I could use EDTA instead of EGTA?

4

Navneet

Actually you cannot replace EGTA with EDTA because EGTA specifically blocks Calcium ions while EDTA is a general divalent ion blocker. In case your experiment is not dependent on other divalent ions, you may use EDTA.

2 Likes