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Striving for gigaseal with force-controlled patch clamp on contracting cardiomyocyte
We aim to determine the key parameters to achieve an electric gigaseal between the cell membrane and the internal wall of a microfabricated pipette. Indeed, we are developing a force-controlled patch clamp using FluidFM to study the electrophysiology of single cells in a gentler and more controlled way.
FluidFM was first developed in 2006 combining the force-sensitive AFM cantilever with hollow microfluidic channels. It opens up a new range of possibilities in single-cell manipulations. Among which, the capability to measure pico-ampere ionic currents naturally generates the idea of force-controlled patch clamp. Traditional single-cell patch clamp, using a glass micro-pipette, lacks force information when approaching the membrane with consequent unavoidable damage of the targeted cells. The force-controlled feature of our FluidFM is expected to facilitate the formation of a controlled probe-cell contact, thus increasing the probability of achieving a gigaseal which in its turn is mandatory to measure the small electrophysiological currents (nA) without considerable leakages.
As model system we are studying cardiomyocytes which are particularly challenging for the formation of a gigaseal because of their mechanical contractions.
In this project, the student will learn to operate the FluidFM technique, to use the cylindrical cantilever, to analyse data with with Python, and to harvest cardiomyocyte. The detailed topic will be discussed and tailored according to the candidate’s interest.
FluidFM was first developed in 2006 combining the force-sensitive AFM cantilever with hollow microfluidic channels. It opens up a new range of possibilities in single-cell manipulations. Among which, the capability to measure pico-ampere ionic currents naturally generates the idea of force-controlled patch clamp. Traditional single-cell patch clamp, using a glass micro-pipette, lacks force information when approaching the membrane with consequent unavoidable damage of the targeted cells. The force-controlled feature of our FluidFM is expected to facilitate the formation of a controlled probe-cell contact, thus increasing the probability of achieving a gigaseal which in its turn is mandatory to measure the small electrophysiological currents (nA) without considerable leakages. As model system we are studying cardiomyocytes which are particularly challenging for the formation of a gigaseal because of their mechanical contractions.
In this project, the student will learn to operate the FluidFM technique, to use the cylindrical cantilever, to analyse data with with Python, and to harvest cardiomyocyte. The detailed topic will be discussed and tailored according to the candidate’s interest.