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Department of Molecular Physiology

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patch clamp
The set up

Patch-clamp experiments are done on a microscope which is mounted inside a Faraday cage (top). This is necessary to shield the recording electrode from AC fields originating from lamps, electronic equipment and wires of the AC supply. A micropipette is directed by a micromanipulator towards the cell, and specialized amplifyiers are used for high-resolution current recording. Test solutions can be applied to cells and membrane patches using a perfusion system.

In our lab, patch-clamp recording is often combined with recording of fluorescence from single cells. Using ion-sensitive fluorescent dyes enables us to relate currents conducted by ion channels with changes in intracellular ion concentration caused by that current - a technique that is particularly important for investigating calcium-permeable channels or calcium-gated channels. For this method, the set up contains lamps, filter wheels, photomultipliers and a camera, which control and record the fluorescence signals

The micropipettes that contain the recording electrode are made from glass tubes using a micropipette puller and a microforge (bottom). The tip of the pipette has an opening diameter of less than 1 Ám. Its rim is smoothed so that it can touch a plasma membrane without cutting into it.

  



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