Patch-clamp recording cell-attached

Figure 1 Schematic diagrams illustrating the patch-clamp technique. (A) Overall setup for isolating single ionic channels in an intact patch of cell membrane. P = patch pipet R = reference microelectrode I = intracellular microelectrode Vp = applied patch potential Em = membrane potential Vm = Em — Vp = potential across the patch A = patch-clamp amplifier. (From Ref. 90.) (B) Five different recording configurations, and procedures used to establish them, (i) Cell attached or intact patch (ii) open cell attached patch (iii) whole cell recording (iv) excised outside-out patch (v) excised inside-out patch. Key i = inside of the cell o = outside of the cell. (Adapted from Ref. 283.)...

A patch-clamp (recording) pipette is attached to an individual cell and a gigaohm seal is formed between the cell and the patch-clamp pipette by applying suction through the pipette. 

Fig. 21.4. Vesicle formation and patch-clamp techniques used to record levamisole receptor channel currents from Ascaris muscle. (A) Muscle membrane vesicles bud-off from the bag membrane following a 10 min collagenase treatment and incubation for 1 h at 37°C in Ascaris saline. (B) Levamisole is applied to the outside surface of the membrane to activate receptor channels cell-attached patches are usually used but it is also possible to make inside-out and outside-out patch recordings.

In the outside-out model, the pipette is attached to the entire cell as in the whole cell model, followed by a sharp pull that causes the cell membrane to break and reseal with the pipette tip (Fig. 3b). With the extracellular region exposed, channel activity as a response to different external stimuli can be probed. This configuration is less common than the inside-out method. Using an outside-out method, single-channel opening activity has been recorded while various neurotransmitters were released. For example, this patch clamp method was used as a detector for capillary electrophoresis separations of GABA, glutamate, and NMDA (7). 

Figure 16.14 Original Kca1-1 (BK) single channel recordings and the inhibitory effect of iberiotoxin. Currents were recorded from a guinea pig urinary bladder smooth muscle cell using the whole cell (cell-attached), perforated patch-clamp technique at 0 mV holding potential (Section...

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