Potentiometric-scanning ion conductance microscopy for measurement at tight junctions.
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abstract
Scanning Ion Conductance Microscopy (SICM) has been developed originally for high-resolution imaging of topographic features. Recently, we have described a hybrid voltage scanning mode of SICM, termed Potentiometric-SICM (P-SICM) for recording transmembrane ionic conductance at specific nanostructures of synthetic and biological interfaces. With this technique, paracellular conductance through tight junctions - a subcellular structure that has been difficult to interrogate previously - has been realized. P-SICM utilizes a dual-barrel pipet to differentiate paracellular from transcellular transport processes with nanoscale spatial resolution. The unique combination of voltage scanning and topographic imaging enables P-SICM to capture paracellular conductance within a nominal radius of several hundred nanometers. This review summarizes recent advances in paracellular conductance recording with an emphasis on the P-SICM based approach, which is applied to detect claudin-2 mediated permeability changes at the tight junction.