Literature DB >> 25023316

Pressure-polished borosilicate pipettes are "universal sealer" yielding low access resistance and efficient intracellular perfusion.

Marco Aquila1, Mascia Benedusi, Anna Fasoli, Giorgio Rispoli.   

Abstract

Whole-cell recording is the most widely used configuration of the patch recording technique, mainly because it allows to manipulate the intracellular environment while recording membrane current. However, the patch pipette tapered shank and the small tip opening give high access resistances and preclude efficient exchange between pipette solution and cell cytosol. Independently by the recording configuration, another problem of this technique is to gain consistently tight seals.Here we describe a method to enlarge the pipette shank without affecting the tip opening diameter, through the calibrated combination of heat and air pressure, with a custom-made inexpensive setup. These pressure-polished pipettes give small access resistances and allow for the accommodation of pulled quartz or plastic perfusion tubes very close to the pipette tip (to deliver exogenous molecules into the cytosol with a controlled timing). Finally, we describe a method to consistently attain seals with pipettes made from just one glass type, for a wide variety of cell types, isolated from different amphibian, reptilian, fish, and mammalian tissues, and on artificial membranes composed of many different lipid mixtures.

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Year:  2014        PMID: 25023316     DOI: 10.1007/978-1-4939-1096-0_18

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  2 in total

1.  Characterization of Zebrafish Green Cone Photoresponse Recorded with Pressure-Polished Patch Pipettes, Yielding Efficient Intracellular Dialysis.

Authors:  Marco Aquila; Mascia Benedusi; Anna Fasoli; Giorgio Rispoli
Journal:  PLoS One       Date:  2015-10-29       Impact factor: 3.240

Review 2.  Advanced real-time recordings of neuronal activity with tailored patch pipettes, diamond multi-electrode arrays and electrochromic voltage-sensitive dyes.

Authors:  Bernd Kuhn; Federico Picollo; Valentina Carabelli; Giorgio Rispoli
Journal:  Pflugers Arch       Date:  2020-10-13       Impact factor: 3.657

  2 in total

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