Fabrication of patch pipets.
Patch clamping refers to a wide range of electrophysiological measurements, all of which have in common the use of patch pipets and the formation of gigaohm seals. The purpose of this unit is to describe the fabrication of patch pipets. The aspects of the pipet geometry that are important to different applications and the different procedures that have been found to most reliably and simply achieve these results are described. Parameters for glass selection are detailed in the beginning of the unit. Pulling patch and whole-cell pipets, elastomer coating, fire polishing, pipet filling, and pipet testing in an experimental setup are highlighted. Additional support protocols describe alternative ways to optimize pipet geometry and cleaning the glass before pulling. Considerations for noise and dynamic performance are emphasized as these two requirements for single-channel and whole-cell current measurements dictate how the pipets must be fabricated.
Rae
J L
JL
Mayo Foundation, Rochester, Minnesota, USA.
Levis
R A
RA
D000818:Animals; D035843:Biomedical Research / Q000295:instrumentation* / Q000379:methods; D004594:Electrophysiology / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D006801:Humans; D009482:Neurophysiology / Q000295:instrumentation* / Q000379:methods; D018408:Patch-Clamp Techniques / Q000295:instrumentation*