Pressure polishing: a method for re-shaping patch pipettes during fire polishing.
The resolution of patch-clamp recordings is limited by the geometrical and electrical properties of patch pipettes. The ideal whole-cell patch pipette has a blunt, cone-shaped tip and a low resistance. The best glasses for making patch pipettes are low noise, low capacitance glasses such as borosilicate and aluminasilicate glasses. Regrettably, nearly all borosilicate glasses form pipettes with sharp, cone-shaped tips and relatively high resistance. It is possible, however, to reshape the tip during fire polishing by pressurizing the pipette lumen during fire polishing, a technique we call 'pressure polishing.' We find that this technique works with pipettes made from virtually any type of glass, including thick-walled aluminasilicate glass. We routinely use this technique to make pipettes suitable for whole-cell patch-clamp recording of tiny neurons (1-3 microm in diameter). Our pipettes are made from thick-walled, borosilicate glass and have submicron tip openings and resistances <10 MOmega. Similar pipettes could be used to record from subcellular neuronal structures such as axons, dendrites and dendritic spines. Pressure polishing should also be useful in patch-clamp applications that benefit from using pipettes with blunt tips, such as perforated-patch whole-cell recordings, low-noise single channel recordings and experiments that require internal perfusion of the pipette.
Goodman
M B
MB
Department of Biological Sciences, Columbia University, New York, NY 10027, USA. mg289@columbia.edu
Lockery
S R
SR
D000818:Animals; D005390:Fires; D005898:Glass; D008839:Microelectrodes; D009474:Neurons / Q000502:physiology; D009482:Neurophysiology / Q000295:instrumentation* / Q000379:methods; D018408:Patch-Clamp Techniques / Q000295:instrumentation*; D011312:Pressure