1Department of Molecular and Cellular Physiology, Stanford University, 2Department of Molecular and Cellular Physiology, Stanford University School of Medicine
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L. Brown, A., E. Johnson, B., B. Goodman, M. Making Patch-pipettes and Sharp Electrodes with a Programmable Puller. J. Vis. Exp. (20), e939, doi:10.3791/939 (2008).
Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades. Today, such pipettes are made from glass capillaries using a programmable puller. Such instruments heat the capillary using either a metal filament or a laser and draw out the glass using gravity, a motor or both. Pipettes for patch-clamp recording are formed using only heat and gravity, while sharp electrodes for intracellular recording use a combination of heat, gravity, and a motor. The procedure used to make intracellular recording pipettes is similar to that used to make injection needles for a variety of applications, including cRNA injection into Xenopus oocytes. In general, capillary glass <1.2 mm in diameter is used to make pipettes for patch clamp recording, while narrower glass is used for intracellular recording (outer diameter = 1.0 mm). For each tool, the puller is programmed slightly differently. This video shows how to make both kinds of recording pipettes using pre-established puller programs.
Using a microelectrode puller such as the Sutter P-97 Flaming/Brown, pull a set of approximately 10-20 pipettes.
Fire polishing pipettes
The protocol illustrated here is in daily use in electrophysiology laboratories and is also used to make injection needles for cells and animals. With a programmable puller, it is easy to make pipettes for a variety of uses. With attention and care, the filament on your puller will last for one year or more. Good luck with your experiments.
We thank the following funding agencies and foundations for support: National Institutes of Health, National Science Foundation, American Heart Association, Muscular Dystrophy Association, the Donald B. and Delia E. Baxter Foundation, the Klingenstein Fund and the McKnight Endowment for Neuroscience.
|Micropipette Puller||Instrument||Sutter Instrument Co.||P-97||Or similar instrument (e.g. Sutter P-87 or P-2000)|
|Glass Capillaries||Reagent||Sutter Instrument Co.||BF150-86-10||Or, similar capillary glass. To make filling the pipette easier, use a capillary with a glass filament.|
1. Sutter Instrument, P-97 Pipette Cookbook, 2008 (rev. D) http://www.sutter.com/contact/faqs/pipette_cookbook.pdf