Name: making patch-pipettes and sharp electrodes with a programmable puller
Uploaded: 2008-10-08T00:00:00
Description: Watch this Scientific Journal Video about Making Patch-pipettes and Sharp Electrodes with a Programmable Puller at JoVE.com

<p class="jove_content">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.</p>

<p class="jove_step">Pulling pipettes</p>
<p class="jove_content">Using a microelectrode puller such as the Sutter P-97 Flaming/Brown, pull a set of approximately 10-20 pipettes.</p>
<ol>
<li>Select your capillary glass. We use borosilicate capillaries (Sutter BF150-110-10, 1.5mm outer diameter, 1.1mm inner diameter, 10cm long). Store the glass carefully so it remains clean and dust free. </li>
<li>Design a pulling program. We use a 5 step program, with descending heat and velocity at each step, and a small pull on the final step. Sutter's pipe...

<p class="jove_content">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.&nbsp; With attention and care, the filament on your puller will last for one year or more.&nbsp; Good luck with your experiments.</p>

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Micropipette Puller</td>
<td>Instrument</td>
<td>Sutter Instruments</td>
<td>P-97</td>
<td>Or similar instrument (e.g. Sutter P-87 or P-2000)</td>
</tr>
<tr>
<td>Glass Capillaries</td>
<td>Reagent</td>
<td>Sutter Instruments</td>
<td>BF150-86-10</td>
<td>Or, similar capillary glass.  To make filling the pipette easier, use a capillary with a glass filament.</td>
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making patch-pipettes and sharp electrodes with a programmable puller

<p class="jove_content">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 &lt;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.</p>

Watch this Scientific Journal Video about Making Patch-pipettes and Sharp Electrodes with a Programmable Puller at JoVE.com

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller

<p class="jove_content">This video shows how to use a programmable puller to make patch pipettes and sharp electrodes for electrophysiology.  The same procedure can be used to make a variety of glass tools, including injection needles.</p>

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades.  Today, such pipettes are made from glass capillaries ...

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