Micro Pipette Pulling
Micro Pipette Pulling - Related Videos
07:51
Pull-down of Calmodulin-binding Proteins
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Cited by 19 •
2012
Calmodulin (CaM) pull-down assay is an effective way to investigate the interaction of CaM with various proteins. This method uses CaM-sepharose beads for efficient and specific analysis of CaM-binding proteins. This provides an important tool to explore CaM signaling in cellular function.
05:30
Making Patch-pipettes and Sharp Electrodes with a Programmable Puller
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Cited by 22 •
2008
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.
08:08
Co-Immunoprecipitation and Pull-Down Assays
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2023
Co-immunoprecipitation (CoIP) and pull-down assays are closely related methods to identify stable protein-protein interactions. These methods are related to immunoprecipitation, a method for separating a target protein bound to an antibody from unbound proteins. In CoIP, an antibody-bound protein is itself bound to another protein that does not bind with the antibody, this is followed by a separation process that preserves the protein-protein complex. The difference in pull-down assays is that...
06:26
Pulling Membrane Nanotubes from Giant Unilamellar Vesicles
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Cited by 42 •
2017
Many proteins in the cell sense and induce membrane curvature. We describe a method to pull membrane nanotubes from lipid vesicles to study the interaction of proteins or any curvature-active molecule with curved membranes in vitro.
05:12
Pressure-polishing Pipettes for Improved Patch-clamp Recording
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Cited by 13 •
2008
This is a guide to modifying the shape of glass micropipettes. Specifically, by using heat and air pressure the taper is widened without increasing the tip opening, leading to lower pipette resistance. This is critical to obtain low noise recordings of small cells but is useful in many applications.
FAQs
These parameters determine how the softened glass separates and therefore shape the resulting taper. Heat controls when the capillary becomes workable, while pull strength and velocity influence how rapidly the glass is drawn apart. Adjusting their balance changes the tip diameter and geometry, allowing researchers to produce pipettes suited to different cellular-scale handling or measurement tasks.
Cooling conditions affect how the glass solidifies after separation, helping establish the final tip shape and dimensions. Because the tip geometry depends on the interaction between heating, pulling, and cooling, inconsistent cooling can contribute to variation between pipettes. Controlled conditions support more reproducible interfaces for experiments that depend on stable contact with individual neurons.
Tip geometry affects how the pipette interfaces with a cell membrane and contributes to the pipette’s electrical resistance. Consistent dimensions can improve seal formation, which is important for obtaining stable electrical measurements. In neuroscience experiments, reproducible geometry therefore helps reduce variation in recordings and supports more dependable analysis of neuronal signaling and membrane properties.
The workflow begins with glass capillary tubing positioned in a pipette puller. The instrument locally heats the glass until it softens, then applies controlled pulling force to separate the capillary into tapered sections. Researchers regulate heat, pull strength, velocity, and cooling conditions to obtain the desired tip diameter, shape, and resistance for the intended experiment.
Pulled micropipettes support several cellular neuroscience applications, including patch-clamp recording, intracellular stimulation, and microinjection. Their fine, tapered geometry provides a controlled interface with individual neurons. Depending on the experimental goal, the same fabrication approach can therefore support electrical measurement, stimulation of neuronal activity, or delivery of material into a cell.
Reproducible pipette fabrication helps keep tip geometry and resistance consistent across experiments. That consistency can improve seal formation and the stability of electrical measurements, making results easier to compare between cells or experimental preparations. It also strengthens studies of neuronal signaling and membrane properties by reducing variation introduced by the pipette itself.