Giant Patch Technique
Giant Patch Technique - Related Videos
09:03
Giant Liposome Preparation for Imaging and Patch-Clamp Electrophysiology
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Cited by 23 •
2013
Reconstituting functional membrane proteins into giant liposomes of defined composition is a powerful approach when combined with patch-clamp electrophysiology. However, conventional giant liposome production may be incompatible with protein stability. We describe protocols for producing giant liposomes from pure lipids or small liposomes containing ion channels.
11:42
Reconstitution of a Transmembrane Protein, the Voltage-gated Ion Channel, KvAP, into Giant Unilamellar Vesicles for Microscopy and Patch Clamp Studies
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Cited by 43 •
2015
The reconstitution of the transmembrane protein, KvAP, into giant unilamellar vesicles (GUVs) is demonstrated for two dehydration-rehydration methods — electroformation, and gel-assisted swelling. In both methods, small unilamellar vesicles containing the protein are fused together to form GUVs that can then be studied by fluorescence microscopy and patch-clamp electrophysiology.
12:53
Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster
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Cited by 17 •
2011
The Giant Fiber System is a simple neuronal circuit of adult Drosophila melanogaster containing the largest neurons in the fly. We describe the protocol for monitoring synaptic transmission through this pathway by recording post synaptic potentials in dorsal longitudinal (DLM) and tergotrochanteral (TTM) muscles following direct stimulation of the Giant Fiber interneurons.
11:56
Slice Patch Clamp Technique for Analyzing Learning-Induced Plasticity
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Cited by 19 •
2017
The slice patch clamp technique is an effective method for analyzing learning-induced changes in the intrinsic properties and plasticity of excitatory or inhibitory synapses.
08:45
Forming Giant-sized Polymersomes Using Gel-assisted Rehydration
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Cited by 5 •
2016
We present a protocol to rapidly form giant polymer vesicles (pGVs). Briefly, polymer solutions are dehydrated on dried agarose films adhered to coverslips. Rehydration of the polymer films results in rapid formation of pGVs. This method greatly advances the preparation of synthetic giant vesicles for direct applications in biomimetic studies.
FAQs
Defined voltage conditions allow researchers to relate changes in membrane potential to measured ion currents. By controlling the voltage across the membrane patch, they can examine channel kinetics, meaning the timing and pattern of channel activity, under reproducible conditions. This makes it possible to characterize how channels transport ions and how their activity changes during membrane signaling.
Controlling the solutions bathing each membrane surface lets researchers expose the membrane to precisely selected ionic or regulatory conditions. They can then evaluate ionic selectivity, pharmacological regulation, and intracellular modulation while measuring the resulting currents. Repeated solution exchange is especially valuable because it allows comparisons between conditions without changing the fundamental membrane preparation.
The expanded membrane area provides more membrane material for analyzing ion transport and signaling while retaining electrical measurement under defined conditions. This creates an intermediate scale between single-channel recordings and whole-cell physiology. The approach can therefore connect detailed channel behavior with broader membrane responses, while also supporting biochemical manipulation of the recorded membrane.
Channel regulation can be examined by changing the chemical environment around the membrane and observing corresponding current changes. The method supports tests of pharmacological regulation and intracellular modulation, while solution exchange permits repeated comparisons across experimental conditions. These measurements help distinguish changes in channel activity from differences caused by the surrounding ionic environment or regulatory factors.
A recording begins when a patch pipette forms a seal with the cell membrane. The attached membrane patch is then excised or expanded, after which researchers control the solution on each membrane surface. They apply defined voltage conditions, exchange solutions when needed, and measure the resulting ion currents to analyze transport and signaling properties.
The technique is suited to questions about channel kinetics, ionic selectivity, pharmacological regulation, and intracellular modulation. Researchers can assess how membrane channels respond to voltage and controlled chemical environments, then compare current patterns across conditions. Its combination of electrical recording, solution control, and biochemical accessibility makes it useful for connecting molecular channel properties with membrane-level physiology.