Manual Patch-clamp Technique
Patch-clamp, albeit technically difficult, remains the most reliable and resolutive method to study ionic channels. Manual patch-clamp is the "gold-standard" for the investigation of ion channel activity. It can be applied in:
cell lines
primary and stem cell-derived cells
tissue slices
Creative Bioarray offers routine assays to study drug effects on voltage-gated and ligand-gated ion channels as well as customer-tailored solutions and assay development.
Manual patch-clamp is a versatile, high-resolution and low-throughput method which measures the currents generated by a single cell upon stimulation. Thanks to the flexibility of the patch-clamp technique, virtually all ionic channels in the body: ligand-gated, voltage-gated, calcium-gated, fast/slow activating and deactivating - can be studied with unmatched resolution using well-designed electrophysiology protocols.
In addition to confirming the activity of potential hits from high or medium throughput screens, manual patch-clamping can be used to assess mechanism of action of compounds and to determine the effects of compounds on the biophysical properties of a channel. Manual patch-clamp can be used for safety evaluation in drug development and lead optimization (eg, manual patch clamp of the hERG potassium channel to evaluate potential cardiac liability).
With manual patch-clamp, very precise physiological and pharmacological questions could be addressed for understanding of mechanisms of action. Both voltage-gated and ligand-gated channels can be tested using manual patch-clamping. This system utilizes stable cell lines or native cells (neurons, cardiac myocytes, etc.). It is the most accurate way of analyzing ion channel activities.
In neuroscience research, manual patch-clamp technology provides the highest resolution for electrophysiological recordings from single neuron down to single channel thanks to the different recording configurations (whole-cell, perforated-patch, cell-attached, outside-out).
Technique details
Patch-Clamp recording is performed with borosilicate electrodes of 1-2 μm tip diameter. A pipette containing electrolyte solution is tightly sealed onto the neuronal membrane and isolates a part of the membrane (patch) electrically. Currents fluxing through the channels in this patch hence flow into the pipette and can be recorded by an electrode that is connected to a highly sensitive differential amplifier. The pipette tip makes a giga-ohm seal contact with the cell membrane. Recording this current allows conclusions about the membrane conductance.
Applications
Here in Creative Bioarray, we provide you with
in situ Patch-clamp assays
Optimized for molecular screening and target finding for ion channels and receptors
Conducted at physiological temperature
Cells are recorded from multiple animal models (rat, mouse, non-primate), human tissues (healthy/diseased), iPSCs…
- Researches can be performed on Pain-related ion channels (Nav1.7,1.8…); Thermo-sensation (TRPV1, M8…) Chloride homeostasis (KCC/NKCC…)
In Vitro / Ex Vivo Patch Clamp
Direct measurement of neuronal activity of a single cell or channel (receptor) and is also used to quantify synaptic inputs from excitatory and inhibitory circuits in tissue obtained from living animals.
Perforated patch and intracellular recordings
Test for G-protein coupled receptors and other signal transduction pathways
For a more detailed discussion on your specific requirements, please contact us now and discover how our electrophysical assay services can assist you!
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