Electrophysiology

Mission
The primary objective of the NUS Medicine Electrophysiology Core is to provide a service and expertise to researchers who plan to conduct experiments requiring electrophysiological techniques, such as patch-clamp electrophysiology, for functional or physiological studies.

What is patch-clamp electrophysiology?
Patch-clamp electrophysiology is the study of electrical activity across biological membrane such as plasma, mitochondria and lysosomal membrane. Movement of ions across the membrane, can in principle be monitored via recordings as current or potential change. To perform whole cell patch-clamp, a micropipette is tightly sealed on the cell membrane forming high resistance gigaseal. This small patch of membrane could then be ruptured with more intense suction to allow for electrode electrical access to the interior of the whole cell. Voltage stimulus is subsequently applied to elicit membrane current. . Alternatively, change in membrane potential, for example action potential which is an important feature of excitable cells such as neurons or muscle fibers could also be recorded

1. Functional characterization and Pharmacological screening of ion channel blockers/activators
a. Voltage gated calcium channels (CaV1.2, CaV1.3, CaV2.1)
b. Voltage gated sodium channels (NaV1.5)
c. Voltage gated potassium channels (KV1.1, KV1.2, KV7.1, KV11.1)
d. Two-pore domain potassium channels (TASK-2)
e. Calcium-activated potassium channels (SKCa, BKCa)
f. Lysosomal ion channels (TPC1, TRPML1, BKCa)
g. Acid sensitive ion channels (ASIC2a)
h. TRP Channels (TRPV1, TRPA1)
2. Whole cell recording of primary or hIPSC-derived cardiomyocytes or neurons.
3. Whole cell recording of neurons within acutely isolated brain slice.
4. Whole endolysosomal patch clamp electrophysiology of cultured cells
5. Electrophysiological analysis of endplate potentials in acutely isolated phrenic nerve-diaphragm preparation

The Core will purchase to provide for the general consumables and reagents that will be proportionately charged to the users. However, users are responsible to supply any materials or reagents that are unique to their particular projects.

1. Nikon Ti inverted fluorescent microscope
2. Nikon Upright Patch Clamp microscope Model: FN1 with Hammamatsu Orca Flash 4
3. Manipulator MP-285/ MPC-325-2 (Sutter Instrument Company)
4. MultiClamp 700B (Axon instruments)
5. Digidata 1550 Low-Noise Data Acquisition System (Axon instruments)
6. Vapor pressure osmometer 5520 (VAPRO)
7. Flaming/brown micropipette puller model P-97 (Sutter Instrument Company)
8. Micro forge MF-900 (NARISHIGE)
9. Single Stage Glass Microelectrode Puller MODEL PP-830 (NARISHIGE)
10. SH27B In-line solution heaters and CC-28 Thermistor (Warner Instruments)
11. A365 Stimulus Isolator (World Precision Instruments)
12. Olympus IX81 microscope with Olympus DP71 camera