Whole-cell patch clamp
The patch-clamp technique involves a glass micropipette forming a tight gigaohm seal with the cell membrane. The Patch-clamp technique is a versatile electrophysiological tool for understanding ion channel behavior. Every cell expresses ion channels, but the most common cells to study with patch-clamp techniques include neurons, muscle fibers, cardiomyocytes, and oocytes overexpressing single ion channels.
To evaluate single ion channel conductance, a glass micropipette forms a tight gigaohm seal with the cell membrane. The micropipette contains an electrode bathed in an electrolytic solution to conduct ions. To measure single ion channels, i.e. sodium (Nav) or potassium (Kv) channels, a “patch” of membrane is pulled away from the cell after forming a gigaohm seal, giving the electrode electrical access to the whole cell. Voltage is then applied, forming a voltage clamp, and membrane current is measured. Voltage change within cell membranes can be altered by applying compounds to block or open channels. These techniques enable researchers to understand how ion channels behave both in normal and disease states and how different drugs, ions, or other analytes can modify these conditions.
Two-electrode voltage clamp
Two-electrode voltage clamp (TEVC) is a conventional electrophysiological technique used to artificially control the membrane potential (Vm) of large cells to study the properties of electrogenic membrane proteins, especially ion channels. It makes use of two intracellular electrodes-a voltage electrode as Vm sensor and a current electrode for current injection to adjust the Vm, thus setting the membrane potential at desired values and recording the membrane current to analyze ion channel activities. We use the TEVC setup to measure ion channel kinetics expressed in Xenopus laevis oocytes.
Our molecular biology expertise includes DNA cloning, cut and paste DNA, bacterial transformation , transfection, cellular screening, cellular culture, extraction of DNA, DNA polymerase DNA dependent, reading and writing DNA, DNA sequencing, RNA synthesis, Western blot, rewriting DNA: mutations, random mutagenesis, point mutation, chromosome mutation. Most important techniques are Polymerase Chain Reaction (PCR), Expression cloning, Gel electrophoresis and Sanger sequencing.
Rodent behavior research
We are analyzing scratching behavior in mice.