flyion Technology
With its Flip-the-Tip™ technology, flyion offers a patch clamp screening method that is significantly easier to use and more efficient than other systems. Automation of the patch clamp method presents various challenges:
- finding a suitable material with a clean and inert surface that is capable of achieving the Gigaseals required for measuring low currents and at the same time avoids non-specific ligand binding to the walls of the measuring vessel
- using as small a measuring chamber as possible so that a significant excess of passive cells acting as ligand catchers and a large drug consumption can be avoided
- facilitating rapid and laminar substance wash-in so that even rapidly-switching ligand-activated channels can be measured
- positioning of micron-sized individual mammalian cells without microscopes or micromanipulators
- implementing the complex testing protocol for a patch clamp experiment in a robust and user-friendly way
Since glass has become established as the best and also cheapest material, flyion is taking a new approach to the way the experiment is conducted: By flushing a cell suspension into a simple glass micropipette and towards its tip, a cell is firmly pressed into the tip; true Gigaseals are created, with the seal forming between the outside of the cell and the smooth inside of the pipette. The Gigaseal’s resistance with this method is as high as with the classic method (1-20 GOhm).
The frequently-used whole cell configuration, where the cell membrane is torn on one side of the seal, can be created with vacuum pulses. The suction from the tip of the pipette tears the area of the cell membrane that is facing the tip aperture. The Flip-the-Tip™ technology also facilitates a perforated patch configuration, where Amphotericin, for example, causes the membrane to perforate within 200s and thus provides electrical access to the inside of the cell.
Another advantage of the Flip-the-Tip™ method is the straightforward addition of substances through a fine glass capillary inside the pipette, which automatically focuses the solution stream onto the cell. Thanks to the pipette’s special geometry and the extremely small extracellular volume, a very rapid solution exchange takes place in less than 10 milliseconds (see Rapid Solution Exchange). This means that even ligand-activated channels that rapidly become desensitized can be measured and it is possible to simulate kinetics as they would be observed in natural synapses.
flyion combines these findings in the f11™ patch clamp instrument. This device allows the tests required for the development of new drugs to be carried out on a wide range of ion channel targets quickly, cheaply and at an early stage in the development process.
Video: Flip-the-Tip patch clamp technology [ 4.839 kB ]
(This Video requires a DivX Codec, which is available free of charge from the DivX website.)