FAQ - CYTOPATCH™
Want to know more about our services? Please contact us to discuss your needs. In the meantime, our FAQ may help.
Can the CytoPatch™ Instrument perform real gigaseals comparable to conventional patch clamping?
How does CytoPatch™ meet the special demands of screening ligand gated ion channels?
Can I test liquid handling of my assay definition without consuming chips?
Can the CytoPatch™ Instrument perform real gigaseals comparable to conventional patch clamping?
Yes. The structure and the material - quarz - of the CytoPatch™ Pipette effectively equals the micropipettes used for conventional patch clamping. As a result, the CytoPatch™ Chip produces an extreme tight, gigaohm seal between the CytoPatch™ Pipette and the cell surface after application of a gentle suction.How does CytoPatch™ meet the special demands of screening ligand gated ion channels?
Ligand gated ion channels, like the neuronal GABA receptors, are opened by biochemical compounds rather than membrane voltage changes. In conventional patch clamping setups, some standards have evolved over the years that are regarded as prerequisites for successful and valid research on this important channel class. The most important of these prerequisites is a fast and reliable compound application, at a predefined and exact time during recording of a pulse protocol. Equally important, but even harder to achieve in automated patch clamping, is the requirement of continuous laminar buffer flow around the cell throughout the whole experiment to avoid unstirred layer effects. Last but not least, complex assay definitions are common in ligand gated channels: In a typical experiment, an agonist (the channel opener) is applied first to check the cells response. Following a complete washout of this control, an antagonist is applied for some time, followed without interruption by a mix of both, agonist and antagonist. The robotics, software and perfusion system of the CytoPatch™ have been designed to meet the special demands of ligand gated channel research. In contrast to the many designs on the market today, CytoPatch™ offers a principle of compound application that equals the well-proven perfusion/superfusion design of manual patch setups (for more details, see question #11). As a consequence, CytoPatch™ delivers continuous laminar buffer flow throughout the experiment, fast compound application and unlimited continuous washout without compromise. At the same time, CytoPatch™ software offers pre-configured assay definitions for ligand gated ion channels. The combination of all these features makes CytoPatch™ the superior tool for ligand gated channel research.Is the chip reusable?
No. It should be handled like a conventional patch pipette. We would therefore recommend using it just once.Can I test liquid handling of my assay definition without consuming chips?
Yes. During assay development and testing, it is possible to test liquid handling sequences in a testing mode that uses only one CytoPatch™ site and does not consume chips. This makes the assay development faster and more cost-efficient.Why is there only one "patch pipette" on each CytoPatch™ Chip? Why is this better than using "patch plates" which use many cells at a time?
Using chips with only one tip ensures the quality of each individual measurement and is at the same time the most cost-efficient solution. The reason for this is that each patched cell has its individual cycle times, a fact that requires each patch process to be fully independent from the others. Imagine ten manual patch-clampers sitting in one room and being forced to start their experiments always in synchrony, whoever fails has to wait. You soon will have a postdoc rebellion! The "BatchSize ONE" principle simply means getting most out of your resources.Is cell quality monitored throughout the experiment?
Yes. All important cell quality parameters, like the resting potential of the cell, its membrane capacity and membrane resistance are measured online throughout the experiment. If the recording is irregular, the measurement on this cell will be stopped automatically and a new cell will be CYTOCENTERED to repeat the particular measurement.Is it possible to perform a complete wash-out?
The wash out speed generally depends on the dissociation constant of the compound and the ability of the perfusion system to assure zero compound concentration at the cell surface during wash out. Our perfusion system is able to generate a continuous laminar buffer flow, which assures that the drug concentration at the cell is effectively zeroed during wash-out. Moreover, there is no limit in the possible wash out time, so that even very "sticky" compounds can be 100% washed off. For more details please download our study.What is the advantage of the CytoPatch™ perfusion system?
The CytoPatch™ perfusion system combines a continuous flow-through of fresh buffer fluid with the ability to apply compounds to the cell in form of a "liquid filament" injected into the buffer flow from close vicinity of the cell. Hence, continuous perfusion, compound application and washout are implemented as laminar flow passes through the recording chamber. As a consequence, CytoPatch™ delivers continuous laminar buffer flow throughout the experiment, very fast compound application and unlimited continuous washout without compromise. Compared to a "mix and read" type compound application (which is also possible with the CytoPatch™), the system is much faster and offers better wash-out times. With this features, the system is ready for measurement of fast acting ion channels.Are internal controls possible during the assays?
The CytoPatch™ Graphical Assay Definition Language, with easy to handle drag and drop icons is flexible and allows a direct and separate application of the control substance to the cell at any time. This system enables the user to add the control substance before, during or after the compound application, on the same cell. The most important advantage of this feature is that this type of control can be analysed using paired statistics, which will deliver far better Z' values for your assays compared to "unpaired" controls done on separate cells.Is the cannula for drug pipetting disposable?
No. There is no need to change it, because the Teflon-coated cannula will be completely washed out between different compound applications. The washing of the cannula is achieved by a continuous flow of washing solutions that are delivered from the back of the device to the tip, just like it is common in many industrial standard pipetting robots. In addition, washing of the cannula is assisted by ultrasonic vibration. However, if a new drug cannula is necessary for any reason, the cannula and the corresponding tube can be quickly and easily exchanged.Is a lot of scripting necessary to perform a run?
No. Our sophisticated software is easy to understand: The Graphical Assay Definition Language displays the steps of your assay protocol in form of pictographs. A simple click on each pictograph will bring you to a page with details of the particular assay step. We deliver the CytoPatch™ instrument with a set of predefined standard assays, which you can customise with a few mouse clicks to fit your needs. Before you now start the run of an assay, you just have to do one more simple thing: tell the software where on the microplates you have placed your compounds. You will do that by registering the so called "platemap", which is nothing more than a simple spreadsheet containing the necessary information.How can the data be analysed?
The Inflow-Analysis makes it possible to define parameters to your needs. The data evaluation occurs automatically during the run. However it is possible to change your evaluation mode (for example your Region Of Interest ) afterwards.Can the data be exported to electrophysiological software?
Yes. The raw data and results will be automatically generated as ASCII (.txt)-files organized by compounds or microplates. These files can be easily imported into "Clamp Fit", databases or other statistical programs.