Publication date: Available online 23 August 2017
Source:Journal of Neuroscience Methods
Author(s): Pedro J. Resto, Abhishek Bhat, Eric Stava, Chong Lor, Elliot Merriam, Ruben E. Diaz-Rivera, Robert Pearce, Robert Blick, Justin C. Williams
BackgroundSurface tension passive pumping is a way to actuate flow without the need for pumps, tubing or valves by using the pressure inside small drop to move liquid via a microfluidic channel. These types of tubeless devices have typically been used in cell biology. Herein we present the use of tubeless devices as a fluid exchange platform for patch clamp electrophysiology.New MethodInertia from high-speed droplets and jets is used to create flow and perform on-the-fly mixing of solutions. These are then flowed over GABA transfected HEK cells under patch in order to perform a dose response analysis.ResultsTIRF imaging and electrical recordings are used to study the fluid exchange properties of the microfluidic device, resulting in 0–90% fluid exchange times of hundreds of milliseconds. COMSOL is used to model flow and fluid exchange within the device. Patch-clamping experiments show the ability to use high-speed passive pumping and its derivatives for studying peak dose responses, but not for studying ion channel kinetics.Comparison with Existing Method(s)Our system results in fluid exchange times slower than when using a standard 12-barrel application system and is not as stable as traditional methods, but it offers a new platform with added functionality.ConclusionsSurface tension passive pumping and tubeless devices can be used in a limited fashion for electrophysiology. Users may obtain peak dose responses but the system, in its current form, is not capable of fluid exchange fast enough to study the kinetics of most ion channels.
from #ORL-AlexandrosSfakianakis via ola Kala on Inoreader http://ift.tt/2vhTCIM
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