Abstract

A simple device for microinjections, manipulations and measurements with microinterferometric monitoring of the membrane / interface processes at the thickness range of 5-1000nm at different angles

Author(s): O.V. Gradov

A conventional technique for experimental morphological and membrane electrophysiological studies at one cell and membrane surface level is Micromanipulations, perfusions and measurements performed using glass microelectrodes crammed with an electrolyte. The typical (effective) diameter of the top of the glass microelectrode is from 500 up to but 100 nm, which prevents one from observing it employing a standard optical microscope in accordance with the optical resolution criteria, since the diameter but 500 nm is indistinguishable within the interference zone. Microprocessor programming of the puller (microforge) that gives pulling and tearing allows to get in certain regimes the adjusted diameter and shape of the micropipette tip, although this result is not fully controlled due to the above limitations. In this connection it's necessary to style the control devices for the micropipette tips both at the preparation and operation stages (intracellular or extracellular insertion). This method also should provide visualization of the processes occurring upon interaction of the microelectrode tip with the cell in real time, counting on the electrode type and state, which allows to level the artifacts arising with the systematic error frequency from the uncontrolled operation of the micropipette tip after alternative ways of the microelectrode filling with the electrolyte. We are using an installation scheme that is used to solve the above problems by means of introducing an interferometric device for microscopic control of the microelectrode and micromanipulator or microperfusor, for the first time for a given type of optical instruments combined with the interferometric optical scheme [A simple device for microinjections, manipulations and measurements using an electromorphological chip under microinterferometric control of the interface and membrane processes at the thickness range of 5-1000 nm at different angles;