Massachusetts Institute of Technology

Georgia Tech Research Corporation

Brian Douglas Allen

Edward Stuart Boyden

Clifton Fonstad, Jr

Craig Richard Forest

Anthony Zorzos

Jorg Scholvin

Ho-Jun Suk

Suhasa Bangalore Kodandaramaiah

Ingrid Van Welie

Jacob Bernstein

Giovanni Talei Franzesi

Christian T Wentz

Michael Baratta

In an automated methodology for in vivo image-guided cell patch clamping, a cell patch clamping device is moved into position and targeted to a specific cell using automated image-guided techniques. Cell contact is determined by analyzing the temporal series of measured resistance levels at the clamping device as it is moved. The difference between successive resistance levels is compared to a threshold, which must be exceeded before cell contact is assumed. Pneumatic control methods are used to achieve gigaseal formation and cell break-in, leading to whole-cell patch clamp formation. An automated robotic system capable of performing this methodology automatically performs patch clamping in vivo, automatically locating cells through image guidance and by analyzing the temporal sequence of electrode impedance changes. By continuously monitoring the patching process and rapidly executing actions triggered by specific measurements, the robot can rapidly find target cells in vivo and establish patch-clamp recordings.

Image-guided closed-loop robotic system for automated whole-cell patch clamping electrophysiology of neurons in vivo

In exemplary implementations of this invention, high-throughput screening of a mammalian brain is performed to locate neural circuit targets of interest. A variety of search patterns may be used for this neural screening, including (a) iterative subdivision, (b) serial search, and (c) combinatorial. To perform this neural screening, an array of optical fibers (or an array of waveguides) is inserted into the brain. Alternately, the array is positioned adjacent to the brain. Each fiber or waveguide in the array is coupled to a light source (LED or laser). The brain has been previously sensitized to light, using genetically encoded optical neural control reagents, which are delivered either using viruses or via transgenic means. In the screening, the array is used to optically perturb the brain. For example, the neurons of the brain may be activated by one color of light, and/or silenced by another color of light.

Methods and apparatus for high-throughput neural screening

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Cambridge, MA, United States of America

Brian Douglas Allen