Entries by Academic Web Pages

Deep Learning for Patch Clamp Electrophysiology

With the recent advances in deep learning enabling rapid and accurate identification of complex structures, it’s application to in vitro patch-clamp electrophysiology was inevitable. We are exploring the application of these techniques to enhance the capabilities of our fully-automatic patch clamp robots. For example, we have demonstrated fully automatic, real-time detection of healthy neurons within traditional DIC images and vision-based techniques to correct for hardware error stack-up during pipette localization.

Autopatching: Interneurons, Synapses, and Dendrites

In vivo patch-clamp is the gold standard for intracellular recordings, but it is a very manual and highly skilled technique. We have created the most automated in vivo patch-clamp robot to date, by enabling production of multiple, serial intracellular recordings without human intervention. Our robot automates pipette filling, Ag/AgCl wire threading, pipette positioning, neuron hunting, break-in, delivering sensory stimulus, and recording quality control, enabling in vivo cell-type characterization.

Cleaning Patch Pipettes Enables Fully Automated Neuroscience

Whole-cell patch clamp is one of the most sensitive techniques in all of neuroscience. This technology has enabled wide ranging discoveries, such as measurements of single ion channels in neurons and the recording of electrically active cells in the living brain. However, it takes a lot of skill and time to perform – neuroscientists can […]

Legendary Graduation Cap Wall

A unique tradition of the Precision Biosystems Laboratory is decorating a graduation cap and wagon geared towards the research and hobbies of each graduate. Immediatley after sucessfully defending their thesis, the recent graduate is given their decorated graduation gown, and told to sit in their graduation wagon so that Craig can pull them around campus […]

Lab Jiu Jitsu Reunion in Ashburn, VA

Former lab members Ilya and Will met up with Colby and Mighten this summer in Asburn, VA to hang out and even did a little jiu jitsu. Ilya (post-doc) and Mighten (visiting summer graduate student) were at Howard Hughes Medical Institute, Will just started his post-doc at Columbia University, and Colby was completing his 3rd […]

Autopatching: Ultra-High Throughput

Intracellular patch-clamp electrophysiology, one of the most ubiquitous, high-fidelity techniques in biophysics, remains laborious and low-throughput. While previous efforts have succeeded at automating some steps of the technique, we have created a robotic ‘PatcherBot’ system that can perform many patch-clamp recordings sequentially, fully unattended. Comprehensive automation is accomplished by outfitting the robot with machine vision, and cleaning pipettes instead of manually exchanging them. The PatcherBot can obtain data at a rate of 16 cells per hour and work with no human intervention for up to 3 h. We have demonstrated the broad applicability and scalability of this system by performing hundreds of recordings in tissue culture cells and mouse brain slices with no human supervision. The system is potentially transformative for applications that depend on many high-quality measurements of single cells, such as drug screening, protein functional characterization, and multimodal cell type investigations.

Neuroanatomical Imaging

Serial section electron microscopy (ssEM), a technique where volumes of tissue can be anatomically reconstructed by imaging consecutive tissue slices, has proven to be a powerful tool for the investigation of brain anatomy. Between the process of cutting the slices—or “sections”—and imaging them, however, handling 100-106 delicate sections remains a bottleneck in ssEM, especially for batches in the “mesoscale” regime, i.e.,102-103 sections. Our lab is developing a tissue section handling device that transports and positions sections accurately and repeatably for automated, robotic section pick-up and placement onto an imaging substrate.

Congrats Dr. Timothy Lee

Dr. Timothy Lee successfully defended his thesis this summer. His thesis focused on automating the collection of serial nano-sections sections used to image and analyze up to 1 cubic milimieter of tissue!

Epithelial Electrophysiology

Recent advancements utilizing induced pluripotent stem cell-derived (iPSC) epithelia have made disease modeling and cell therapy for many, previously untreatable diseases possible. However, current electrophysiology techniques – used to validate epithelial function – are painstaking and require a highly trained user; limiting experimental throughput. We are developing new techniques and devices that enable both high-throughput and high-quality electrophysiological measurements. Our lab has built a robot that automates intracellular electrophysiology of epithelia that automates the pipette insertion process and, simultaneously improves throughput. We are also exploring new electrochemical impedance spectroscopy (EIS)-based techniques to extract high resolution, membrane-specific properties non invasively. These techniques could be used as the basis for at-line functional characterization of epithelia in all future iPSC-based therapies.