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.
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Landry, C. R., Yip, M. C., Kolb, I., Stoy, W. M., Gonzalez, M. M. and Forest, C. R. (2021). Method for Rapid Enzymatic Cleaning for Reuse of Patch Clamp Pipettes: Increasing Throughput by Eliminating Manual Pipette Replacement between Patch Clamp Attempts. Bio-protocol 11(14): e4085. DOI: 10.21769/BioProtoc.4085.[PDF]
M.M. Gonzalez, M.C. Yip, C.F. Lewallen, M.J. Rowan, C.R. Forest, Machine learning-based pipette correction for automated patch clamp in vitro, SfN Global Connectome, Virtual Conference, Jan 11-13, 2021.
Kolb, I., Landry, C. R., Yip, M. C., Lewallen, C. F., Stoy, W. A., Lee, J., Felouzis, A., Yang, B., Boyden, E. S., Rozell, C. J., & Forest, C. R. (2019). PatcherBot: a single-cell electrophysiology robot for adherent cells and brain slices. Journal of neural engineering, 16(4), 046003. https://doi.org/10.1088/1741-2552/ab1834[PDF]
C. Landry, I. Kolb, W. Stoy, M. Yip, C. Lewallen, C. Forest, Pipette cleaning enables hundreds of automated patch attempts with a single pipette, Proceedings of the Annual Meeting of the Society for Neuroscience (SfN), Sand Diego, CA, Nov. 3-7, 2018.
L. Li, B. Ouellette, W. Stoy, E. Garren, T.L. Daigle, C.R. Forest, C. Koch, H. Zeng, A robot for high yield electrophysiology and morphology of single neurons in vivo, Nature Communications, 15604 (2017), doi:10.1038/ncomms15604[PDF]
M.A. Stockslager*, C.M. Capocasale* (*co-first authors), G.L. Holst, M.D. Simon, Y. Li, D.J. McGruder, E.B. Rousseau, W.A. Stoy, T. Sulchek, C.R. Forest, Optical method for automated measurement of glass micropipette tip geometry, Precision Engineering, Vol 46, p. 88-95, October 2016.
I. Kolb, W.A. Stoy, E. Rousseau, O.A. Moody, A. Jenkins, C.R. Forest, Cleaning patch-clamp pipettes for immediate reuse, Scientific Reports 6: 35001, 2016. doi:10.1038/srep35001 (over 10 articles in popular press, example below in D.21, Altmetric score in 97th percentile of 133,000 articles of similar age in all journals)
Q. Wu*, I. Kolb* (*co-first authors), B.M. Callahan, Z. Su, W. Stoy, S.B. Kodandaramaiah, R. Neve, H. Zeng, E.S. Boyden, C.R. Forest#, A.A. Chubykin# (#co-corresponding authors), Integration of autopatching with automated pipette and cell detection in vitro, J. Neurophysiology (in press). doi: 10.1152/jn.00386.2016
R.R. Harrison, I. Kolb, S.B. Kodandaramaiah, A.A. Chubykin, A. Yang, M.F. Bear, E.S. Boyden, C.R. Forest, Microchip amplifier for in vitro, in vivo, and automated whole-cell patch-clamp recording, J. Neurophysiology
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