Abstract
Whole-cell patch clamping in vivo is an important neuroscience technique that uniquely provides access to both suprathreshold spiking and subthreshold synaptic events of single neurons in the brain. This article describes how to set up and use the autopatcher, which is a robot for automatically obtaining high-yield and high-quality whole-cell patch clamp recordings in vivo. By following this protocol, a functional experimental rig for automated whole-cell patch clamping can be set up in 1 week. High-quality surgical preparation of mice takes ∼1 h, and each autopatching experiment can be carried out over periods lasting several hours. Autopatching should enable in vivo intracellular investigations to be accessible by a substantial number of neuroscience laboratories, and it enables labs that are already doing in vivo patch clamping to scale up their efforts by reducing training time for new lab members and increasing experimental durations by handling mentally intensive tasks automatically.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 634-654 |
| Number of pages | 21 |
| Journal | Nature Protocols |
| Volume | 11 |
| Issue number | 4 |
| DOIs | |
| State | Published - Apr 1 2016 |
| Externally published | Yes |
Bibliographical note
Funding Information:acknoWleDGMents We thank B.D. Allen and H.-J. Suk for feedback on the manuscript. C.R.F. acknowledges the National Institutes of Health (NIH) BRAIN Initiative (National Eye Institute (NEI) and National Institute of Mental Health (NIMH) 1-U01-MH106027-01), an NIH Single Cell Grant 1 R01 EY023173, the National Science Foundation (NSF) (Education and Human Resources (Her) 0965945 and Computer and Information Science and Engineering (CISE) 1110947), an NIH Computational Neuroscience Training grant (no. 5T90DA032466), the Georgia Tech Translational Research Institute for Biomedical Engineering & Science (TRIBES) Seed Grant Awards Program, the Georgia Tech Fund for Innovation in Research and Education (GT-FIRE), the Wallace H. Coulter Translational/Clinical Research Grant Program and support from Georgia Tech through the Institute for Bioengineering and Biosciences Junior Faculty Award, the Technology Fee Fund, Invention Studio, and the George W. Woodruff School of Mechanical Engineering. E.S.B. acknowledges NIH 1R01EY023173, the New York Stem Cell Foundation-Robertson Award, a NIH Director’s Pioneer Award 1DP1NS087724, an NIH Director’s Transformative Award (NIH 1R01MH103910) and an NIH BRAIN initiative grant (NIH 1R24MH106075). G.T.F. acknowledges a Friends of the McGovern Institute Fellowship.
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