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.
At a glance
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- Supplementary Text and Figures (180 KB)
- Supplementary Data 1: File archive consisting of software required for running the Autopatcher. (2,115 KB)
Includes two Labview library files – ‘Autopatcher 2000.llb’ and ‘Hardware.llb’ that can be opened using Labview installed in Step 8 of the protocol. Also included is a corresponding ‘Autopatcher software configuration manual.pdf’ that provides detailed instructions on installation of software and configuring the software settings to control the autopatcher control box.
- Supplementary Data 2: File archive consisting of mechanical drawings and computer aided design (CAD) files for making the custom head fixation base and headplate. (365 KB)
‘Headfixation fixation base CAD.pdf’ is a mechanical drawing of the headfixation base, while ‘Head fixation base CAD.SLDPRT’ is the 3D drawing that can be opened in Solidworks software. ‘Head Plate CAD.pdf’ is a mechanical drawing of headplate implant, and ‘Head Plate CAD.SLDPRT’ is the 3D drawing that can be opened in Solidworks software.
- Supplementary Data 3: File archive consisting of mechanical drawings and computer aided design (CAD) files and instructions for assembling the autopatcher pipette actuator assembly. (3,022 KB)
Assembly instructions are provided in ‘Autopatcher Robotic Arm Assembly Manual.pdf’. ‘adapter plate 1.PDF’ and ‘adapter plate 1.SLDDRW’ are mechanical drawings of the adaptor plate used for mounting programmable linear stage onto Sutter manipulator. ‘adapter plate 1.SLDPRT’ is the corresponding 3D CAD file that can be opened in Solidworks. ‘adapter plate 2.PDF’ and ‘adapter plate 2.SLDDRW’ are mechanical drawings of the adaptor plate used to mount the amplifier headstage onto the programmable linear stage. ‘adapter plate 2.SLDPRT’ is the corresponding 3D CAD file that can be opened in Solidworks.
- Supplementary Data 4: File archive consisting of mechanical drawings, computer aided design (CAD) files and instructions for assembling the autopatcher control box. (10,129 KB)
Assembly instructions are provided in the ‘Autopatcher control box assembly manual.pdf’ while ‘Autopatcher control box parts list.xlsx’ provides complete list of parts required for assembling the control box. Details of each part include description, name of vendor, catalog number, price/unit (as on Aug 2015), and quantity of each part. The sub-folder ‘Laser cutter files’ contains the ‘Autopatcher panels front & back.ai’ and ‘Autopatcher structural base, platform, & manometer clamp.ai’ files which can be used to cut two structural elements used for control box assembly (See the ‘Autopatcher control box assembly manual.pdf’). The sub-folder ‘Circuit board files’ contains: ‘Autopatcher PCB parts list.xlsx’ – a full parts list of all components on the pressure control printed circuit board (PCB) and valve relay PCB. ‘pressure_board.brd’ and ‘pressure_board.sch’ are the pressure control PCB CAD files, while ‘valve-relay_board.brd’ and ‘valve-relay_board.sch’ are the valve relay PCB CAD files.