Closed-Loop Real-Time Imaging Enables Fully Automated Cell-Targeted Patch-Clamp Neural Recording In Vivo

Abstract

In our original Figure 1B, the appearance of Figures 1Bii and 1Biii incorrectly made it look like the pipette descended into the brain in the downward direction (implying that it would cause brain damage), and furthermore in the original Figure 1Biv the black arrows incorrectly implied that the pipette descends into the brain further downward. In reality, the pipette enters the brain at a diagonal angle, along the axis of the pipette, and furthermore as the pipette approaches the cell, it does so in staggered downward and lateral steps that simulate diagonal movement. We have now corrected Figure 1B to reflect the real configuration of the system. In particular, the pipette enters into the brain (Figure 1Biii) at a diagonal, and toward the end of the process, the pipette moves in staggered downward and lateral steps to approach the cell (we have removed the downward black arrows from Figure 1Biv, so that the lower left of Figure 1B provides the full explanation of the final stages of pipette movement). The figure has now been corrected online. The authors apologize for any confusion this error may have caused.

An animal-actuated rotational head-fixation system for 2-photon imaging during 2-d navigation

“…Our approach could therefore facilitate multiple patch-clamp recordings 73,[76][77][78] and next-generation silicon laminar probes 57,72 (Janelia/IMEC, Neuropixels consortium). In both cases, the electrophysiology system could be driven from commercially available small form factor desktop computers (Intel NUC or similar) that can mount to the rotating headpost frame, powered by a battery and controlled via WiFi.…”

“…The majority of methods, especially those capable of recording from large populations of neurons, or subcellular activity, require head-fixation. Canonical examples of this are conventional [64][65][66] and meso-scale 67 2-photon Ca2 + imaging 64 of genetically encoded fluorescent indicators 68 , cell-targeted optogenetic interventions [69][70][71] , and large-scale extracellular recordings of neural populations 72 , or whole-cell recordings of single [73][74][75] or multiple neurons [76][77][78][79] .…”

Robotic platform for microinjection into single cells in intact tissue

“…Robotic systems have enabled the automation of difficult laboratory techniques that require precise micromanipulation such as in vivo patch clamping of single [50][51][52] as well as multiple neurons in vivo 53 . Additionally, previous work relied on camera images to guide automated patch clamping systems to specific locations in tissue 11,54 . These applications resulted in significant improvement in the success of patch clamping and enabled neuroscientists to perform complex experiments previously limited by technical difficulties.…”

“…2A, Supplementary Note 1). Once the calibration step is completed, the Autoinjector can guide the injection micropipette to specific locations in the FOV using the micromanipulator, similar to previous algorithms 10,11 . The user then draws a line along the desired path of microinjection on the microscope image using the graphical user interface (GUI) ( Fig.…”

Reconstruction of in-vivo subthreshold activity of single neurons from large-scale spiking recordings

“…Such recordings have been done in anaesthetized animals (Kodandaramaiah et al 2012, Yu & Ferster 2010, Volgushev et al 2006, Lampl et al 1999, head fixed awake animals (Zhao et al 2016, Gentet et al 2010, and even in freely moving animals (Lee et al 2006). To patch multiple cells in vivo is difficult and may in the future be facilitated by patch "robots" (Kodandaramaiah et al 2018, Suk et al 2017. Imaging methods might be more suitable for addressing the membrane potential of multiple cells simultaneously (Bando et al 2019), but may still represent a smoothing of activity, owing to limitations on scanning and shuttering speed when compared to electrophysiological sampling rates.…”