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3 Janelia Publications

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    04/03/17 | Efficient method for whole-cell recording in freely moving rodents using ultraviolet-cured collar-based pipette stabilization.
    Lee D, Lee AK
    Cold Spring Harbor Protocols. 2017 Apr 03;2017(4):pdb.prot095810. doi: 10.1101/pdb.prot095810

    Whole-cell recording is a key technique for investigating synaptic and cellular mechanisms underlying various brain functions. However, because of its high sensitivity to mechanical disturbances, applying the whole-cell recording method to freely moving animals has been challenging. Here, we describe a technique for obtaining such recordings in freely moving, drug-free animals with a high success rate. This technique involves three major steps: obtaining a whole-cell recording from awake head-fixed animals, reliable and efficient stabilization of the pipette with respect to the animal's head using an ultraviolet (UV)-transparent collar and UV-cured adhesive, and rapid release of the animal from head fixation without loss of the recording. This technique has been successfully applied to obtain intracellular recordings from the hippocampus of freely moving rats and mice exploring a spatial environment, and should be generally applicable to other brain areas in animals engaged in a variety of natural behaviors.

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    04/03/17 | In vivo patch-clamp recording in awake head-fixed rodents.
    Lee D, Lee AK
    Cold Spring Harbor Protocols. 2017 Apr 03;2017(4):pdb.prot095802. doi: 10.1101/pdb.prot095802

    Whole-cell recording has been used to measure and manipulate a neuron's spiking and subthreshold membrane potential, allowing assessment of the cell's inputs and outputs as well as its intrinsic membrane properties. This technique has also been combined with pharmacology and optogenetics as well as morphological reconstruction to address critical questions concerning neuronal integration, plasticity, and connectivity. This protocol describes a technique for obtaining whole-cell recordings in awake head-fixed animals, allowing such questions to be investigated within the context of an intact network and natural behavioral states. First, animals are habituated to sit quietly with their heads fixed in place. Then, a whole-cell recording is obtained using an efficient, blind patching protocol. We have successfully applied this technique to rats and mice.

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    04/03/17 | Whole-cell recording in the awake brain.
    Lee D, Lee AK
    Cold Spring Harbor Protocols. 2017 Apr 03;2017(4):pdb.top087304. doi: 10.1101/pdb.top087304

    Intracellular recording is an essential technique for investigating cellular mechanisms underlying complex brain functions. Despite the high sensitivity of the technique to mechanical disturbances, intracellular recording has been applied to awake, behaving, and even freely moving, animals. Here we summarize recent advances in these methods and their application to the measurement and manipulation of membrane potential dynamics for understanding neuronal computations in behaving animals.

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