Acute Brain Slice Methods for Adult and Aging Animals: Application of Targeted Patch Clamp Analysis and Optogenetics

  • Jonathan T. TingEmail author
  • Tanya L. Daigle
  • Qian Chen
  • Guoping Feng
Part of the Methods in Molecular Biology book series (MIMB, volume 1183)


The development of the living acute brain slice preparation for analyzing synaptic function roughly a half century ago was a pivotal achievement that greatly influenced the landscape of modern neuroscience. Indeed, many neuroscientists regard brain slices as the gold-standard model system for detailed cellular, molecular, and circuitry level analysis and perturbation of neuronal function. A critical limitation of this model system is the difficulty in preparing slices from adult and aging animals, and over the past several decades few substantial methodological improvements have emerged to facilitate patch clamp analysis in the mature adult stage. In this chapter we describe a robust and practical protocol for preparing brain slices from mature adult mice that are suitable for patch clamp analysis. This method reduces swelling and damage in superficial layers of the slices and improves the success rate for targeted patch clamp recordings, including recordings from fluorescently labeled populations in slices derived from transgenic mice. This adult brain slice method is suitable for diverse experimental applications, including both monitoring and manipulating neuronal activity with genetically encoded calcium indicators and optogenetic actuators, respectively. We describe the application of this adult brain slice platform and associated methods for screening kinetic properties of Channelrhodopsin (ChR) variants expressed in genetically defined neuronal subtypes.

Key words

Acute brain slice Adult animals Patch clamp recording Protective recovery method NMDG aCSF Optogenetics GCaMP Channelrhodopsin 



This work was supported in part by a National Alliance for Research on Schizophrenia and Depression: The Brain and Behavior Research Foundation Young Investigator Award to J.T.T., and US National Institutes of Health Ruth L. Kirschstein National Research Service Awards to J.T.T. (F32-MH084460).


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Jonathan T. Ting
    • 1
    Email author
  • Tanya L. Daigle
    • 2
    • 3
  • Qian Chen
    • 3
  • Guoping Feng
    • 3
  1. 1.Human Cell Types DepartmentAllen Institute for Brain ScienceSeattleUSA
  2. 2.Department of Cell BiologyDuke University Medical CenterDurhamUSA
  3. 3.McGovern Institute for Brain Research and Department of Brain and Cognitive SciencesMITCambridgeUSA

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