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Related Experiment Videos

Two-photon targeted patching (TPTP) in vivo.

Shoji Komai1, Winfried Denk, Pavel Osten

  • 1Graduate School of Biological Sciences, Nara Institute of Science and Technology, Takayama 8916-5, Ikoma, Nara 630-0192, Japan.

Nature Protocols
|April 5, 2007
PubMed
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Two-photon targeted patching (TPTP) enables precise electrophysiological recordings from specific cells in vivo using advanced microscopy. This method allows detailed study of cellular function in intact organisms, applicable to various cell types.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Biophysics

Background:

  • Two-photon-excited fluorescence laser-scanning microscopy (2PLSM) is crucial for imaging biological processes in vivo.
  • Nonlinear optical methods like 2PLSM allow deep tissue imaging.
  • Combining microscopy with electrophysiology is essential for understanding cellular function.

Purpose of the Study:

  • To describe a novel protocol for targeted electrophysiological recordings in vivo.
  • To enable recording from specifically identified and labeled cells within intact biological tissues.
  • To demonstrate the utility of two-photon targeted patching (TPTP) for cellular analysis.

Main Methods:

  • The study introduces the two-photon targeted patching (TPTP) protocol.
  • TPTP utilizes 2PLSM to visualize and target fluorescently labeled cells.

Related Experiment Videos

  • Cells are identified using optical and genetic markers, such as green fluorescent protein (GFP).

    • Main Results:

    • TPTP successfully obtained electrophysiological recordings from specific cells in vivo.
    • The technique allows targeting of genetically identified neurons and their compartments, including dendrites.
    • Recordings can be repeated over several hours in the same preparation.

    Conclusions:

    • TPTP is a versatile method for in vivo cellular electrophysiology.
    • This technique integrates advanced microscopy with patch-clamp recording for precise cellular analysis.
    • TPTP is applicable to both neuronal and non-neuronal cell types.