Quantitative Measurements of the Activity of Signaling Molecules by Two-Photon Fluorescence Lifetime Imaging Microscopy

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  • Murakoshi Hideji
    自然科学研究機構　生理学研究所，脳機能計測・支援センター

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Other Title
  • 二光子蛍光寿命イメージング顕微鏡法によるシグナル分子活性計測
  • ニ ミツコ ケイコウ ジュミョウ イメージング ケンビキョウホウ ニ ヨル シグナル ブンシ カッセイ ケイソク

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Description

<p>Recent popularization of 2-photon fluorescence microscopy has enabled us to image cellular structures such as synapses located in deep tissue at the high spatial resolution. However, signal transduction mechanisms in synapse has been still elusive because of the lack of techniques to visualize protein activities or protein-protein interactions. Recently, the improvement of 2-photon fluorescence lifetime imaging microscopy (2pFLIM) to visualize the Förster resonance energy transfer (FRET) has overcame such a difficulty and has enabled us to visualize biochemical reactions. Using this technique, we have recently succeeded in imaging the activity of small GTPases. Here I introduce the principle of the 2pFLIM for monitoring intracellular protein activities and protein-protein interactions with an example: detecting small GTPase (Cdc42 and RhoA) activity in dendrites and synapses of hippocampal neurons in brain slices.</p>

Journal

  • KENBIKYO

    KENBIKYO 50 (2), 106-110, 2015-08-30

    The Japanese Society of Microscopy

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