装置と技術

蛍光寿命イメージングシステムの製作と生体試料および高分子試料への応用

Construction of a Fluorescence Lifetime Imaging System and its Application to Biological Systems and Polymer Materials

中林 孝和・飯森 俊文・金城 政孝・太田 信廣
Takakazu NAKABAYASHI, Toshifumi IIMORI, Masataka KINJO, and Nobuhiro OHTA

細胞内での蛋白質間の相互作用や光励起ダイナミクスを明らかにすることを目的として，蛍光寿命イメージングシステムおよび細胞内の時間分解蛍光スペルトル測定システムの製作を行なった．時間ゲート法を用いることにより，数分で生細胞の蛍光寿命イメージングを測定できることがわかった．またピコ秒時間分解能における時間相関光子計数システムを組み合わせることにより，特定の部位における蛍光寿命の定量的な値を求めることができるような構成にした．

We have constructed a fluorescence lifetime imaging (FLIM) system to study the photo-induced dynamics of living cells. The output from a femtosecond mode-locked Ti:sapphire laser is frequency-doubled and focused onto the sample with an objective lens of a confocal microscope. The fluorescence decay of the sample is measured at each pixel of a scanning image. Each fluorescence lifetime is evaluated quickly using time-gating electronics, and the fluorescence lifetime image can be obtained within several minutes. We have combined the FLIM system with a picosecond time-correlated single-photon counting system in which a microchannel-plate photomultiplier is used for detection, so that the value of the fluorescence lifetime at a position can be analyzed quantitatively. A high contrast light-scattering image of a polymer film is also found to be obtained with the FLIM system.

Keywords: Fluorescence; Lifetime; Imaging; Living Cell; Polymer Film