Application of fluorescence lifetime imaging microscopy to the investigation of intracellular PDT mechanisms

A D Scully, R B Ostler, D Phillips, P O'Neill, K M S Townsend, A W Parker, A J MacRobert
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Abstract

The potential for the application of fluorescence lifetime imaging (FLIM) microscopy to studies of photosensitization mechanisms in photodynamic therapy (PDT) has been investigated. The fluorescence microscope incorporates a standard inverted optical microscope, a picosecond pulsed dye-laser excitation source, and an intensified CCD camera detector capable of being gated on a sub-nanosecond timescale. Fluorescence lifetime images resulting from multi-component analysis of sub-nanosecond gated fluorescence images of monolayer V79-4 Chinese hamster lung fibroblasts stained with disulphonated aluminium phthalocyanine (AlPcS2), a photosensitizer used in PDT, are presented. The results of these measurements are discussed in terms of the intracellular localization of the sensitizer. Preliminary results from multi-component FLIM of V79-4 cells multiply stained with AlPcS2 and a potential intracellular pH lifetime probe, 5(+6)-carboxynaphthofluorescein, are also presented.

荧光寿命成像显微镜在细胞内PDT机制研究中的应用
研究了荧光寿命成像(FLIM)显微技术在光动力治疗(PDT)中光敏化机制研究中的应用潜力。该荧光显微镜包括一个标准的倒置光学显微镜,一个皮秒脉冲染料激光激发源,以及一个增强的CCD相机探测器,能够在亚纳秒的时间尺度上进行门控。本文报道了用PDT光敏剂二磺化酞菁铝(AlPcS2)染色的单层V79-4中国仓鼠肺成纤维细胞亚纳秒门控荧光图像的多组分分析。这些测量的结果在细胞内敏化剂的定位方面进行了讨论。用AlPcS2和潜在的细胞内pH寿命探针5(+6)-carboxynaphthofluorescein对V79-4细胞进行多组分FLIM染色的初步结果也被提出。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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