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System for fluorescence lifetime imaging microscopy

机译：荧光寿命成像显微镜系统

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Abstract: One of the promising recent developments in fluorescence microscopy is fluorescence lifetime imaging microscopy (FLIM). In this technique the fluorescence lifetime (ns range) of molecules is expressed in the image rather than the intensity of the light emitted by these molecules. This physical property is of interest as it gives information about the local environment of the molecule, such as molecular concentration of O$-2$/, Ca$+2$PLU$/, pH, and conjugation. We develop an affordable, robust and easy-to-use FLIM workstation which is completely automated and does not need any difficult calibration procedure. The system consists of a standard fluorescence microscope, a modulated excitation light source, a camera, a modulation signal generator and acquisition/processing software. The camera contains an Intensified CCD of which the image intensifier gain is modulated. Depending on the application different light sources can be selected. The current light source contains a 12 mW modulated laser-diode emitting at 635 nm. A homodyne detection scheme with modulation frequencies of 1 to 100 MHz is applied, aiming at a resolution of 0.1 ns or better. High level image acquisition strategies are implemented in software, along with the low level image processing routines for lifetime estimation, calibration and correction. An evaluation of the system and its critical components will be presented in this paper. !9
机译：摘要：荧光寿命成像显微镜（FLIM）是荧光显微镜领域最近有希望的发展之一。在这种技术中，分子的荧光寿命（ns范围）在图像中表示，而不是这些分子发出的光的强度。该物理性质令人关注，因为它提供了有关分子局部环境的信息，例如O $ -2 $ /，Ca $ + 2 $ PLU $ /的分子浓度，pH和结合。我们开发了价格合理，功能强大且易于使用的FLIM工作站，该工作站是完全自动化的，不需要任何困难的校准过程。该系统由标准荧光显微镜，调制激发光源，照相机，调制信号发生器和采集/处理软件组成。照相机包含一个增强型CCD，其图像增强器增益被调制。根据应用，可以选择不同的光源。当前光源包含一个在635 nm发射的12 mW调制激光二极管。针对0.1 ns或更佳的分辨率，采用了调制频率为1至100 MHz的零差检测方案。高级图像采集策略在软件中实现，同时还有用于寿命估算，校准和校正的低级图像处理例程。本文将对系统及其关键组件进行评估。 ！9

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