Delft University of Technology, Lorentzweg 1, 2628 CJ, Delft, The Netherlands.
J Biomed Opt. 2012 Dec;17(12):126020. doi: 10.1117/1.jbo.17.12.126020.
We have built an all-solid-state camera that is directly modulated at the pixel level for frequency-domain fluorescence lifetime imaging microscopy (FLIM) measurements. This novel camera eliminates the need for an image intensifier through the use of an application-specific charge coupled device design in a frequency-domain FLIM system. The first stage of evaluation for the camera has been carried out. Camera characteristics such as noise distribution, dark current influence, camera gain, sampling density, sensitivity, linearity of photometric response, and optical transfer function have been studied through experiments. We are able to do lifetime measurement using our modulated, electron-multiplied fluorescence lifetime imaging microscope (MEM-FLIM) camera for various objects, e.g., fluorescein solution, fixed green fluorescent protein (GFP) cells, and GFP-actin stained live cells. A detailed comparison of a conventional microchannel plate (MCP)-based FLIM system and the MEM-FLIM system is presented. The MEM-FLIM camera shows higher resolution and a better image quality. The MEM-FLIM camera provides a new opportunity for performing frequency-domain FLIM.
我们构建了一种直接在像素级进行调制的全固态相机，用于频域荧光寿命成像显微镜 (FLIM) 测量。该新型相机通过在频域 FLIM 系统中使用特定于应用的电荷耦合器件设计，无需图像增强器。该相机的第一阶段评估已经完成。通过实验研究了相机的噪声分布、暗电流影响、相机增益、采样密度、灵敏度、光度响应线性度和光传递函数等特性。我们能够使用我们调制的、电子倍增荧光寿命成像显微镜 (MEM-FLIM) 相机对各种物体进行寿命测量，例如荧光素溶液、固定绿色荧光蛋白 (GFP) 细胞和 GFP-肌动蛋白染色的活细胞。本文详细比较了传统微通道板 (MCP)-based FLIM 系统和 MEM-FLIM 系统。MEM-FLIM 相机具有更高的分辨率和更好的图像质量。MEM-FLIM 相机为进行频域 FLIM 提供了新的机会。
