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用于光谱分辨各向异性成像的共聚焦显微镜的设计与应用

Design and application of a confocal microscope for spectrally resolved anisotropy imaging.

作者信息

Esposito Alessandro, Bader Arjen N, Schlachter Simon C, van den Heuvel Dave J, Schierle Gabriele S Kaminski, Venkitaraman Ashok R, Kaminski Clemens F, Gerritsen Hans C

机构信息

The Medical Research Council Cancer Cell Unit, Hutchison/MRC Research Centre, Hills Road, Cambridge CB2 0XZ, UK.

出版信息

Opt Express. 2011 Jan 31;19(3):2546-55. doi: 10.1364/OE.19.002546.

Abstract

Biophysical imaging tools exploit several properties of fluorescence to map cellular biochemistry. However, the engineering of a cost-effective and user-friendly detection system for sensing the diverse properties of fluorescence is a difficult challenge. Here, we present a novel architecture for a spectrograph that permits integrated characterization of excitation, emission and fluorescence anisotropy spectra in a quantitative and efficient manner. This sensing platform achieves excellent versatility of use at comparatively low costs. We demonstrate the novel optical design with example images of plant cells and of mammalian cells expressing fluorescent proteins undergoing energy transfer.

摘要

生物物理成像工具利用荧光的多种特性来绘制细胞生物化学图谱。然而,设计一种经济高效且用户友好的检测系统来感知荧光的多样特性是一项艰巨的挑战。在此,我们展示了一种新型光谱仪架构,它能够以定量且高效的方式对激发光谱、发射光谱和荧光各向异性光谱进行综合表征。这个传感平台以相对较低的成本实现了出色的通用性。我们通过植物细胞以及表达经历能量转移的荧光蛋白的哺乳动物细胞的示例图像来展示这种新型光学设计。

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