Fluorescence emission patterns near glass and metal-coated surfaces investigated with back focal plane imaging
Fluorescence emission patterns near glass and metal-coated surfaces investigated with back focal plane imaging
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DOI：
10.1117/1.2052867
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发表时间：
2005-09-01
影响因子：
3.5
通讯作者：
Axelrod, D
中科院分区：
文献类型：
--
作者：
Mattheyses, AL;Axelrod, D
显微镜观察到的荧光团通常位于靠近平面的地方，如在全内反射显微镜或单分子研究中。从经典电磁理论的观点出发，可以很好地理解在两个介质之间的平面（可能是金属薄膜）附近的荧光激发和发射的光学性质。我们提出了一种实验方法，该方法可以用显微镜光学测量在裸表面或支持表面等离子体共振的金属薄膜涂层表面附近的荧光手机发射模式的角依赖性。该技术涉及改变显微镜光学器件，直接记录（在数字CCD相机上）物镜后焦平面上的强度模式，从而直接映射荧光的角度发射模式。玻璃表面和镀铝表面的实验发射谱都是各向异性的，在临界角处或在临界角和表面等离子体角处都有峰值。观察到的剖面与计算机计算结果一致，并提出了在细胞生物物理学中可能有用的一些光学修饰。(c) 2005年光学仪器工程师学会。
Often fluorophores observed by microscopy are located close to a planar surface as in total internal reflection microscopy or single molecule studies. The optical properties of fluorescence exci tation and emission near a planar surface (possibly metal film coated) between two dielectrics are well understood from the viewpoint of classical electromagnetic theory. We present an experimental method whereby the angular dependence of the emission pattern of a fluoro phore near a bare surface or a metal film coated surface that supports surface plasmon resonance can be measured with microscope optics. and The technique involves an alteration of the microscope optics to directly record (on a digital CCD camera) the intensity pattern at the objective's back focal plane, which directly maps the angular emission pattern of fluorescence. The experimental emission profile on both glass and aluminum-coated surfaces is anisotropic with a peak at either the critical angle or both the critical angle and the surface plasmon angle. The observed profiles agree well with computer calculations and suggest some optical modifications that are potentially useful in cell biophysics. (c) 2005 Society of Photo-Optical Instrumentation Engineers.