Optimal resolution in Fresnel incoherent correlation holographic fluorescence microscopy.
Optimal resolution in Fresnel incoherent correlation holographic fluorescence microscopy.
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DOI：
10.1364/oe.19.005047
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发表时间：
2011-03-14
期刊：
影响因子：
3.8
通讯作者：
Rosen J
中科院分区：
文献类型：
--
作者：
Brooker G;Siegel N;Wang V;Rosen J
菲涅耳非相干相关全息术（FINCH）使全息图和3D图像能够仅用相机和空间光调制器（SLM）从非相干光中创建。我们先前描述了它的应用，显微非相干荧光，其中一个复杂的全息图包含所有的3D信息在显微镜领域，避免了扫描或连续切片的需要。我们现在报告的实验，导致最佳的光学，电光，和计算条件所需的全息图，产生高品质的3D图像从荧光显微镜标本。从我们以前的FINCH配置的一个重要改进利用SLM的偏振灵敏度，使相同的SLM像素，创建模拟显微镜管透镜的球面波，也通过从无限远校正显微镜物镜的平面波，使两种波类型之间的干涉在相机创建全息图。这一进步极大地提高了FINCH系统的分辨率。从成像的荧光USAF模式和花粉粒幻灯片的结果显示分辨率接近瑞利极限，通过这种简单的方法进行三维荧光显微成像。
Fresnel Incoherent Correlation Holography (FINCH) enables holograms and 3D images to be created from incoherent light with just a camera and spatial light modulator (SLM). We previously described its application to microscopic incoherent fluorescence wherein one complex hologram contains all the 3D information in the microscope field, obviating the need for scanning or serial sectioning. We now report experiments which have led to the optimal optical, electro-optic, and computational conditions necessary to produce holograms which yield high quality 3D images from fluorescent microscopic specimens. An important improvement from our previous FINCH configurations capitalizes on the polarization sensitivity of the SLM so that the same SLM pixels which create the spherical wave simulating the microscope tube lens, also pass the plane waves from the infinity corrected microscope objective, so that interference between the two wave types at the camera creates a hologram. This advance dramatically improves the resolution of the FINCH system. Results from imaging a fluorescent USAF pattern and a pollen grain slide reveal resolution which approaches the Rayleigh limit by this simple method for 3D fluorescent microscopic imaging.