Digital inline holographic microscopy (DIHM) of weakly-scattering subjects.
Digital inline holographic microscopy (DIHM) of weakly-scattering subjects.
复制标题
弱散射对象的数字在线全息显微镜 (DIHM)。
DOI：
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复制
发表时间：
2014
期刊：
影响因子：
--
通讯作者：
L. Wilson
中科院分区：
文献类型：
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作者：
C. Giuliano;Rongjing Zhang;L. Wilson
弱散射物体，如小胶体颗粒和大多数生物细胞，在显微镜中经常遇到。事实上，已经开发了一系列技术来更好地可视化这些相位对象；相衬和DIC是增强对比度最常用的方法。然而，在成像平面外方向记录位置和形状仍然具有挑战性。本文介绍了一种简单的实验方法，利用数字内联全息显微镜（DIHM）在三维空间中精确确定物体的位置和几何形状。一般来说，可访问的样本量由相机传感器在横向上的尺寸和轴向上的照明相干度来定义。典型的样品体积范围从200 μ m x 200 μ m x 200 μ m使用LED照明，到5mm x 5mm x 5mm或更大使用激光照明。这种照明光被配置成使平面波入射到样品上。然后，样品体积中的物体散射光，散射光与未散射光相互干扰，形成垂直于照明方向的干涉图案。该图像（全息图）包含三维重建所需的深度信息，可以在CMOS或CCD相机等标准成像设备上捕获。采用Rayleigh-Sommerfeld反向传播法对显微镜图像进行数值重聚焦，并采用基于Gouy相位异常的简单成像启发式方法识别重建体内的散射目标。这种简单但稳健的方法可以对微观样品中物体的位置和形状进行明确、无模型的测量。
Weakly-scattering objects, such as small colloidal particles and most biological cells, are frequently encountered in microscopy. Indeed, a range of techniques have been developed to better visualize these phase objects; phase contrast and DIC are among the most popular methods for enhancing contrast. However, recording position and shape in the out-of-imaging-plane direction remains challenging. This report introduces a simple experimental method to accurately determine the location and geometry of objects in three dimensions, using digital inline holographic microscopy (DIHM). Broadly speaking, the accessible sample volume is defined by the camera sensor size in the lateral direction, and the illumination coherence in the axial direction. Typical sample volumes range from 200 µm x 200 µm x 200 µm using LED illumination, to 5 mm x 5 mm x 5 mm or larger using laser illumination. This illumination light is configured so that plane waves are incident on the sample. Objects in the sample volume then scatter light, which interferes with the unscattered light to form interference patterns perpendicular to the illumination direction. This image (the hologram) contains the depth information required for three-dimensional reconstruction, and can be captured on a standard imaging device such as a CMOS or CCD camera. The Rayleigh-Sommerfeld back propagation method is employed to numerically refocus microscope images, and a simple imaging heuristic based on the Gouy phase anomaly is used to identify scattering objects within the reconstructed volume. This simple but robust method results in an unambiguous, model-free measurement of the location and shape of objects in microscopic samples.
影响因子：
8.6
作者：
Besseling, R.;Weeks, Eric R.;Poon, W. C. K.
通讯作者：
Poon, W. C. K.
细菌鞭毛旋转的动力学和能量学。
DOI：
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发表时间：
1982
期刊：
Symposia of the Society for Experimental Biology
影响因子：
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作者：
Berg,HC;Manson,MD;Conley,MP
通讯作者：
Conley,MP