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首页> 外文会议>Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XIII; Progress in Biomedical Optics and Imaging; vol.7 no.13 >Quantitative phase-contrast microscopy by angular spectrum digital holography
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Quantitative phase-contrast microscopy by angular spectrum digital holography

机译：角谱数字全息定量相差显微镜

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摘要

Techniques of digital holography are improved in order to obtain high-resolution, high-fidelity images of quantitative phase-contrast microscopy. In particular, the angular spectrum method of calculating the holographic optical field is seen to have several advantages over the more commonly used Fresnel transformation or Huygens convolution method. Spurious noise and interference components can be tightly controlled through the analysis and filtering of the angular spectrum. The reconstruction distance does not have a lower limit and the off-axis angle between the object and reference can be lower than the Fresnel requirement and still be able to cleanly separate out the zero-order background. Holographic phase images are largely immune from the coherent noise common in amplitude images. Together with the use of a miniature pulsed laser, the resulting images have 0.5 μm diffraction-limited lateral resolution and the phase profile is accurate to about 30 nm of optical path length. SKOV-3 (ovarian cancer cells) and HUVEC (human umbilical vein endothelial cells) are imaged that display intra-cellular and intra-nuclear organelles with clarity and quantitative accuracy. The technique clearly exceeds currently available methods in phase-contrast optical microscopy in the level of resolution and detail, and provides a new modality for imaging morphology of cellular and intracellular structures that is not currently available.
机译：为了获得定量，相衬显微镜的高分辨率，高保真图像，数字全息技术得到了改进。尤其是，与更常用的菲涅耳变换或惠更斯卷积方法相比，计算全息光学场的角谱方法具有多个优点。杂散噪声和干扰分量可以通过对角谱进行分析和过滤来严格控制。重建距离没有下限，并且对象与参考之间的离轴角可以小于菲涅耳要求，并且仍然能够清晰地分离出零阶背景。全息相位图像很大程度上不受振幅图像中常见的相干噪声的影响。与微型脉冲激光一起使用时，所得图像具有0.5μm的衍射极限横向分辨率，并且相位轮廓精确到大约30 nm的光程长度。对SKOV-3（卵巢癌细胞）和HUVEC（人脐静脉内皮细胞）进行成像，显示细胞内和核内细胞器的清晰度和定量准确性。该技术在分辨率和细节上明显超过了目前在相衬光学显微镜中可用的方法，并且为细胞和细胞内结构的成像形态学提供了一种目前尚不可用的新形式。

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