Significance: Differential interference contrast (DIC) microscopes allow noninvasive in vivo observation of transparent microstructures in tissue without the use of fluorescent dyes or genetic modification. We show how to modify a DIC microscope to measure the sample phase distribution accurately and in real-time even deep inside sample tissue. Aim: Our aim is to improve the DIC microscope’s phase measurement to remove the phase bias that occurs in the presence of strong scattering. Approach: A quarter-wave plate was added in front of the polarization camera, allowing a modified phase calculation to incorporate all four polarization orientation angles (0 deg, 45 deg, 90 deg, and 135 deg) captured simultaneously by the polarization camera, followed by deconvolution. Results: We confirm that the proposed method reduces phase measurement error in the presence of scattering and demonstrate the method using in vivo imaging of a beating heart inside a medaka egg and the whole-body blood circulation in a young medaka fish. Conclusions: Modifying a polarization-camera DIC microscope with a quarter-wave plate allows users to image deep inside samples without phase bias due to scattering effects.

中文翻译：

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最小化微分干涉对比显微镜中散射引起的相位误差

意义：微分干涉对比 (DIC) 显微镜允许在不使用荧光染料或基因改造的情况下对组织中的透明微结构进行无创体内观察。我们展示了如何修改 DIC 显微镜以准确实时地测量样品相位分布，甚至深入样品组织内部。目标：我们的目标是改进 DIC 显微镜的相位测量，以消除存在强散射时出现的相位偏差。方法：在偏振相机前面添加一个四分之一波片，允许修改相位计算以包含偏振相机同时捕获的所有四个偏振方向角（0 度、45 度、90 度和 135 度），然后通过反卷积。结果：我们确认所提出的方法在存在散射的情况下减少了相位测量误差，并使用鳉卵内跳动的心脏的活体成像和年轻鳉鱼的全身血液循环证明了该方法。结论：使用四分之一波片修改偏振相机 DIC 显微镜允许用户在没有由于散射效应产生的相位偏差的情况下对样品内部深处进行成像。