van Rooij Jos, Kalkman Jeroen
Department of Imaging Physics, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands.
Biomed Opt Express. 2019 Mar 11;10(4):1782-1793. doi: 10.1364/BOE.10.001782. eCollection 2019 Apr 1.
In this work we demonstrate large-scale high-sensitivity optical diffraction tomography (ODT) of zebrafish. We make this possible by three improvements. First, we obtain a large field of view while still maintaining a high resolution by using a high magnification over numerical aperture ratio digital holography set-up. With the inclusion of phase shifting we operate close to the optimum magnification over numerical aperture ratio. Second, we decrease the noise in the reconstructed images by implementing off-axis sample placement and numerical focus tracking in combination with the acquisition of a large number of projections. Although both techniques lead to an increase in sensitivity independently, we show that combining them is necessary in order to make optimal use of the potential gain offered by each respective method and obtain a refractive index (RI) sensitivity of . Third, we optimize the optical clearing procedure to prevent scattering and refraction to occur. We demonstrate our technique by imaging a zebrafish larva over 13 field of view with 4 micrometer resolution. Finally, we demonstrate a clinical application of our technique by imaging an entire adult cryoinjured zebrafish heart.
在这项工作中，我们展示了对斑马鱼的大规模高灵敏度光学衍射层析成像（ODT）。我们通过三项改进实现了这一点。首先，我们使用高放大倍率与数值孔径比的数字全息装置，在保持高分辨率的同时获得了大视野。通过加入相移，我们在接近最佳放大倍率与数值孔径比的条件下操作。其次，我们通过实施离轴样本放置和数值焦点跟踪，并结合采集大量投影，来降低重建图像中的噪声。虽然这两种技术各自都会提高灵敏度，但我们表明，将它们结合起来是必要的，以便充分利用每种方法提供的潜在增益，并获得 的折射率（RI）灵敏度。第三，我们优化光学清除程序，以防止散射和折射的发生。我们通过对一只斑马鱼幼虫在13 视野上进行4微米分辨率的成像来展示我们的技术。最后，我们通过对一条成年冷冻损伤斑马鱼心脏进行整体成像，展示了我们技术的临床应用。
