Wu Jimin, Boominathan Vivek, Veeraraghavan Ashok, Robinson Jacob T
Department of Bioengineering, Rice University, Houston, Texas 77005, USA.
Department of Electrical and Computer Engineering, Rice University, Houston, Texas 77005, USA.
Biomed Opt Express. 2023 Jul 10;14(8):4037-4051. doi: 10.1364/BOE.490199. eCollection 2023 Aug 1.
Traditional miniaturized fluorescence microscopes are critical tools for modern biology. Invariably, they struggle to simultaneously image with a high spatial resolution and a large field of view (FOV). Lensless microscopes offer a solution to this limitation. However, real-time visualization of samples is not possible with lensless imaging, as image reconstruction can take minutes to complete. This poses a challenge for usability, as real-time visualization is a crucial feature that assists users in identifying and locating the imaging target. The issue is particularly pronounced in lensless microscopes that operate at close imaging distances. Imaging at close distances requires shift-varying deconvolution to account for the variation of the point spread function (PSF) across the FOV. Here, we present a lensless microscope that achieves real-time image reconstruction by eliminating the use of an iterative reconstruction algorithm. The neural network-based reconstruction method we show here, achieves more than 10000 times increase in reconstruction speed compared to iterative reconstruction. The increased reconstruction speed allows us to visualize the results of our lensless microscope at more than 25 frames per second (fps), while achieving better than 7 µm resolution over a FOV of 10 mm. This ability to reconstruct and visualize samples in real-time empowers a more user-friendly interaction with lensless microscopes. The users are able to use these microscopes much like they currently do with conventional microscopes.
传统的小型化荧光显微镜是现代生物学的关键工具。它们总是难以同时实现高空间分辨率和大视场(FOV)成像。无透镜显微镜为这一局限性提供了解决方案。然而,无透镜成像无法对样本进行实时可视化,因为图像重建可能需要数分钟才能完成。这对可用性构成了挑战,因为实时可视化是帮助用户识别和定位成像目标的关键特性。在近距离成像的无透镜显微镜中,这个问题尤为突出。近距离成像需要采用变移反卷积来考虑点扩散函数(PSF)在整个视场中的变化。在此,我们展示了一种无透镜显微镜,它通过摒弃迭代重建算法实现了实时图像重建。我们在此展示的基于神经网络的重建方法,与迭代重建相比,重建速度提高了一万多倍。重建速度的提升使我们能够以每秒超过25帧(fps)的速度可视化无透镜显微镜的结果,同时在10毫米的视场内实现优于7微米的分辨率。这种实时重建和可视化样本的能力使无透镜显微镜的交互更加用户友好。用户使用这些显微镜的方式与他们目前使用传统显微镜的方式非常相似。