论文标题

低剂量3D体积荧光显微镜成像使用压缩传感

Low dosage 3D volume fluorescence microscopy imaging using compressive sensing

论文作者

Mannam, Varun, Brandt, Jacob, Smith, Cody J., Howard, Scott

论文摘要

荧光显微镜已成为观察胚胎(体内)长期成像的重要工具。但是,累积暴露是对这种敏感的活样品的光毒。虽然诸如荧光荧光显微镜(LSFM)之类的技术可减少暴露,但它不适合深层成像模型。其他计算技术在计算上是昂贵的,并且通常缺乏恢复质量。为了应对这一挑战,可以使用各种低剂量成像技术,这些技术是使用轴向方向(z轴)的几个切片来实现3D体积重建的;但是,他们通常缺乏恢复质量。同样,在轴向方向上获取密集的图像(以小步骤)在计算上也很昂贵。为了应对这一挑战,我们提出了一种基于压缩感应的方法(CS)方法,用于以相同的信噪比(SNR)(少于一半的激发剂量)重建3D体积。我们介绍了理论并实验验证方法。为了证明我们的技术,我们使用共聚焦显微镜捕获了斑马鱼胚胎脊髓(30UM厚度)中RFP标记的神经元的3D体积。从结果来看,我们观察到基于CS的方法可从不到20%的整个堆栈光学部分获得准确的3D体积重建。这项工作中开发的基于CS的方法可以轻松地应用于其他深层成像方式,例如两光子和灯表显微镜,其中降低样品的光吸毒性是一个至关重要的挑战。

Fluorescence microscopy has been a significant tool to observe long-term imaging of embryos (in vivo) growth over time. However, cumulative exposure is phototoxic to such sensitive live samples. While techniques like light-sheet fluorescence microscopy (LSFM) allow for reduced exposure, it is not well suited for deep imaging models. Other computational techniques are computationally expensive and often lack restoration quality. To address this challenge, one can use various low-dosage imaging techniques that are developed to achieve the 3D volume reconstruction using a few slices in the axial direction (z-axis); however, they often lack restoration quality. Also, acquiring dense images (with small steps) in the axial direction is computationally expensive. To address this challenge, we present a compressive sensing (CS) based approach to fully reconstruct 3D volumes with the same signal-to-noise ratio (SNR) with less than half of the excitation dosage. We present the theory and experimentally validate the approach. To demonstrate our technique, we capture a 3D volume of the RFP labeled neurons in the zebrafish embryo spinal cord (30um thickness) with the axial sampling of 0.1um using a confocal microscope. From the results, we observe the CS-based approach achieves accurate 3D volume reconstruction from less than 20% of the entire stack optical sections. The developed CS-based methodology in this work can be easily applied to other deep imaging modalities such as two-photon and light-sheet microscopy, where reducing sample photo-toxicity is a critical challenge.

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