基于凸优化的无荧光光场介观三维重建方法

Convex optimization-based mesoscopic 3D reconstruction method for fluorescence-free light fields

  • 摘要: 当前,显微三维观测多采用特异性荧光染色,会对样本造成不可逆的损伤,存在成本高、光毒性等问题。提出了无荧光标记、介观高分辨率三维重建的方法,即RGB-MesoLFM。首先,搭建了无荧光标记、白光介观光路,解耦了RGB三波段光场数据;然后,根据波动光学点扩散函数,计算出RGB三波段任意深度下的系统点脉冲响应;最后,基于凸优化理论构建目标函数,将RGB三波段光场数据、点脉冲响应作为输入,进行3D反卷积迭代,可重建出任意深度下高分辨率目标物体的切片图像。该三维重建实验是基于鸡蛋胚胎切片的光场采样结果,在重建切片序列间隔为4 µm的情况下,获得0~−20 µm范围下切片图像;与传统荧光重建方法相比,本文所提方法无需荧光,采用白光即可成像;其重建任意深度切片的峰值信噪比(PSNR)数值,较传统方法提升约10%。

     

    Abstract: Currently, the microscopic 3D observation primarily relies on specific fluorescence staining, which will cause irreversible damage to samples and presents issues such as high cost and phototoxicity. A novel method for mesoscopic high-resolution 3D reconstruction without fluorescence was proposed, known as RGB-MesoLFM. First, a fluorescence-free labeled white-light mesoscopic circuit was constructed, where the RGB three-band light field data were decoupled. Subsequently, based on fluctuating optics point spread function, the point impulse response of the system at any depth within the three bands was computed. Finally, an objective function was developed using convex optimization theory, considering the RGB three-band light field data and point impulse response as input for 3D deconvolution iteration, which can reconstruct the slice image of the high-resolution target object at any depth. The 3D reconstruction experiment was based on the light field sampling results of egg embryo slices, and could obtain slice images in the range of 0~−20 µm with a sequence interval of 4 µm. Compared with the traditional fluorescence reconstruction method, the proposed method does not require fluorescence, but uses white light for imaging. The reconstructed peak signal to noise ratio (PSNR) value of arbitrary depth slices is about 10% higher than that of traditional methods.

     

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