Chung Jaebum, Kim Jinho, Ou Xiaoze, Horstmeyer Roarke, Yang Changhuei
Department of Electrical Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Biomed Opt Express. 2016 Jan 7;7(2):352-68. doi: 10.1364/BOE.7.000352. eCollection 2016 Feb 1.
This paper presents a method to simultaneously acquire an aberration-corrected, wide field-of-view fluorescence image and a high-resolution coherent bright-field image using a computational microscopy method. First, the procedure applies Fourier ptychographic microscopy (FPM) to retrieve the amplitude and phase of a sample, at a resolution that significantly exceeds the cutoff spatial frequency of the microscope objective lens. At the same time, redundancy within the set of acquired FPM bright-field images offers a means to estimate microscope aberrations. Second, the procedure acquires an aberrated fluorescence image, and computationally improves its resolution through deconvolution with the estimated aberration map. An experimental demonstration successfully improves the bright-field resolution of fixed, stained and fluorescently tagged HeLa cells by a factor of 4.9, and reduces the error caused by aberrations in a fluorescence image by up to 31%, over a field of view of 6.2 mm by 9.3 mm. For optimal deconvolution, we show the fluorescence image needs to have a signal-to-noise ratio of at least ~18.
本文提出了一种使用计算显微镜方法同时获取像差校正后的宽视场荧光图像和高分辨率相干明场图像的方法。首先,该过程应用傅里叶叠层显微镜(FPM)以显著超过显微镜物镜截止空间频率的分辨率检索样品的幅度和相位。同时,采集的FPM明场图像集内的冗余提供了一种估计显微镜像差的方法。其次,该过程获取一幅像差荧光图像,并通过与估计的像差图进行去卷积计算来提高其分辨率。实验证明,在6.2毫米×9.3毫米的视场内,成功将固定、染色和荧光标记的HeLa细胞的明场分辨率提高了4.9倍,并将荧光图像中由像差引起的误差降低了多达31%。为了实现最佳去卷积,我们表明荧光图像的信噪比至少需要达到~18。