Wide-field anti-aliased quantitative differential phase contrast microscopy

Published on 2018-09-13T18:36:19Z (GMT) by
Differential phase contrast (DPC) microscopy is a popular methodology to recover quantitative phase information of thin transparent sample under multi-axis asymmetric illumination patterns. Based on spatially partially coherent illuminations, DPC provides high-quality, speckle-free 3D reconstructions with lateral resolution up to twice the coherent diffraction limit, under the precondition that the pixel size of the imaging sensor is small enough to prevent spatial aliasing/undersampling. However, microscope cameras are in general designed to have a large pixel size so that the intensity information transmitted by the optical system cannot be adequately sampled or digitized. On the other hand, using a image sensor with smaller pixel size or adding a magnification camera adapter to the camera can resolve the undersampling at the expense of a reduced field of view (FOV). To solve this tradeoff, we introduce a new variation of quantitative DPC approach, termed anti-aliased DPC (AADPC), which uses several aliased intensity images under asymmetric illuminations to recover wide-field aliasing-free phase images. AADPC starts from an initial phase estimate obtained by a DPC-like deconvolution based on the system’s weak phase transfer function. Then the obtained initial phase map is further refined by the iterative de-multiplexing algorithm to overcome pixel-aliasing and improve the imaging resolution. The data redundancy requirement as well as the optimal illumination scheme of AADPC are analyzed and discussed based on several simulations, suggesting the spatial undersampling can be mitigated through the iterative algorithm that uses only 4 images, yielding a nearly 4-fold increase in the space-bandwidth product (SBP) compared to conventional DPC approach. We experimentally verify that AADPC can achieve a half-pitch imaging resolution of 345 nm, corresponding to 1.88× of the theoretical Nyquist-Shannon sampling resolution limit imposed by the sensor pixel size. The high-speed, high-throughput quantitative phase imaging capabilities of AADPC are also demonstrated by imaging HeLa cells mitosis in vitro, achieving a full-pitch lateral resolution of 665 nm across a wide FOV of 1.77mm2 at 25 fps.

Cite this collection

Fan, Yao; Sun, Jiasong; Zhang, Jianqin; Zuo, Chao (2018): Wide-field anti-aliased quantitative differential

phase contrast microscopy. The Optical Society. Collection.