RESUMEN

Quantitative phase microscopy (QPM) is indispensable in biomedical research due to its advantages in unlabeled transparent sample thickness quantification and obtaining refractive index information. Fourier ptychographic microscopy (FPM) is among the most promising QPM methods, incorporating multi-angle illumination and iterative phase recovery for high-resolution quantitative phase imaging (QPI) of large cell populations over a wide field of-view (FOV) in a single pass. However, FPM is limited by data redundancy and sequential acquisition strategies, resulting in low imaging efficiency, which in turn limits its real-time application in in vitro label-free imaging. Here, we report a fast QPM based on Fourier ptychography (FQP-FPM), which uses an optimized annular downsampling and parallel acquisition strategy to minimize the amount of data required in the front end and reduce the iteration time of the back-end algorithm (3.3% and 4.4% of conventional FPM, respectively). Theoretical and data redundancy analyses show that FQP-FPM can realize high-throughput quantitative phase reconstruction at thrice the resolution of the coherent diffraction limit by acquiring only ten raw images, providing a precondition for in vitro label-free real-time imaging. The FQP-FPM application was validated for various in vitro label-free live-cell imaging. Cell morphology and subcellular phenomena in different periods were observed with a synthetic aperture of 0.75 NA at a 10× FOV, demonstrating its advantages and application potential for fast high-throughput QPI.

RESUMEN

Fourier ptychographic microscopy (FPM) is used to achieve high resolution and a large field of view. However, traditional FPM image reconstruction methods often yield poor image quality when encountering out-of-focus issues during reconstruction. Therefore, this study proposes a defocus-distance regression network based on convolutional neural networks. In an experimental validation, the root-mean-square error calculated from 1000 sets of predicted and true values was approximately 6.2 µm. The experimental results suggest that the proposed method has good generalization, maintains high accuracy in predicting defocus distances even for different biological samples, and extends the imaging depth-of-field of the FPM system by a factor of more than 3.

RESUMEN

Significance: Fourier ptychographic microscopy (FPM) is a new, developing computational imaging technology. It can realize the quantitative phase imaging of a wide field of view and high-resolution (HR) simultaneously by means of multi-angle illumination via a light emitting diode (LED) array, combined with a phase recovery algorithm and the synthetic aperture principle. However, in the FPM reconstruction process, LED position misalignment affects the quality of the reconstructed image, and the reconstruction efficiency of the existing LED position correction algorithms needs to be improved. Aim: This study aims to improve the FPM correction method based on simulated annealing (SA) and proposes a position misalignment correction method (AA-C algorithm) using an improved phase recovery strategy. Approach: The spectrum function update strategy was optimized by adding an adaptive control factor, and the reconstruction efficiency of the algorithm was improved. Results: The experimental results show that the proposed method is effective and robust for position misalignment correction of LED arrays in FPM, and the convergence speed can be improved by 21.2% and 54.9% compared with SC-FPM and PC-FPM, respectively. Conclusions: These results can reduce the requirement of the FPM system for LED array accuracy and improve robustness.

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RESUMEN

Significance: Fourier ptychographic microscopy (FPM) enables quantitative phase imaging with a large field-of-view and high resolution by acquiring a series of low-resolution intensity images corresponding to different spatial frequencies stitched together in the Fourier domain. However, the presence of various aberrations in an imaging system can significantly degrade the quality of reconstruction results. The imaging performance and efficiency of the existing embedded optical pupil function recovery (EPRY-FPM) aberration correction algorithm are low due to the optimization strategy. Aim: An aberration correction method (AA-P algorithm) based on an improved phase recovery strategy is proposed to improve the reconstruction image quality. Approach: This algorithm uses adaptive modulation factors, which are added while updating iterations to optimize the spectral function and optical pupil function updates of the samples, respectively. The effectiveness of the proposed algorithm is verified through simulations and experiments using an open-source biological sample dataset. Results: Experimental results show that the proposed AA-P algorithm in an optical system with hybrid aberrations, recovered complex amplitude images with clearer contours and higher phase contrast. The image reconstruction quality was improved by 82.6% when compared with the EPRY-FPM algorithm. Conclusions: The proposed AA-P algorithm can reconstruct better results with faster convergence, and the recovered optical pupil function can better characterize the aberration of the imaging system. Thus, our method is expected to reduce the strict requirements of wavefront aberration for the current FPM.

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RESUMEN

Fourier ptychographic microscopy (FPM) is a promising super-resolution computational imaging technology. It stitches a series of low-resolution (LR) images in the Fourier domain by an iterative method. Thus, it obtains a large field of view and high-resolution quantitative phase images. Owing to its capability to perform high-spatial bandwidth product imaging, FPM is widely used in the reconstruction of conventional static samples. However, the influence of the FPM imaging mechanism limits its application in high-speed dynamic imaging. To solve this problem, an adaptive-illumination FPM scheme using regional energy estimation is proposed. Starting with several captured real LR images, the energy distribution of all LR images is estimated, and select the measurement images with large information to perform FPM reconstruction. Simulation and experimental results show that the method produces efficient imaging performance and reduces the required volume of data to more than 65% while ensuring the quality of FPM reconstruction.

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