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1.
Opt Express ; 32(1): 339-354, 2024 Jan 01.
Article in English | MEDLINE | ID: mdl-38175060

ABSTRACT

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.

2.
Biomed Opt Express ; 15(1): 95-113, 2024 Jan 01.
Article in English | MEDLINE | ID: mdl-38223174

ABSTRACT

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.

3.
J Biomed Opt ; 28(11): 116503, 2023 Nov.
Article in English | MEDLINE | ID: mdl-38078152

ABSTRACT

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.


Subject(s)
Lighting , Microscopy , Microscopy/methods , Fourier Analysis , Algorithms
4.
Microvasc Res ; 150: 104593, 2023 11.
Article in English | MEDLINE | ID: mdl-37582460

ABSTRACT

Nailfold capillary density is an essential physiological parameter for analyzing nailfold health; however, clinical images of the nailfold are taken in many situations, and most clinicians subjectively analyze nailfold images. Therefore, based on the improved "you only look once v5" (YOLOv5) algorithm, this study proposes an automated method for measuring nailfold capillary density. The improved technique can effectively and rapidly detect distal capillaries by incorporating methods or structures such as 9mosaic, spatial pyramid pooling cross-stage partial construction, bilinear interpolation, and efficient intersection over union. First, the modified YOLOv5 algorithm was used to detect nailfold capillaries. Subsequently, the number of distal capillaries was filtered using the 90° method. Finally, the capillary density was calculated. The results showed that the Average Precision (AP)@0.5 value of the proposed approach reached 85.2 %, which was an improvement of 4.93 %, 5.24 %, and 107 % compared with the original YOLOv5, YOLOv6, and simple-faster rapid-region convolutional network (R-CNN), respectively. For different nailfold images, using the density calculated by nailfold experts as a benchmark, the calculated results of the proposed method were consistent with the manually calculated results and superior to those of the original YOLOv5.


Subject(s)
Capillaries , Nails , Nails/blood supply , Microscopic Angioscopy/methods , Algorithms
5.
J Biomed Opt ; 28(3): 036006, 2023 03.
Article in English | MEDLINE | ID: mdl-36923986

ABSTRACT

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.


Subject(s)
Microscopy , Optical Devices , Microscopy/methods , Microscopy, Phase-Contrast , Light
6.
Opt Express ; 31(3): 4367-4378, 2023 Jan 30.
Article in English | MEDLINE | ID: mdl-36785407

ABSTRACT

Coupling between exiting wavefront error of space gravitational wave telescopes and tilt-to-length (TTL) noise affects the measurement accuracy. Using the LISA Pathfinder signal, we analyzed cancellation and superposition of TTL coupling noise under various optical aberrations. We proposed proportion requirements of any two aberrations amplitude when noise was cancelled and an aberration amplitude control requirement when noise was superposed. Taking them as the aberration control requirements of gravitational wave telescope optical system, the exiting wavefront error requirements was reduced while suppressing the TTL coupling noise. A 40× optical telescope system with detection aperture φ=200 mm was designed. The exiting wavefront error was relaxed from 0.02 λ to 0.0496 λ. The maximum coupling coefficient value did not exceed 6.9448 pm/µrad within a pointing jitter angle of ±300 µrad. The proposed approach should be useful in future telescope design.

7.
Microvasc Res ; 146: 104472, 2023 03.
Article in English | MEDLINE | ID: mdl-36572207

ABSTRACT

Collecting and analyzing human nailfold images is an important component of studying human microcirculation. However, the large-field-of-view and high-resolution nailfold images captured by research microscopes introduce issues such as uneven brightness, low imaging contrast, and unclear vascular contours. To overcome these issues, this paper proposes a hybrid enhancement algorithm for nailfold images with large fields of view. First, adaptive histogram equalization with limited contrast (Clahe) is used to redistribute gray levels to enhance the brightness and contrast of images. Next, nonlocal means denoising (NL-means) is used to remove the noise amplified by Clahe algorithm. Finally, unsharp masking (Usm) is used to enhance the edge contour information of nailfold blood vessels. Comparing the enhanced images reveals that the hybrid enhancement algorithm improves the brightness and contrast of the nailfold image, makes the nailfold vessel contour more obvious, and the image noise continues to remain small, and it obtains the best visual effect. It is superior to other algorithms in terms of objective indicators and subjective evaluation.


Subject(s)
Algorithms , Image Enhancement , Humans , Image Enhancement/methods , Microcirculation
8.
J Biophotonics ; 16(3): e202200240, 2023 03.
Article in English | MEDLINE | ID: mdl-36366908

ABSTRACT

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.


Subject(s)
Lighting , Microscopy , Microscopy/methods , Fourier Analysis , Algorithms , Image Processing, Computer-Assisted
9.
Opt Express ; 30(11): 18505-18517, 2022 May 23.
Article in English | MEDLINE | ID: mdl-36221650

ABSTRACT

Fourier ptychographic microscopy (FPM) imaging is a computational imaging technology that can reconstruct wide-field high-resolution (HR) images. It uses a series of low-resolution images captured by a camera under different illumination angles. The images are stitched in the Fourier domain to expand their spectral range. Under high-angle illumination, a dark-field image is noisy with a low signal-to-noise ratio, which significantly reduces the reconstruction quality of FPM. Conventional reconstruction algorithms often have low FPM imaging performance and efficiency due to optimization strategies. In response to these problems, this paper proposes an FPM imaging method based on an improved phase recovery strategy to optimize the alternating iterative algorithm. The technique uses an improved threshold method to reduce noise in the image preprocessing stage to maximize the retention of high-frequency sample information. Moreover, an adaptive control factor is added in the subsequent iterative update process to balance the sample spectrum function. This study verifies the effectiveness of the proposed method on both simulation and experimental images. The results show that the proposed method can effectively suppress image background noise and has a faster convergence speed and higher robustness. In addition, it can be used to reconstruct HR complex amplitude images of objects under wide field-of-view conditions.

10.
J Biophotonics ; 12(5): e201800329, 2019 05.
Article in English | MEDLINE | ID: mdl-30315638

ABSTRACT

A large-depth-of-field full-field optical angiography (LD-FFOA) method is developed to expand the depth-of-field (DOF) using a contrast pyramid fusion algorithm (CPFA). The absorption intensity fluctuation modulation effect is utilized to obtain full-field optical angiography (FFOA) images at different focus positions. The CPFA is used to process these FFOA images with different focuses. By selecting high-contrast areas, the CPFA can highlight the characteristics and details of blood vessels to obtain LD-FFOA images. In the optimal case of the proposed method, the DOF for FFOA is more than tripled using 10 differently focused FFOA images. Both the phantom and animal experimental results show that the LD-FFOA resolves FFOA defocusing issues induced by surface and thickness inhomogeneities in biological samples. The proposed method can be potentially applied to practical biological experiments.


Subject(s)
Angiography/methods , Optical Imaging/methods
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