Using symmetric illumination and color camera to achieve high throughput Fourier ptychographic microscopy

Zhang, Ke; Gilley, Patrik; Abdoli, Neman; Chen, Xuxin; Fung, Kar‐Ming; Qiu, Yuchen

doi:10.1002/jbio.202200303

Cited by 3 publications

(2 citation statements)

References 36 publications

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“…Thus, FPM simultaneously achieves both high-resolution and large FOV imaging. Since a substantial number of raw images are needed in FPM, different strategies [5][6][7][8][9][10][11] have been investigated to reduce the load of image acquisition. Among them, multiplexed FPM [5][6][7][8] has demonstrated its effectiveness by adopting multiple LEDs and/or multiple wavelengths in per image capturing.…”

Section: Introductionmentioning

confidence: 99%

Evaluating the imaging quality of multiplexed illumination Fourier ptychography microscopy

Gilley,

Zhang,

Abdoli

et al. 2024

Biophotonics and Immune Responses XIX

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Fourier ptychography microscopy (FPM) is a computational imaging technique that enables high resolution and large FOV simultaneously. For FPM, multiplexed LED illumination can significantly improve the efficiency of image data acquisition at the cost of deteriorated quality in the reconstructed images. In this study, we aim to evaluate the imaging quality of multiplexed FPM with different illumination configurations. For this purpose, a prototype FPM microscope was developed, which was equipped with a 4×/0.1 NA objective lens. This prototype was used to test 1 LED conventional, 2 LED multiplexed, and 4 LED multiplexed FPM illumination configurations on a standard USAF 1951 resolution target and a cytology sample. Modulation transfer function (MTF) curves were generated from the reconstructed images to quantitatively compare the performance of different LED combinations. The results demonstrated that the resolution target image reconstructed using 1 LED illumination raw images can resolve up to 912.3 lp/mm, but it decreased to 812.7 lp/mm and 724.1 lp/mm when 2 LED and 4 LED illumination were adopted, respectively. The corresponding MTF curves indicate decreased contrast on most spatial frequencies when comparing reconstructed results between multiplexed (2/4 LED) and conventional illumination configurations. Accordingly, the quality of reconstructed clinical cytology sample images decreases as the number of LEDs per image increases. However, all of them have satisfactory quality for most clinical applications. This preliminary study provides useful information to facilitate the development of multiplexed illumination FPM imaging systems in the future.

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Section: Introductionmentioning

confidence: 99%

Evaluating the imaging quality of multiplexed illumination Fourier ptychography microscopy

Gilley,

Zhang,

Abdoli

et al. 2024

Biophotonics and Immune Responses XIX

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show abstract

“…However, there are very few studies focused on experimental assessment of the DOF of FPM systems. Accordingly, in this study we estimated the DOF of our FPM prototype equipped with a 4×/0.13 NA objective lens, under both conventional single LED illumination and symmetric illumination [8][9][10] . The modulation transfer function (MTF) of the FPM system was first estimated using the standard USAF1951 resolution target, and the spatial frequency at which the MTF value drops to 0.5 was determined.…”

Section: Introductionmentioning

confidence: 99%

Evaluating the depth of field for Fourier ptychography microscopy

Zhang¹,

Gilley

Abdoli³

et al. 2023

Biophotonics and Immune Responses XVIII

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Fourier Ptychography Microscopy (FPM) is considered as one emerging technology for the development of high efficiency and low-cost microscopic scanners. One of the major advantages of FPM is its large depth of field (DOF), which significantly reduces the mechanical accuracy requirement of the scanning stages. In this study, we experimentally measured the DOF for our FPM prototype under different illumination conditions. The measurements were based on the theory that the DOF is considered as the range along optical axis for which the contrast is above 80% of the maximum when adjusting the focus location. Accordingly, the contrast is estimated using the bar pattern on the standard resolution target USAF1951 where the modulation transfer function (MTF) curve value drops to 0.5. During the experiment, the FPM prototype is equipped with a 4×/0.13 NA objective lens, and the DOF measurement was conducted with conventional single LED illumination and symmetric illumination. The results demonstrate that the DOF of the single LED illumination FPM is 15.3 µm, which is close to the DOF of the objective lens (14.5 µm). The DOF increases to 22.7 µm when symmetric illumination is adopted, which agrees with the theoretical conclusion. This investigation provides meaningful information for the future optimization of the FPM-based microscopic digitizers.

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Development and Assessment of Multiple Illumination Color Fourier Ptychographic Microscopy for High Throughput Sample Digitization

Gilley,

Zhang,

Abdoli

et al. 2024

Sensors

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In this study, we proposed a multiplexed color illumination strategy to improve the data acquisition efficiency of Fourier ptychography microscopy (FPM). Instead of sequentially lighting up one single channel LED, our method turns on multiple white light LEDs for each image acquisition via a color camera. Thus, each raw image contains multiplexed spectral information. An FPM prototype was developed, which was equipped with a 4×/0.13 NA objective lens to achieve a spatial resolution equivalent to that of a 20×/0.4 NA objective lens. Both two- and four-LED illumination patterns were designed and applied during the experiments. A USAF 1951 resolution target was first imaged under these illumination conditions, based on which MTF curves were generated to assess the corresponding imaging performance. Next, H&E tissue samples and analyzable metaphase chromosome cells were used to evaluate the clinical utility of our strategy. The results show that the single and multiplexed (two- or four-LED) illumination results achieved comparable imaging performance on all the three channels of the MTF curves. Meanwhile, the reconstructed tissue or cell images successfully retain the definition of cell nuclei and cytoplasm and can better preserve the cell edges as compared to the results from the conventional microscopes. This study initially validates the feasibility of multiplexed color illumination for the future development of high-throughput FPM scanning systems.

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Using symmetric illumination and color camera to achieve high throughput Fourier ptychographic microscopy

Cited by 3 publications

References 36 publications

Evaluating the imaging quality of multiplexed illumination Fourier ptychography microscopy

Evaluating the imaging quality of multiplexed illumination Fourier ptychography microscopy

Evaluating the depth of field for Fourier ptychography microscopy

Development and Assessment of Multiple Illumination Color Fourier Ptychographic Microscopy for High Throughput Sample Digitization

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