Depth-multiplexed ptychographic microscopy for high-throughput imaging of stacked bio-specimens on a chip

Guo, Chengfei; Jiang, Shaowei; Yang, Liming; Song, Pengming; Pirhanov, Azady; Wang, Ruihai; Wang, Tianbo; Shao, Xiaopeng; Qian, Wenyuan; Cho, Yong Ku; Zheng, Guoan

doi:10.1016/j.bios.2022.115049

Cited by 18 publications

(8 citation statements)

References 46 publications

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“…In general, the performance of the FP method is influenced by the thickness of the tissue slide, i.e. thicker samples exhibit greater light scattering and absorption processes [18]. In Fig.…”

Section: Results and Conclusionmentioning

confidence: 99%

Towards kidney histophysiology by using Fourier ptychography

Valentino,

Bianco,

Miccio

et al. 2023

Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI

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Tissue physiology and especially pathology need a cutting-edge microscopic technique capable of imaging tissue morphology and functioning over a wide view with high-resolution details. Fourier Ptychography (FP), a quantitative phase image (QPI) method, overcomes this trade-off relying on the synthetic numerical aperture (NA) principle. FP uses low NA microscope objectives for having huge FoV and multiple illumination angles to synthetize a big NA. Moreover, FP furnishes phase contrast images in a stain-free modality after applying phase retrieval algorithms. The retrieved phase maps allow the possibility of extracting quantitative information about the sample, e.g. refractive index variation, physical thickness and consequently the 3D profile. Here, we apply FP to the analysis of kidney tissues. Both stain-free and stained slides with different dyes and thicknesses are considered. FP phase images are obtained and compared to light-microscopy images of the same slides to discuss the capability of QPI in describing the morphology of kidney samples. Usually, haematoxylin and eosin (H&E) stain is considered a gold standard to make pathological inference on sick tissues because it marks cell nuclei. We propose a new way of observing renal inner structures such as glomeruli and tubules, which are clearly visible without staining. This work could lead physicians to use stainfree phase-contrast images, paving the way to extract FP diagnostic and prognostic biomarkers to enrich pathomic. We reach ~ 0.5 µm of resolution over an area of 3.3 mm2 for each sample up to 10 µm of tissue thickness.

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Section: Results and Conclusionmentioning

confidence: 99%

Towards kidney histophysiology by using Fourier ptychography

Valentino,

Bianco,

Miccio

et al. 2023

Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI

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“…Figure 8 illustrates the versatility of the coded thin film, extending its application from lens-based settings to lensless coded ptychography. [60][61][62][63][64][65][66][67][68][69] Figure 8a showcases the experimental setup where the coded thin film is attached to the coverglass of an image sensor, effectively serving as a computational scattering lens. The use of coded thin film replaces the use of blood cell layers or microbeads as coded surfaces, [30,60,63] which previously required permanent attachment to a sensor's coverglass, a process often marred by inconsistencies in uniformity.…”

Section: Experimental Demonstrationsmentioning

confidence: 99%

Spatially‐Coded Fourier Ptychography: Flexible and Detachable Coded Thin Films for Quantitative Phase Imaging with Uniform Phase Transfer Characteristics

Wang,

Yang,

Lee

et al. 2024

Advanced Optical Materials

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Fourier ptychography (FP) is an enabling imaging technique that produces high‐resolution complex‐valued images with extended field coverages. However, when FP images a phase object with any specific spatial frequency, the captured images contain only constant values, rendering the recovery of the corresponding linear phase ramp impossible. This challenge is not unique to FP but also affects other common microscopy techniques — a rather counterintuitive outcome given their widespread use in phase imaging. The underlying issue originates from the non‐uniform phase transfer characteristic inherent in microscope systems, which impedes the conversion of object wavefields into discernible intensity variations. To address this challenge, spatially‐coded Fourier ptychography (scFP) is presented for true quantitative phase imaging. In scFP, a flexible and detachable coded thin film is attached atop the image sensor in a regular FP setup. The spatial modulation of this thin film ensures a uniform phase response across the entire synthetic bandwidth. It improves reconstruction quality, corrects refractive index underestimation issues prevalent in conventional FP, and adds a new dimension of measurement diversity in spatial domain. The development of scFP is expected to catalyze new research directions and applications for phase imaging, emphasizing the need for true quantitative accuracy with uniform frequency response.

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“…FP achieves this goal by capturing a set of low-resolution images at different illumination angles and then by computationally combining them to reconstruct one high-resolution image. This enables the visualization of fine local details of individual cells and the global organization of the tissue, allowing the detection of malignant cells and metastasis in histopathological screenings [9][10][11][12][13][14][15][16]. Moreover, being a non-destructive imaging, FP allows for repeated measurements of the same sample over time without the ambiguities related to photobleaching.…”

Section: ! Introductionmentioning

confidence: 99%

Deep learning assisted Fourier ptychography for cells and tissue analysis

Bianco,

Valentino,

Behal

et al. 2023

Optical Methods for Inspection, Characterization, and Imaging of Biomaterials VI

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Fourier Ptychography (FP) is a quantitative phase imaging technique that overcomes the trade-off between lateral resolution and field of view. Typically, low numerical aperture (NA) microscope objectives are used, and super-resolution is obtained by illuminating specimens at different angles, thus generating a larger synthetic NA. These features are pivotal in both cells and tissue slide analysis. FP provides phase contrast images by applying phase retrieval algorithms and the quantitative information can be used to describe and characterize stain-free cells and tissues. To make FP microscopy viable for clinical practice, the issues arising from misalignments of the optical system or the presence of scattering structures to be imaged should be considered though. These non-ideal recording conditions result in phase artefacts in the recovered highresolution FP reconstruction. To tackle this problem, we propose a blind multi-look FP (ML-FP) algorithm that directly minimizes the artefacts while ensuring correct phase retrieval and can be used by unskilled operators in clinical practice. Here, we show how ML-FP allows the analysis of cells seeded onto micropatterned substrates (for mechanobiology applications) and tissues (for physiology and diagnostic applications). In order to improve robustness in the presence of misalignments, we use the ML-FP outcome as a ground truth and train a GAN architecture to emulate the phase retrieval process. The GAN receives as input the complex amplitude at the first iteration of the FP phase retrieval algorithm and returns in real-time the high-resolution FP complex amplitude.

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Depth-multiplexed ptychographic microscopy for high-throughput imaging of stacked bio-specimens on a chip

Cited by 18 publications

References 46 publications

Towards kidney histophysiology by using Fourier ptychography

Towards kidney histophysiology by using Fourier ptychography

Spatially‐Coded Fourier Ptychography: Flexible and Detachable Coded Thin Films for Quantitative Phase Imaging with Uniform Phase Transfer Characteristics

Deep learning assisted Fourier ptychography for cells and tissue analysis

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