Efficient quantitative phase microscopy using programmable annular LED illumination

Li, Jiaji; Chen, Qian; Zhang, Jialin; Zhang, Yan; Lu, Linpeng; Zuo, Chao

doi:10.1364/boe.8.004687

Cited by 52 publications

(26 citation statements)

References 51 publications

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“…Quantitative phase imaging (QPI) [1,4] is one such technique that measures an incident field's phase shifts induced by the sample to recover the sample's physical properties without staining or labeling. Both interferometry [5][6][7][8] and non-interferometry based [9][10][11][12][13] QPI techniques have been developed to recover a sample's phase map in two-dimensions (2D). A single 2D integrated phase image, however, is insufficient for characterizing 3D heterogeneous samples.…”

Section: Introductionmentioning

confidence: 99%

High-speed in vitro intensity diffraction tomography

Li¹,

Matlock²,

Li³

et al. 2019

Advanced Optical Imaging Technologies II

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We demonstrate a label-free, scan-free intensity diffraction tomography technique utilizing annular illumination (aIDT) to rapidly characterize large-volume 3D refractive index distributions in vitro. By optimally matching the illumination geometry to the microscope pupil, our technique reduces the data requirement by 60× to achieve highspeed 10 Hz volume rates. Using 8 intensity images, we recover ∼ 350 × 100 × 20µm 3 volumes with near diffraction-limited lateral resolution of 487 nm and axial resolution of 3.4 µm. Our technique's large volume rate and high resolution enables 3D quantitative phase imaging of complex living biological samples across multiple length scales. We demonstrate aIDT's capabilities on unicellular diatom microalgae, epithelial buccal cell clusters with native bacteria, and live Caenorhabditis elegans specimens. Within these samples, we recover macro-scale cellular structures, subcellular organelles, and dynamic micro-organism tissues with minimal motion artifacts. Quantifying such features has significant utility in oncology, immunology, and cellular pathophysiology, where these morphological features are evaluated for changes in the presence of disease, parasites, and new drug treatments. Finally, we simulate our aIDT system to highlight the accuracy and sensitivity of our technique. aIDT shows promise as a powerful high-speed, label-free computational microscopy technique applications where natural imaging is required to evaluate environmental effects on a sample in real-time. We provide example datasets and an open source implementation of aIDT at https://github.com/bu-cisl/IDT-using-Annular-Illumination.

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Section: Introductionmentioning

confidence: 99%

High-speed in vitro intensity diffraction tomography

Li¹,

Matlock²,

Li³

et al. 2019

Advanced Optical Imaging Technologies II

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show abstract

“…fluorescence, DIC, phase contrast). More recently, it has been found that the shape of the illumination aperture has a significant impact on the lateral resolution and noise sensitivity of TIE reconstruction [160][161][162][163], and by simply replacing the conventional circular illumination aperture with an annular one, high-resolution low-noise phase imaging can be achieved by using only 3-plane intensity measurements [160,161]. Figure 18.…”

Section: Samplementioning

confidence: 99%

3D Imaging based on Depth Measurement Technologies

Ni¹,

Zuo²,

Lam

et al. 2018

Preprint

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Three-dimensional (3D) imaging has attracted more and more interests because of its widespread applications, especially in information and life science. These techniques can be broadly divided into two types: ray-based and wavefront-based 3D imaging. Issues such as imaging quality and system complexity of these techniques limit the applications significantly, and therefore many investigations have focused on 3D imaging from depth measurements. This paper presents an overview of 3D imaging from depth measurements, and provides a summary of the connection between these the ray-based and wavefront-based 3D imaging techniques.

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“…Quantitative phase imaging (QPI) 1,4 is one such technique that measures an incident field's phase shifts induced by the sample to recover the sample's physical properties without staining or labeling. Both interferometry [5][6][7][8] and intensity-only [9][10][11][12][13] QPI techniques have been developed to recover a sample's phase map in two dimensions. A single two-dimensional (2-D) integrated phase image, however, is insufficient for characterizing 3-D heterogeneous samples.…”

Section: Introductionmentioning

confidence: 99%

High-speed in vitro intensity diffraction tomography

Matlock

et al. 2019

Adv. Photon.

Self Cite

116

View full text Add to dashboard Cite

We demonstrate a label-free, scan-free intensity diffraction tomography technique utilizing annular illumination (aIDT) to rapidly characterize large-volume three-dimensional (3-D) refractive index distributions in vitro. By optimally matching the illumination geometry to the microscope pupil, our technique reduces the data requirement by 60 times to achieve high-speed 10-Hz volume rates. Using eight intensity images, we recover volumes of ∼350 μm × 100 μm × 20 μm, with near diffraction-limited lateral resolution of ∼487 nm and axial resolution of ∼3.4 μm. The attained large volume rate and high-resolution enable 3-D quantitative phase imaging of complex living biological samples across multiple length scales. We demonstrate aIDT's capabilities on unicellular diatom microalgae, epithelial buccal cell clusters with native bacteria, and live Caenorhabditis elegans specimens. Within these samples, we recover macroscale cellular structures, subcellular organelles, and dynamic microorganism tissues with minimal motion artifacts. Quantifying such features has significant utility in oncology, immunology, and cellular pathophysiology, where these morphological features are evaluated for changes in the presence of disease, parasites, and new drug treatments. Finally, we simulate the aIDT system to highlight the accuracy and sensitivity of the proposed technique. aIDT shows promise as a powerful high-speed, label-free computational microscopy approach for applications where natural imaging is required to evaluate environmental effects on a sample in real time.

show abstract

Efficient quantitative phase microscopy using programmable annular LED illumination

Cited by 52 publications

References 51 publications

High-speed in vitro intensity diffraction tomography

High-speed in vitro intensity diffraction tomography

3D Imaging based on Depth Measurement Technologies

High-speed in vitro intensity diffraction tomography

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