Optofluidic time-stretch quantitative phase microscopy

Guo, Baoping; Lei, Cheng; Wu, Yi; Kobayashi, Hirofumi; Ito, Takuro; Yalikun, Yaxiaer; Lee, Sangwook; Isozaki, Akihiro; Li, Ming; Jiang, Yiyue; Yasumoto, Atsushi; Carlo, Dino Di; Tanaka, Yo; Yatomi, Yutaka; Ozeki, Yasuyuki; Goda, Keisuke

doi:10.1016/j.ymeth.2017.10.004

Cited by 40 publications

(20 citation statements)

References 35 publications

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“…Another benefit from the interferometry‐free operation is that the quantitative phase retrieval in multi‐ATOM bypasses the need for the iterative phase unwrapping algorithms commonly used in QPI . Not only can it reduce the computational complexity which favors ultrafast real‐time image processing, but also, more importantly, eliminates the phase unwrapping error which is susceptible to noise and low fringe visibility . Hence, phase retrieval in multi‐ATOM is essentially error‐free compared to that in iTM (Figure E,F).…”

Section: Resultsmentioning

confidence: 99%

Multi‐ATOM: Ultrahigh‐throughput single‐cell quantitative phase imaging with subcellular resolution

Lee

Lau

Tang

et al. 2019

Journal of Biophotonics

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A growing body of evidence has substantiated the significance of quantitative phase imaging (QPI) in enabling cost‐effective and label‐free cellular assays, which provides useful insights into understanding the biophysical properties of cells and their roles in cellular functions. However, available QPI modalities are limited by the loss of imaging resolution at high throughput and thus run short of sufficient statistical power at the single‐cell precision to define cell identities in a large and heterogeneous population of cells—hindering their utility in mainstream biomedicine and biology. Here we present a new QPI modality, coined multiplexed asymmetric‐detection time‐stretch optical microscopy (multi‐ATOM) that captures and processes quantitative label‐free single‐cell images at ultrahigh throughput without compromising subcellular resolution. We show that multi‐ATOM, based upon ultrafast phase‐gradient encoding, outperforms state‐of‐the‐art QPI in permitting robust phase retrieval at a QPI throughput of >10 000 cell/sec, bypassing the need for interferometry which inevitably compromises QPI quality under ultrafast operation. We employ multi‐ATOM for large‐scale, label‐free, multivariate, cell‐type classification (e.g. breast cancer subtypes, and leukemic cells vs peripheral blood mononuclear cells) at high accuracy (>94%). Our results suggest that multi‐ATOM could empower new strategies in large‐scale biophysical single‐cell analysis with applications in biology and enriching disease diagnostics.

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Section: Resultsmentioning

confidence: 99%

Multi‐ATOM: Ultrahigh‐throughput single‐cell quantitative phase imaging with subcellular resolution

Lee

Lau

Tang

et al. 2019

Journal of Biophotonics

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show abstract

“…Another benefit from the interferometry-free operation is that the quantitative phase retrieval in multi-ATOM bypasses the need for the iterative phase unwrapping algorithms commonly used in QPI [29][30][31]. Not only can it reduce the computational complexity which favours ultrafast real-time image processing, but also more importantly, eliminates the phase unwrapping error which is susceptible to noise and low fringe visibility [22,29]. Hence, phase retrieval in multi-ATOM is essentially error-free compared to that in iTM (Figure 3d-e).…”

Section: Imaging Performance Of Multi-atommentioning

confidence: 99%

“…By contrast, optical time-stretch imaging offers a unique speed advantage over the conventional camera technologies, by achieving continuous frame rate of megahertz or beyond in real-time for high-throughput single-cell imaging in flow [17][18][19][20][21]. Further empowered by interferometry, this technology enables ultrafast QPI for label-free, cancer cell classification [22,23]. However, revealing sub-cellular features remains challenging in interferometric timestretch QPI.…”

Section: Introductionmentioning

confidence: 99%

Multi-ATOM: Ultrahigh-throughput single-cell quantitative phase imaging with subcellular resolution

Lee

Lau

Tang

et al. 2019

Preprint

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A growing body of evidence has substantiated the significance of quantitative phase imaging (QPI) in enabling cost-effective and label-free cellular assay, which provides useful insights into understanding biophysical properties of cells and their roles in cellular functions. However, available QPI modalities are limited by the loss of imaging resolution at high throughput and thus run short of sufficient statistical power at the single cell precision to define cell identities in a large and heterogeneous population of cellshindering their utility in mainstream biomedicine and biology. Here we present a new QPI modality, coined multi-ATOM that captures and processes quantitative label-free single-cell images at ultra-high throughput without compromising sub-cellular resolution. We show that multi-ATOM, based upon ultrafast phase-gradient encoding, outperforms state-of-the-art QPI in permitting robust phase retrieval at a QPI throughput of >10,000 cell/sec, bypassing the need for interferometry which inevitably compromises QPI quality under ultrafast operation. We employ multi-ATOM for large-scale, label-free, multi-variate, cell-type classification (e.g. breast cancer sub-types, and leukemic cells versus peripheral blood mononuclear cells) at high accuracy (>94%). Our results suggest that multi-ATOM could empower new strategies in large-scale biophysical single-cell analysis with applications in biology and enriching disease diagnostics.

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“…Subsequently, the system was modified and advanced to image fluorescently-labelled organelles in live worms (C. elegans) [15]. In the past few years, lightsheet imaging cytometry (LIC) has gained popularity and many research groups have started exploring LIC to incorporate live organism imaging [21], time-stretch imaging [22], phase-contrast [23], high-speed interrogation [24], fast sectioning [25], depth-penetration [26], determination of protein concentration [27], measurement of intracellular content [11] and label-free cell identification [28]. The new volume flow cytometry (parallel-iLIFE) is the next generation technology for applications in diverse fields.…”

Section: Introductionmentioning

confidence: 99%

High Throughput Volume Flow Cytometry (parallel-iLIFE) Resolves Mitochondrial Network On the Go

Kumar

Joshi

Basumatary

et al. 2020

Preprint

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Cell screening and viability studies are paramount to access cell morphology and intracellular molecular variations within large heterogeneous populations of cells. This forms the basis for diagnosis of infections, evaluating immunohistochemistry and routine histopathology. The proposed volume flow cytometry (also termed as, parallel Integrated Light-sheet imaging and flow-based enquiry (parallel-iLIFE)) is a powerful method that adds new capabilities ( 3D volume visualization, organelle-level resolution and multi-organelle screening) powered by light sheet based illumination. Unlike state-of-the-art point-illumination based imaging cytometry techniques, light sheet based parallel-iLIFE technique is capable of screening species with high throughput and near diffraction-limited resolution. The flow system was realized on a multichannel (Y-type) microfluidic chip that enables visualization of mitochondrial network of several cells in-parallel at a relatively high flow-rate of 2000 nl/min. The calibration of system requires study of point emitters (fluorescent beads) at physiologically relevant flow-rates (50 - 2000 nl/min) for determining flow-induced optical aberration in the system point spread function (PSF). Subsequently, recorded raw images and volumes were deconvolved with flow-variant PSF to reconstruct cellular mitocondrial network. High throughput investigation of HeLa cells were carried out at sub-cellular resolution in real-time and critical parameters (mitochondria count and size distribution, morphology and cell strain statistics) are determined on-the-go. These parameters determine the physiological state of cells and help determine changes in mitochondrial distribution over-time that may have consequences in disease diagnosis. The development of volume flow cytometry system (parallel-iLIFE ) and its suitability to study sub-cellular components at high-throughput high-content capacity with organelle-level resolution may enable disease diagnosis on a single microfluidic chip.

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Optofluidic time-stretch quantitative phase microscopy

Cited by 40 publications

References 35 publications

Multi‐ATOM: Ultrahigh‐throughput single‐cell quantitative phase imaging with subcellular resolution

Multi‐ATOM: Ultrahigh‐throughput single‐cell quantitative phase imaging with subcellular resolution

Multi-ATOM: Ultrahigh-throughput single-cell quantitative phase imaging with subcellular resolution

High Throughput Volume Flow Cytometry (parallel-iLIFE) Resolves Mitochondrial Network On the Go

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