A light sheet based high throughput 3D-imaging flow cytometer for phytoplankton analysis

Wu, Jianglai; Li, Jianping; Chan, Robert K. Y.

doi:10.1364/oe.21.014474

Cited by 75 publications

(54 citation statements)

References 17 publications

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“…An inverted fluorescence microscope is positioned to take the perfectly focused image of the LS-illuminated section. This allows continuous recording of images sequentially as the sample travels Tutorial through this plane [124,125]. Such systems have been used for visualizing phytoplankton in water quality control studies.…”

Section: 2d Fluidics Approachmentioning

confidence: 99%

Light-sheet microscopy: a tutorial

et al. 2018

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This paper is intended to give a comprehensive review of light-sheet (LS) microscopy from an optics perspective. As such, emphasis is placed on the advantages that LS microscope configurations present, given the degree of freedom gained by uncoupling the excitation and detection arms. The new imaging properties are first highlighted in terms of optical parameters and how these have enabled several biomedical applications. Then, the basics are presented for understanding how a LS microscope works. This is followed by a presentation of a tutorial for LS microscope designs, each working at different resolutions and for different applications. Then, based on a numerical Fourier analysis and given the multiple possibilities for generating the LS in the microscope (using Gaussian, Bessel, and Airy beams in the linear and nonlinear regimes), a systematic comparison of their optical performance is presented. Finally, based on advances in optics and photonics, the novel optical implementations possible in a LS microscope are highlighted.

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Section: 2d Fluidics Approachmentioning

confidence: 99%

Light-sheet microscopy: a tutorial

et al. 2018

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“…Since Huisken and his colleagues applied the light-sheet technique for reconstruction and visualization of developmental embryo [26], light-sheet fluorescent microscopy has been shown with wide applications in biomedicine such as the long-time imaging in developmental biology [27], and the real-time tracking of molecule or cell dynamics in living tissues [28,29]. In flow cytometry, Chan et al have shown the sectioning of fluorescent ocean phytoplankton by adopting the light-sheet illumination [30]. The light-sheet method may be applied to 2D light scattering cytometry to enhance the SNR as a controllable illumination for label-free cell analysis.…”

Section: Introductionmentioning

confidence: 99%

Light-sheet-based 2D light scattering cytometry for label-free characterization of senescent cells

Lin

et al. 2016

Biomed. Opt. Express

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A light-sheet-based 2D light scattering cytometer is developed for label-free characterization of senescent cells. The light-sheet provides an illumination beam with controlled thickness for single cell excitation, and 2D light scattering patterns are obtained by using a defocused imaging method. The principle of this cytometer is validated by distinguishing microspheres with submicron resolution. Automatic classification of senescent and normal cells is achieved at single cell level by using the support vector machine (SVM) algorithm, where a sensitivity of 89.1% and a specificity of 96.4% are obtained. Our results suggest that the light-sheet-based 2D light scattering label-free cytometry has the capability to perform size differentiation of beads with submicron resolution and to classify different groups of cells without fluorescent labeling, showing the potential for clinical diagnosis of senescence-related diseases.

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“…These imaging systems are, in general, referred to as Imaging Flow Cytometry (IFC) systems. [18][19][20][21][22][23][24][25] While IFC systems offer the advantages of both microscopy (acquisition of morphological features from images and high spatial resolution) and flow cytometry (automation, high sensitivity, and high-throughput), 26 one of the major limitation is the requirement of using high speed cameras. Most of the reported IFC systems in the literature employed cameras with frame rates of few to several thousand frames per second.…”

Section: Introductionmentioning

confidence: 99%

Automated cell viability assessment using a microfluidics based portable imaging flow analyzer

2015

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In this work, we report a system-level integration of portable microscopy and microfluidics for the realization of optofluidic imaging flow analyzer with a throughput of 450 cells/s. With the use of a cellphone augmented with off-the-shelf optical components and custom designed microfluidics, we demonstrate a portable optofluidic imaging flow analyzer. A multiple microfluidic channel geometry was employed to demonstrate the enhancement of throughput in the context of low frame-rate imaging systems. Using the cell-phone based digital imaging flow analyzer, we have imaged yeast cells present in a suspension. By digitally processing the recorded videos of the flow stream on the cellphone, we demonstrated an automated cell viability assessment of the yeast cell population. In addition, we also demonstrate the suitability of the system for blood cell counting.

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A light sheet based high throughput 3D-imaging flow cytometer for phytoplankton analysis

Cited by 75 publications

References 17 publications

Light-sheet microscopy: a tutorial

Light-sheet microscopy: a tutorial

Light-sheet-based 2D light scattering cytometry for label-free characterization of senescent cells

Automated cell viability assessment using a microfluidics based portable imaging flow analyzer

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