High-speed microparticle isolation unlimited by Poisson statistics

Iino, Takanori; Okano, Kazunori; Lee, Sang Wook; Yamakawa, Takeshi; Hagihara, Hiroaki; Hong, Zhen Yi; Maeno, Takanori; Kasai, Yusuke; Sakuma, Shinya; Hayakawa, Takeshi; Arai, Fumihito; Ozeki, Yasuyuki; Goda, Keisuke; Hosokawa, Yoichiroh

doi:10.1039/c9lc00324j

Cited by 24 publications

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“…For example, flow cytometry employs two sheath fluids to focus cells for the detection and measurements of physical and chemical characteristics of cells 9 . Moreover, particle focusing has been integrated with other on-chip functional components in microfluidic devices for the manipulation and analysis of different types of synthetic and biological particles, such as micelles, bacteria, microalgae, normal and cancer cells [10][11][12] .A variety of microfluidic techniques have been developed for the focusing of particles, which can be categorized into two groups: active and passive techniques 13 . Active techniques, such as thermophoresis 14 , dielectrophoresis (DEP) 15 , optical trapping 16 and acoustophoresis 17 , apply external forces to achieve particle focusing in microchannels.…”

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Hydrodynamic particle focusing enhanced by femtosecond laser deep grooving at low Reynolds numbers

Zhang

Namoto

Okano

et al. 2021

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Microfluidic focusing of particles (both synthetic and biological), which enables precise control over the positions of particles in a tightly focused stream, is a prerequisite step for the downstream processing, such as detection, trapping and separation. In this study, we propose a novel hydrodynamic focusing method by taking advantage of open v-shaped microstructures on a glass substrate engraved by femtosecond pulse (fs) laser. The fs laser engraved microstructures were capable of focusing polystyrene particles and live cells in rectangular microchannels at relatively low Reynolds numbers (Re). Numerical simulations were performed to explain the mechanisms of particle focusing and experiments were carried out to investigate the effects of groove depth, groove number and flow rate on the performance of the groove-embedded microchannel for particle focusing. We found out that 10-µm polystyrene particles are directed toward the channel center under the effects of the groove-induced secondary flows in low-Re flows, e.g. Re < 1. Moreover, we achieved continuous focusing of live cells with different sizes ranging from 10 to 15 µm, i.e. human T-cell lymphoma Jurkat cells, rat adrenal pheochromocytoma PC12 cells and dog kidney MDCK cells. The glass grooves fabricated by fs laser are expected to be integrated with on-chip detection components, such as contact imaging and fluorescence lifetime-resolved imaging, for various biological and biomedical applications, where particle focusing at a relatively low flow rate is desirable.

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Hydrodynamic particle focusing enhanced by femtosecond laser deep grooving at low Reynolds numbers

Zhang

Namoto

Okano

et al. 2021

Sci Rep

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“…The time distribution of the particles aligned in the focused sample flow entering the sorting region is random. The time intervals between two successive particles follow a Poisson distribution, expressed as 17 , 33 , where N is the total throughput. Figure S8 proves that the experimental result is coincident with the Poisson distribution.…”

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Rapid switching and durable on-chip spark-cavitation-bubble cell sorter

et al. 2022

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Precise and high-speed sorting of individual target cells from heterogeneous populations plays an imperative role in cell research. Although the conventional fluorescence-activated cell sorter (FACS) is capable of rapid and accurate cell sorting, it occupies a large volume of the instrument and inherently brings in aerosol generation as well as cross-contamination among samples. The sorting completed in a fully enclosed and disposable microfluidic chip has the potential to eliminate the above concerns. However, current microfluidic cell sorters are hindered by the high complexities of the fabrication procedure and the off-chip setup. In this paper, a spark-cavitation-bubble-based fluorescence-activated cell sorter is developed to perform fast and accurate sorting in a microfluidic chip. It features a simple structure and an easy operation. This microfluidic sorter comprises a positive electrode of platinum and a negative electrode of tungsten, which are placed on the side of the main channel. By applying a high-voltage discharge on the pair of electrodes, a single spark cavitation bubble is created to deflect the target particle into the downstream collection channel. The sorter has a short switching time of 150 μs and a long lifespan of more than 100 million workable actions. In addition, a novel control strategy is proposed to dynamically adjust the discharge time to stabilize the size of the cavitation bubble for continuous sorting. The dynamic control of continuously triggering the sorter, the optimal delay time between fluorescence detection and cell sorting, and a theoretical model to predict the ideal sorting recovery and purity are studied to improve and evaluate the sorter performance. The experiments demonstrate that the sorting rate of target particles achieves 1200 eps, the total analysis throughput is up to 10,000 eps, the particles sorted at 4000 eps exhibit a purity greater than 80% and a recovery rate greater than 90%, and the sorting effect on the viability of HeLa cells is negligible.

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“…Surface acoustic waves, dielectrophoresis, and membrane pumps have been reported as cell sorting methodologies. Recently, cell sorting with the use of laser-induced cavitation bubbles demonstrated an unprecedentedly narrow sorting window down to ∼10 μm at a flow speed of 1 m/s . Additionally, some sorting methods can compromise cell viability .…”

Section: Future Opportunities and Challenges Of Coherent Raman Flow C...mentioning

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High-Throughput Raman Flow Cytometry and Beyond

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Metrics & MoreArticle Recommendations CONSPECTUS: Flow cytometry is a powerful tool with applications in diverse fields such as microbiology, immunology, virology, cancer biology, stem cell biology, and metabolic engineering. It rapidly counts and characterizes large heterogeneous populations of cells in suspension (e.g., blood cells, stem cells, cancer cells, and microorganisms) and dissociated solid tissues (e.g., lymph nodes, spleen, and solid tumors) with typical throughputs of 1,000−100,000 events per second (eps). By measuring cell size, cell granularity, and the expression of cell surface and intracellular molecules, it provides systematic insights into biological processes. Flow cytometers may also include cell sorting capabilities to enable subsequent additional analysis of the sorted sample (e.g., electron microscopy and DNA/RNA sequencing), cloning, and directed evolution. Unfortunately, traditional flow cytometry has several critical limitations as it mainly relies on fluorescent labeling for cellular phenotyping, which is an indirect measure of intracellular molecules and surface antigens. Furthermore, it often requires time-consuming preparation protocols and is incompatible with cell therapy. To overcome these difficulties, a different type of flow cytometry based on direct measurements of intracellular molecules by Raman spectroscopy, or "Raman flow cytometry" for short, has emerged. Raman flow cytometry obtains a chemical fingerprint of the cell in a nondestructive manner, allowing for single-cell metabolic phenotyping. However, its slow signal acquisition due to the weak light−molecule interaction of spontaneous Raman scattering prevents the throughput necessary to interrogate large cell populations in reasonable time frames, resulting in throughputs of about 1 eps. The remedy to this throughput limit lies in coherent Raman scattering methods such as stimulated Raman scattering (SRS) and coherent anti-Stokes Raman scattering (CARS), which offer a significantly enhanced light−sample interaction and hence enable high-throughput Raman flow cytometry, Raman imaging flow cytometry, and even Raman image-activated cell sorting (RIACS). In this Account, we outline recent advances, technical challenges, and emerging opportunities of coherent Raman flow cytometry. First, we review the principles of various types of SRS and CARS and introduce several techniques of coherent Raman flow cytometry such as CARS, multiplex CARS, Fourier-transform CARS, SRS, SRS imaging flow cytometry, and RIACS. Next, we discuss a unique set of applications enabled by coherent Raman flow cytometry, from microbiology and lipid biology to cancer detection and cell therapy. Finally, we describe future opportunities and challenges of coherent Raman flow cytometry including increasing sensitivity and throughput, integration with droplet microfluidics, utilizing machine learning techniques, or achieving in vivo flow cytometry. This Account summarizes the growing field of high-throughput Raman flow cytometry and the bright future it ca...

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High-speed microparticle isolation unlimited by Poisson statistics

Abstract: We demonstrate an on-chip microparticle sorter with an ultrashort switching window using femtosecond laser pulses to overcome the fundamental limitation of the sorting performance described by Poisson statistics.

Cited by 24 publications

References 40 publications

Hydrodynamic particle focusing enhanced by femtosecond laser deep grooving at low Reynolds numbers

Hydrodynamic particle focusing enhanced by femtosecond laser deep grooving at low Reynolds numbers

Rapid switching and durable on-chip spark-cavitation-bubble cell sorter

High-Throughput Raman Flow Cytometry and Beyond

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