Publications by authors named "Akihiro Isozaki"

Animals navigate their environment by manipulating their movements and adjusting their trajectory which requires a sophisticated integration of sensory data with their current motor status. Here, we utilize the nematode to explore the neural mechanisms of processing the sensory and motor information for navigation. We developed a microfluidic device which allows animals to freely move their heads while receiving temporal NaCl stimuli.

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  • Scientists are using artificial intelligence (AI) to help analyze and sort images of cells in a faster way than before.
  • They compared different methods, like old-school techniques and newer AI methods, to see which was better for identifying specific types of cells.
  • The study showed that using deep learning (a type of AI) improved their ability to find the right cells much better than traditional methods, even though it took longer to process the images.
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  • * A new method called VIFFI flow cytometry combines virtual-freezing imaging and 5-ALA stimulation to enhance detection and classification of CTCs in blood samples, even in the absence of traditional biomarkers.
  • * Using this technique, researchers successfully identified CTCs in blood samples from breast cancer patients, which could improve cancer prognosis, treatment guidance, and strategy design in clinical settings.
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Intelligent image-activated cell sorting (iIACS) has enabled high-throughput image-based sorting of single cells with artificial intelligence (AI) algorithms. This AI-on-a-chip technology combines fluorescence microscopy, AI-based image processing, sort-timing prediction, and cell sorting. Sort-timing prediction is particularly essential due to the latency on the order of milliseconds between image acquisition and sort actuation, during which image processing is performed.

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Organelle positioning in cells is associated with various metabolic functions and signaling in unicellular organisms. Specifically, the microalga Chlamydomonas reinhardtii repositions its mitochondria, depending on the levels of inorganic carbon. Mitochondria are typically randomly distributed in the Chlamydomonas cytoplasm, but relocate toward the cell periphery at low inorganic carbon levels.

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Imaging flow cytometry (IFC) has become a powerful tool for diverse biomedical applications by virtue of its ability to image single cells in a high-throughput manner. However, there remains a challenge posed by the fundamental trade-off between throughput, sensitivity, and spatial resolution. Here we present deep-learning-enhanced imaging flow cytometry (dIFC) that circumvents this trade-off by implementing an image restoration algorithm on a virtual-freezing fluorescence imaging (VIFFI) flow cytometry platform, enabling higher throughput without sacrificing sensitivity and spatial resolution.

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  • Droplet microfluidics is a versatile technique used in biomedical and industrial fields for tasks like single-cell analysis and metabolic engineering, with droplet sorting being key for isolating specific small droplets.
  • Recent efforts focus on sorting larger droplets to leverage their size, but achieving high sorting throughput has been challenging.
  • A new upgraded fluorescence-activated droplet sorting system, featuring more electrodes and a slanted microchannel, successfully sorted 1 nL droplets at a record rate of 1752 droplets per second, doubling the previous maximum throughput.
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Single-cell analysis has become one of the main cornerstones of biotechnology, inspiring the advent of various microfluidic compartments for cell cultivation such as microwells, microtrappers, microcapillaries, and droplets. A fundamental assumption for using such microfluidic compartments is that unintended stress or harm to cells derived from the microenvironments is insignificant, which is a crucial condition for carrying out unbiased single-cell studies. Despite the significance of this assumption, simple viability or growth tests have overwhelmingly been the assay of choice for evaluating culture conditions while empirical studies on the sub-lethal effect on cellular functions have been insufficient in many cases.

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Raman optical activity (ROA) is effective for studying the conformational structure and behavior of chiral molecules in aqueous solutions and is advantageous over X-ray crystallography and nuclear magnetic resonance spectroscopy in sample preparation and cost performance. However, ROA signals are inherently minuscule; 3-5 orders of magnitude weaker than spontaneous Raman scattering due to the weak chiral light-matter interaction. Localized surface plasmon resonance on metallic nanoparticles has been employed to enhance ROA signals, but suffers from detrimental spectral artifacts due to its photothermal heat generation and inability to efficiently transfer and enhance optical chirality from the far field to the near field.

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In the past few decades, microalgae-based bioremediation methods for treating heavy metal (HM)-polluted wastewater have attracted much attention by virtue of their environment friendliness, cost efficiency, and sustainability. However, their HM removal efficiency is far from practical use. Directed evolution is expected to be effective for developing microalgae with a much higher HM removal efficiency, but there is no non-invasive or label-free indicator to identify them.

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Technological advances in image-based platelet analysis or platelet morphometry are critical for a better understanding of the structure and function of platelets in biological research as well as for the development of better clinical strategies in medical practice. Recently, the advent of high-throughput optical imaging and deep learning has boosted platelet morphometry to the next level by providing a new set of capabilities beyond what is achievable with traditional platelet morphometry, shedding light on the unexplored domain of platelet analysis. This Opinion article introduces emerging opportunities in 'intelligent' platelet morphometry, which are expected to pave the way for a new class of diagnostics, pharmacometrics, and therapeutics.

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  • Recent advancements in imaging technologies, like image-activated cell sorting and imaging-based cell picking, have enhanced our understanding of biological systems over the past decade.
  • Traditional methods often depend on fluorescent labeling for identifying cellular characteristics, which can be limited and indirect.
  • The new approach demonstrated involves Raman image-activated cell sorting that uses ultrafast stimulated Raman scattering (SRS) microscopy to directly analyze single live cells without fluorescent labels, allowing for real-time sorting of various cell types at high speeds.
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Artificial intelligence (AI) has dramatically changed the landscape of science, industry, defence, and medicine in the last several years. Supported by considerably enhanced computational power and cloud storage, the field of AI has shifted from mostly theoretical studies in the discipline of computer science to diverse real-life applications such as drug design, material discovery, speech recognition, self-driving cars, advertising, finance, medical imaging, and astronomical observation, where AI-produced outcomes have been proven to be comparable or even superior to the performance of human experts. In these applications, what is essentially important for the development of AI is the data needed for machine learning.

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The advent of intelligent image-activated cell sorting (iIACS) has enabled high-throughput intelligent image-based sorting of single live cells from heterogeneous populations. iIACS is an on-chip microfluidic technology that builds on a seamless integration of a high-throughput fluorescence microscope, cell focuser, cell sorter, and deep neural network on a hybrid software-hardware data management architecture, thereby providing the combined merits of optical microscopy, fluorescence-activated cell sorting (FACS), and deep learning. Here we report an iIACS machine that far surpasses the state-of-the-art iIACS machine in system performance in order to expand the range of applications and discoveries enabled by the technology.

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An amendment to this paper has been published and can be accessed via a link at the top of the paper.

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  • Imaging flow cytometry merges flow cytometry with fluorescence imaging and digital analysis, making it valuable for research in areas like cancer biology and drug discovery.
  • A major limitation is its reliance on fluorescent labeling for identifying cell types.
  • The study introduces a label-free chemical imaging approach using advanced laser technology, allowing for rapid analysis of living cells, with applications in studying microalgal metabolism and detecting cancer markers without labels.
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Intelligent image-activated cell sorting (iIACS) is a machine-intelligence technology that performs real-time intelligent image-based sorting of single cells with high throughput. iIACS extends beyond the capabilities of fluorescence-activated cell sorting (FACS) from fluorescence intensity profiles of cells to multidimensional images, thereby enabling high-content sorting of cells or cell clusters with unique spatial chemical and morphological traits. Therefore, iIACS serves as an integral part of holistic single-cell analysis by enabling direct links between population-level analysis (flow cytometry), cell-level analysis (microscopy), and gene-level analysis (sequencing).

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A fundamental challenge of biology is to understand the vast heterogeneity of cells, particularly how cellular composition, structure, and morphology are linked to cellular physiology. Unfortunately, conventional technologies are limited in uncovering these relations. We present a machine-intelligence technology based on a radically different architecture that realizes real-time image-based intelligent cell sorting at an unprecedented rate.

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We present on-chip fluorescence imaging flow cytometry by light-sheet excitation on a mirror-embedded microfluidic chip. The method allows us to obtain microscopy-grade fluorescence images of cells flowing at a high speed of 1 m/s, which is comparable to the flow speed of conventional non-imaging flow cytometers. To implement the light-sheet excitation of flowing cells in a microchannel, we designed and fabricated a mirror-embedded PDMS-based microfluidic chip.

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  • Rapidly assessing cellular variations in large populations is crucial for understanding cellular diversity, and optofluidic time-stretch microscopy offers a solution through high-throughput imaging flow cytometry.
  • This method allows for the analysis of various cell types, supporting applications in biology, medicine, pharmaceuticals, and renewable energy.
  • The provided protocol includes detailed instructions on building the necessary equipment, acquiring high-resolution images of over 10,000 cells per second, and utilizing computational tools for efficient data analysis, with an estimated completion time of 1 month for a small research team.
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Innovations in optical microscopy have opened new windows onto scientific research, industrial quality control, and medical practice over the last few decades. One of such innovations is optofluidic time-stretch quantitative phase microscopy - an emerging method for high-throughput quantitative phase imaging that builds on the interference between temporally stretched signal and reference pulses by using dispersive properties of light in both spatial and temporal domains in an interferometric configuration on a microfluidic platform. It achieves the continuous acquisition of both intensity and phase images with a high throughput of more than 10,000 particles or cells per second by overcoming speed limitations that exist in conventional quantitative phase imaging methods.

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We propose a reconfigurable terahertz (THz) metamaterial that can control the transmittance by out-of-plane actuation with changing the sub-micron gap distance between electrically coupled metamaterial elements. By using the out-of-plane actuation, it was possible to avoid contact between the coupled metamaterial elements across the small initial gap during the adjustment of the gap size. THz spectroscopy was performed during actuation, and the transmission dip frequency was confirmed to be tunable from 0.

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Active modulation of the polarization states of terahertz light is indispensable for polarization-sensitive spectroscopy, having important applications such as non-contact Hall measurements, vibrational circular dichroism measurements and anisotropy imaging. In the terahertz region, the lack of a polarization modulator similar to a photoelastic modulator in the visible range hampers expansion of such spectroscopy. A terahertz chiral metamaterial has a huge optical activity unavailable in nature; nevertheless, its modulation is still challenging.

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We propose a method to measure light transmittance of layered metamaterials by placing the metamaterials directly on a Si photodiode. Our measurement method enables the direct detection of transmitted light that appears as an evanescent wave in natural materials. Here, we report the transmittance measurements of a typical metamaterial using this method.

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