Sadao Ota

Ghost imaging is a technique used to produce an object's image without using a spatially resolving detector. Here we develop a technique we term "ghost cytometry," an image-free ultrafast fluorescence "imaging" cytometry based on a single-pixel detector. Spatial information obtained from the motion of cells relative to a static randomly patterned optical structure is compressively converted into signals that arrive sequentially at a single-pixel detector. Combinatorial use of the temporal wavefo

Gently down the stream: A microfluidic technique uses a continuous fluid stream to generate monodisperse unilamellar phospholipid vesicles from a single bilayer (see picture). Since the vesicles are robust and efficiently encapsulate high concentrations of various molecules, they are useful as delivery vehicles and as model cellular systems.

We present a simple method to form free-standing lipid membranes on arrayed microchambers (>100). The formed membranes are perpendicular to an imaging plane with control of solute concentration on each side of the membranes. This platform let us quantitatively detect membrane transport of non-charged fluorescent molecules, induced by membrane proteins.

Remotely manipulating a large number of microscopic objects is important to soft-condensed matter physics, biophysics, and nanotechnology. Optical tweezers and optoelectronic tweezers have been widely used for this purpose but face critical challenges when applied to nanoscale objects, including severe photoinduced damages, undesired ionic convections, or irreversible particle immobilization on surfaces. We report here the first demonstration of a lipid bilayer-integrated optoelectronic tweezers

In this review, we focus on the applications of machine learning methods for analyzing image data acquired in imaging flow cytometry technologies. We propose that the analysis approaches can be categorized into two groups based on the type of data, raw imaging signals or features explicitly extracted from images, being analyzed by a trained model. We hope that this categorization is helpful for understanding uniqueness, differences and opportunities when the machine learning-based analysis is im

Sanft dem Strom hinab: Mit einer Mikrofluidiktechnik werden in einem kontinuierlichen Fluidstrom monodisperse unilamellare Phospholipidvesikel aus einer einzigen Doppelschicht erzeugt (siehe Bild). Da die Vesikel robust sind und effizient eine Vielzahl an Molekülen in hohen Konzentrationen einkapseln, eignen sie sich als Transportvehikel und als Modelle für Zellsysteme. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed,

We present a simple microfluidic method to generate high-density femotoliter-sized microreactor arrays within microfluidic channels. In general, we designed a main channel with many small chambers built into its walls. After sequentially infusing aqueous solution and organic solvent from a single tube into the device, aqueous droplets are confined in the chambers by the solvent flow. The generated reactors are small and stable enough for carrying out ultrasensitive biochemical assays at single m

Abstract Imaging flow cytometry shows significant potential for increasing our understanding of heterogeneous and complex life systems and is useful for biomedical applications. Ghost cytometry is a recently proposed approach for directly analyzing compressively measured signals of cells, thereby relieving a computational bottleneck for real‐time data analysis in high‐throughput imaging cytometry. In our previous work, we demonstrated that this image‐free approach could distinguish cells from tw

Early diagnosis and prompt initiation of appropriate treatment are critical for improving the prognosis of acute leukemia. Acute leukemia is diagnosed by microscopic morphological examination of bone marrow smears and flow cytometric immunophenotyping of bone marrow cells stained with fluorophore-conjugated antibodies. However, these diagnostic processes require trained professionals and are time and resource-intensive. Here, we present a novel diagnostic approach using ghost cytometry, a recent

Recent advancements in image-based pooled CRISPR screening have facilitated the mapping of diverse genotype-phenotype associations within mammalian cells. However, the rapid enrichment of cells based on morphological information continues to pose a challenge, constraining the capacity for large-scale gene perturbation screening across diverse high-content cellular phenotypes. In this study, we demonstrate the applicability of multimodal ghost cytometry-based cell sorting, including both fluoresc

Brownian motion of slender particles near a boundary is ubiquitous in biological systems and in nanomaterial assembly, but the complex hydrodynamic interaction in those systems is still poorly understood. Here, we report experimental and computational studies of the Brownian motion of silicon nanowires tethered on a substrate. An optical interference method enabled direct observation of microscopic rotations of the slender bodies in three dimensions with high angular and temporal resolutions. Th

We present a highly parallel and reproducible method for reconstituting an array of lipid bilayers to analyze membrane transport. We infuse buffer/lipid/buffer solutions sequentially into a microchannel with numerous microchambers in its walls and seal each chamber by a lipid bilayer containing membrane proteins. Due to the small volume of the chamber (2 pL), membrane transport of confined fluorescent molecules across the bilayer through the proteins is readily observed as changes in fluorescent

Di Carlo et al . comment that our original results were insufficient to prove that the ghost cytometry technique is performing a morphologic analysis of cells in flow. We emphasize that the technique is primarily intended to acquire and classify morphological information of cells in a computationally efficient manner without reconstructing images. We provide additional supporting information, including images reconstructed from the compressive waveforms and a discussion of current and future thr

Abstract Adipocyte spheroids are a promising three-dimensional (3D) cell culture model for obesity research because they reproduce 3D adipose tissue structures and cell-cell interactions better than 2D cultures. However, current methods fail to produce uniformly sized, small adipocyte spheroids at large scales, significantly limiting their use in analysis such as large-scale drug screening. Here, we develop a scalable method that combines simple microfluidics with templated emulsification to gen