Drug screening at single-organoid resolution via bioprinting and interferometry

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: June 6, 2023

Abstract High throughput drug screening is an established approach to investigate tumor biology and identify therapeutic leads. Traditional platforms use two-dimensional cultures which do not accurately reflect the of human tumors. More clinically relevant model systems such as three-dimensional organoids can be difficult scale screen. Manually seeded coupled destructive endpoint assays allow for characterization treatment response, but capture transitory changes intra-sample heterogeneity underlying observed resistance therapy. We present a pipeline generate bioprinted linked label-free, time-resolved imaging via high-speed live cell interferometry (HSLCI) machine learning-based quantitation individual organoids. Bioprinting cells gives rise 3D structures with unaltered histology gene expression profiles. HSLCI in tandem segmentation classification tools enables accurate, label-free parallel mass measurements thousands demonstrate that this strategy identifies transiently or persistently sensitive resistant specific therapies, information could used guide rapid therapy selection.

Language: Английский

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Quantitative phase imaging based on holography: trends and new perspectives

Light Science & Applications, Journal Year: 2024, Volume and Issue: 13(1)

Published: June 27, 2024

Abstract In 1948, Dennis Gabor proposed the concept of holography, providing a pioneering solution to quantitative description optical wavefront. After 75 years development, holographic imaging has become powerful tool for wavefront measurement and phase imaging. The emergence this technology given fresh energy physics, biology, materials science. Digital holography (DH) possesses advantages wide-field, non-contact, precise, dynamic capability complex-waves. DH unique capabilities propagation fields by measuring light scattering with information. It offers visualization refractive index thickness distribution weak absorption samples, which plays vital role in pathophysiology various diseases characterization materials. provides possibility bridge gap between disciplines. is described complex amplitude. complex-value complex-domain reconstructed from intensity-value camera real-domain. Here, we regard process recording reconstruction as transformation real-domain, discuss mathematics physical principles reconstruction. We review underlying principles, technical approaches, breadth applications. conclude emerging challenges opportunities based on combining other methodologies that expand scope utility even further. multidisciplinary nature brings application experts together label-free cell analytical chemistry, clinical sciences, sensing, semiconductor production.

Language: Английский

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Imagining the future of optical microscopy: everything, everywhere, all at once

Communications Biology, Journal Year: 2023, Volume and Issue: 6(1)

Published: Oct. 28, 2023

The optical microscope has revolutionized biology since at least the 17

Language: Английский

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Single-protein optical holography

Nature Photonics, Journal Year: 2024, Volume and Issue: 18(4), P. 388 - 395

Published: March 13, 2024

Abstract Light scattering by nanoscale objects is a fundamental physical property defined their cross-section and thus polarizability. Over the past decade, number of studies have demonstrated single-molecule sensitivity imaging interference between from object interest reference field. This approach has enabled mass measurement single biomolecules in solution owing to linear scaling image contrast with molecular Nevertheless, all implementations so far are based on common-path interferometer cannot separate independently tune scattered light fields, thereby prohibiting access rich toolbox available holographic imaging. Here we demonstrate comparable using non-common-path geometry dark-field microscope, similar Mach–Zehnder interferometer. We into four parallel, inherently phase-stable detection channels, delivering five orders magnitude boost terms over state-of-the-art methods. detection, resolution proteins below 100 kDa. Separate amplitude phase measurements also yield direct information sample identity experimental determination polarizability biomolecules.

Language: Английский

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Quantitative phase microscopies: accuracy comparison

Light Science & Applications, Journal Year: 2024, Volume and Issue: 13(1)

Published: Oct. 11, 2024

Abstract Quantitative phase microscopies (QPMs) play a pivotal role in bio-imaging, offering unique insights that complement fluorescence imaging. They provide essential data on mass distribution and transport, inaccessible to techniques. Additionally, QPMs are label-free, eliminating concerns of photobleaching phototoxicity. However, navigating through the array available QPM techniques can be complex, making it challenging select most suitable one for particular application. This tutorial review presents thorough comparison main techniques, focusing their accuracy terms measurement precision trueness. We focus 8 namely digital holographic microscopy (DHM), cross-grating wavefront (CGM), which is based QLSI (quadriwave lateral shearing interferometry), diffraction (DPM), differential phase-contrast (DPC) microscopy, phase-shifting interferometry (PSI) imaging, Fourier (FPM), spatial light interference (SLIM), transport-of-intensity equation (TIE) For this purpose, we used home-made numerical toolbox discrete dipole approximation (IF-DDA). designed compute electromagnetic field at sample plane microscope, irrespective object’s complexity or illumination conditions. upgraded enable model any type QPM, take into account shot noise. In nutshell, results show DHM PSI inherently free from artefacts rather suffer coherent noise; CGM, DPC, DPM TIE, there trade-off between trueness, balanced by varying experimental parameter; FPM SLIM inherent cannot discarded experimentally cases, not quantitative especially large objects covering part view, such as eukaryotic cells.

Language: Английский

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Polyelectrolyte-multivalent molecule complexes: physicochemical properties and applications

Soft Matter, Journal Year: 2023, Volume and Issue: 19(11), P. 2013 - 2041

Published: Jan. 1, 2023

The complexation of polyelectrolytes with other oppositely charged structures gives rise to a great variety functional materials potential applications in wide spectrum technological fields. Depending on the assembly conditions, polyelectrolyte complexes can acquire different macroscopic configurations such as dense precipitates, nanosized colloids and liquid coacervates. In past 50 years, much progress has been achieved understand principles behind phase separation induced by interaction two aqueous solutions, especially for symmetric systems (systems which both polyions have similar molecular weight concentration). However, recent alternative building blocks small molecules (multivalent inorganic species, oligopeptides, oligoamines, among others) gained attention areas. this review, we discuss physicochemical characteristics formed multivalent molecules, putting special emphasis their similarities well-known polycation-polyanion complexes. addition, analyze these act versatile platforms various fields, biomedicine advanced engineering.

Language: Английский

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Advances in Microfluidics for Single Red Blood Cell Analysis

Biosensors, Journal Year: 2023, Volume and Issue: 13(1), P. 117 - 117

Published: Jan. 9, 2023

The utilizations of microfluidic chips for single RBC (red blood cell) studies have attracted great interests in recent years to filter, trap, analyze, and release erythrocytes various applications. Researchers this field highlighted the vast potential developing micro devices industrial academia usages, including lab-on-a-chip organ-on-a-chip systems. This article critically reviews current state-of-the-art advances microfluidics analyses, integrated sensors platforms microscopic/tomographic/spectroscopic trapping arrays (including bifurcating channels), dielectrophoretic agglutination/aggregation studies, as well clinical implications covering cancer, sepsis, prenatal, Sickle Cell diseases. Microfluidics based microarrays, sorting/counting techniques acoustic, dielectrophoretic, hydrodynamic, magnetic, optical techniques) are also reviewed. Lastly, organs on chips, multi-organ drug discovery involving described. limitations drawbacks each technology addressed future prospects discussed.

Language: Английский

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Label‐Free Intracellular Multi‐Specificity in Yeast Cells by Phase‐Contrast Tomographic Flow Cytometry

Small Methods, Journal Year: 2023, Volume and Issue: 7(11)

Published: Sept. 5, 2023

Abstract In‐flow phase‐contrast tomography provides a 3D refractive index of label‐free cells in cytometry systems. Its major limitation, as with any quantitative phase imaging approach, is the lack specificity compared to fluorescence microscopy, thus restraining its huge potentialities single‐cell analysis and diagnostics. Remarkable results introducing are obtained through artificial intelligence (AI), but only for adherent cells. However, accessing ground truth obtaining accurate voxel‐level co‐registration image pairs AI training not viable high‐throughput cytometry. The recent statistical inference approach significant step forward remains limited cells’ nuclei. Here, generalized computational strategy based on self‐consistent achieve intracellular multi‐specificity shown. Various subcellular compartments (i.e., nuclei, cytoplasmic vacuoles, peri‐vacuolar membrane area, cytoplasm, vacuole‐nucleus contact site) can be identified characterized quantitatively at different phases life cycle by using yeast biological model. Moreover, first time, virtual reality introduced handling information content single Full fruition proofed exploring interacting biophysical parameters demand, opening route metaverse microscopy.

Language: Английский

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Wavefront Microscopy Using Quadriwave Lateral Shearing Interferometry: From Bioimaging to Nanophotonics

ACS Photonics, Journal Year: 2023, Volume and Issue: 10(2), P. 322 - 339

Published: Jan. 17, 2023

Common cameras are only sensitive to the intensity of light, discarding an essential feature a light wave: its phase profile or, equivalently, wavefront profile. This Review focuses on rising imaging technique called quadriwave lateral shearing interferometry (QLSI), based simple use 2-dimensional diffraction grating, aka cross-grating, in front regular camera. We detail working principle QLSI and implementation optical microscope. highlight microscopy applications bioimaging nanophotonics, particular for characterization living cells, nanoparticles, 2D materials, metasurfaces, microscale temperature gradients, surface topography. Finally, we draw critical comparison with current quantitative techniques, namely, digital holography (DHM), spatial interference (SLIM), (DPM).

Language: Английский

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Motion-resolved, reference-free holographic imaging via spatiotemporally regularized inversion

Optica, Journal Year: 2023, Volume and Issue: 11(1), P. 32 - 32

Published: Nov. 29, 2023

Holography is a powerful technique that records the amplitude and phase of an optical field simultaneously, enabling variety applications such as label-free biomedical analysis coherent diffraction imaging. Holographic recording without reference wave has been long pursued because it obviates high experimental requirements conventional interferometric methods. However, due to ill-posed nature underlying retrieval problem, reference-free holographic imaging faced with inherent tradeoff between fidelity temporal resolution. Here, we propose general computational framework, termed spatiotemporally regularized inversion (STRIVER), achieve motion-resolved, fidelity. Specifically, STRIVER leverages signal priors in spatiotemporal domain jointly eliminate ambiguities motion artifacts, and, when combined diversity measurement schemes, produces physically reliable, time-resolved video from series intensity-only measurements. We experimentally demonstrate near-field ptychography, where dynamic freely swimming paramecia performed at framerate-limited speed 112 fps. The proposed method can be potentially extended other spectral regimes, modalities, pushing resolution toward higher limits.

Language: Английский

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