Browse
Submit Data
Databases
API
Help

Dataset Information

22 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Correlative imaging of the spatio-angular dynamics of biological systems with multimodal instant polarization microscope.

ABSTRACT: The spatial and angular organization of biological macromolecules is a key determinant, as well as informative readout, of their function. Correlative imaging of the dynamic spatio-angular architecture of cells and organelles is valuable, but remains challenging with current methods. Correlative imaging of spatio-angular dynamics requires fast polarization-, depth-, and wavelength-diverse measurement of intrinsic optical properties and fluorescent labels. We report a multimodal instant polarization microscope (miPolScope) that combines a broadband polarization-resolved detector, automation, and reconstruction algorithms to enable label-free imaging of phase, retardance, and orientation, multiplexed with fluorescence imaging of concentration, anisotropy, and orientation of molecules at diffraction-limited resolution and high speed. miPolScope enabled multimodal imaging of myofibril architecture and contractile activity of beating cardiomyocytes, cell and organelle architecture of live HEK293T and U2OS cells, and density and anisotropy of white and grey matter of mouse brain tissue across the visible spectrum. We anticipate these developments in joint quantitative imaging of density and anisotropy to enable new studies in tissue pathology, mechanobiology, and imaging-based screens.

SUBMITTER: Ivanov IE

PROVIDER: S-EPMC9203109 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Json Xml

Similar Datasets

Construction of an instant structured illumination microscope.

Project description:A challenge in biological imaging is to capture high-resolution images at fast frame rates in live cells. The "instant structured illumination microscope" (iSIM) is a system designed for this purpose. Similarly to standard structured illumination microscopy (SIM), an iSIM provides a twofold improvement over widefield microscopy, in x, y and z, but also allows much faster image acquisition, with real-time display of super-resolution images. The assembly of an iSIM is reasonably complex, involving the combination and alignment of many optical components, including three micro-optics arrays (two lenslet arrays and an array of pinholes, all with a pitch of 222 ?m) and a double-sided scanning mirror. In addition, a number of electronic components must be correctly controlled. Construction of the system is therefore not trivial, but is highly desirable, particularly for live-cell imaging. We report, and provide instructions for, the construction of an iSIM, including minor modifications to a previous design in both hardware and software. The final instrument allows us to rapidly acquire fluorescence images at rates faster than 100 fps, with approximately twofold improvement in resolution in both x-y and z; sub-diffractive biological features have an apparent size (full width at half maximum) of 145 nm (lateral) and 320 nm (axial), using a 1.49 NA objective and 488 nm excitation.

| S-EPMC4641873 | biostudies-other

An adaptive microscope for the imaging of biological surfaces.

Project description:Scanning fluorescence microscopes are now able to image large biological samples at high spatial and temporal resolution. This comes at the expense of an increased light dose which is detrimental to fluorophore stability and cell physiology. To highly reduce the light dose, we designed an adaptive scanning fluorescence microscope with a scanning scheme optimized for the unsupervised imaging of cell sheets, which underly the shape of many embryos and organs. The surface of the tissue is first delineated from the acquisition of a very small subset (~0.1%) of sample space, using a robust estimation strategy. Two alternative scanning strategies are then proposed to image the tissue with an improved photon budget, without loss in resolution. The first strategy consists in scanning only a thin shell around the estimated surface of interest, allowing high reduction of light dose when the tissue is curved. The second strategy applies when structures of interest lie at the cell periphery (e.g. adherens junctions). An iterative approach is then used to propagate scanning along cell contours. We demonstrate the benefit of our approach imaging live epithelia from Drosophila melanogaster. On the examples shown, both approaches yield more than a 20-fold reduction in light dose -and up to more than 80-fold- compared to a full scan of the volume. These smart-scanning strategies can be easily implemented on most scanning fluorescent imaging modality. The dramatic reduction in light exposure of the sample should allow prolonged imaging of the live processes under investigation.

| S-EPMC8497591 | biostudies-literature

Multimodal spectral imaging of cells using a transmission diffraction grating on a light microscope.

Project description:A multimodal methodology for spectral imaging of cells is presented. The spectral imaging setup uses a transmission diffraction grating on a light microscope to concurrently record spectral images of cells and cellular organelles by fluorescence, darkfield, brightfield, and differential interference contrast (DIC) spectral microscopy. Initially, the setup was applied for fluorescence spectral imaging of yeast and mammalian cells labeled with multiple fluorophores. Fluorescence signals originating from fluorescently labeled biomolecules in cells were collected through triple or single filter cubes, separated by the grating, and imaged using a charge-coupled device (CCD) camera. Cellular components such as nuclei, cytoskeleton, and mitochondria were spatially separated by the fluorescence spectra of the fluorophores present in them, providing detailed multi-colored spectral images of cells. Additionally, the grating-based spectral microscope enabled measurement of scattering and absorption spectra of unlabeled cells and stained tissue sections using darkfield and brightfield or DIC spectral microscopy, respectively. The presented spectral imaging methodology provides a readily affordable approach for multimodal spectral characterization of biological cells and other specimens.

| S-EPMC3163165 | biostudies-literature

Multimodal correlative imaging and modelling of phosphorus uptake from soil by hyphae of mycorrhizal fungi.

Project description:Phosphorus (P) is essential for plant growth. Arbuscular mycorrhizal fungi (AMF) aid its uptake by acquiring P from sources distant from roots in return for carbon. Little is known about how AMF colonise soil pore-space, and models of AMF-enhanced P-uptake are poorly validated. We used synchrotron X-ray computed tomography to visualize mycorrhizas in soil and synchrotron X-ray fluorescence/X-ray absorption near edge structure (XRF/XANES) elemental mapping for P, sulphur (S) and aluminium (Al) in combination with modelling. We found that AMF inoculation had a suppressive effect on colonisation by other soil fungi and identified differences in structure and growth rate between hyphae of AMF and nonmycorrhizal fungi. Our results showed that AMF co-locate with areas of high P and low Al, and preferentially associate with organic-type P species over Al-rich inorganic P. We discovered that AMF avoid Al-rich areas as a source of P. Sulphur-rich regions were found to be correlated with higher hyphal density and an increased organic-associated P-pool, whilst oxidized S-species were found close to AMF hyphae. Increased S oxidation close to AMF suggested the observed changes were microbiome-related. Our experimentally-validated model led to an estimate of P-uptake by AMF hyphae that is an order of magnitude lower than rates previously estimated - a result with significant implications for the modelling of plant-soil-AMF interactions.

| S-EPMC9307049 | biostudies-literature

Targetable Tetrazine-Based Dynamic Nuclear Polarization Agents for Biological Systems.

Project description:Dynamic nuclear polarization (DNP) has shown great promise as a tool to enhance the nuclear magnetic resonance signals of proteins in the cellular environment. As sensitivity increases, the ability to select and efficiently polarize a specific macromolecule over the cellular background has become desirable. Herein, we address this need and present a tetrazine-based DNP agent that can be targeted selectively to proteins containing the unnatural amino acid (UAA) norbornene-lysine. This UAA can be introduced efficiently into the cellular milieu by genetic means. Our approach is bio-orthogonal and easily adaptable to any protein of interest. We illustrate the scope of our methodology and investigate the DNP transfer mechanisms in several biological systems. Our results shed light on the complex polarization-transfer pathways in targeted DNP and ultimately pave the way to selective DNP-enhanced NMR spectroscopy in both bacterial and mammalian cells.

| S-EPMC7445144 | biostudies-literature

All-dielectric polarization-independent optical angular filter.

Project description:We report on an all-dielectric, polarization-independent angular filter with one-dimension (1D) photonic crystal (PC) composed of semiconductor compatible Si/SiO2 pairs. The near-symmetric directional band gap of p- and s-polarized components and Fabry-Pérot (F-P) resonances are utilized to realize efficient polarization-independent angular filtering for normal incidence. The proposed angular filter is designed and experimentally demonstrated in a large area (5?cm?×?5?cm) with multilayer sputtering depositions. Experimental measurements show that a divergence angle of the polarization-independently transmitted beam through the angular filtering sample at 1550?nm is 2.2° only and the transmission is as high as 0.8 at normal incidence. The proposed and demonstrated angular filter suggests an effective way to design and implement semiconductor compatible, all-dielectric and polarization-independent angular filters in a fashion of simple structure and easy-fabrication, which is expected to have great applications in lighting, beam manipulation, optical coupling and optical communications.

| S-EPMC5707375 | biostudies-literature

Fluorescence-detected linear dichroism imaging in a re-scan confocal microscope equipped with differential polarization attachment.

Project description:Confocal laser scanning microscopy is probably the most widely used and one of the most powerful techniques in basic biology, medicine and material sciences that is employed to elucidate the architecture of complex cellular structures and molecular macro-assemblies. It has recently been shown that the information content, signal-to-noise ratio and resolution of such microscopes (LSMs) can be improved significantly by adding different attachments or modifying their design, while retaining their user-friendly features and relatively moderate costs. Differential polarization (DP) attachments, using high-frequency modulation/demodulation circuits, have made LSMs capable of high-precision 2D and 3D mapping of the anisotropy of microscopic samples-without interfering with their 'conventional' fluorescence or transmission imaging (Steinbach et al. in Methods Appl Fluoresc 2:015005, 2014). The resolution and the quality of fluorescence imaging have been enhanced in the recently constructed Re-scan confocal microscopy (RCM) (De Luca et al. in Biomed Opt Express 4:2644-2656, 2013). In this work, we developed the RCM technique further, by adding a DP-attachment modulating the exciting laser beam via a liquid crystal (LC) retarder synchronized with the data acquisition system; by this means, and with the aid of a software, fluorescence-detected linear dichroism (FDLD), characteristic of the anisotropic molecular organization of the sample, could be recorded in parallel with the confocal fluorescence imaging. For demonstration, we show FDLD images of a plant cell wall (Ginkgo biloba) stained with Etzold's staining solution.

| S-EPMC6647120 | biostudies-literature

Polarization-Encrypted Orbital Angular Momentum Multiplexed Metasurface Holography.

Project description:Metasurface holography has the advantage of realizing complex wavefront modulation by thin layers together with the progressive technique of computer-generated holographic imaging. Despite the well-known light parameters, such as amplitude, phase, polarization, and frequency, the orbital angular momentum (OAM) of a beam can be regarded as another degree of freedom. Here, we propose and demonstrate orbital angular momentum multiplexing at different polarization channels using a birefringent metasurface for holographic encryption. The OAM selective holographic information can only be reconstructed with the exact topological charge and a specific polarization state. By using an incident beam with different topological charges as erasers, we mimic a super-resolution case for the reconstructed image, in analogy to the well-known STED technique in microscopy. The combination of multiple polarization channels together with the orbital angular momentum selectivity provides a higher security level for holographic encryption. Such a technique can be applied for beam shaping, optical camouflage, data storage, and dynamic displays.

| S-EPMC7254835 | biostudies-literature

Photonic polarization gears for ultra-sensitive angular measurements.

Project description:Quantum metrology bears a great promise in enhancing measurement precision, but is unlikely to become practical in the near future. Its concepts can nevertheless inspire classical or hybrid methods of immediate value. Here we demonstrate NOON-like photonic states of m quanta of angular momentum up to m=100, in a setup that acts as a 'photonic gear', converting, for each photon, a mechanical rotation of an angle ? into an amplified rotation of the optical polarization by m?, corresponding to a 'super-resolving' Malus' law. We show that this effect leads to single-photon angular measurements with the same precision of polarization-only quantum strategies with m photons, but robust to photon losses. Moreover, we combine the gear effect with the quantum enhancement due to entanglement, thus exploiting the advantages of both approaches. The high 'gear ratio' m boosts the current state of the art of optical non-contact angular measurements by almost two orders of magnitude.

| S-EPMC3791460 | biostudies-literature