uPTI: uniaxial permittivity tensor imaging of intrinsic density and anisotropy

Yeh, Li-Hao; Ivanov, Ivan E.; Guo, Syuan-Ming; Chhun, Bryant B.; Hashemi, Ezzat; Han, May; Mehta, Shalin B.

doi:10.1364/ntm.2021.nm3c.4

Cited by 9 publications

(33 citation statements)

References 7 publications

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“…We employ the Stokes formalism [10, 16, 17] to calibrate the polarization response of the light path which is represented by an “instrument tensor” A (discussed in fig. 2).…”

Section: Resultsmentioning

confidence: 99%

“…Quantitative data reconstruction requires calibration of the microscope and correction of background introduced by the sample chamber, as we have previously described [10, 16, 17]. Here we extend these methods to address challenges inherent in the design of the miPolScope detector.…”

Section: Resultsmentioning

confidence: 99%

“…The detection path of miPolScope is calibrated by transmitting light of known Stokes states of polarization through the microscope and measuring the resulting intensity. An “instrument matrix” is then computed, which transforms the known input polarization state into intensity measurements [10, 17, 31, 32]. During data reconstruction, the inverse of the instrument matrix is used to determine how the sample perturbs the polarization state of the illumination light based on the recorded intensity images.…”

Section: Resultsmentioning

confidence: 99%

“…During data reconstruction, the inverse of the instrument matrix is used to determine how the sample perturbs the polarization state of the illumination light based on the recorded intensity images. Previously we averaged the intensity of the recorded images before computing the instrument matrix [16, 17]. Here we extend this approach by calibrating the microscope for every pixel in image space to compute an instrument tensor, which now also accounts for position-dependent polarization distortions over the field of view (fig.…”

Section: Resultsmentioning

confidence: 99%

“…Using uniaxial permittivity tensor imaging (uPTI) we measured phase, principal retardance, 3D orientation, and optic sign of multiple organelles and structures in cells and tissues at high resolution. QLIPP and uPTI are readily compatible with other imaging modalities such as fluorescence or H&E staining and can be multiplexed into existing imaging pipelines [16, 17]. They are, however, limited in the acquisition speed they can achieve, as they sequentially illuminate the sample with light of different polarization or at different angles of incidence.…”

Section: Introductionmentioning

confidence: 99%

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Correlative imaging of spatio-angular dynamics of molecular assemblies and cells with multimodal instant polarization microscope

Ivanov

Yeh

Pérez-Bermejo

et al. 2022

Preprint

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Biological function depends on the spatio-angular architecture of macromolecules - for example, functions of lipid membrane and cytoskeletal polymers arise from both the spatial and the angular organization of the constituent molecules. Correlative imaging of cellular and molecular architecture is valuable across cell biology and pathology. However, current live imaging methods primarily focus on spatial component of the architecture. Imaging the dynamic angular architecture of cells and organelles requires fast polarization-, depth-, and wavelength-diverse measurement of intrinsic optical properties and fluorophore concentration, but remains challenging with current designs. 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.

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“…We employ the Stokes formalism [10, 16, 17] to calibrate the polarization response of the light path which is represented by an “instrument tensor” A (discussed in fig. 2).…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Correlative imaging of spatio-angular dynamics of molecular assemblies and cells with multimodal instant polarization microscope

Ivanov

Yeh

Pérez-Bermejo

et al. 2022

Preprint

Self Cite

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Polarization-sensitive intensity diffraction tomography

et al. 2023

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Optical anisotropy, which is an intrinsic property of many materials, originates from the structural arrangement of molecular structures, and to date, various polarization-sensitive imaging (PSI) methods have been developed to investigate the nature of anisotropic materials. In particular, the recently developed tomographic PSI technologies enable the investigation of anisotropic materials through volumetric mappings of the anisotropy distribution of these materials. However, these reported methods mostly operate on a single scattering model, and are thus not suitable for three-dimensional (3D) PSI imaging of multiple scattering samples. Here, we present a novel reference-free 3D polarization-sensitive computational imaging technique—polarization-sensitive intensity diffraction tomography (PS-IDT)—that enables the reconstruction of 3D anisotropy distribution of both weakly and multiple scattering specimens from multiple intensity-only measurements. A 3D anisotropic object is illuminated by circularly polarized plane waves at various illumination angles to encode the isotropic and anisotropic structural information into 2D intensity information. These information are then recorded separately through two orthogonal analyzer states, and a 3D Jones matrix is iteratively reconstructed based on the vectorial multi-slice beam propagation model and gradient descent method. We demonstrate the 3D anisotropy imaging capabilities of PS-IDT by presenting 3D anisotropy maps of various samples, including potato starch granules and tardigrade.

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DaXi—high-resolution, large imaging volume and multi-view single-objective light-sheet microscopy

et al. 2022

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The promise of single-objective light-sheet microscopy is to combine the convenience of standard single-objective microscopes with the speed, coverage, resolution and gentleness of light-sheet microscopes. We present DaXi, a single-objective light-sheet microscope design based on oblique plane illumination that achieves: (1) a wider field of view and high-resolution imaging via a custom remote focusing objective; (2) fast volumetric imaging over larger volumes without compromising image quality or necessitating tiled acquisition; (3) fuller image coverage for large samples via multi-view imaging and (4) higher throughput multi-well imaging via remote coverslip placement. Our instrument achieves a resolution of 450 nm laterally and 2 μm axially over an imaging volume of 3,000 × 800 × 300 μm. We demonstrate the speed, field of view, resolution and versatility of our instrument by imaging various systems, including Drosophila egg chamber development, zebrafish whole-brain activity and zebrafish embryonic development – up to nine embryos at a time.

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uPTI: uniaxial permittivity tensor imaging of intrinsic density and anisotropy

Abstract: We present a label-free computational microscopy method, uPTI, which reports components of uniaxial permittivity tensor, including phase and 3D anisotropy across volumes. We report data from mouse brain tissue to demonstrate how uPTI enables discovery of intrinsic biological architecture.

Cited by 9 publications

References 7 publications

Correlative imaging of spatio-angular dynamics of molecular assemblies and cells with multimodal instant polarization microscope

Correlative imaging of spatio-angular dynamics of molecular assemblies and cells with multimodal instant polarization microscope

Polarization-sensitive intensity diffraction tomography

DaXi—high-resolution, large imaging volume and multi-view single-objective light-sheet microscopy

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