skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

Explore Research Products in the PAR
It may take a few hours for recently added research products to appear in PAR search results.


Title: Optical projection tomography of fluorescent microscopic specimens using lateral translation of tube lens
Optical projection tomography (OPT) is a three-dimensional (3D) fluorescence imaging technique, in which projection images are acquired for varying orientations of a sample using a large depth of field. OPT is typically applied to a millimeter-sized specimen, because the rotation of a microscopic specimen is challenging and not compatible with live cell imaging. In this Letter, we demonstrate fluorescence optical tomography of a microscopic specimen by laterally translating the tube lens of a wide-field optical microscope, which allows for high-resolution OPT without rotating the sample. The cost is the reduction of the field of view to about halfway along the direction of the tube lens translation. Using bovine pulmonary artery endothelial cells and 0.1 µm beads, we compare the 3D imaging performance of the proposed method with that of the conventional objective-focus scan method.  more » « less
Award ID(s):
1808331
PAR ID:
10412138
Author(s) / Creator(s):
Publisher / Repository:
Optical Society of America
Date Published:
Journal Name:
Optics Letters
Volume:
48
Issue:
10
ISSN:
0146-9592; OPLEDP
Format(s):
Medium: X Size: Article No. 2623
Size(s):
Article No. 2623
Sponsoring Org:
National Science Foundation
More Like this
  1. ; ; (, Sensors)
    null (Ed.)
    Hyperspectral three-dimensional (3D) imaging can provide both 3D structural and functional information of a specimen. The imaging throughput is typically very low due to the requirement of scanning mechanisms for different depths and wavelengths. Here we demonstrate hyperspectral 3D imaging using Snapshot projection optical tomography (SPOT) and Fourier-transform spectroscopy (FTS). SPOT allows us to instantaneously acquire the projection images corresponding to different viewing angles, while FTS allows us to perform hyperspectral imaging at high spectral resolution. Using fluorescent beads and sunflower pollens, we demonstrate the imaging performance of the developed system. 
    more » « less
  2. ; ; ; ; ; (, Journal of the Optical Society of America A)
    Illuminating or imaging samples from a broad angular range is essential in a wide variety of computational 3D imaging and resolution-enhancement techniques, such as optical projection tomography, optical diffraction tomography, synthetic aperture microscopy, Fourier ptychographic microscopy, structured illumination microscopy, photogrammetry, and optical coherence refraction tomography. The wider the angular coverage, the better the resolution enhancement or 3D-resolving capabilities. However, achieving such angular ranges is a practical challenge, especially when approaching ±<#comment/> 90 ∘<#comment/> or beyond. Often, researchers resort to expensive, proprietary high numerical aperture (NA) objectives or to rotating the sample or source-detector pair, which sacrifices temporal resolution or perturbs the sample. Here, we propose several new strategies for multiangle imaging approaching 4pi steradians using concave parabolic or ellipsoidal mirrors and fast, low rotational inertia scanners, such as galvanometers. We derive theoretically and empirically relations between a variety of system parameters (e.g.,  NA, wavelength, focal length, telecentricity) and achievable fields of view (FOVs) and importantly show that intrinsic tilt aberrations donotrestrict FOV for many multiview imaging applications, contrary to conventional wisdom. Finally, we present strategies for avoiding spherical aberrations at obliquely illuminated flat boundaries. Our simple designs allow for high-speed multiangle imaging for microscopic, mesoscopic, and macroscopic applications. 
    more » « less
  3. ; ; ; (, Proceedings of the National Academy of Sciences)
    na (Ed.)
    Fast, nondestructive three-dimensional (3D) imaging of live suspension cells remains challenging without substrate treatment or fixation, precluding scalable single-cell morphometry with minimal alterations. While optical sectioning techniques achieve 3D live cell imaging, lateral versus depth resolution differences further complicate analysis. We present a scalable microfluidic method capable of 3D fluorescent isotropic imaging of live, nonadherent cells suspended inside picoliter droplets with high-speed single-cell volumetric readout (800 to 1,200 slices in 5 to 8 s) and near-diffraction limit resolution (~216 nm). The platform features a droplet trap array that leverages flow-induced droplet interfacial shear to generate intradroplet microvortices, which rotate single cells on their axis to enable optical projection tomography (OPT)-based imaging. This allows gentle (~1 mPa shear stress) observation of cells encapsulated inside nontoxic isotonic buffer droplets, facilitating scalable OPT acquisition by simultaneous spinning of hundreds of cells. We demonstrate 3D imaging of live myeloid and lymphoid cells in suspension, including K562 cells, as well as naive and activated T cells—small cells prone to movement in their suspended phenotype. Our fully suspended, orientation-independent cell morphometry, driven by isotropic imaging and spherical harmonic analysis, enabled the study of primary T cells across various immunological activation states. This approach unveiled six distinct nuclear content distributions, contrasting with conventional 2D images that typically portray spheroid and bean-like nuclear shapes associated with lymphocytes. Our arrayed-droplet OPT technology is capable of isotropic, single live-cell 3D imaging, with the potential to perform large-scale morphometry of immune cell effector function states while providing compatibility with microfluidic droplet operations. 
    more » « less
  4. ; ; ; ; ; ; (, Optics Express)
    Stimulated Raman projection tomography is a label-free volumetric chemical imaging technology allowing three-dimensional (3D) reconstruction of chemical distribution in a biological sample from the angle-dependent stimulated Raman scattering projection images. However, the projection image acquisition process requires rotating the sample contained in a capillary glass held by a complicated sample rotation stage, limiting the volumetric imaging speed, and inhibiting the study of living samples. Here, we report a tilt-angle stimulated Raman projection tomography (TSPRT) system which acquires angle-dependent projection images by utilizing tilt-angle beams to image the sample from different azimuth angles sequentially. The TSRPT system, which is free of sample rotation, enables rapid scanning of different views by a tailor-designed four-galvo-mirror scanning system. We present the design of the optical system, the theory, and calibration procedure for chemical tomographic reconstruction. 3D vibrational images of polystyrene beads and C. elegans are demonstrated in the C-H vibrational region. 
    more » « less
  5. ; ; ; (, Sensors)
    Spectroscopic microtomography provides the ability to perform 4D (3D structural and 1D chemical) imaging of a thick microscopic specimen. Here, we demonstrate spectroscopic microtomography in the short-wave infrared (SWIR) wavelength using digital holographic tomography, which captures both the absorption coefficient and refractive index. A broadband laser in tandem with a tunable optical filter allows us to scan the wavelength from 1100 to 1650 nm. Using the developed system, we measure human hair and sea urchin embryo samples. The resolution estimated with gold nanoparticles is 1.51 μm (transverse) and 1.57 μm (axial) for the field of view of 307 × 246 μm2. The developed technique will enable accurate and efficient analyses of microscopic specimens that have a distinctive absorption or refractive index contrast in the SWIR range. 
    more » « less
  • Citation Formats
  • MLA
  • APA
  • Chicago
  • BibTeX
  • Save / Share this Record
  • Send to Email