1,766 research outputs found
3D correlative single-cell imaging utilizing fluorescence and refractive index tomography
Cells alter the path of light, a fact that leads to well-known aberrations in
single cell or tissue imaging. Optical diffraction tomography (ODT) measures
the biophysical property that causes these aberrations, the refractive index
(RI). ODT is complementary to fluorescence imaging and does not require any
markers. The present study introduces RI and fluorescence tomography with
optofluidic rotation (RAFTOR) of suspended cells, quantifying the intracellular
RI distribution and colocalizing it with fluorescence in 3D. The technique is
validated with cell phantoms and used to confirm a lower nuclear RI for HL60
cells. Furthermore, the nuclear inversion of adult mouse photoreceptor cells is
observed in the RI distribution. The applications shown confirm predictions of
previous studies and illustrate the potential of RAFTOR to improve our
understanding of cells and tissues.Comment: 15 pages, 5 figure
Four-dimensional dynamic flow measurement by holographic particle image velocimetry
The ultimate goal of holographic particle image velocimetry (HPIV) is to provide space- and time-resolved measurement of complex flows. Recent new understanding of holographic imaging of small particles, pertaining to intrinsic aberration and noise in particular, has enabled us to elucidate fundamental issues in HPIV and implement a new HPIV system. This system is based on our previously reported off-axis HPIV setup, but the design is optimized by incorporating our new insights of holographic particle imaging characteristics. Furthermore, the new system benefits from advanced data processing algorithms and distributed parallel computing technology. Because of its robustness and efficiency, for the first time to our knowledge, the goal of both temporally and spatially resolved flow measurements becomes tangible. We demonstrate its temporal measurement capability by a series of phase-locked dynamic measurements of instantaneous three-dimensional, three-component velocity fields in a highly three-dimensional vortical flow--the flow past a tab
Maskless imaging of dense samples using pixel super-resolution based multi-height lensfree on-chip microscopy.
Lensfree in-line holographic microscopy offers sub-micron resolution over a large field-of-view (e.g., ~24 mm2) with a cost-effective and compact design suitable for field use. However, it is limited to relatively low-density samples. To mitigate this limitation, we demonstrate an on-chip imaging approach based on pixel super-resolution and phase recovery, which iterates among multiple lensfree intensity measurements, each having a slightly different sample-to-sensor distance. By digitally aligning and registering these lensfree intensity measurements, phase and amplitude images of dense and connected specimens can be iteratively reconstructed over a large field-of-view of ~24 mm2 without the use of any spatial masks. We demonstrate the success of this multi-height in-line holographic approach by imaging dense Papanicolaou smears (i.e., Pap smears) and blood samples
High-resolution transport-of-intensity quantitative phase microscopy with annular illumination
For quantitative phase imaging (QPI) based on transport-of-intensity equation
(TIE), partially coherent illumination provides speckle-free imaging,
compatibility with brightfield microscopy, and transverse resolution beyond
coherent diffraction limit. Unfortunately, in a conventional microscope with
circular illumination aperture, partial coherence tends to diminish the phase
contrast, exacerbating the inherent noise-to-resolution tradeoff in TIE
imaging, resulting in strong low-frequency artifacts and compromised imaging
resolution. Here, we demonstrate how these issues can be effectively addressed
by replacing the conventional circular illumination aperture with an annular
one. The matched annular illumination not only strongly boosts the phase
contrast for low spatial frequencies, but significantly improves the practical
imaging resolution to near the incoherent diffraction limit. By incorporating
high-numerical aperture (NA) illumination as well as high-NA objective, it is
shown, for the first time, that TIE phase imaging can achieve a transverse
resolution up to 208 nm, corresponding to an effective NA of 2.66. Time-lapse
imaging of in vitro Hela cells revealing cellular morphology and subcellular
dynamics during cells mitosis and apoptosis is exemplified. Given its
capability for high-resolution QPI as well as the compatibility with widely
available brightfield microscopy hardware, the proposed approach is expected to
be adopted by the wider biology and medicine community.Comment: This manuscript was originally submitted on 20 Feb. 201
Automated Three-Dimensional Microbial Sensing and Recognition Using Digital Holography and Statistical Sampling
We overview an approach to providing automated three-dimensional (3D) sensing and recognition of biological micro/nanoorganisms integrating Gabor digital holographic microscopy and statistical sampling methods. For 3D data acquisition of biological specimens, a coherent beam propagates through the specimen and its transversely and longitudinally magnified diffraction pattern observed by the microscope objective is optically recorded with an image sensor array interfaced with a computer. 3D visualization of the biological specimen from the magnified diffraction pattern is accomplished by using the computational Fresnel propagation algorithm. For 3D recognition of the biological specimen, a watershed image segmentation algorithm is applied to automatically remove the unnecessary background parts in the reconstructed holographic image. Statistical estimation and inference algorithms are developed to the automatically segmented holographic image. Overviews of preliminary experimental results illustrate how the holographic image reconstructed from the Gabor digital hologram of biological specimen contains important information for microbial recognition
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Computational cytometer based on magnetically modulated coherent imaging and deep learning.
Detecting rare cells within blood has numerous applications in disease diagnostics. Existing rare cell detection techniques are typically hindered by their high cost and low throughput. Here, we present a computational cytometer based on magnetically modulated lensless speckle imaging, which introduces oscillatory motion to the magnetic-bead-conjugated rare cells of interest through a periodic magnetic force and uses lensless time-resolved holographic speckle imaging to rapidly detect the target cells in three dimensions (3D). In addition to using cell-specific antibodies to magnetically label target cells, detection specificity is further enhanced through a deep-learning-based classifier that is based on a densely connected pseudo-3D convolutional neural network (P3D CNN), which automatically detects rare cells of interest based on their spatio-temporal features under a controlled magnetic force. To demonstrate the performance of this technique, we built a high-throughput, compact and cost-effective prototype for detecting MCF7 cancer cells spiked in whole blood samples. Through serial dilution experiments, we quantified the limit of detection (LoD) as 10 cells per millilitre of whole blood, which could be further improved through multiplexing parallel imaging channels within the same instrument. This compact, cost-effective and high-throughput computational cytometer can potentially be used for rare cell detection and quantification in bodily fluids for a variety of biomedical applications
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