Abstract Since its first demonstration over 100 years ago, scattering‐based light‐sheet microscopy has recently re‐emerged as a key modality in label‐free tissue imaging and cellular morphometry; however, scattering‐based light‐sheet imaging with subcellular resolution remains an unmet target. This is because related approaches inevitably superimpose speckle or granular intensity modulation on to the native subcellular features. Here, we addressed this challenge by deploying a time‐averaged pseudo‐thermalized light‐sheet illumination. While this approach increased the lateral dimensions of the illumination sheet, we achieved subcellular resolving power after image deconvolution. We validated this approach by imaging cytosolic carbon depots in yeast and bacteria with increased specificity, no staining, and ultralow irradiance levels. Overall, we expect this scattering‐based light‐sheet microscopy approach will advance single, live cell imaging by conferring low‐irradiance and label‐free operation towards eradicating phototoxicity.
more »
« less
Virtual‐point‐based deconvolution for optical‐resolution photoacoustic microscopy
Abstract Optical‐resolution photoacoustic microscopy (OR‐PAM) has been increasingly utilized for in vivo imaging of biological tissues, offering structural, functional, and molecular information. In OR‐PAM, it is often necessary to make a trade‐off between imaging depth, lateral resolution, field of view, and imaging speed. To improve the lateral resolution without sacrificing other performance metrics, we developed a virtual‐point‐based deconvolution algorithm for OR‐PAM (VP‐PAM). VP‐PAM has achieved a resolution improvement ranging from 43% to 62.5% on a single‐line target. In addition, it has outperformed Richardson‐Lucy deconvolution with 15 iterations in both structural similarity index and peak signal‐to‐noise ratio on an OR‐PAM image of mouse brain vasculature. When applied to an in vivo glass frog image obtained by a deep‐penetrating OR‐PAM system with compromised lateral resolution, VP‐PAM yielded enhanced resolution and contrast with better‐resolved microvessels.
more »
« less
- Award ID(s):
- 2144788
- PAR ID:
- 10576335
- Publisher / Repository:
- Wiley-VCH GmbH
- Date Published:
- Journal Name:
- Journal of Biophotonics
- Volume:
- 17
- Issue:
- 8
- ISSN:
- 1864-063X
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
Intraoperative imaging of slide-free specimens is crucial for oncology surgeries, allowing surgeons to quickly identify tumor margins for precise surgical guidance. While high-resolution ultraviolet photoacoustic microscopy has been demonstrated for slide-free histology, the imaging speed is insufficient, due to the low laser repetition rate and the limited depth of field. To address these challenges, we present parallel ultraviolet photoacoustic microscopy (PUV-PAM) with simultaneous scanning of eight optical foci to acquire histology-like images of slide-free fresh specimens, improving the ultraviolet PAM imaging speed limited by low laser repetition rates. The PUV-PAM has achieved an imaging speed of 0.4 square millimeters per second (i.e., 4.2 minutes per square centimeter) at 1.3-micrometer resolution using a 50-kilohertz laser. In addition, we demonstrated the PUV-PAM with eight needle-shaped beams for an extended depth of field, allowing fast imaging of slide-free tissues with irregular surfaces. We believe that the PUV-PAM approach will enable rapid intraoperative photoacoustic histology and provide prospects for ultrafast optical-resolution PAM.more » « less
-
Abstract By fitting observed data with predicted seismograms, least‐squares migration (LSM) computes a generalized inverse for a subsurface reflectivity model, which can improve image resolution and reduce artifacts caused by incomplete acquisition. However, the large computational cost of LSM required for simulations and migrations limits its wide applications for large‐scale imaging problems. Using point‐spread function (PSF) deconvolution, we present an efficient and stable high‐resolution imaging method. The PSFs are first computed on a coarse grid using local ray‐based Gaussian beam Born modeling and migration. Then, we interpolate the PSFs onto a fine‐image grid and apply a high‐dimensional Gaussian function to attenuate artifacts far away from the PSF centers. With 2D/3D partition of unity, we decompose the traditional adjoint migration results into local images with the same window size as the PSFs. Then, these local images are deconvolved by the PSFs in the wavenumber domain to reduce the effects of the band‐limited source function and compensate for irregular subsurface illumination. The final assembled image is obtained by applying the inverse of the partitions for the deconvolved local images. Numerical examples for both synthetic and field data demonstrate that the proposed PSF deconvolution can significantly improve image resolution and amplitudes for deep structures, while not being sensitive to velocity errors as the data‐domain LSM.more » « less
-
The human sense of smell plays an important role in appetite and food intake, detecting environmental threats, social interactions, and memory processing. However, little is known about the neural circuity supporting its function. The olfactory tracts project from the olfactory bulb along the base of the frontal cortex, branching into several striae to meet diverse cortical regions. Historically, using diffusion magnetic resonance imaging (dMRI) to reconstruct the human olfactory tracts has been prevented by susceptibility and motion artifacts. Here, we used a dMRI method with readout segmentation of long variable echo-trains (RESOLVE) to minimize image distortions and characterize the human olfactory tracts in vivo . We collected high-resolution dMRI data from 25 healthy human participants (12 male and 13 female) and performed probabilistic tractography using constrained spherical deconvolution (CSD). At the individual subject level, we identified the lateral, medial, and intermediate striae with their respective cortical connections to the piriform cortex and amygdala (AMY), olfactory tubercle (OT), and anterior olfactory nucleus (AON). We combined individual results across subjects to create a normalized, probabilistic atlas of the olfactory tracts. We then investigated the relationship between olfactory perceptual scores and measures of white matter integrity, including mean diffusivity (MD). Importantly, we found that olfactory tract MD negatively correlated with odor discrimination performance. In summary, our results provide a detailed characterization of the connectivity of the human olfactory tracts and demonstrate an association between their structural integrity and olfactory perceptual function. SIGNIFICANCE STATEMENT This study provides the first detailed in vivo description of the cortical connectivity of the three olfactory tract striae in the human brain, using diffusion magnetic resonance imaging (dMRI). Additionally, we show that tract microstructure correlates with performance on an odor discrimination task, suggesting a link between the structural integrity of the olfactory tracts and odor perception. Lastly, we generated a normalized probabilistic atlas of the olfactory tracts that may be used in future research to study its integrity in health and disease.more » « less
-
Endoscopy, enabling high-resolution imaging of deep tissues and internal organs, plays an important role in basic research and clinical practice. Recent advances in photoacoustic microscopy (PAM), demonstrating excellent capabilities in high-resolution functional imaging, have sparked significant interest in its integration into the field of endoscopy. However, there are challenges in achieving functional PAM in the endoscopic setting. This Perspective article discusses current progress in the development of endoscopic PAM and the challenges related to functional measurements. Then, it points out potential directions to advance endoscopic PAM for functional imaging by leveraging fiber optics, microfabrication, optical engineering, and computational approaches. Finally, it highlights emerging opportunities for functional endoscopic PAM in basic and translational biomedicine.more » « less
An official website of the United States government

