The artery wall is equipped with a water permeation barrier that allows blood to flow at high pressure without significant water leak. The precise location of this barrier is unknown despite its importance in vascular function and its contribution to many vascular complications when it is compromised. Herein we map the water permeability in intact arteries, using coherent anti-Stokes Raman scattering (CARS) microscopy and isotopic perfusion experiments. Generation of the CARS signal is optimized for water imaging with broadband excitation. We identify the water permeation barrier as the endothelial basolateral membrane and show that the apical membrane is highly permeable. This is confirmed by the distribution of the AQP1 water channel within endothelial membranes. These results indicate that arterial pressure equilibrates within the endothelium and is transmitted to the supporting basement membrane and internal elastic lamina macromolecules with minimal deformation of the sensitive endothelial cell. Disruption of this pressure transmission could contribute to endothelial cell dysfunction in various pathologies.
[1] Millimeter-wave interferometric synthetic aperture imagers are currently being developed for short-range applications such as concealed weapons detection. In contrast to the traditional snapshot imaging approach, we investigate the potential of mechanical scanning between the scene and the array in order to reduce the number of antennas and correlators. We assess the trade-off between this hardware reduction, the radiometric sensitivity and the imaging frame rate of the system. We show that rotational scanning achieves a more uniform coverage of the (u, v) plane than the more conventional linear scanning. We use a genetic algorithm to optimize two-dimensional arrays for maximum uniform (u, v) coverage after a rotational mechanical scan and demonstrates improvements in the array point spread function. Imaging performance is assessed with simulated millimeter-wave scenes. Results show an increased image quality is achieved with the optimized array compared with a conventional power law Y-shaped array. Finally we discuss the increased demands on system stability and calibration that the increased acquisition time of the proposed technique places.Citation: Lucotte, B. M., B. Grafulla-González, and A. R. Harvey (2009), Array rotation aperture synthesis for short-range imaging at millimeter wavelengths, Radio Sci., 44, RS1006,
SummaryUsing the intrinsic optical properties of collagen and elastin, two-photon microscopy was applied to evaluate the threedimensional (3D) macromolecular structural development of the mouse thoracic aorta from birth to 60 days old. Baseline development was established in the Scavenger Receptor Class B Type I-Deficient, Hypomorphic Apolipoprotein ER61 (SR-BI KO/ApoeR61 h/h ) mouse in preparation for modeling atherosclerosis. Precise dissection enabled direct observation of the artery wall in situ. En-face, optical sectioning of the aorta provided a novel assessment of the macromolecular structural development. During aortic development, the undulating lamellar elastin layers compressed consistent with the increases in mean aortic pressure with age. In parallel, a net increase in overall wall thickness (p<0.05, in day 60 compared with day 1 mice) occurred with age whereas the ratio of the tunicas adventitia and media to full aortic thickness remained nearly constant across age groups (~1:2.6, respectively). Histochemical analyses by brightfield microscopy and ultrastructure validated structural proteins and lipid deposition findings derived from two-photon microscopy. Development was associated with decreased decorin but not biglycan proteoglycan expression. This non-destructive 3D in situ approach revealed the aortic wall microstructure development. Coupling this approach with the intrinsic optical properties of the macromolecules may provide unique vascular wall 3D structure in many pathological conditions, including aortic atherosclerosis, dissections and aneurysms. (J Histochem Cytochem 63:8-21, 2015)
SUMMARY In conventional multi-probe fluorescence microscopy, narrow bandwidth filters on detectors are used to avoid bleed-through artifacts between probes. The limited bandwidth reduces the signal-to-noise ratio (SNR) of the detection, often severely compromising one or more channels. Herein, we describe a process of using independent component analysis (ICA) to discriminate the position of different probes using only a dichroic mirror to differentiate the signals directed to the detectors. ICA was particularly effective in samples where the spatial overlap between the probes is minimal, a very common case in cellular microscopy. This imaging scheme collects nearly all of the emitted light, significantly improving the image SNR. In this study, we focused on the detection of two fluorescence probes used in vivo, NAD(P)H and ANEPPS. The optimal dichroic mirror cutoff frequency was determined with simulations using the probes spectral emissions. A quality factor, defined as the cross-channel contrast-to-noise ratio, was optimized to maximize signals while maintaining spatial discrimination between the probes after ICA post-processing. Simulations indicate that a ~3 fold increase in SNR using the ICA approach can be achieved over the conventional narrow-band filtering approach without loss of spatial discrimination. We confirmed this predicted performance from experimental imaging of NAD(P)H and ANEPPS in mouse skeletal muscle,in vivo. For many multi-probe studies, the increased sensitivity of this “full bandwidth” approach will lead to improved image quality and/or reduced excitation power requirements.
In this review we focus on the impact of tissue motion on attempting to conduct sub-cellular resolution optical microscopy, in vivo. Our position is that tissue motion is one of the major barriers to conducting these studies along with light induced damage, optical probe loading as well as absorbance and scattering effects on the excitation point spread function and collection of emitted light. Recent developments in the speed of image acquisition have reached the limit, in most cases, where the signal from a sub-cellular voxel limits the speed and not the scanning rate of the microscope. Different schemes for compensating for tissue displacements due to rigid body and deformation are presented from tissue restriction, gating, adaptive gating and active tissue tracking. We argue that methods that minimally impact the natural physiological motion of the tissue are desirable since the major reason to perform in vivo studies is to evaluate normal physiological functions. Towards this goal, the methods for active tracking using the optical imaging data itself to monitor tissue displacement and actively move the FOV of the microscope to match the tissue deformation in near real time. Critical for this development was the implementation of near real time image processing in conjunction with the control of the microscope imaging parameters. Clearly the continuing development of methods of motion compensation as well as significant technological solutions to the other barriers to tissue sub-cellular optical imaging in vivo, including optical aberrations and overall signal to noise, will make major contributions to the understanding of cell biology within the body.
Hard x-ray lenses are useful elements in x-ray microscopy and in creating focused illumination for analytical applications such as x-ray fluorescence imaging. Recently, polymer compound refractive lenses for focused illumination in the soft x-ray regime (< 10 keV) have been created with nano-printing. However, there are no such lenses yet for hard x-rays, particularly of short focal lengths for benchtop microscopy. We report the first instance of a nano-printed lens for hard x-ray microscopy, and evaluate its imaging performance. The lens consists of a spherically focusing compound refractive lens designed for 22 keV photon energy, with a tightly packed structure to provide a short total length of 1.8 mm and a focal length of 21.5 mm. The resulting lens technology was found to enable benchtop microscopy at 74x magnification and 1.1 μm de-magnified image pixel size at the object plane. It was used to image and evaluate the focal spots of tungsten-anode micro-focus x-ray sources. The overall system resolution with broadband illumination from a tungsten-anode x-ray tube at 30 kV and 10 mm focal distance was measured to be 2.30±0.22 μm.
SUMMARY This paper investigates a post-processing approach to correct spatial distortion in two-photon fluorescence microscopy images for vascular network reconstruction. It is aimed at in vivo imaging of large field-of-view, deep-tissue studies of vascular structures. Based on simple geometric modeling of the object-of-interest, a distortion function is directly estimated from the image volume by deconvolution analysis. Such distortion function is then applied to sub volumes of the image stack to adaptively adjust for spatially varying distortion and reduce the image blurring through blind deconvolution. The proposed technique was first evaluated in phantom imaging of fluorescent microspheres that are comparable in size to the underlying capillary vascular structures. The effectiveness of restoring three-dimensional spherical geometry of the microspheres using the estimated distortion function was compared with empirically measured point-spread function. Next, the proposed approach was applied to in vivo vascular imaging of mouse skeletal muscle to reduce the image distortion of the capillary structures. We show that the proposed method effectively improve the image quality and reduce spatially varying distortion that occurs in large field-of-view deep-tissue vascular dataset. The proposed method will help in qualitative interpretation and quantitative analysis of vascular structures from fluorescence microscopy images.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.