scholarly journals Optical Imaging of the Small Intestine Immune Compartments Across Scales

2021 ◽  
Author(s):  
Arielle Planchette ◽  
Cédric Schmidt ◽  
Olivier Burri ◽  
Mercedes Gomez de Agüero ◽  
Aleksandra Radenovic ◽  
...  
Keyword(s):  
Spatial Distribution ◽  
Small Intestine ◽  
High Resolution ◽  
Imaging Modality ◽  
Regions Of Interest ◽  
Label Free ◽  
3 Dimensional ◽  
Wide Range ◽  
Mouse Small Intestine ◽  
Made In

Abstract The limitations of 2D microscopy constrain our ability to observe and understand tissue-wide networks that are, by nature, 3-dimensional. Optical projection tomography enables the acquisition of large volumes (ranging from micrometres to centimetres) in various tissues, with label-free capacities for the observation of auto-fluorescent signals as well fluorescent-labelled targets of interest in multiple channels. We present a multi-modal workflow for the characterization of both structural and quantitative parameters of the mouse small intestine. As proof of principle, we evidence its applicability for imaging the mouse intestinal immune compartment and surrounding mucosal structures. We quantify the volumetric size and spatial distribution of Isolated Lymphoid Follicles (ILFs) and quantify density of villi throughout centimetre long segments of intestine. Furthermore, we exhibit the age- and microbiota-dependence for ILF development, and leverage a technique that we call reverse-OPT for identifying and homing in on regions of interest. Several quantification capabilities are displayed, including villous density in the autofluorescent channel and the size and spatial distribution of the signal of interest at millimetre-scale volumes. The concatenation of 3D image acquisition with the reverse-OPT sample preparation and a 2D high-resolution imaging modality adds value to interpretations made in 3D. This cross-modality referencing technique is found to provide accurate localisation of ROIs and to add value to interpretations made in 3D. Importantly, OPT may be used to identify sparsely-distributed regions of interest in large volumes whilst retaining compatibility with high-resolution microscopy modalities, including confocal microscopy. We believe this pipeline to be approachable for a wide-range of specialties, and to provide a new method for characterisation of the mouse intestinal immune compartment.

scholarly journals Waterproof Galvanometer Scanner-Based Handheld Photoacoustic Microscopy Probe for Wide-Field Vasculature Imaging In Vivo

Photonics ◽  
2021 ◽  
Vol 8 (8) ◽  
pp. 305
Author(s):  
Daewoon Seong ◽  
Sangyeob Han ◽  
Jaeyul Lee ◽  
Euimin Lee ◽  
Yoonseok Kim ◽  
...  
Keyword(s):  
Imaging Modality ◽  
Wide Field ◽  
Photoacoustic Microscopy ◽  
Label Free ◽  
Experimental Conditions ◽  
Sample Shape ◽  
Mouse Tissues ◽  
Wide Range ◽  
Galvanometer Scanner

Photoacoustic imaging (PAI) is a hybrid non-invasive imaging technique used to merge high optical contrast and high acoustic resolution in deep tissue. PAI has been extensively developed by utilizing its advantages that include deep imaging depth, high resolution, and label-free imaging. As a representative implementation of PAI, photoacoustic microscopy (PAM) has been used in preclinical and clinical studies for its micron-scale spatial resolution capability with high optical absorption contrast. Several handheld and portable PAM systems have been developed that improve its applicability to several fields, making it versatile. In this study, we developed a laboratory-customized, two-axis, waterproof, galvanometer scanner-based handheld PAM (WP-GVS-HH-PAM), which provides an extended field of view (14.5 × 9 mm2) for wide-range imaging. The fully waterproof handheld probe enables free movement for imaging regardless of sample shape, and volume rate and scanning region are adjustable per experimental conditions. Results of WP-GVS-HH-PAM-based phantom and in vivo imaging of mouse tissues (ear, iris, and brain) confirm the feasibility and applicability of our system as an imaging modality for various biomedical applications.

Development of a conical anode Fe-gun for low voltage SEM

1988 ◽  
Vol 46 ◽  
pp. 978-979
Author(s):  
T. Miyokawa ◽  
S. Norioka ◽  
S. Goto
Keyword(s):  
High Resolution ◽  
Field Emission ◽  
Low Voltage ◽  
Irradiation Damage ◽  
Cold Cathode ◽  
Virtual Source ◽  
Source Position ◽  
Electrostatic Lens ◽  
Electrostatic Lenses ◽  
Wide Range

Field emission SEMs (FE-SEMs) are becoming popular due to their high resolution needs. In the field of semiconductor product, it is demanded to use the low accelerating voltage FE-SEM to avoid the electron irradiation damage and the electron charging up on samples. However the accelerating voltage of usual SEM with FE-gun is limited until 1 kV, which is not enough small for the present demands, because the virtual source goes far from the tip in lower accelerating voltages. This virtual source position depends on the shape of the electrostatic lens. So, we investigated several types of electrostatic lenses to be applicable to the lower accelerating voltage. In the result, it is found a field emission gun with a conical anode is effectively applied for a wide range of low accelerating voltages.A field emission gun usually consists of a field emission tip (cold cathode) and the Butler type electrostatic lens.

High resolution spot scan imaging of frozen, hydrated actin bundles with 400kV electrons

1992 ◽  
Vol 50 (1) ◽  
pp. 512-513
Author(s):  
J. Jakana ◽  
M.F. Schmid ◽  
P. Matsudaira ◽  
W. Chiu
Keyword(s):  
High Resolution ◽  
Binding Proteins ◽  
Actin Binding ◽  
Eukaryotic Cells ◽  
Dimensional Structure ◽  
Structural Basis ◽  
Actin Bundle ◽  
Actin Bundles ◽  
3 Dimensional ◽  
Macromolecular Assembly

Actin is a protein found in all eukaryotic cells. In its polymerized form, the cells use it for motility, cytokinesis and for cytoskeletal support. An example of this latter class is the actin bundle in the acrosomal process from the Limulus sperm. The different functions actin performs seem to arise from its interaction with the actin binding proteins. A 3-dimensional structure of this macromolecular assembly is essential to provide a structural basis for understanding this interaction in relationship to its development and functions.

Functional information from magnetic resonance imaging

1995 ◽  
Vol 53 ◽  
pp. 84-85
Author(s):  
Alan P. Koretsky ◽  
Afonso Costa e Silva ◽  
Yi-Jen Lin
Keyword(s):  
Magnetic Resonance Imaging ◽  
Magnetic Resonance ◽  
High Resolution ◽  
Cancer Biology ◽  
Imaging Modality ◽  
Magnetic Resonance Images ◽  
Great Success ◽  
Functional Information ◽  
Resonance Imaging ◽  
High Resolution Mri

Magnetic resonance imaging (MRI) has become established as an important imaging modality for the clinical management of disease. This is primarily due to the great tissue contrast inherent in magnetic resonance images of normal and diseased organs. Due to the wide availability of high field magnets and the ability to generate large and rapidly switched magnetic field gradients there is growing interest in applying high resolution MRI to obtain microscopic information. This symposium on MRI microscopy highlights new developments that are leading to increased resolution. The application of high resolution MRI to significant problems in developmental biology and cancer biology will illustrate the potential of these techniques.In combination with a growing interest in obtaining high resolution MRI there is also a growing interest in obtaining functional information from MRI. The great success of MRI in clinical applications is due to the inherent contrast obtained from different tissues leading to anatomical information.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

Three-fold astigmatism: An important TEM aberration

1993 ◽  
Vol 51 ◽  
pp. 972-973 ◽  
Author(s):  
O.L. Krivanek ◽  
M.L. Leber
Keyword(s):  
High Resolution ◽  
Spatial Frequency ◽  
Field Emission ◽  
Azimuthal Angle ◽  
Hollow Cone ◽  
Small Probe ◽  
Wide Range ◽  
High Resolution Images ◽  
Image Position

Three-fold astigmatism resembles regular astigmatism, but it has 3-fold rather than 2-fold symmetry. Its contribution to the aberration function χ(q) can be written as:where A3 is the coefficient of 3-fold astigmatism, λ is the electron wavelength, q is the spatial frequency, ϕ the azimuthal angle (ϕ = tan-1 (qy/qx)), and ϕ3 the direction of the astigmatism.Three-fold astigmatism is responsible for the “star of Mercedes” aberration figure that one obtains from intermediate lenses once their two-fold astigmatism has been corrected. Its effects have been observed when the beam is tilted in a hollow cone over a wide range of angles, and there is evidence for it in high resolution images of a small probe obtained in a field emission gun TEM/STEM instrument. It was also expected to be a major aberration in sextupole-based Cs correctors, and ways were being developed for dealing with it on Cs-corrected STEMs.

Scanning tunneling microscopy of polymers: A status report

1989 ◽  
Vol 47 ◽  
pp. 330-331
Author(s):  
P.E. Russell ◽  
I.H. Musselman
Keyword(s):  
High Resolution ◽  
Scanning Tunneling Microscopy ◽  
Sample Surface ◽  
Atomic Scale ◽  
Constant Current ◽  
Scanning Tunneling ◽  
Status Report ◽  
Tunneling Microscopy ◽  
Research Issues ◽  
Wide Range

Scanning tunneling microscopy (STM) has evolved rapidly in the past few years. Major developments have occurred in instrumentation, theory, and in a wide range of applications. In this paper, an overview of the application of STM and related techniques to polymers will be given, followed by a discussion of current research issues and prospects for future developments. The application of STM to polymers can be conveniently divided into the following subject areas: atomic scale imaging of uncoated polymer structures; topographic imaging and metrology of man-made polymer structures; and modification of polymer structures. Since many polymers are poor electrical conductors and hence unsuitable for use as a tunneling electrode, the related atomic force microscopy (AFM) technique which is capable of imaging both conductors and insulators has also been applied to polymers.The STM is well known for its high resolution capabilities in the x, y and z axes (Å in x andy and sub-Å in z). In addition to high resolution capabilities, the STM technique provides true three dimensional information in the constant current mode. In this mode, the STM tip is held at a fixed tunneling current (and a fixed bias voltage) and hence a fixed height above the sample surface while scanning across the sample surface.

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