scholarly journals Three-Dimensional High-Resolution Digital Inline Hologram Reconstruction with a Volumetric Deconvolution Method

Sensors ◽  
2018 ◽  
Vol 18 (9) ◽  
pp. 2918 ◽  
Author(s):  
Junseong Eom ◽  
Sangjun Moon
Keyword(s):  
High Resolution ◽  
Whole Blood ◽  
Tissue Sample ◽  
Three Dimensional ◽  
Kidney Tissue ◽  
Wide Field ◽  
Deconvolution Technique ◽  
Micron Size ◽  
Volumetric Images ◽  
Focus Image

The digital in-line holographic microscope (DIHM) was developed for a 2D imaging technology and has recently been adapted to 3D imaging methods, providing new approaches to obtaining volumetric images with both a high resolution and wide field-of-view (FOV), which allows the physical limitations to be overcome. However, during the sectioning process of 3D image generation, the out-of-focus image of the object becomes a significant impediment to obtaining evident 3D features in the 2D sectioning plane of a thick biological sample. Based on phase retrieved high-resolution holographic imaging and a 3D deconvolution technique, we demonstrate that a high-resolution 3D volumetric image, which significantly reduces wave-front reconstruction and out-of-focus artifacts, can be achieved. The results show a 3D volumetric image that is more finely focused compared to a conventional 3D stacked image from 2D reconstructed images in relation to micron-size polystyrene beads, a whole blood smear, and a kidney tissue sample. We believe that this technology can be applicable for medical-grade images of smeared whole blood or an optically cleared tissue sample for mobile phytological microscopy and laser sectioning microscopy.

scholarly journals Quantitative morphometric measurements using site selective image cytometry of intact tissue

2008 ◽  
Vol 6 (suppl_1) ◽  
Author(s):  
Hyuk-Sang Kwon ◽  
Yoon Sung Nam ◽  
Dominika M Wiktor-Brown ◽  
Bevin P Engelward ◽  
Peter T.C So
Keyword(s):  
High Resolution ◽  
Three Dimensional ◽  
Image Cytometry ◽  
Regions Of Interest ◽  
Wide Field ◽  
Detailed Measurement ◽  
Cell Clusters ◽  
Two Photon ◽  
Recombinant Cell ◽  
Site Selective

Site selective two-photon tissue image cytometry has previously been successfully applied to measure the number of rare cells in three-dimensional tissue specimens up to cubic millimetres in size. However, the extension of this approach for high-throughput quantification of cellular morphological states has not been demonstrated. In this paper, we report the use of site-selective tissue image cytometry for the study of homologous recombination (HR) events during cell division in the pancreas of transgenic mice. Since HRs are rare events, recombinant cells distribute sparsely inside the organ. A detailed measurement throughout the whole tissue is thus not practical. Instead, the site selective two-photon tissue cytometer incorporates a low magnification, wide field, one-photon imaging subsystem that rapidly identifies regions of interest containing recombinant cell clusters. Subsequently, high-resolution three-dimensional assays based on two-photon microscopy can be performed only in these regions of interest. We further show that three-dimensional morphology extraction algorithms can be used to analyse the resultant high-resolution two-photon image stacks providing information not only on the frequency and the distribution of these recombinant cell clusters and their constituent cells, but also on their morphology.

scholarly journals Widefield light sheet microscopy using an Airy beam combined with deep-learning super-resolution

2020 ◽  
Author(s):  
Stella Corsetti ◽  
Philip Wijesinghe ◽  
Persephone B. Poulton ◽  
Shuzo Sakata ◽  
Khushi Vyas ◽  
...  
Keyword(s):  
Deep Learning ◽  
High Resolution ◽  
Dynamic Range ◽  
Tissue Sample ◽  
Super Resolution ◽  
Light Sheet ◽  
Wide Field ◽  
Light Sheet Microscopy ◽  
Airy Beam ◽  
Model Of Alzheimer’S Disease

AbstractImaging across length scales and in depth has been an important pursuit of widefield optical imaging. This promises to reveal fine cellular detail within a widefield snapshot of a tissue sample. Current advances often sacrifice resolution through selective sub-sampling to provide a wide field of view in a reasonable time scale. We demonstrate a new avenue for recovering high-resolution images from sub-sampled data in light-sheet microscopy using deep-learning super-resolution. We combine this with the use of a widefield Airy beam to achieve high-resolution imaging over extended fields of view and depths. We characterise our method on fluorescent beads as test targets. We then demonstrate improvements in imaging amyloid plaques in a cleared brain from a mouse model of Alzheimer’s disease, and in excised healthy and cancerous colon and breast tissues. This development can be widely applied in all forms of light sheet microscopy to provide a two-fold increase in the dynamic range of the imaged length scale. It has the potential to provide further insight into neuroscience, developmental biology and histopathology.

scholarly journals Multi-fluorescence high-resolution episcopic microscopy (MF-HREM) for three dimensional imaging of adult murine organs

2020 ◽  
Author(s):  
Claire Walsh ◽  
Natalie Holroyd ◽  
Eoin Finnerty ◽  
Sean G. Ryan ◽  
Paul W. Sweeney ◽  
...  
Keyword(s):  
High Resolution ◽  
3D Imaging ◽  
Three Dimensional ◽  
Image Resolution ◽  
Biological Research ◽  
Wide Field ◽  
Vascular Networks ◽  
Optical Sectioning ◽  
Trade Offs ◽  
Xenograft Models

AbstractThree-dimensional microscopy of large biological samples (>0.5 cm3) is transforming biological research. Many existing techniques require trade-offs between image resolution and sample size, require clearing or use optical sectioning. These factors complicate the implementation of large volume 3D imaging. Here we present Multi-fluorescent High Resolution Episcopic Microscopy (MF-HREM) which allows 3D imaging of large samples without the need for clearing or optical sectioning.MF-HREM uses serial-sectioning and block-facing wide-field fluorescence, without the need for tissue clearing or optical sectioning. We detail developments in sample processing including stain penetration, resin embedding and imaging. In addition, we describe image post-processing methods needed to segment and further quantify these data. Finally, we demonstrate the wide applicability of MF-HREM by: 1) quantifying adult mouse glomeruli. 2) identifying injected cells and vascular networks in tumour xenograft models; 3) quantifying vascular networks and white matter track orientation in mouse brain.

High Resolution Microscopy of Multi-Layered Biological Crystals

1982 ◽  
Vol 40 ◽  
pp. 80-83
Author(s):  
H.A. Cohen ◽  
T.W. Jeng ◽  
W. Chiu
Keyword(s):  
High Resolution ◽  
Low Dose ◽  
Three Dimensional ◽  
Data Interpretation ◽  
Two Dimensional ◽  
Biological Objects ◽  
Density Maps ◽  
Density Map ◽  
Complex Crystal ◽  
High Resolution Microscopy

This tutorial will discuss the methodology of low dose electron diffraction and imaging of crystalline biological objects, the problems of data interpretation for two-dimensional projected density maps of glucose embedded protein crystals, the factors to be considered in combining tilt data from three-dimensional crystals, and finally, the prospects of achieving a high resolution three-dimensional density map of a biological crystal. This methodology will be illustrated using two proteins under investigation in our laboratory, the T4 DNA helix destabilizing protein gp32*I and the crotoxin complex crystal.

Electron crystallographic determination of the 3-D structure of staurolite at atomic resolution

1991 ◽  
Vol 49 ◽  
pp. 540-541
Author(s):  
Kenneth H. Downing ◽  
Hu Meisheng ◽  
Hans-Rudolf Went ◽  
Michael A. O'Keefe
Keyword(s):  
High Resolution ◽  
Three Dimensional ◽  
High Resolution Electron Microscopy ◽  
Atomic Resolution ◽  
Dimensional Structure ◽  
Structure Information ◽  
Resolution Electron ◽  
Scattering Effects ◽  
High Resolution Images ◽  
Effects Of Radiation

With current advances in electron microscope design, high resolution electron microscopy has become routine, and point resolutions of better than 2Å have been obtained in images of many inorganic crystals. Although this resolution is sufficient to resolve interatomic spacings, interpretation generally requires comparison of experimental images with calculations. Since the images are two-dimensional representations of projections of the full three-dimensional structure, information is invariably lost in the overlapping images of atoms at various heights. The technique of electron crystallography, in which information from several views of a crystal is combined, has been developed to obtain three-dimensional information on proteins. The resolution in images of proteins is severely limited by effects of radiation damage. In principle, atomic-resolution, 3D reconstructions should be obtainable from specimens that are resistant to damage. The most serious problem would appear to be in obtaining high-resolution images from areas that are thin enough that dynamical scattering effects can be ignored.

Three-Dimensional Immunoelectron Microscopy of Yeast Cells by a New High-Resolution Replica Method

1985 ◽  
Vol 43 ◽  
pp. 562-563
Author(s):  
Hirano T. ◽  
M. Yamaguchi ◽  
M. Hayashi ◽  
Y. Sekiguchi ◽  
A. Tanaka
Keyword(s):  
High Resolution ◽  
Plasma Polymerization ◽  
Three Dimensional ◽  
Freeze Fracture ◽  
Replica Method ◽  
Yeast Cells ◽  
Dynamic Aspect ◽  
Replica Technique ◽  
Gaseous Hydrocarbon ◽  
Transmission Electron

A plasma polymerization film replica method is a new high resolution replica technique devised by Tanaka et al. in 1978. It has been developed for investigation of the three dimensional ultrastructure in biological or nonbiological specimens with the transmission electron microscope. This method is based on direct observation of the single-stage replica film, which was obtained by directly coating on the specimen surface. A plasma polymerization film was deposited by gaseous hydrocarbon monomer in a glow discharge.The present study further developed the freeze fracture method by means of a plasma polymerization film produces a three dimensional replica of chemically untreated cells and provides a clear evidence of fine structure of the yeast plasma membrane, especially the dynamic aspect of the structure of invagination (Figure 1).

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