Effect of optical tissue clearing on spatial resolution and sensitivity of bioluminescence imaging

2006 ◽  
Vol 11 (4) ◽  
pp. 041119 ◽  
Author(s):  
E. Duco Jansen ◽  
Patrick M. Pickett ◽  
Mark A. Mackanos ◽  
John Virostko
Keyword(s):  
Spatial Resolution ◽  
Bioluminescence Imaging ◽  
Tissue Clearing

scholarly journals A versatile tiling light sheet microscope for cleared tissues imaging

2019 ◽  
Author(s):  
Yanlu Chen ◽  
Xiaoliang Li ◽  
Dongdong Zhang ◽  
Chunhui Wang ◽  
Ruili Feng ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Phase Modulation ◽  
3D Imaging ◽  
Tissue Expansion ◽  
Light Sheet ◽  
Nanometer Scale ◽  
Tissue Clearing ◽  
Working Principle ◽  
Tissue Size ◽  
Imaging Speed

AbstractWe present a tiling light sheet microscope compatible with all tissue clearing methods for rapid multicolor 3D imaging of centimeter-scale cleared tissues with micron-scale (4×4×10 μm3) to nanometer-scale (70×70×200 nm3) spatial resolution. The microscope uses tiling light sheets to achieve a more advanced multicolor 3D imaging ability than conventional light sheet microscopes. The illumination light is phase modulated to adjust the 3D imaging ability of the microscope based on the tissue size and the desired spatial resolution and imaging speed, and also to keep the microscope aligned. The ability of the microscope to align via phase modulation alone also makes the microscope reliable and easy to operate. Here we describe the working principle and design of the microscope. We demonstrate its imaging ability by imaging various cleared tissues prepared by different tissue clearing and tissue expansion techniques.

scholarly journals Use of an Image Restoration Process to Improve Spatial Resolution in Bioluminescence Imaging

2011 ◽  
Vol 10 (6) ◽  
pp. 7290.2011.00012 ◽  
Author(s):  
Sabrina Pesnel ◽  
Smaïl Akkoul ◽  
Roger Ledée ◽  
Rémy Leconge ◽  
Arnaud Pillon ◽  
...  
Keyword(s):  
Image Restoration ◽  
Spatial Resolution ◽  
Bioluminescence Imaging ◽  
Peritoneal Tumor ◽  
Scattering Correction ◽  
Before And After ◽  
In Vivo Bioluminescence Imaging ◽  
Restoration Method ◽  
Reference Measurements

To improve spatial resolution in in vivo bioluminescence imaging, a photon scattering correction, image restoration method was tested. The chosen algorithm was tested on in vivo bioluminescent images acquired on three representative tumor models: subcutaneous, pulmonary, and disseminated peritoneal. Tumor size was chosen as a quantitative criterion, such that the tumor reference measurements (determined photographically or by computed tomography) were compared to those derived from bioluminescent images, before and after restoration. This technique allowed a significant reduction to be achieved in the relative error between reference measurements and dimensions derived from bioluminescent images. In addition, improved delineation of the tumor foci was achieved. The restoration method allows spatial resolution in bioluminescence imaging to be improved, with interesting perspectives in terms of staging and quantitation in experimental oncology.

Spatial resolution of X-ray microanalysis in thin foils

1978 ◽  
Vol 36 (1) ◽  
pp. 544-545
Author(s):  
R. Hutchings ◽  
I.P. Jones ◽  
M.H. Loretto ◽  
R.E. Smallman
Keyword(s):  
Spatial Resolution ◽  
Electron Probe ◽  
Beam Divergence ◽  
Inelastic Interaction ◽  
Specimen Thickness ◽  
High Current ◽  
Beam Spreading ◽  
X Ray ◽  
Thin Foils ◽  
Complex Calculation

There is increasing interest in X-ray microanalysis of thin specimens and the present paper attempts to define some of the factors which govern the spatial resolution of this type of microanalysis. One of these factors is the spreading of the electron probe as it is transmitted through the specimen. There will always be some beam-spreading with small electron probes, because of the inevitable beam divergence associated with small, high current probes; a lower limit to the spatial resolution is thus 2αst where 2αs is the beam divergence and t the specimen thickness.In addition there will of course be beam spreading caused by elastic and inelastic interaction between the electron beam and the specimen. The angle through which electrons are scattered by the various scattering processes can vary from zero to 180° and it is clearly a very complex calculation to determine the effective size of the beam as it propagates through the specimen.

Precipitation in silicon: Microanalysis

1988 ◽  
Vol 46 ◽  
pp. 474-475
Author(s):  
R.W. Carpenter
Keyword(s):  
Transition Metals ◽  
Spatial Resolution ◽  
Thin Foil ◽  
Precipitation Reaction ◽  
High Current ◽  
Reaction Products ◽  
Carbon And Nitrogen ◽  
Dielectric Layers ◽  
Precipitation Processes ◽  
Areas Of Interest

Interest in precipitation processes in silicon appears to be centered on transition metals (for intrinsic and extrinsic gettering), and oxygen and carbon in thermally aged materials, and on oxygen, carbon, and nitrogen in ion implanted materials to form buried dielectric layers. A steadily increasing number of applications of microanalysis to these problems are appearing. but still far less than the number of imaging/diffraction investigations. Microanalysis applications appear to be paced by instrumentation development. The precipitation reaction products are small and the presence of carbon is often an important consideration. Small high current probes are important and cryogenic specimen holders are required for consistent suppression of contamination buildup on specimen areas of interest. Focussed probes useful for microanalysis should be in the range of 0.1 to 1nA, and estimates of spatial resolution to be expected for thin foil specimens can be made from the curves shown in Fig. 1.

High spatial resolution x-ray microanalysis of interfaces

1995 ◽  
Vol 53 ◽  
pp. 284-285
Author(s):  
J. R. Michael
Keyword(s):  
Spatial Resolution ◽  
Electron Probe ◽  
Solid Angle ◽  
Collection Efficiency ◽  
Spatial Scales ◽  
Energy Dispersive Spectrometer ◽  
X Rays ◽  
X Ray ◽  
Probe Size ◽  
Brightness Field

X-ray microanalysis in the analytical electron microscope (AEM) refers to a technique by which chemical composition can be determined on spatial scales of less than 10 nm. There are many factors that influence the quality of x-ray microanalysis. The minimum probe size with sufficient current for microanalysis that can be generated determines the ultimate spatial resolution of each individual microanalysis. However, it is also necessary to collect efficiently the x-rays generated. Modern high brightness field emission gun equipped AEMs can now generate probes that are less than 1 nm in diameter with high probe currents. Improving the x-ray collection solid angle of the solid state energy dispersive spectrometer (EDS) results in more efficient collection of x-ray generated by the interaction of the electron probe with the specimen, thus reducing the minimum detectability limit. The combination of decreased interaction volume due to smaller electron probe size and the increased collection efficiency due to larger solid angle of x-ray collection should enhance our ability to study interfacial segregation.

Scanning x-ray fluorescence microscopy and principal component analysis

1992 ◽  
Vol 50 (2) ◽  
pp. 1752-1753
Author(s):  
Brian Cross
Keyword(s):  
Principal Component Analysis ◽  
Fluorescence Microscopy ◽  
Spatial Resolution ◽  
High Speed ◽  
Image Acquisition ◽  
Principal Component ◽  
Fluorescence Microscope ◽  
Acquisition Rate ◽  
X Ray ◽  
Speed Up

A relatively new entry, in the field of microscopy, is the Scanning X-Ray Fluorescence Microscope (SXRFM). Using this type of instrument (e.g. Kevex Omicron X-ray Microprobe), one can obtain multiple elemental x-ray images, from the analysis of materials which show heterogeneity. The SXRFM obtains images by collimating an x-ray beam (e.g. 100 μm diameter), and then scanning the sample with a high-speed x-y stage. To speed up the image acquisition, data is acquired "on-the-fly" by slew-scanning the stage along the x-axis, like a TV or SEM scan. To reduce the overhead from "fly-back," the images can be acquired by bi-directional scanning of the x-axis. This results in very little overhead with the re-positioning of the sample stage. The image acquisition rate is dominated by the x-ray acquisition rate. Therefore, the total x-ray image acquisition rate, using the SXRFM, is very comparable to an SEM. Although the x-ray spatial resolution of the SXRFM is worse than an SEM (say 100 vs. 2 μm), there are several other advantages.

Quantitative parallel EELS spectrum imaging developed as a new tool in AEM

1992 ◽  
Vol 50 (2) ◽  
pp. 1202-1203
Author(s):  
Gianluigi Botton ◽  
Gilles L'espérance
Keyword(s):  
Statistical Analysis ◽  
Spatial Resolution ◽  
Personal Computer ◽  
Systematic Study ◽  
Present Author ◽  
Chemical Elements ◽  
Detection Limits ◽  
Research Groups ◽  
Quantitative Performance ◽  
Spectrum Imaging

As interest for parallel EELS spectrum imaging grows in laboratories equipped with commercial spectrometers, different approaches were used in recent years by a few research groups in the development of the technique of spectrum imaging as reported in the literature. Either by controlling, with a personal computer both the microsope and the spectrometer or using more powerful workstations interfaced to conventional multichannel analysers with commercially available programs to control the microscope and the spectrometer, spectrum images can now be obtained. Work on the limits of the technique, in terms of the quantitative performance was reported, however, by the present author where a systematic study of artifacts detection limits, statistical errors as a function of desired spatial resolution and range of chemical elements to be studied in a map was carried out The aim of the present paper is to show an application of quantitative parallel EELS spectrum imaging where statistical analysis is performed at each pixel and interpretation is carried out using criteria established from the statistical analysis and variations in composition are analyzed with the help of information retreived from t/γ maps so that artifacts are avoided.

Cristal-growth dynamics of n-doped gaAs

1992 ◽  
Vol 50 (2) ◽  
pp. 1544-1545
Author(s):  
Pham V. Huong ◽  
Stéphanie Bouchet ◽  
Jean-Claude Launay
Keyword(s):  
Crystal Growth ◽  
Spatial Resolution ◽  
Epitaxial Layer ◽  
Outer Surface ◽  
Gaas Substrate ◽  
Structural Modification ◽  
Growth Dynamics ◽  
Argon Laser ◽  
High Anisotropy ◽  
Crystal Gaas

Microstructure of epitaxial layers of doped GaAs and its crystal growth dynamics on single crystal GaAs substrate were studied by Raman microspectroscopy with a Dilor OMARS instrument equipped with a 1024 photodiode multichannel detector and a ion-argon laser Spectra-Physics emitting at 514.5 nm.The spatial resolution of this technique, less than 1 μm2, allows the recording of Raman spectra at several spots in function of thickness, from the substrate to the outer deposit, including areas around the interface (Fig.l).The high anisotropy of the LO and TO Raman bands is indicative of the orientation of the epitaxial layer as well as of the structural modification in the deposit and in the substrate at the interface.With Sn doped, the epitaxial layer also presents plasmon in Raman scattering. This fact is already very well known, but we additionally observed that its frequency increases with the thickness of the deposit. For a sample with electron density 1020 cm-3, the plasmon L+ appears at 930 and 790 cm-1 near the outer surface.

Practical Importance of Spatial Resolution and Analytical Sensitivity in AEM X-ray Microanalysis

1990 ◽  
Vol 48 (2) ◽  
pp. 432-433
Author(s):  
J. Zhang ◽  
D.B. Williams ◽  
J.I. Goldstein
Keyword(s):  
Spatial Resolution ◽  
Practical Importance ◽  
Beam Current ◽  
Specimen Thickness ◽  
Analytical Sensitivity ◽  
Related Factors ◽  
X Ray ◽  
Probe Size ◽  
Excitation Volume ◽  
Two Parameters

Analytical sensitivity and spatial resolution are important and closely related factors in x-ray microanalysis using the AEM. Analytical sensitivity is the ability to distinguish, for a given element under given conditions, between two concentrations that are nearly equal. The analytical sensitivity is directly related to the number of x-ray counts collected and, therefore, to the probe current, specimen thickness and counting time. The spatial resolution in AEM analysis is determined by the probe size and beam broadening in the specimen. A finer probe and a thinner specimen give a higher spatial resolution. However, the resulting lower beam current and smaller X-ray excitation volume degrade analytical sensitivity. A compromise must be made between high spatial resolution and an acceptable analytical sensitivity. In this paper, we show the necessity of evaluating these two parameters in order to determine the low temperature Fe-Ni phase diagram.A Phillips EM400T AEM with an EDAX/TN2000 EDS/MCA system and a VG HB501 FEG STEM with a LINK AN10 EDS/MCA system were used.

High spatial resolution AEM observations of yttrium segregation in chromia scales grown on Co-45%Cr

1986 ◽  
Vol 44 ◽  
pp. 518-519
Author(s):  
K. Przybylski ◽  
A. J. Garratt-Reed ◽  
G. J. Yurek
Keyword(s):  
Spatial Resolution ◽  
Outer Surface ◽  
Maximum Concentration ◽  
High Spatial Resolution ◽  
Reactive Elements ◽  
Chromia Scales

The addition of so-called “reactive” elements such as yttrium to alloys is known to enhance the protective nature of Cr2O3 or Al2O3 scales. However, the mechanism by which this enhancement is achieved remains unclear. An A.E.M. study has been performed of scales grown at 1000°C for 25 hr. in pure O2 on Co-45%Cr implanted at 70 keV with 2x1016 atoms/cm2 of yttrium. In the unoxidized alloys it was calculated that the maximum concentration of Y was 13.9 wt% at a depth of about 17 nm. SIMS results showed that in the scale the yttrium remained near the outer surface.

Sign in / Sign up

Export Citation Format

Share Document