CNN-based leukocyte detection for microscopy imaging at 10x magnification objective lens

2020 ◽  
Author(s):  
Qiwei Wang ◽  
Shusheng Bi ◽  
Minglei Sun ◽  
Shaobao Yang ◽  
Lingkun Chen ◽  
...  
Keyword(s):  
Objective Lens ◽  
Microscopy Imaging

scholarly journals 2.7 Å cryo-EM structure of vitrified M. musculus H-chain apoferritin from 200 keV “screening microscope”

2019 ◽  
Author(s):  
Farzad Hamdi ◽  
Christian Tüting ◽  
Dmitry A. Semchonok ◽  
Fotis L. Kyrilis ◽  
Annette Meister ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mechanical Stability ◽  
Objective Lens ◽  
List Type ◽  
Constant Power ◽  
Contrast Transfer Function ◽  
Microscopy Imaging ◽  
Detection Technology ◽  
High Emission ◽  
Solvent Molecules

AbstractHere we present the structure of mouse H-chain apoferritin at 2.7 Å (FSC=0.143) solved by single particle cryogenic electron microscopy (cryo-EM) using a 200 kV device. Data were collected using a compact, two-lens illumination system with a constant power objective lens, without the use of energy filters or aberration correctors. Coulomb potential maps reveal clear densities for main chain carbonyl oxygens, residue side chains (including alternative conformations) and bound solvent molecules. We argue that the advantages offered by (a) the high electronic and mechanical stability of the microscope, (b) the high emission stability and low beam energy spread of the high brightness Field Emission Gun (x-FEG), (c) direct electron detection technology and (d) particle-based Contrast Transfer Function (CTF) refinement have contributed to achieving resolution close to the Rayleigh limit. Overall, we show that basic electron optical settings for automated cryo-electron microscopy imaging, widely thought of as a “screening cryo-microscope”, can be used to determine structures approaching atomic resolution.HighlightsThe 2.7 Å structure of mouse apoferritin was solved using a 200 keV screening cryo-microscopeThe apoferritin reconstruction was resolved without an energy filter, aberration correctors, or constant-power condenser lensesComparison to available crystallographic and cryo-EM structures from high-end cryo-microscopes demonstrates consistency in resolved water molecules, metals and side chain orientationsAlthough radiation damage is more prominent at 200 keV compared to 300 keV, this type of instrumentation is more accessible to research laboratories due to its compactness and simplicity

Photoacoustic microscopy imaging – From a single erythrocyte to a vascular network/Bildgebung mittels photoakustischer Mikroskopie – Vom einzelnen Erythrozyt zum Gefäßnetz

2013 ◽  
Vol 2 (1) ◽  
Author(s):  
Yi Yuan ◽  
Zhongjiang Chen ◽  
Sihua Yang ◽  
Da Xing
Keyword(s):  
Focused Ultrasound ◽  
Vascular Network ◽  
Ultrasound Transducer ◽  
Objective Lens ◽  
Photoacoustic Microscopy ◽  
Two Dimensional ◽  
Label Free ◽  
Microscopy Imaging ◽  
Water Container

AbstractA label-free angiography photoacoustic microscopy system is proposed which was produced by the integration of a two-dimensional scanning galvanometer, an objective lens, a focused ultrasound transducer, a water container and a sample supporter. Using this system,

Three Dimensional Light Microscopy: Imaging & Corrections for Quantitative Analysis

1999 ◽  
Vol 5 (S2) ◽  
pp. 522-523
Author(s):  
J.N. Turner ◽  
W. hain ◽  
D.H. Szarowski ◽  
S. Lasek ◽  
L. Kam ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Light Microscopy ◽  
Three Dimensional ◽  
Image Resolution ◽  
Index Of Refraction ◽  
Three Dimensions ◽  
Objective Lens ◽  
Microscopy Imaging ◽  
Data Set ◽  
Isotropic Resolution

There are several forms of three-dimensional (3-D) light microscopy but all utilize the principle of optical section recording, i.e. the 3-D image is a sequence of two-dimensional (2-D) images called optical sections. The optical sections are particular focal planes formed within the thick specimen and usually correspond to the conventional image projections recorded in a light microscope, referred to as x,y projections. The optical sections are recorded for a sequence of focus- or z-positions. This “stack” of 2-D images is the data set for the 3-D image. If quantitative analysis is to be performed on the 3-D images, the choice of the z-dimension increment between 2-D images is especially important, and its value may be more or less critical depending on the analysis algorithm used. A reasonable starting value for this dimension is the depth-of-field of the objective lens, but the actual value may have to be smaller to optimize the image analysis or larger to decrease the influence of photobleaching. The most photostable dyes should be selected and the specimen should be mounted in index-of-refraction matching media with an antioxidant.The image resolution in all three-dimensions is determined by the 3-D point-spread-function (psf), and as a rough rule of thumb the z-resolution is degraded by a factor of 3 relative to the x,y resolution. To achieve or at least approach isotropic resolution the 3-D image can be deconvolved. Figure 1 shows the 2-D maximum value projection of a 3-D image of a cultured glial cell dual labeled for actin and vinculin before and after deconvolution. The actin fibers and vinculin focal contacts are more clearly resolved after deconvolution. Although a single cultured cell might traditionally be considered a thin object, it is really a thick object if the desired spatial resolution is less than the thickness of the cell. It is desirable to image as deep into a thick object as possible to maximize the tissue volume sampled. However, it has been shown that the image signal decreases with depth into the specimen. Figure 2 demonstrates this effect in a 3-D image of the nuclei of the rat hippocampus that have been labeled with the fluorescent Schiffs reagent acriflavine. In the x,y projection, it is not clear why some nuclei are dimmer than others, but the x,z projection shows that the dimmer ones tend to be deeper in the section. It has been shown that this depth dependent signal attenuation follows the form of an exponential function.

Design of objective lens for 200-kV high-resolution electron microscopes

1983 ◽  
Vol 41 ◽  
pp. 314-315
Author(s):  
K. Tsuno ◽  
T. Honda ◽  
Y. Harada ◽  
M. Naruse
Keyword(s):  
Optical Properties ◽  
Computer Technology ◽  
Axial Magnetic Field ◽  
Chromatic Aberration ◽  
Objective Lens ◽  
Resolution Electron ◽  
Correction Of Astigmatism ◽  
Three Stages ◽  
Electron Microscopes ◽  
Stage 1

Developement of computer technology provides much improvements on electron microscopy, such as simulation of images, reconstruction of images and automatic controll of microscopes (auto-focussing and auto-correction of astigmatism) and design of electron microscope lenses by using a finite element method (FEM). In this investigation, procedures for simulating the optical properties of objective lenses of HREM and the characteristics of the new lens for HREM at 200 kV are described.The process for designing the objective lens is divided into three stages. Stage 1 is the process for estimating the optical properties of the lens. Firstly, calculation by FEM is made for simulating the axial magnetic field distributions Bzc of the lens. Secondly, electron ray trajectory is numerically calculated by using Bzc. And lastly, using Bzc and ray trajectory, spherical and chromatic aberration coefficients Cs and Cc are numerically calculated. Above calculations are repeated by changing the shape of lens until! to find an optimum aberration coefficients.

A Liquid Nitrogen Cooled Stage for STEM

1974 ◽  
Vol 32 ◽  
pp. 444-445
Author(s):  
Louis T. Germinario
Keyword(s):  
Electron Microscope ◽  
High Resolution ◽  
Liquid Nitrogen ◽  
Room Temperature ◽  
Objective Lens ◽  
Thermal Drift ◽  
Specimen Holder ◽  
Resolution Capability ◽  
Focal Position

A liquid nitrogen stage has been developed for the JEOL JEM-100B electron microscope equipped with a scanning attachment. The design is a modification of the standard JEM-100B SEM specimen holder with specimen cooling to any temperatures In the range ~ 55°K to room temperature. Since the specimen plane is maintained at the ‘high resolution’ focal position of the objective lens and ‘bumping’ and thermal drift la minimized by supercooling the liquid nitrogen, the high resolution capability of the microscope is maintained (Fig.4).

Observability of Very Thick Specimen by Using Transmission Scanning Electron Microscope

1971 ◽  
Vol 29 ◽  
pp. 26-27
Author(s):  
K. Shibatomi ◽  
T. Yamanoto ◽  
H. Koike
Keyword(s):  
Electron Microscope ◽  
Image Quality ◽  
Scanning Electron Microscope ◽  
Chromatic Aberration ◽  
Image Contrast ◽  
Objective Lens ◽  
Thick Specimen ◽  
Transmission Electron ◽  
Scanning Electron

In the observation of a thick specimen by means of a transmission electron microscope, the intensity of electrons passing through the objective lens aperture is greatly reduced. So that the image is almost invisible. In addition to this fact, it have been reported that a chromatic aberration causes the deterioration of the image contrast rather than that of the resolution. The scanning electron microscope is, however, capable of electrically amplifying the signal of the decreasing intensity, and also free from a chromatic aberration so that the deterioration of the image contrast due to the aberration can be prevented. The electrical improvement of the image quality can be carried out by using the fascionating features of the SEM, that is, the amplification of a weak in-put signal forming the image and the descriminating action of the heigh level signal of the background. This paper reports some of the experimental results about the thickness dependence of the observability and quality of the image in the case of the transmission SEM.

Resolving Crystallites in Zirconium Oxide Films

1969 ◽  
Vol 27 ◽  
pp. 140-141
Author(s):  
R.A. Ploc
Keyword(s):  
Electron Microscope ◽  
Inelastic Scattering ◽  
Zirconium Oxide ◽  
Oxide Films ◽  
Optic Axis ◽  
Objective Lens ◽  
Microscope Objective ◽  
Linear Optic ◽  
A Current ◽  
Microscope Objective Lens

The optic axis of an electron microscope objective lens is usually assumed to be straight and co-linear with the mechanical center. No reason exists to assume such perfection and, indeed, simple reasoning suggests that it is a complicated curve. A current centered objective lens with a non-linear optic axis when used in conjunction with other lenses, leads to serious image errors if the nature of the specimen is such as to produce intense inelastic scattering.

Electronic Cone Illumination with the Conventional Transmission Electron Microscope

1977 ◽  
Vol 35 ◽  
pp. 72-73
Author(s):  
William Krakow
Keyword(s):  
Electronic Device ◽  
Chromatic Aberration ◽  
Objective Lens ◽  
Hollow Cone ◽  
Transmission Electron ◽  
Lissajous Figures ◽  
High Resolution Images ◽  
Imaging Mode ◽  
Diffraction Patterns ◽  
Transmission Microscope

An electronic device has been constructed which manipulates the primary beam in the conventional transmission microscope to illuminate a specimen under a variety of virtual condenser aperture conditions. The device uses the existing tilt coils of the microscope, and modulates the D.C. signals to both x and y tilt directions simultaneously with various waveforms to produce Lissajous figures in the back-focal plane of the objective lens. Electron diffraction patterns can be recorded which reflect the manner in which the direct beam is tilted during exposure of a micrograph. The device has been utilized mainly for the hollow cone imaging mode where the device provides a microscope transfer function without zeros in all spatial directions and has produced high resolution images which are also free from the effect of chromatic aberration. A standard second condenser aperture is employed and the width of the cone annulus is readily controlled by defocusing the second condenser lens.

High Resolution Specimen Area Imaged Under Negligible Field Aberrations

1970 ◽  
Vol 28 ◽  
pp. 540-541 ◽  
Author(s):  
T. Yanaka ◽  
K. Shirota
Keyword(s):  
High Resolution ◽  
Field Distribution ◽  
Image Space ◽  
Beam Deflection ◽  
Objective Lens ◽  
Resolution Limit ◽  
Leakage Flux ◽  
Specimen Area ◽  
Points Of View ◽  
Aberration Theory

It is significant to note field aberrations (chromatic field aberration, coma, astigmatism and blurring due to curvature of field, defined by Glaser's aberration theory relative to the Blenden Freien System) of the objective lens in connection with the following three points of view; field aberrations increase as the resolution of the axial point improves by increasing the lens excitation (k2) and decreasing the half width value (d) of the axial lens field distribution; when one or all of the imaging lenses have axial imperfections such as beam deflection in image space by the asymmetrical magnetic leakage flux, the apparent axial point has field aberrations which prevent the theoretical resolution limit from being obtained.

The Reduction of Chromatic Aberrations Due to Instrumental Instabilities

1971 ◽  
Vol 29 ◽  
pp. 14-15
Author(s):  
J. S. Lally ◽  
R. Evans
Keyword(s):  
Electron Microscope ◽  
High Voltage ◽  
Focal Length ◽  
Chromatic Aberration ◽  
Objective Lens ◽  
Usual Practice ◽  
Voltage Electron ◽  
Short Focal Length ◽  
Chromatic Aberrations ◽  
Electron Microscopes

One of the instrumental factors often limiting the resolution of the electron microscope is image defocussing due to changes in accelerating voltage or objective lens current. This factor is particularly important in high voltage electron microscopes both because of the higher voltages and lens currents required but also because of the inherently longer focal lengths, i.e. 6 mm in contrast to 1.5-2.2 mm for modern short focal length objectives.The usual practice in commercial electron microscopes is to design separately stabilized accelerating voltage and lens supplies. In this case chromatic aberration in the image is caused by the random and independent fluctuations of both the high voltage and objective lens current.

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