Chromatic aberration-based phase and fluorescence microscope for cell cycle study (Conference Presentation)

Author(s):  
Ondrej Mandula ◽  
Jean-Philippe Kleman ◽  
Francoise Lacroix ◽  
Dainel Fiole ◽  
Cédric Allier ◽  
...  
Author(s):  
Ondrej Mandula ◽  
Cédric Allier ◽  
Dainel Fiole ◽  
Jean-Philippe Kleman ◽  
Francoise Lacroix ◽  
...  

Author(s):  
K. Tsuno ◽  
T. Honda ◽  
Y. Harada ◽  
M. Naruse

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.


Author(s):  
L. E. Thomas ◽  
J. S. Lally ◽  
R. M. Fisher

In addition to improved penetration at high voltage, the characteristics of HVEM images of crystalline materials are changed markedly as a result of many-beam excitation effects. This leads to changes in optimum imaging conditions for dislocations, planar faults, precipitates and other features.Resolution - Because of longer focal lengths and correspondingly larger aberrations, the usual instrument resolution parameter, CS174 λ 374 changes by only a factor of 2 from 100 kV to 1 MV. Since 90% of this change occurs below 500 kV any improvement in “classical” resolution in the MVEM is insignificant. However, as is widely recognized, an improvement in resolution for “thick” specimens (i.e. more than 1000 Å) due to reduced chromatic aberration is very large.


Author(s):  
D. L. Misell

In the electron microscopy of biological sections the adverse effect of chromatic aberration on image resolution is well known. In this paper calculations are presented for the inelastic and elastic image intensities using a wave-optical formulation. Quantitative estimates of the deterioration in image resolution as a result of chromatic aberration are presented as an alternative to geometric calculations. The predominance of inelastic scattering in the unstained biological and polymeric materials is shown by the inelastic to elastic ratio, I/E, within an objective aperture of 0.005 rad for amorphous carbon of a thickness, t=50nm, typical of biological sections; E=200keV, I/E=16.


Author(s):  
Tai-Te Chao ◽  
John Sullivan ◽  
Awtar Krishan

Maytansine, a novel ansa macrolide (1), has potent anti-tumor and antimitotic activity (2, 3). It blocks cell cycle traverse in mitosis with resultant accumulation of metaphase cells (4). Inhibition of brain tubulin polymerization in vitro by maytansine has also been reported (3). The C-mitotic effect of this drug is similar to that of the well known Vinca- alkaloids, vinblastine and vincristine. This study was carried out to examine the effects of maytansine on the cell cycle traverse and the fine struc- I ture of human lymphoblasts.Log-phase cultures of CCRF-CEM human lymphoblasts were exposed to maytansine concentrations from 10-6 M to 10-10 M for 18 hrs. Aliquots of cells were removed for cell cycle analysis by flow microfluorometry (FMF) (5) and also processed for transmission electron microscopy (TEM). FMF analysis of cells treated with 10-8 M maytansine showed a reduction in the number of G1 cells and a corresponding build-up of cells with G2/M DNA content.


Author(s):  
K. Shibatomi ◽  
T. Yamanoto ◽  
H. Koike

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.


Author(s):  
Irwin I. Singer

Our previous results indicate that two types of fibronectin-cytoskeletal associations may be formed at the fibroblast surface: dorsal matrixbinding fibronexuses generated in high serum (5% FBS) cultures, and ventral substrate-adhering units formed in low serum (0.3% FBS) cultures. The substrate-adhering fibronexus consists of at least vinculin (VN) and actin in its cytoplasmic leg, and fibronectin (FN) as one of its major extracellular components. This substrate-adhesion complex is localized in focal contacts, the sites of closest substratum approach visualized with interference reflection microscopy, which appear to be the major points of cell-tosubstrate adhesion. In fibroblasts, the latter substrate-binding complex is characteristic of cultures that are arrested at the G1 phase of the cell cycle due to the low serum concentration in their medium. These arrested fibroblasts are very well spread, flattened, and immobile.


Author(s):  
William Krakow

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.


Author(s):  
J. S. Lally ◽  
R. Evans

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.


Author(s):  
S. Takashima ◽  
H. Hashimoto ◽  
S. Kimoto

The resolution of a conventional transmission electron microscope (TEM) deteriorates as the specimen thickness increases, because chromatic aberration of the objective lens is caused by the energy loss of electrons). In the case of a scanning electron microscope (SEM), chromatic aberration does not exist as the restrictive factor for the resolution of the transmitted electron image, for the SEM has no imageforming lens. It is not sure, however, that the equal resolution to the probe diameter can be obtained in the case of a thick specimen. To study the relation between the specimen thickness and the resolution of the trans-mitted electron image obtained by the SEM, the following experiment was carried out.


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