<title>Edge control in computer-controlled optical polishing</title>

Electron crystallography is an emerging field for structure determination as evidenced by a number of membrane proteins that have been solved to near-atomic resolution. Advances in specimen preparation and in data acquisition with a 400kV microscope by computer controlled spot scanning mean that our ability to record electron image data will outstrip our capacity to analyze it. The computed fourier transform of these images must be processed in order to provide a direct measurement of amplitudes and phases needed for 3-D reconstruction.In anticipation of this processing bottleneck, we have written a program that incorporates a menu-and mouse-driven procedure for auto-indexing and refining the reciprocal lattice parameters in the computed transform from an image of a crystal. It is linked to subsequent steps of image processing by a system of data bases and spawned child processes; data transfer between different program modules no longer requires manual data entry. The progress of the reciprocal lattice refinement is monitored visually and quantitatively. If desired, the processing is carried through the lattice distortion correction (unbending) steps automatically.

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Applications of CCEM to Environmental Health Problems

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100117558 ◽

1985 ◽

Vol 43 ◽

pp. 102-103

Author(s):

R. J. Lee ◽

J. S. Walker

Keyword(s):

Electron Microscopy ◽

Image Analysis ◽

Environmental Health ◽

Computer Control ◽

External Agent ◽

External Agents ◽

Computer Controlled ◽

Primitive State ◽

Textural Changes ◽

General Aspects

Electron microscopy (EM), with the advent of computer control and image analysis techniques, is rapidly evolving from an interpretative science into a quantitative technique. Electron microscopy is potentially of value in two general aspects of environmental health: exposure and diagnosis.In diagnosis, electron microscopy is essentially an extension of optical microscopy. The goal is to characterize cellular changes induced by external agents. The external agent could be any foreign material, chemicals, or even stress. The use of electron microscopy as a diagnostic tool is well- developed, but computer-controlled electron microscopy (CCEM) has had only limited impact, mainly because it is fairly new and many institutions lack the resources to acquire the capability. In addition, major contributions to diagnosis will come from CCEM only when image analysis (IA) and processing algorithms are developed which allow the morphological and textural changes recognized by experienced medical practioners to be quantified. The application of IA techniques to compare cellular structure is still in a primitive state.

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High-speed brightfield 3-D imaging and 3-D image analysis of thick and overlapped cell clusters in cytological preparations

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100168827 ◽

1994 ◽

Vol 52 ◽

pp. 218-219

Author(s):

Robert W. Mackin

Keyword(s):

Image Analysis ◽

High Speed ◽

Three Dimensional ◽

Image Data ◽

Ccd Camera ◽

Objective Lens ◽

Array Processor ◽

Vaginal Smears ◽

Low Contrast ◽

Computer Controlled

This paper presents two advances towards the automated three-dimensional (3-D) analysis of thick and heavily-overlapped regions in cytological preparations such as cervical/vaginal smears. First, a high speed 3-D brightfield microscope has been developed, allowing the acquisition of image data at speeds approaching 30 optical slices per second. Second, algorithms have been developed to detect and segment nuclei in spite of the extremely high image variability and low contrast typical of such regions. The analysis of such regions is inherently a 3-D problem that cannot be solved reliably with conventional 2-D imaging and image analysis methods.High-Speed 3-D imaging of the specimen is accomplished by moving the specimen axially relative to the objective lens of a standard microscope (Zeiss) at a speed of 30 steps per second, where the stepsize is adjustable from 0.2 - 5μm. The specimen is mounted on a computer-controlled, piezoelectric microstage (Burleigh PZS-100, 68/μm displacement). At each step, an optical slice is acquired using a CCD camera (SONY XC-11/71 IP, Dalsa CA-D1-0256, and CA-D2-0512 have been used) connected to a 4-node array processor system based on the Intel i860 chip.

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The new CM-Series TEMs: integration of a five-axis motorized, fully computer-controlled goniometer

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100147132 ◽

1993 ◽

Vol 51 ◽

pp. 258-259

Author(s):

Marc J.C. de Jong ◽

P. Emile S.J. Asselbergs ◽

Max T. Otten

Keyword(s):

Mechanical Design ◽

Computer Control ◽

Objective Lens ◽

Access Port ◽

Vibration Stability ◽

Transmission Electron ◽

Computer Controlled ◽

High Degree ◽

Five Axis ◽

Five Axes

A new step forward in Transmission Electron Microscopy has been made with the introduction of the CompuStage on the CM-series TEMs: CM120, CM200, CM200 FEG and CM300. This new goniometer has motorization on five axes (X, Y, Z, α, β), all under full computer control by a dedicated microprocessor that is in communication with the main CM processor. Positions on all five axes are read out directly - not via a system counting motor revolutions - thereby providing a high degree of accuracy. The CompuStage enters the octagonal block around the specimen through a single port, allowing the specimen stage to float freely in the vacuum between the objective-lens pole pieces, thereby improving vibration stability and freeing up one access port. Improvements in the mechanical design ensure higher stability with regard to vibration and drift. During stage movement the holder O-ring no longer slides, providing higher drift stability and positioning accuracy as well as better vacuum.

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Computer-Controlled Hrem Alignment Using Automated Diffractogram Analysis

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100181415 ◽

1990 ◽

Vol 48 (1) ◽

pp. 532-533

Author(s):

G.Y. Fan ◽

O.L. Krivanek

Keyword(s):

Electron Microscope ◽

High Resolution ◽

The Other ◽

Full Information ◽

Resolution Electron ◽

Skilled Operator ◽

Automatic Alignment ◽

High Resolution Electron Microscope ◽

Computer Controlled ◽

Recorded Image

Full alignment of a high resolution electron microscope (HREM) requires five parameters to be optimized: the illumination angle (beam tilt) x and y, defocus, and astigmatism magnitude and orientation. Because neither voltage nor current centering lead to the correct illumination angle, all the adjustments must be done on the basis of observing contrast changes in a recorded image. The full alignment can be carried out by a computer which is connected to a suitable image pick-up device and is able to control the microscope, sometimes with greater precision and speed than even a skilled operator can achieve. Two approaches to computer-controlled (automatic) alignment have been investigated. The first is based on measuring the dependence of the overall contrast in the image of a thin amorphous specimen on the relevant parameters, the other on measuring the image shift. Here we report on our progress in developing a new method, which makes use of the full information contained in a computed diffractogram.

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