The localization of the defects in semiconductor devices using a new laser scanning

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.

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Algorithms for automated montage synthesis of images from laser-scanning confocal microscopes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100139627 ◽

1995 ◽

Vol 53 ◽

pp. 650-651

Author(s):

D. E. Becker

Keyword(s):

Image Analysis ◽

High Resolution ◽

Laser Scanning ◽

Cell Counting ◽

Wide Area ◽

Data Set ◽

Computational Synthesis ◽

Automated Cell Counting ◽

Partial View ◽

The Individual

An efficient, robust, and widely-applicable technique is presented for computational synthesis of high-resolution, wide-area images of a specimen from a series of overlapping partial views. This technique can also be used to combine the results of various forms of image analysis, such as segmentation, automated cell counting, deblurring, and neuron tracing, to generate representations that are equivalent to processing the large wide-area image, rather than the individual partial views. This can be a first step towards quantitation of the higher-level tissue architecture. The computational approach overcomes mechanical limitations, such as hysterisis and backlash, of microscope stages. It also automates a procedure that is currently done manually. One application is the high-resolution visualization and/or quantitation of large batches of specimens that are much wider than the field of view of the microscope.The automated montage synthesis begins by computing a concise set of landmark points for each partial view. The type of landmarks used can vary greatly depending on the images of interest. In many cases, image analysis performed on each data set can provide useful landmarks. Even when no such “natural” landmarks are available, image processing can often provide useful landmarks.

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Formation of defects in implanted hipox-structures

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100131668 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1406-1407

Author(s):

Peter Pegler ◽

N. David Theodore ◽

Ming Pan

Keyword(s):

High Pressure ◽

Cross Section ◽

Semiconductor Devices ◽

Silicon Substrates ◽

Processing Conditions ◽

Pressure Oxidation ◽

Deep Trench ◽

Local Oxidation ◽

Trench Isolation ◽

And Behavior

High-pressure oxidation of silicon (HIPOX) is one of various techniques used for electrical-isolation of semiconductor-devices on silicon substrates. Other techniques have included local-oxidation of silicon (LOCOS), poly-buffered LOCOS, deep-trench isolation and separation of silicon by implanted oxygen (SIMOX). Reliable use of HIPOX for device-isolation requires an understanding of the behavior of the materials and structures being used and their interactions under different processing conditions. The effect of HIPOX-related stresses in the structures is of interest because structuraldefects, if formed, could electrically degrade devices.This investigation was performed to study the origin and behavior of defects in recessed HIPOX (RHIPOX) structures. The structures were exposed to a boron implant. Samples consisted of (i) RHlPOX'ed strip exposed to a boron implant, (ii) recessed strip prior to HIPOX, but exposed to a boron implant, (iii) test-pad prior to HIPOX, (iv) HIPOX'ed region away from R-HIPOX edge. Cross-section TEM specimens were prepared in the <110> substrate-geometry.

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3-Dimensional immunolabeling and in situ hybridization detection using fluorescence photooxidation and intermediate-voltage Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100168554 ◽

1994 ◽

Vol 52 ◽

pp. 164-165

Author(s):

Thomas J. Deerinck ◽

Maryann E. Martone ◽

Varda Lev-Ram ◽

David P. L. Green ◽

Roger Y. Tsien ◽

...

Keyword(s):

Laser Scanning ◽

Reactive Oxygen ◽

Confocal Laser Scanning Microscope ◽

Fluorescent Dye ◽

Confocal Laser ◽

3 Dimensional ◽

Voltage Electron ◽

Dimensional Distribution ◽

Electron Microscopes ◽

New Generation

The confocal laser scanning microscope has become a powerful tool in the study of the 3-dimensional distribution of proteins and specific nucleic acid sequences in cells and tissues. This is also proving to be true for a new generation of high contrast intermediate voltage electron microscopes (IVEM). Until recently, the number of labeling techniques that could be employed to allow examination of the same sample with both confocal and IVEM was rather limited. One method that can be used to take full advantage of these two technologies is fluorescence photooxidation. Specimens are labeled by a fluorescent dye and viewed with confocal microscopy followed by fluorescence photooxidation of diaminobenzidine (DAB). In this technique, a fluorescent dye is used to photooxidize DAB into an osmiophilic reaction product that can be subsequently visualized with the electron microscope. The precise reaction mechanism by which the photooxidation occurs is not known but evidence suggests that the radiationless transfer of energy from the excited-state dye molecule undergoing the phenomenon of intersystem crossing leads to the formation of reactive oxygen species such as singlet oxygen. It is this reactive oxygen that is likely crucial in the photooxidation of DAB.

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Five-Dimensional Microscopy using Widefield-Deconvolution: Practical Considerations and Biological Applications

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100163800 ◽

1996 ◽

Vol 54 ◽

pp. 268-269

Author(s):

W.F. Marshall ◽

K. Oegema ◽

J. Nunnari ◽

A.F. Straight ◽

D.A. Agard ◽

...

Keyword(s):

Confocal Microscopy ◽

High Speed ◽

Laser Scanning ◽

Ccd Camera ◽

Image Plane ◽

Time Lapse ◽

Laser Scanning Confocal Microscopy ◽

Three Dimensions ◽

Excitation Light ◽

Cooled Ccd

The ability to image cells in three dimensions has brought about a revolution in biological microscopy, enabling many questions to be asked which would be inaccessible without this capability. There are currently two major methods of three dimensional microscopy: laser-scanning confocal microscopy and widefield-deconvolution microscopy. The method of widefield-deconvolution uses a cooled CCD to acquire images from a standard widefield microscope, and then computationally removes out of focus blur. Using such a scheme, it is easy to acquire time-lapse 3D images of living cells without killing them, and to do so for multiple wavelengths (using computer-controlled filter wheels). Thus, it is now not only feasible, but routine, to perform five dimensional microscopy (three spatial dimensions, plus time, plus wavelength).Widefield-deconvolution has several advantages over confocal microscopy. The two main advantages are high speed of acquisition (because there is no scanning, a single optical section is acquired at a time by using a cooled CCD camera) and the use of low excitation light levels Excitation intensity can be much lower than in a confocal microscope for three reasons: 1) longer exposures can be taken since the entire 512x512 image plane is acquired in parallel, so that dwell time is not an issue, 2) the higher quantum efficiently of a CCD detect over those typically used in confocal microscopy (although this is expected to change due to advances in confocal detector technology), and 3) because no pinhole is used to reject light, a much larger fraction of the emitted light is collected. Thus we can typically acquire images with thousands of photons per pixel using a mercury lamp, instead of a laser, for illumination. The use of low excitation light is critical for living samples, and also reduces bleaching. The high speed of widefield microscopy is also essential for time-lapse 3D microscopy, since one must acquire images quickly enough to resolve interesting events.

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