Use Of Laser Scanning Microscopy Research Articles

Laser Scanning Microscopy of Yersinia pestis Infected Tissues.

Laser scanning microscopy (LSM) is a technology that allows for direct observations of host-pathogen interactions during infection. Two of the most available forms of LSM are confocal and two-photon LSM. In addition to high resolution and contrast, these two technologies also provide high excitation penetrance in unsectioned samples. High penetrance allows for imaging of layers of tissue that are difficult to image with other more conventional microscopy approaches. Thus, confocal and two-photon LSM open the possibility of observing infection in a three-dimensional context, where the natural architecture of a tissue is preserved. Few studies have used LSM technology to gain insights into Yersinia pestis pathogenesis in the mammalian host. The use of LSM in the plague field has an enormous potential for the discovery of the mechanisms that lie behind key aspects of pathogenesis such as colonization, dissemination, and tissue damage. This chapter provides guidance for the implementation of confocal or two-photon LSM to study Y. pestis interactions with the host in unsectioned tissues. This document provides specific instructions applied to imaging of Y. pestis, and also discusses relevant aspects of imaging, such as the operation of laser scanning microscopes and the use of fluorescent probes.

Surface Topography Quantification of Super Hard Abrasive Tools by Laser Scanning Microscopy

Abstract Non-conventional super hard abrasive tools are made of composite materials containing super hard grains, e.g., diamond or cubic boron nitride (CBN) grains, bound by a metallic constitutive phase. These tools are usually produced by means of sintering, and are widely applied in the abrasive machining processes of modern manufacturing, especially in precision machining. The abrasive grains, which induce the material removal processes, are embedded in the metallic binder. They emerge as a consequence of self-dressing, resulting in a self-sharping effect. Therefore, the cutting surface of the tool displays an irregular topography. Quantification of surface topography scenario may supply valuable information to evaluate and understand its correlation to wear mechanisms. In this study, an experimental protocol consisting of five steps: specimen preparation, surface scanning, image assembly, image digital processing and surface quantification, was proposed and validated by characterizing two CBN honing tools used for precision machining: B151/L2/2010/50 (B151) and B91/128/x44/35 (B91) CBN honing stones. It involved the use of laser scanning microscopy and digital imaging processing for assessing significant dimensional, geometrical, and positional properties of CBN grains at the surface of super hard abrasive tools. It was shown that surface topography quantification is an effective method to evaluate and obtain the defined parameters. However, smaller grains may require images with higher resolution; thus, scanning must be refined. Finally, a critical comparative analysis of the experimental results attained for the studied tools pointed out honing stone B91 as more appropriated than B151 one for achieving a higher machining quality of the workpiece.

Methodology of evaluation of abrasive tool wear with the use of laser scanning microscopy

Grinding is one of the basic precise material removal methods. Abrasive and shape wear, as well as smearing of the tools' active surface handicap the processing results. The loss of cutting capacity in abrasive tools or alteration of their shape influences the surface quality and precision of the workpiece dimensions and its shape. Evaluation of the abrasive tool surface is the basic criterion of forecasting the tools' durability and the process results. The applied method of laser scanning made determination of the surface coordinates and subsequently of its geometric features with micrometric accuracy possible. Using the information on the abrasive tool surface geometric structure, a methodology of evaluation of the level of changes in geometric features of the tool during the grinding process was developed. Criteria for evaluation of the level of abrasive grains attritious wear, the degree of smearing of the abrasive tool surface and evaluation of the cutting capability of the abrasive tools were determined. The developed method allowed for evaluation of the level of abrasive tools' wear, and subsequently formed foundations for assessment of the influence of the grinding parameters on the durability of abrasive tools, evaluation of the influence of the parameters of the process of shaping the abrasive tools' active surfaces on their geometric characteristics and evaluation of the level of correlation between the monitored process parameters and the degree of the abrasive tools' wear.

Konfokale Laserscan-Mikroskopie

The diagnosis of skin diseases relies strongly on clinical inspection and the use of invasive diagnostic procedures. Routine histology requires the removal of tissue for microscopic evaluation, which is associated with pain, and risk of infection as well as scar formation. In the past 15 years a number of non-invasive diagnostic devices have been evaluated for use in clinical and investigative dermatology. A number of studies have assessed the applicability of confocal laser scanning microscopy (LSM) as an optical diagnostic device in dermatology. By correlating LSM images with established features of routine histology, it was possible to define diagnostic LSM parameters for a number of selected skin diseases. Present data supports the use of LSM as an adjunct diagnostic device for selected skin conditions in clinical as well as investigative dermatology. Since LSM examinations may repeatedly be performed, LSM is particularly suited for evaluation of dynamic, neoplastic and regenerative skin processes as well as the definition of disease extent and response to therapy.

Laser Scanning Microscopy of the Human Larynx Mucosa: A Preliminary, Ex Vivo Study

Laser scanning microscopy (LSM) supplies in vivo information from epithelia up to depths between 0.1 to 0.5 mm. The aim of this ex vivo prospective pilot study was to investigate the potential use of LSM for the diagnosis of laryngeal cancer and its precursors. Forty-three larynx specimens of 26 patients (age 35-61 years, mean age 51.9+/-9.5 years; 7 women and 19 men) with laryngeal lesions were investigated with LSM. The LSM findings were compared with histopathologic sections. The following criteria were used for characterization of cancerous lesions: enlarged nuclei, enlarged cells with variable shapes, cluster of cells, increased nucleus/cytoplasm ratio, irregular cell architecture, and loss of cellular junctions characterized by lack of visualization of the cell membrane. LSM enables the visualization of epithelium up to the basement membrane, Reincke space, the subepithelial vessels, and the fibers of the subepithelial space. In contrast to the squamous epithelium, the respiratory epithelium bears kinocilia. The beat of the cilia and the directed mucous transport can be observed ex vivo. With the use of the presented malignancy criteria, a sensitivity of 72.7% and a specificity of 82.9% for differentiation of dysplasia and benign laryngeal lesions from cancer were reached. LSM in an ex vivo manner supplies microscopic images up to the subepithelial space. LSM could represent a new technology in laryngology to visualize larynx epithelia. In the next step, in vivo LSM will be applied to evaluate laryngeal lesion in vivo.