Novel Mechanism for Driving Amoeboid-Like Motility of Human Neutrophils under an Electric Field, Based on Intracellular Proton Currents and Cytoplasm Streaming

In photoelectron microscopy (PEM), also called photoemission electron microscopy (PEEM), the image is formed by electrons which have been liberated from the specimen by ultraviolet light. The electrons are accelerated by an electric field before being imaged by an electron lens system. The specimen is supported on a planar electrode (or the electrode itself may be the specimen), and the accelerating field is applied between the specimen, which serves as the cathode, and an anode. The accelerating field is essentially uniform except for microfields near the surface of the specimen and a diverging field near the anode aperture. The uniform field forms a virtual image of the specimen (virtual specimen) at unit lateral magnification, approximately twice as far from the anode as is the specimen. The diverging field at the anode aperture in turn forms a virtual image of the virtual specimen at magnification 2/3, at a distance from the anode of 4/3 the specimen distance. This demagnified virtual image is the object for the objective stage of the lens system.

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Neutrophil Myeloperoxidase Destruction by Ultraviolet Irradiation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102778 ◽

1988 ◽

Vol 46 ◽

pp. 138-139

Author(s):

J. Hanker ◽

B. Giammara ◽

G. Strauss

Keyword(s):

Uv Radiation ◽

Myeloid Leukemia ◽

Stratospheric Ozone ◽

Psoriasis Patient ◽

Whole Body ◽

Human Neutrophils ◽

Cupric Nitrate ◽

The Earth ◽

Routine Handling ◽

Uvb Phototherapy

Only a fraction of the UV radiation emitted by the sun reaches the earth; most of the UVB (290-320nm) is eliminated by stratospheric ozone. There is increasing concern, however, that man-made chemicals are damaging this ozone layer. Although the effects of UV on DNA or as a carcinogen are widely known, preleukemia and acute myeloid leukemia (AML) have only rarely been reported in psoriasis patients treated with 8-methoxypsoralen and UV (PUVA). It was therefore of interest to study the effects of UV on the myeloperoxidase (MP) activity of human neutrophils. The peroxidase activity of enriched leukocyte preparations on coverslips was shown cytochemically with a diaminobenzidine medium and cupric nitrate intensification.Control samples (Figs. 1,4,5) of human bloods that were not specifically exposed to UV radiation or light except during routine handling were compared with samples which had been exposed in one of several different ways. One preparation (Fig. 2) was from a psoriasis patient who had received whole-body UVB phototherapy repeatedly.

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Field-assisted ionization theory for microscopists

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100171833 ◽

1994 ◽

Vol 52 ◽

pp. 820-821

Author(s):

Patrick P. Camus

Keyword(s):

Electric Field ◽

Electron Tunneling ◽

Ionization Probability ◽

Critical Distance ◽

Tunneling Probability ◽

Field Ionization ◽

Radius Of Curvature ◽

Field Evaporation ◽

A Value ◽

Ion Emission

The theory of field ion emission is the study of electron tunneling probability enhanced by the application of a high electric field. At subnanometer distances and kilovolt potentials, the probability of tunneling of electrons increases markedly. Field ionization of gas atoms produce atomic resolution images of the surface of the specimen, while field evaporation of surface atoms sections the specimen. Details of emission theory may be found in monographs.Field ionization (FI) is the phenomena whereby an electric field assists in the ionization of gas atoms via tunneling. The tunneling probability is a maximum at a critical distance above the surface,xc, Fig. 1. Energy is required to ionize the gas atom at xc, I, but at a value reduced by the appliedelectric field, xcFe, while energy is recovered by placing the electron in the specimen, φ. The highest ionization probability occurs for those regions on the specimen that have the highest local electric field. Those atoms which protrude from the average surfacehave the smallest radius of curvature, the highest field and therefore produce the highest ionizationprobability and brightest spots on the imaging screen, Fig. 2. This technique is called field ion microscopy (FIM).

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Application of high-resolution field-emission LVSEM, backscatter imaging, immunogold staining to definition of the spatial distributions of two human neutrophil adherence receptors

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100146023 ◽

1993 ◽

Vol 51 ◽

pp. 36-37

Author(s):

Robert D. Nelson ◽

Sharon R. Hasslen ◽

Stanley L. Erlandsen

Keyword(s):

Cell Surface ◽

Surface Membrane ◽

Three Dimensional ◽

Human Neutrophils ◽

Spatial Distributions ◽

Immunogold Staining ◽

Functional Control ◽

Neutrophil Adherence ◽

Definition Of ◽

Mode Of Attachment

Receptors are commonly defined in terms of number per cell, affinity for ligand, chemical structure, mode of attachment to the cell surface, and mechanism of signal transduction. We propose to show that knowledge of spatial distribution of receptors on the cell surface can provide additional clues to their function and components of functional control.L-selectin and Mac-1 denote two receptor populations on the neutrophil surface that mediate neutrophil-endothelial cell adherence interactions and provide for targeting of neutrophil recruitment to sites of inflammation. We have studied the spatial distributions of these receptors using LVSEM and backscatter imaging of isolated human neutrophils stained with mouse anti-receptor (primary) antibody and goat anti-mouse (secondary) antibody conjugated to 12 nm colloidal gold. This combination of techniques provides for three-dimensional analysis of the expression of these receptors on different surface membrane domains of the neutrophil: the ruffles and microvilli that project from the cell surface, and the cell body between these projecting structures.

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