Imaging mammalian cells with soft x rays: The importance of specimen preparation

1997 ◽  
Author(s):  
J T Brown ◽  
W Meyer-Ilse
Keyword(s):  
Mammalian Cells ◽  
Specimen Preparation ◽  
X Rays

Studies of metaphase chromosomes in the scanning transmission x-ray microscope: Image fidelity, radiation damage and specimen preparation

1992 ◽  
Vol 50 (2) ◽  
pp. 1038-1039
Author(s):  
Shawn Williams ◽  
Xiaodong Zhang ◽  
Susan Lamm ◽  
Jack Van’t Hof
Keyword(s):  
Visible Light ◽  
Radiation Damage ◽  
Cross Section ◽  
Imaging Techniques ◽  
Dose Range ◽  
Specimen Preparation ◽  
X Rays ◽  
Metaphase Chromosomes ◽  
X Ray ◽  
Scanning Transmission

The Scanning Transmission X-ray Microscope (STXM) is well suited for investigating metaphase chromosome structure. The absorption cross-section of soft x-rays having energies between the carbon and oxygen K edges (284 - 531 eV) is 6 - 9.5 times greater for organic specimens than for water, which permits one to examine unstained, wet biological specimens with resolution superior to that attainable using visible light. The attenuation length of the x-rays is suitable for imaging micron thick specimens without sectioning. This large difference in cross-section yields good specimen contrast, so that fewer soft x-rays than electrons are required to image wet biological specimens at a given resolution. But most imaging techniques delivering better resolution than visible light produce radiation damage. Soft x-rays are known to be very effective in damaging biological specimens. The STXM is constructed to minimize specimen dose, but it is important to measure the actual damage induced as a function of dose in order to determine the dose range within which radiation damage does not compromise image quality.

An x-ray microprobe based upon a close-coupled focal-spot / glass capillary arrangement

1992 ◽  
Vol 50 (2) ◽  
pp. 1758-1759
Author(s):  
D. A. Carpenter ◽  
M. A. Taylor
Keyword(s):  
Imaging Techniques ◽  
Fluorescence Analysis ◽  
High Sensitivity ◽  
Specimen Preparation ◽  
X Rays ◽  
Glass Capillary ◽  
Close Coupling ◽  
X Ray ◽  
Point To Point ◽  
Beam Damage

The development of intense sources of x rays has led to renewed interest in the use of microbeams of x rays in x-ray fluorescence analysis. Sparks pointed out that the use of x rays as a probe offered the advantages of high sensitivity, low detection limits, low beam damage, and large penetration depths with minimal specimen preparation or perturbation. In addition, the option of air operation provided special advantages for examination of hydrated systems or for nondestructive microanalysis of large specimens.The disadvantages of synchrotron sources prompted the development of laboratory-based instrumentation with various schemes to maximize the beam flux while maintaining small point-to-point resolution. Nichols and Ryon developed a microprobe using a rotating anode source and a modified microdiffractometer. Cross and Wherry showed that by close-coupling the x-ray source, specimen, and detector, good intensities could be obtained for beam sizes between 30 and 100μm. More importantly, both groups combined specimen scanning with modern imaging techniques for rapid element mapping.

X-ray Gabor holography

1995 ◽  
Vol 53 ◽  
pp. 612-613
Author(s):  
Steve Lindaas ◽  
Chris Jacobsen ◽  
Alex Kalinovsky ◽  
Malcolm Howells
Keyword(s):  
Surface Relief ◽  
Biological Imaging ◽  
Specimen Preparation ◽  
X Rays ◽  
X Ray ◽  
Water Window ◽  
Biological Objects ◽  
Transmission Imaging ◽  
Atomic Force ◽  
Electron Microscopes

Soft x-ray microscopy offers an approach to transmission imaging of wet, micron-thick biological objects at a resolution superior to that of optical microscopes and with less specimen preparation/manipulation than electron microscopes. Gabor holography has unique characteristics which make it particularly well suited for certain investigations: it requires no prefocussing, it is compatible with flash x-ray sources, and it is able to use the whole footprint of multimode sources. Our method serves to refine this technique in anticipation of the development of suitable flash sources (such as x-ray lasers) and to develop cryo capabilities with which to reduce specimen damage. Our primary emphasis has been on biological imaging so we use x-rays in the water window (between the Oxygen-K and Carbon-K absorption edges) with which we record holograms in vacuum or in air.The hologram is recorded on a high resolution recording medium; our work employs the photoresist poly(methylmethacrylate) (PMMA). Following resist “development” (solvent etching), a surface relief pattern is produced which an atomic force microscope is aptly suited to image.

Reactive oxygen species in cell responses to toxic agents

2002 ◽  
Vol 21 (2) ◽  
pp. 85-90 ◽  
Author(s):  
L E Feinendegen
Keyword(s):  
Radiation Exposure ◽  
Mammalian Cells ◽  
Low Dose ◽  
X Rays ◽  
Low Dose Radiation ◽  
Cell Responses ◽  
Toxic Agents ◽  
Particle Tracks ◽  
Dose Radiation

This review first summarizes experimental data on biological effects of different concentrations of ROS in mammalian cells and on their potential role in modifying cell responses to toxic agents. It then attempts to link the role of steadily produced metabolic ROS at various concentrations in mammalian cells to that of environmentally derived ROS bursts from exposure to ionizing radiation. The ROS from both sources are known to both cause biological damage and change cellular signaling, depending on their concentration at a given time. At low concentrations signaling effects of ROS appear to protect cellular survival and dominate over damage, and the reverse occurs at high ROS concentrations. Background radiation generates suprabasal ROS bursts along charged particle tracks several times a year in each nanogram of tissue, i.e., average mass of a mammalian cell. For instance, a burst of about 200 ROS occurs within less than a microsecond from low-LET irradiation such as X-rays along the track of a Compton electron (about 6 keV, ranging about 1 μm). One such track per nanogram tissue gives about 1 mGy to this mass. The number of instantaneous ROS per burst along the track of a 4-meV ¬-particle in 1 ng tissue reaches some 70000. The sizes, types and sites of these bursts, and the time intervals between them directly in and around cells appear essential for understanding low-dose and low dose-rate effects on top of effects from endogenous ROS. At background and low-dose radiation exposure, a major role of ROS bursts along particle tracks focuses on ROS-induced apoptosis of damage-carrying cells, and also on prevention and removal of DNA damage from endogenous sources by way of temporarily protective, i.e., adaptive, cellular responses. A conclusion is to consider low-dose radiation exposure as a provider of physiological mechanisms for tissue homoeostasis.

Survival of mammalian cells exposed to X rays at ultra-high dose-rates

1969 ◽  
Vol 42 (494) ◽  
pp. 102-107 ◽  
Author(s):  
Roger J. Berry ◽  
Eric J. Hall ◽  
David W. Forster ◽  
Thomas H. Storr ◽  
Michael J. Goodman
Keyword(s):  
Mammalian Cells ◽  
High Dose ◽  
X Rays ◽  
Dose Rates

The Sequential Irradiation of Mammalian Cells with X Rays and Charged Particles of High LET

1983 ◽  
Vol 93 (3) ◽  
pp. 444 ◽  
Author(s):  
R. P. Bird ◽  
M. Zaider ◽  
H. H. Rossi ◽  
E. J. Hall ◽  
S. A. Marino ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Charged Particles ◽  
X Rays ◽  
High Let
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