Investigation of the biological objects by scattered waves

High Resolution Microscopy of Multi-Layered Biological Crystals

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010005319x ◽

1982 ◽

Vol 40 ◽

pp. 80-83

Author(s):

H.A. Cohen ◽

T.W. Jeng ◽

W. Chiu

Keyword(s):

High Resolution ◽

Low Dose ◽

Three Dimensional ◽

Data Interpretation ◽

Two Dimensional ◽

Biological Objects ◽

Density Maps ◽

Density Map ◽

Complex Crystal ◽

High Resolution Microscopy

This tutorial will discuss the methodology of low dose electron diffraction and imaging of crystalline biological objects, the problems of data interpretation for two-dimensional projected density maps of glucose embedded protein crystals, the factors to be considered in combining tilt data from three-dimensional crystals, and finally, the prospects of achieving a high resolution three-dimensional density map of a biological crystal. This methodology will be illustrated using two proteins under investigation in our laboratory, the T4 DNA helix destabilizing protein gp32*I and the crotoxin complex crystal.

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Studies on Water in the Hydration Chamber of a Modified Electron Microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100069715 ◽

1970 ◽

Vol 28 ◽

pp. 544-545

Author(s):

R. C. Moretz ◽

G. G. Hausner ◽

D. F. Parsons

Keyword(s):

Thin Film ◽

Thin Films ◽

Electron Microscope ◽

Steady State ◽

Room Temperature ◽

Small Mass ◽

Equilibrium Pressure ◽

Biological Objects ◽

Mechanical Pump ◽

Differential Pumping

Use of the electron microscope to examine wet objects is possible due to the small mass thickness of the equilibrium pressure of water vapor at room temperature. Previous attempts to examine hydrated biological objects and water itself used a chamber consisting of two small apertures sealed by two thin films. Extensive work in our laboratory showed that such films have an 80% failure rate when wet. Using the principle of differential pumping of the microscope column, we can use open apertures in place of thin film windows.Fig. 1 shows the modified Siemens la specimen chamber with the connections to the water supply and the auxiliary pumping station. A mechanical pump is connected to the vapor supply via a 100μ aperture to maintain steady-state conditions.

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Improved 3-D reconstruction using stereo computer graphics and multiple tilt EM images

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100139524 ◽

1995 ◽

Vol 53 ◽

pp. 630-631

Author(s):

Lee D. Peachey ◽

Lou Fodor ◽

John C. Haselgrove ◽

Stanley M. Dunn ◽

Junqing Huang

Keyword(s):

Computer Graphics ◽

High Performance ◽

Transverse Section ◽

Three Dimensional ◽

Stereo Pair ◽

3D Reconstructions ◽

Video Cameras ◽

Biological Objects ◽

Microscope Images ◽

High Performance Computer

Stereo pairs of electron microscope images provide valuable visual impressions of the three-dimensional nature of specimens, including biological objects. Beyond this one seeks quantitatively accurate models and measurements of the three dimensional positions and sizes of structures in the specimen. In our laboratory, we have sought to combine high resolution video cameras with high performance computer graphics systems to improve both the ease of building 3D reconstructions and the accuracy of 3D measurements, by using multiple tilt images of the same specimen tilted over a wider range of angles than can be viewed stereoscopically. Ultimately we also wish to automate the reconstruction and measurement process, and have initiated work in that direction.Figure 1 is a stereo pair of 400 kV images from a 1 micrometer thick transverse section of frog skeletal muscle stained with the Golgi stain. This stain selectively increases the density of the transverse tubular network in these muscle cells, and it is this network that we reconstruct in this example.

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X-ray Gabor holography

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100139433 ◽

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.

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Prospects for scanned-probe microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100167767 ◽

1994 ◽

Vol 52 ◽

pp. 6-7

Author(s):

Jean-Paul Revel

Keyword(s):

High Vacuum ◽

Physiological Saline ◽

Scanning Probe ◽

Aqueous Environment ◽

Whole Cells ◽

Material Scientist ◽

Biological Objects ◽

Spatially Resolved ◽

Atomic Force ◽

Scanned Probe Microscopy

In the last 50+ years the electron microscope and allied instruments have led the way as means to acquire spatially resolved information about very small objects. For the material scientist and the biologist both, imaging using the information derived from the interaction of electrons with the objects of their concern, has had limitations. Material scientists have been handicapped by the fact that their samples are often too thick for penetration without using million volt instruments. Biologists have been handicapped both by the problem of contrast since most biological objects are composed of elements of low Z, and also by the requirement that sample be placed in high vacuum. Cells consist of 90% water, so elaborate precautions have to be taken to remove the water without losing the structure altogether. We are now poised to make another leap forwards because of the development of scanned probe microscopies, particularly the Atomic Force Microscope (AFM). The scanning probe instruments permit resolutions that electron microscopists still work very hard to achieve, if they have reached it yet. Probably the most interesting feature of the AFM technology, for the biologist in any case, is that it has opened the dream of high resolution in an aqueous environment. There are few restrictions on where the instrument can be used. AFMs can be made to work in high vacuum, allowing the material scientist to avoid contamination. The biologist can be made happy as well. The tips used for detection are made of silicon nitride,(Si3N4), and are essentially unaffected by exposure to physiological saline (about which more below). So here is an instrument which can look at living whole cells and at atoms as well.

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Development of reference materials with the composition of propyl and isopropyl alcohol solutions

Izmeritel`naya Tekhnika ◽

10.32446/0368-1025it.2020-6-66-72 ◽

2020 ◽

pp. 66-72

Author(s):

Irina A. Piterskikh ◽

Svetlana V. Vikhrova ◽

Nina G. Kovaleva ◽

Tatyana O. Barynskaya

Keyword(s):

Reference Materials ◽

Measurement Errors ◽

Isopropyl Alcohol ◽

Certified Reference Materials ◽

Toxic Substances ◽

Biological Objects ◽

Margin Of Error ◽

Measurement Results ◽

And Control ◽

Characteristic Mass

Certified reference materials (CRM) composed of propyl (11383-2019) and isopropyl (11384-2019) alcohols solutions were created for validation of measurement procedures and control of measurement errors of measurement results of mass concentrations of toxic substances (alcohol) in biological objects (urine, blood) and water. Two ways of establishing the value of the certified characteristic – mass consentration of propanol-1 or propanol-2 have been studied. The results obtained by the preparation procedure and comparison with the standard are the same within the margin of error.

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