Methods of Microorganism Immobilization for Dynamic Atomic-Force Studies (Review)

The atomic force microscope (AFM) is capable of imaging electrically conductive and non-conductive surfaces at atomic resolution. When used to image biological samples, however, lateral resolution is often limited to nanometer levels, due primarily to AFM tip/sample interactions. Several approaches to immobilize and stabilize soft or flexible molecules for AFM have been examined, notably, tethering coating, and freezing. Although each approach has its advantages and disadvantages, rapid freezing techniques have the special advantage of avoiding chemical perturbation, and minimizing physical disruption of the sample. Scanning with an AFM at cryogenic temperatures has the potential to image frozen biomolecules at high resolution. We have constructed a force microscope capable of operating immersed in liquid n-pentane and have tested its performance at room temperature with carbon and metal-coated samples, and at 143° K with uncoated ferritin and purple membrane (PM).

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AFM study of the dynamics of α-alumina surface faceting during high-temperature processing

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010013804x ◽

1995 ◽

Vol 53 ◽

pp. 334-335 ◽

Cited By ~ 1

Author(s):

Michael W. Bench ◽

Jason R. Heffelfinger ◽

C. Barry Carter

Keyword(s):

High Temperature ◽

Thin Foil ◽

Sem Analysis ◽

Conductive Coatings ◽

Force Microscopy ◽

Minimal Sample ◽

Atomic Force ◽

Formation And Evolution ◽

High Temperature Processing ◽

Nominal Surface

To gain a better understanding of the surface faceting that occurs in α-alumina during high temperature processing, atomic force microscopy (AFM) studies have been performed to follow the formation and evolution of the facets. AFM was chosen because it allows for analysis of topographical details down to the atomic level with minimal sample preparation. This is in contrast to SEM analysis, which typically requires the application of conductive coatings that can alter the surface between subsequent heat treatments. Similar experiments have been performed in the TEM; however, due to thin foil and hole edge effects the results may not be representative of the behavior of bulk surfaces.The AFM studies were performed on a Digital Instruments Nanoscope III using microfabricated Si3N4 cantilevers. All images were recorded in air with a nominal applied force of 10-15 nN. The alumina samples were prepared from pre-polished single crystals with (0001), , and nominal surface orientations.

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Characterization of photosystem II with the atomic-force microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100130365 ◽

1992 ◽

Vol 50 (2) ◽

pp. 1146-1147

Author(s):

Kathleen M. Marr ◽

Mary K. Lyon

Keyword(s):

Photosystem Ii ◽

Atomic Force Microscope ◽

Three Dimensional ◽

Biologically Active ◽

Atomic Force ◽

Freeze Etching ◽

Image Averaging ◽

Oxygen Evolving ◽

Averaging Techniques

Photosystem II (PSII) is different from all other reaction centers in that it splits water to evolve oxygen and hydrogen ions. This unique ability to evolve oxygen is partly due to three oxygen evolving polypeptides (OEPs) associated with the PSII complex. Freeze etching on grana derived insideout membranes revealed that the OEPs contribute to the observed tetrameric nature of the PSIl particle; when the OEPs are removed, a distinct dimer emerges. Thus, the surface of the PSII complex changes dramatically upon removal of these polypeptides. The atomic force microscope (AFM) is ideal for examining surface topography. The instrument provides a topographical view of individual PSII complexes, giving relatively high resolution three-dimensional information without image averaging techniques. In addition, the use of a fluid cell allows a biologically active sample to be maintained under fully hydrated and physiologically buffered conditions. The OEPs associated with PSII may be sequentially removed, thereby changing the surface of the complex by one polypeptide at a time.

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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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Scanning tunneling microscopy and atomic force microscopy: New tools for biology

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100155864 ◽

1989 ◽

Vol 47 ◽

pp. 778-779

Author(s):

CE Bracker ◽

P. K. Hansma

Keyword(s):

Physical Property ◽

Scanning Probe ◽

Scanning Tunneling ◽

Small Scale ◽

Tunneling Microscopy ◽

Force Microscopy ◽

New Family ◽

Small Probe ◽

Atomic Force ◽

Transmission Electron

A new family of scanning probe microscopes has emerged that is opening new horizons for investigating the fine structure of matter. The earliest and best known of these instruments is the scanning tunneling microscope (STM). First published in 1982, the STM earned the 1986 Nobel Prize in Physics for two of its inventors, G. Binnig and H. Rohrer. They shared the prize with E. Ruska for his work that had led to the development of the transmission electron microscope half a century earlier. It seems appropriate that the award embodied this particular blend of the old and the new because it demonstrated to the world a long overdue respect for the enormous contributions electron microscopy has made to the understanding of matter, and at the same time it signalled the dawn of a new age in microscopy. What we are seeing is a revolution in microscopy and a redefinition of the concept of a microscope.Several kinds of scanning probe microscopes now exist, and the number is increasing. What they share in common is a small probe that is scanned over the surface of a specimen and measures a physical property on a very small scale, at or near the surface. Scanning probes can measure temperature, magnetic fields, tunneling currents, voltage, force, and ion currents, among others.

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Imaging polymers, proteins and dna in aqueous solutions with the atomic force microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100152136 ◽

1989 ◽

Vol 47 ◽

pp. 32-33

Author(s):

S.A.C. Gould ◽

B. Drake ◽

C.B. Prater ◽

A.L. Weisenhorn ◽

S.M. Lindsay ◽

...

Keyword(s):

Amino Acids ◽

Dna Sequencing ◽

Aqueous Solutions ◽

Atomic Force Microscope ◽

Piezoelectric Crystal ◽

Atomic Force ◽

Structure Of Molecules ◽

Optical Lever

The atomic force microscope (AFM) is an instrument that can be used to image many samples of interest in biology and medicine. Images of polymerized amino acids, polyalanine and polyphenylalanine demonstrate the potential of the AFM for revealing the structure of molecules. Images of the protein fibrinogen which agree with TEM images demonstrate that the AFM can provide topographical data on larger molecules. Finally, images of DNA suggest the AFM may soon provide an easier and faster technique for DNA sequencing.The AFM consists of a microfabricated SiO2 triangular shaped cantilever with a diamond tip affixed at the elbow to act as a probe. The sample is mounted on a electronically driven piezoelectric crystal. It is then placed in contact with the tip and scanned. The topography of the surface causes minute deflections in the 100 μm long cantilever which are detected using an optical lever.

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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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Morphology and development of flow-pattern defect with extended nonagitated Secco etching

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100172073 ◽

1994 ◽

Vol 52 ◽

pp. 868-869

Author(s):

Y. Pan

Keyword(s):

Flow Pattern ◽

Silicon Crystal ◽

Gate Oxide ◽

Crystal Growth Rate ◽

Etch Depth ◽

Force Microscopy ◽

Etch Pit ◽

Float Zone ◽

Atomic Force ◽

Multiple Step

The D defect, which causes the degradation of gate oxide integrities (GOI), can be revealed by Secco etching as flow pattern defect (FPD) in both float zone (FZ) and Czochralski (Cz) silicon crystal or as crystal originated particles (COP) by a multiple-step SC-1 cleaning process. By decreasing the crystal growth rate or high temperature annealing, the FPD density can be reduced, while the D defectsize increased. During the etching, the FPD surface density and etch pit size (FPD #1) increased withthe etch depth, while the wedge shaped contours do not change their positions and curvatures (FIG.l).In this paper, with atomic force microscopy (AFM), a simple model for FPD morphology by non-crystallographic preferential etching, such as Secco etching, was established.One sample wafer (FPD #2) was Secco etched with surface removed by 4 μm (FIG.2). The cross section view shows the FPD has a circular saucer pit and the wedge contours are actually the side surfaces of a terrace structure with very small slopes. Note that the scale in z direction is purposely enhanced in the AFM images. The pit dimensions are listed in TABLE 1.

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The atomic-force microscope and its future in biology

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100149350 ◽

1993 ◽

Vol 51 ◽

pp. 704-705

Author(s):

Jean-Paul Revel

Keyword(s):

Silicon Nitride ◽

Laser Beam ◽

Atomic Force Microscope ◽

Scanning Tunneling Microscope ◽

Point Of View ◽

Scanning Tunneling ◽

Close Relative ◽

Tunneling Microscope ◽

Atomic Force ◽

Sharp Point

The last few years have been marked by a series of remarkable developments in microscopy. Perhaps the most amazing of these is the growth of microscopies which use devices where the place of the lens has been taken by probes, which record information about the sample and display it in a spatial from the point of view of the context. From the point of view of the biologist one of the most promising of these microscopies without lenses is the scanned force microscope, aka atomic force microscope.This instrument was invented by Binnig, Quate and Gerber and is a close relative of the scanning tunneling microscope. Today's AFMs consist of a cantilever which bears a sharp point at its end. Often this is a silicon nitride pyramid, but there are many variations, the object of which is to make the tip sharper. A laser beam is directed at the back of the cantilever and is reflected into a split, or quadrant photodiode.

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