A TEM study of electron tunneling in biological macromolecules

Author(s):  
J. A. Panitz

Tunneling is a ubiquitous phenomenon. Alpha particle disintegration, the Stark effect, superconductivity in thin films, field-emission, and field-ionization are examples of electron tunneling phenomena. In the scanning tunneling microscope (STM) electron tunneling is used as an imaging modality. STM images of flat surfaces show structure at the atomic level. However, STM images of large biological species deposited onto flat surfaces are disappointing. For example, unstained virus particles imaged in the STM do not resemble their TEM counterparts.It is not clear how an STM image of a biological species is formed. Most biological species are large compared to the nominal electrode separation of ∼ 1nm that is required for electron tunneling. To form an image of a biological species, the tunneling electrodes must be separated by a distance that would normally be too large for a tunneling current to be observed.

Author(s):  
R. Reifenberger ◽  
A. M. Baro ◽  
L. Vazquez ◽  
A. Bartolami ◽  
N. Garcia ◽  
...  

Scanning tunneling microscopy (STM) is the newest method that allows real space imaging of surfaces on a scale extending to atomic dimensions. The STM technique was developed by C. Binnig, H. Rohrer, and co-workers at IBM Zurich laboratories and is notable for its ability to image a wide range of materials under a variety of different conditions. The technique has received considerable attention for its ability to provide information about postions of individual surface atoms with unprecendented three-dimensional resolution.The physical basis for the STM is a quantum mechanical electron tunneling from a sharp tip to a conducting substrate. The exponential dependence of the tunneling current with the tip-to-sample separation allows a measurement of vertical position differences smaller than ca. 0.01 nm often with a lateral resolution of better than ca. 0.2 nm. By rastering the tip across a sample surface while monitoring the tunneling current, a three dimensional topographic image of the surface can be obtained.


Author(s):  
K. A. Fisher ◽  
M. B. Shattuck ◽  
M. G. L. Gustafsson ◽  
J. Clarke

Monolayer freeze-fracture combined with scanned probe microscopy (SPM) offers unique advantages for studies of biological structure. The freeze-fracture methodology incorporates rapid freezing approaches for sample preservation and stabilization, and the scanned probe microscopies, especially scanning tunneling microscopy (STM) and atomic force microscopy (AFM), allow high resolution examination of surface features including digital mapping, quantification, and display. In routine biological STM a sharp conductive probe is positioned with piezoelectric transducers close to a conductive surface. When electron tunneling begins, the probe is scanned while electronic feedback maintains constant current. Because the tunneling current is logarithmically sensitive to separation between the tip and the sample, the feedback signal can be calibrated to indicate height with sub-Angstrom sensitivity and precision. In routine biological AFM, the sample is scanned while the force between the tip and the sample is kept constant.There are two fundamental problems in STM examinations of biological systems. First, biological samples are soft and are often perturbed by the scanning probe; and second, they are not electrically conductive. Coating samples with metal replicas simultaneously circumvents both these difficulties, conferring sample stability and electrical conductivity. The STM can be used to measure sample heights quite accurately, and has been used to measure the thickness and changes in thickness of metal-coated purple membrane and the depth of surface features of freeze-fracture replicas of synthetic phospholipids.


Author(s):  
Oliver C. Wells ◽  
Mark E. Welland

Scanning tunneling microscopes (STM) exist in two versions. In both of these, a pointed metal tip is scanned in close proximity to the specimen surface by means of three piezos. The distance of the tip from the sample is controlled by a feedback system to give a constant tunneling current between the tip and the sample. In the low-end STM, the system has a mechanical stability and a noise level to give a vertical resolution of between 0.1 nm and 1.0 nm. The atomic resolution STM can show individual atoms on the surface of the specimen.A low-end STM has been put into the specimen chamber of a scanning electron microscope (SEM). The first objective was to investigate technological problems such as surface profiling. The second objective was for exploratory studies. This second objective has already been achieved by showing that the STM can be used to study trapping sites in SiO2.


Author(s):  
W. Lo ◽  
J.C.H. Spence ◽  
M. Kuwabara

Work on the integration of STM with REM has demonstrated the usefulness of this combination. The STM has been designed to replace the side entry holder of a commercial Philips 400T TEM. It allows simultaneous REM imaging of the tip/sample region of the STM (see fig. 1). The REM technique offers nigh sensitivity to strain (<10−4) through diffraction contrast and high resolution (<lnm) along the unforeshortened direction. It is an ideal technique to use for studying tip/surface interactions in STM.The elastic strain associated with tunnelling was first imaged on cleaved, highly doped (S doped, 5 × 1018cm-3) InP(110). The tip and surface damage observed provided strong evidence that the strain was caused by tip/surface contact, most likely through an insulating adsorbate layer. This is consistent with the picture that tunnelling in air, liquid or ordinary vacuum (such as in a TEM) occurs through a layer of contamination. The tip, under servo control, must compress the insulating contamination layer in order to get close enough to the sample to tunnel. The contaminant thereby transmits the stress to the sample. Elastic strain while tunnelling from graphite has been detected by others, but never directly imaged before. Recent results using the STM/REM combination has yielded the first direct evidence of strain while tunnelling from graphite. Figure 2 shows a graphite surface elastically strained by the STM tip while tunnelling (It=3nA, Vtip=−20mV). Video images of other graphite surfaces show a reversible strain feature following the tip as it is scanned. The elastic strain field is sometimes seen to extend hundreds of nanometers from the tip. Also commonly observed while tunnelling from graphite is an increase in the RHEED intensity of the scanned region (see fig.3). Debris is seen on the tip and along the left edges of the brightened scan region of figure 4, suggesting that tip abrasion of the surface has occurred. High resolution TEM images of other tips show what appear to be attached graphite flakes. The removal of contamination, possibly along with the top few layers of graphite, seems a likely explanation for the observed increase in RHEED reflectivity. These results are not inconsistent with the “sliding planes” model of tunnelling on graphite“. Here, it was proposed that the force due to the tunnelling probe acts over a large area, causing shear of the graphite planes when the tip is scanned. The tunneling current is then modulated as the planes of graphite slide in and out of registry. The possiblity of true vacuum tunnelling from the cleaned graphite surface has not been ruled out. STM work function measurements are needed to test this.


Author(s):  
Mircea Fotino ◽  
D.C. Parks

In the last few years scanning tunneling microscopy (STM) has made it possible and easily accessible to visualize surfaces of conducting specimens at the atomic scale. Such performance allows the detailed characterization of surface morphology in an increasing spectrum of applications in a wide variety of fields. Because the basic imaging process in STM differs fundamentally from its equivalent in other well-established microscopies, good understanding of the imaging mechanism in STM enables one to grasp the correct information content in STM images. It thus appears appropriate to explore by STM the structure of amorphous carbon films because they are used in many applications, in particular in the investigation of delicate biological specimens that may be altered through the preparation procedures.All STM images in the present study were obtained with the commercial instrument Nanoscope II (Digital Instruments, Inc., Santa Barbara, California). Since the importance of the scanning tip for image optimization and artifact reduction cannot be sufficiently emphasized, as stressed by early analyses of STM image formation, great attention has been directed toward adopting the most satisfactory tip geometry. The tips used here consisted either of mechanically sheared Pt/Ir wire (90:10, 0.010" diameter) or of etched W wire (0.030" diameter). The latter were eventually preferred after a two-step procedure for etching in NaOH was found to produce routinely tips with one or more short whiskers that are essentially rigid, uniform and sharp (Fig. 1) . Under these circumstances, atomic-resolution images of cleaved highly-ordered pyro-lytic graphite (HOPG) were reproducibly and readily attained as a standard criterion for easily recognizable and satisfactory performance (Fig. 2).


2008 ◽  
Vol 8 (11) ◽  
pp. 5702-5707 ◽  
Author(s):  
Ge-Bo Pan ◽  
Jun Luo ◽  
Qi-Yu Zheng ◽  
Li-Jun Wan

Well-ordered arrays of chiral molecular cavities have been constructed by self-assembly of inherently chiral calix[4]crown on Au(111) in 0.1 M HClO4 solution and investigated by scanning tunneling microscopy (STM). The chiral features are clearly observed in high resolution STM images. It is found that the adsorption of the two enantiomers results in the same ordered structures with upright orientation on Au(111). Moreover, only phase separation has been observed for the racemic mixture of the two enantiomers in the experiment. This is mainly due to the weak molecule-substrate interaction as well as asymmetric geometrical structures of the two enantiomers. The present study provides a simple method for construction of ordered arrays of chiral molecular cavities, which are of potential in chemical sensors, chiral recognition, and nonlinear optics.


1998 ◽  
Vol 05 (01) ◽  
pp. 69-76
Author(s):  
F. P. Netzer ◽  
L. Vitali ◽  
J. Kraft ◽  
M. G. Ramesy

The interaction of vapor phase P2 with the [Formula: see text] monolayer surface at room temperature and elevated temperature has been monitored by scanning tunneling microscopy (STM) and spectroscopy (STS) in conjunction with Auger electron spectroscopy and low-energy electron diffraction (LEED). The surface rection can be readily followed by STM because of the very different contrast of the reacted areas in the STM images. The reaction develops around overlayer defects at room temperature and appears to be diffusion-limited, whereas at 300°C the reaction is initiated at the step edges, from which the reaction front progresses onto the lower terrace areas. At elevated temperature several ordered surface reconstructions, showing different STS fingerprints, are detected on the P–In/Si(111) surfaces, which are associated tentatively with P- and Si-terminated structures and an ordered InP phase.


1994 ◽  
Vol 332 ◽  
Author(s):  
S.M. Lindsay ◽  
J. Pan ◽  
T.W. Jing

ABSTRACTWe use electrochemical methods to control the adsorption of molecules onto an electrode for imaging in-situ by scanning tunneling microscopy. Measurements of the barrier for electron tunneling show that the mechanism of electron transfer differs from vacuum tunneling. Barriers depend upon the direction of electron tunneling, indicating the presence of permanently aligned dipoles in the tunnel gap. We attribute a sharp dip in the barrier near zero field to induced polarization. We propose a ‘tunneling’ process consisting of two parts: One is delocalization of quantum-coherent states in parts of the molecular adlayer that hybridize strongly (interaction ≥ kT) with Bloch states in the metal. This gives rise to a quantum-point-contact conductance, Gc ≤ 2e2/h at a height zo. The other part comes from the exponential decay of the tails of localized states, G = Gc exp{−2K(z − z0)}. Because measured decay lengths, (2K‘)−1, are small (≈ 1 Å), STM contrast is dominated by the contour along which G[z0 (x,y)] = Gc. Measured changes in z0 are used to calculate images which are in reasonable agreement with observations. We illustrate this with images of synthetic DNA oligomers.


2014 ◽  
Vol 10 ◽  
pp. 2783-2788 ◽  
Author(s):  
Stefan-S Jester ◽  
A Vikas Aggarwal ◽  
Daniel Kalle ◽  
Sigurd Höger

Self-assembled monolayers of a molecular spoked wheel (a shape-persistent macrocycle with an intraannular spoke/hub system) and its synthetic precursor are investigated by scanning tunneling microscopy (STM) at the liquid/solid interface of 1-octanoic acid and highly oriented pyrolytic graphite. The submolecularly resolved STM images reveal that the molecules indeed behave as more or less rigid objects of certain sizes and shapes – depending on their chemical structures. In addition, the images provide insight into the multilayer growth of the molecular spoked wheels (MSWs), where the first adlayer acts as a template for the commensurate adsorption of molecules in the second layer.


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