Identification of a Small Probe That Can Be Conjugated to Proteins by Proximity Labeling

2019 ◽  
Vol 15 (1) ◽  
pp. 39-43
Author(s):  
Weiping Sun ◽  
Yinbo Huo ◽  
Yuxuan Mei ◽  
Qingtong Zhou ◽  
Suwen Zhao ◽  
...  
Keyword(s):  
Author(s):  
Richard L. McConville

A second generation twin lens has been developed. This symmetrical lens with a wider bore, yet superior values of chromatic and spherical aberration for a given focal length, retains both eucentric ± 60° tilt movement and 20°x ray detector take-off angle at 90° to the tilt axis. Adjust able tilt axis height, as well as specimen height, now ensures almost invariant objective lens strengths for both TEM (parallel beam conditions) and STEM or nano probe (focused small probe) modes.These modes are selected through use of an auxiliary lens situ ated above the objective. When this lens is on the specimen is illuminated with a parallel beam of electrons, and when it is off the specimen is illuminated with a focused probe of dimensions governed by the excitation of the condenser 1 lens. Thus TEM/STEM operation is controlled by a lens which is independent of the objective lens field strength.


Author(s):  
James Pawley

Operation of the SEM with V0 = l-3kV (LVSEM) was early recognized to reduce charging artefacts and increase topographic contrast. This early promise was not pursued because several theoretical and practical considerations made it difficult to produce a small probe diameter (d0) at low voltage. Recently, the necessity of using low V0 to image uncoated semiconductors without damaging them has prompted a re-evaluation of LVSEM. This re-evaluation has taken the form of efforts to eliminate the practical constraints and to alleviate the theoretical ones. In the process, some heretofore neglected theoretical advantages of LVSEM have emerged. These problems and possibilities will now be discussed in more detail.


Author(s):  
J. Bentley ◽  
E. A. Kenik

Instruments combining a 100 kV transmission electron microscope (TEM) with scanning transmission (STEM), secondary electron (SEM) and x-ray energy dispersive spectrometer (EDS) attachments to give analytical capabilities are becoming increasingly available and useful. Some typical applications in the field of materials science which make use of the small probe size and thin specimen geometry are the chemical analysis of small precipitates contained within a thin foil and the measurement of chemical concentration profiles near microstructural features such as grain boundaries, point defect clusters, dislocations, or precipitates. Quantitative x-ray analysis of bulk samples using EDS on a conventional SEM is reasonably well established, but much less work has been performed on thin metal foils using the higher accelerating voltages available in TEM based instruments.


Author(s):  
J W Steeds

That the techniques of convergent beam electron diffraction (CBED) are now widely practised is evident, both from the way in which they feature in the sale of new transmission electron microscopes (TEMs) and from the frequency with which the results appear in the literature: new phases of high temperature superconductors is a case in point. The arrival of a new generation of TEMs operating with coherent sources at 200-300kV opens up a number of new possibilities.First, there is the possibility of quantitative work of very high accuracy. The small probe will essentially eliminate thickness or orientation averaging and this, together with efficient energy filtering by a doubly-dispersive electron energy loss spectrometer, will yield results of unsurpassed quality. The Bloch wave formulation of electron diffraction has proved itself an effective and efficient method of interpreting the data. The treatment of absorption in these calculations has recently been improved with the result that <100> HOLZ polarity determinations can now be performed on III-V and II-VI semiconductors.


Author(s):  
E. J. Kirkland

In a STEM an electron beam is focused into a small probe on the specimen. This probe is raster scanned across the specimen to form an image from the electrons transmitted through the specimen. The objective lens is positioned before the specimen instead of after the specimen as in a CTEM. Because the probe is focused and scanned before the specimen, accurate annular dark field (ADF) STEM image simulation is more difficult than CTEM simulation. Instead of an incident uniform plane wave, ADF-STEM simulation starts with a probe wavefunction focused at a specified position on the specimen. The wavefunction is then propagated through the specimen one atomic layer (or slice) at a time with Fresnel diffraction between slices using the multislice method. After passing through the specimen the wavefunction is diffracted onto the detector. The ADF signal for one position of the probe is formed by integrating all electrons scattered outside of an inner angle large compared with the objective aperture.


Author(s):  
Judith M. Brock ◽  
Max T. Otten ◽  
Marc. J.C. de Jong

A Field Emission Gun (FEG) on a TEM/STEM instrument provides a major improvement in performance relative to one equipped with a LaB6 emitter. The improvement is particularly notable for small-probe techniques: EDX and EELS microanalysis, convergent beam diffraction and scanning. The high brightness of the FEG (108 to 109 A/cm2srad), compared with that of LaB6 (∼106), makes it possible to achieve high probe currents (∼1 nA) in probes of about 1 nm, whilst the currents for similar probes with LaB6 are about 100 to 500x lower. Accordingly the small, high-intensity FEG probes make it possible, e.g., to analyse precipitates and monolayer amounts of segregation on grain boundaries in metals or ceramics (Fig. 1); obtain high-quality convergent beam patterns from heavily dislocated materials; reliably detect 1 nm immuno-gold labels in biological specimens; and perform EDX mapping at nm-scale resolution even in difficult specimens like biological tissue.The high brightness and small energy spread of the FEG also bring an advantage in high-resolution imaging by significantly improving both spatial and temporal coherence.


Author(s):  
Zhifeng Shao

A small electron probe has many applications in many fields and in the case of the STEM, the probe size essentially determines the ultimate resolution. However, there are many difficulties in obtaining a very small probe.Spherical aberration is one of them and all existing probe forming systems have non-zero spherical aberration. The ultimate probe radius is given byδ = 0.43Csl/4ƛ3/4where ƛ is the electron wave length and it is apparent that δ decreases only slowly with decreasing Cs. Scherzer pointed out that the third order aberration coefficient always has the same sign regardless of the field distribution, provided only that the fields have cylindrical symmetry, are independent of time and no space charge is present. To overcome this problem, he proposed a corrector consisting of octupoles and quadrupoles.


Author(s):  
Z. L. Wang ◽  
J. Bentley

The success of obtaining atomic-number-sensitive (Z-contrast) images in scanning transmission electron microscopy (STEM) has shown the feasibility of imaging composition changes at the atomic level. This type of image is formed by collecting the electrons scattered through large angles when a small probe scans across the specimen. The image contrast is determined by two scattering processes. One is the high angle elastic scattering from the nuclear sites,where ϕNe is the electron probe function centered at bp = (Xp, yp) after penetrating through the crystal; F denotes a Fourier transform operation; D is the detection function of the annular-dark-field (ADF) detector in reciprocal space u. The other process is thermal diffuse scattering (TDS), which is more important than the elastic contribution for specimens thicker than about 10 nm, and thus dominates the Z-contrast image. The TDS is an average “elastic” scattering of the electrons from crystal lattices of different thermal vibrational configurations,


Author(s):  
CE Bracker ◽  
P. K. Hansma

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.


Author(s):  
O.L. Krivanek ◽  
M.L. Leber

Three-fold astigmatism resembles regular astigmatism, but it has 3-fold rather than 2-fold symmetry. Its contribution to the aberration function χ(q) can be written as:where A3 is the coefficient of 3-fold astigmatism, λ is the electron wavelength, q is the spatial frequency, ϕ the azimuthal angle (ϕ = tan-1 (qy/qx)), and ϕ3 the direction of the astigmatism.Three-fold astigmatism is responsible for the “star of Mercedes” aberration figure that one obtains from intermediate lenses once their two-fold astigmatism has been corrected. Its effects have been observed when the beam is tilted in a hollow cone over a wide range of angles, and there is evidence for it in high resolution images of a small probe obtained in a field emission gun TEM/STEM instrument. It was also expected to be a major aberration in sextupole-based Cs correctors, and ways were being developed for dealing with it on Cs-corrected STEMs.


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