scholarly journals A 1.4-nm gold cluster covalently attached to antibodies improves immunolabeling.

1992 ◽  
Vol 40 (2) ◽  
pp. 177-184 ◽  
Author(s):  
J F Hainfeld ◽  
F R Furuya

A large gold cluster (Au1.4nm) was covalently coupled to IgG and Fab' fragments. Its gold core is 1.4 nm in diameter and the Fab'-Au1.4nm immunoconjugate is the smallest gold immunoprobe that can be seen directly in the conventional electron microscope. It is useful in high-resolution immunolabeling, providing a resolution of 7.0 nm. The cluster's visibility can be enhanced with silver development for use in EM or light microscopy for histological purposes, or to detect less than or equal to 0.2 pg of antigen in immunoblots. By using a gold compound with covalent attachment, a number of advantages over colloidal gold probes are realized, including better resolution, stability, uniformity, sensitivity, and complete absence of aggregation; its small size should also improve penetration and more quantitative labeling of antigenic sites.

Author(s):  
James F. Hainfeld ◽  
Frederic R. Furuya ◽  
Kyra Carbone

A new 1.4 nm gold cluster that is stable, water soluble, and can be covalently linked to specific sites has recently been described. Previous applications have coupled this gold to Fab’ fragments to make one of the smallest EM immunolabels yet developed. This gold differs from colloidal gold significantly: it does not aggregate proteins by non-specific binding; binding is covalent and to a specific residue (e.g., thiol, amine, or carboxyl); it is very uniform in size; its small size makes it a high resolution marker.


Author(s):  
A. V. Crewe

The high resolution STEM is now a fact of life. I think that we have, in the last few years, demonstrated that this instrument is capable of the same resolving power as a CEM but is sufficiently different in its imaging characteristics to offer some real advantages.It seems possible to prove in a quite general way that only a field emission source can give adequate intensity for the highest resolution^ and at the moment this means operating at ultra high vacuum levels. Our experience, however, is that neither the source nor the vacuum are difficult to manage and indeed are simpler than many other systems and substantially trouble-free.


Author(s):  
C. T. Nightingale ◽  
S. E. Summers ◽  
T. P. Turnbull

The ease of operation of the scanning electron microscope has insured its wide application in medicine and industry. The micrographs are pictorial representations of surface topography obtained directly from the specimen. The need to replicate is eliminated. The great depth of field and the high resolving power provide far more information than light microscopy.


Author(s):  
Louis T. Germinario

A liquid nitrogen stage has been developed for the JEOL JEM-100B electron microscope equipped with a scanning attachment. The design is a modification of the standard JEM-100B SEM specimen holder with specimen cooling to any temperatures In the range ~ 55°K to room temperature. Since the specimen plane is maintained at the ‘high resolution’ focal position of the objective lens and ‘bumping’ and thermal drift la minimized by supercooling the liquid nitrogen, the high resolution capability of the microscope is maintained (Fig.4).


Author(s):  
Mihir Parikh

It is well known that the resolution of bio-molecules in a high resolution electron microscope depends not just on the physical resolving power of the instrument, but also on the stability of these molecules under the electron beam. Experimentally, the damage to the bio-molecules is commo ly monitored by the decrease in the intensity of the diffraction pattern, or more quantitatively by the decrease in the peaks of an energy loss spectrum. In the latter case the exposure, EC, to decrease the peak intensity from IO to I’O can be related to the molecular dissociation cross-section, σD, by EC = ℓn(IO /I’O) /ℓD. Qu ntitative data on damage cross-sections are just being reported, However, the microscopist needs to know the explicit dependence of damage on: (1) the molecular properties, (2) the density and characteristics of the molecular film and that of the support film, if any, (3) the temperature of the molecular film and (4) certain characteristics of the electron microscope used


Author(s):  
H. Tochigi ◽  
H. Uchida ◽  
S. Shirai ◽  
K. Akashi ◽  
D. J. Evins ◽  
...  

A New High Excitation Objective Lens (Second-Zone Objective Lens) was discussed at Twenty-Sixth Annual EMSA Meeting. A new commercially available Transmission Electron Microscope incorporating this new lens has been completed.Major advantages of the new instrument allow an extremely small beam to be produced on the specimen plane which minimizes specimen beam damages, reduces contamination and drift.


Author(s):  
S. Saito ◽  
H. Todokoro ◽  
S. Nomura ◽  
T. Komoda

Field emission scanning electron microscope (FESEM) features extremely high resolution images, and offers many valuable information. But, for a specimen which gives low contrast images, lateral stripes appear in images. These stripes are resulted from signal fluctuations caused by probe current noises. In order to obtain good images without stripes, the fluctuations should be less than 1%, especially for low contrast images. For this purpose, the authors realized a noise compensator, and applied this to the FESEM.Fig. 1 shows an outline of FESEM equipped with a noise compensator. Two apertures are provided gust under the field emission gun.


Author(s):  
H. Rose

The scanning transmission electron microscope offers the possibility of utilizing inelastically scattered electrons. Use of these electrons in addition to the elastically scattered electrons should reduce the scanning time (dose) Which is necessary to keep the quantum noise below a certain level. Hence it should lower the radiation damage. For high resolution, Where the collection efficiency of elastically scattered electrons is small, the use of Inelastically scattered electrons should become more and more favorable because they can all be detected by means of a spectrometer. Unfortunately, the Inelastic scattering Is a non-localized interaction due to the electron-electron correlation, occurring predominantly at the circumference of the atomic electron cloud.


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