Revealing gross three-dimensional structure in the SEM

Author(s):  
Sterling P. Newberry

The beautiful three dimensional representation of small object surfaces by the SEM leads one to search for ways to open up the sample and look inside. Could this be the answer to a better microscopy for gross biological 3-D structure? We know from X-Ray microscope images that Freeze Drying and Critical Point Drying give promise of adequately preserving gross structure. Can we slice such preparations open for SEM inspection? In general these preparations crush more readily than they slice. Russell and Dagihlian got around the problem by “deembedding” a section before imaging. This some what defeats the advantages of direct dry preparation, thus we are reluctant to accept it as the final solution to our problem. Alternatively, consider fig 1 wherein a freeze dried onion root has a window cut in its surface by a micromanipulator during observation in the SEM.

2003 ◽  
Vol 788 ◽  
Author(s):  
Predrag Kisa ◽  
Patrick Fisher ◽  
Al Olszewski ◽  
Ian Nettleship ◽  
Nicholas G. Eror

ABSTRACTThis study investigated the microstructural characteristics of directionally solidified freeze-dried silica sols. Porous structures were formed by depositing silica sol on silicon (100) single crystals. The deposited sols were unidirectionaly solidified by placing the silicon substrate on a copper block immersed in liquid nitrogen and then subsequently freeze-dried. Freeze drying removal of ice crystals created three-dimensional pore channels ranging from 3 to10 micrometers in diameter aggregated in grain like colonies 50–100 micrometers in diameter. Pore size, spacing, colony size and microstructure were determined using optical microscopy (OM) and scanning electron microscopy (SEM) while the structure of the amorphous SiO2 was characterized by X-ray diffraction (XRD). The microstructure results are compared and contrasted with silica aerogel obtained through conventional processing using supercritical CO2.


Author(s):  
T.A. Ceskaa ◽  
B. McEwena ◽  
S.J. Edelsteina

The three-dimensional structure of zinc-induced tubulin sheets freed from microtubule associated proteins (MAPs) has been reconstructed from a series of electron micrographs of negatively stained specimens at various tilt angles. Digitized images of these sheets were analyzed by computer using a combination of real space correlation techniques and Fourier analysis to obtain a three-dimensional representation of the structure.


Author(s):  
Robert Glaeser ◽  
Thomas Bauer ◽  
David Grano

In transmission electron microscopy, the 3-dimensional structure of an object is usually obtained in one of two ways. For objects which can be included in one specimen, as for example with elements included in freeze- dried whole mounts and examined with a high voltage microscope, stereo pairs can be obtained which exhibit the 3-D structure of the element. For objects which can not be included in one specimen, the 3-D shape is obtained by reconstruction from serial sections. However, without stereo imagery, only detail which remains constant within the thickness of the section can be used in the reconstruction; consequently, the choice is between a low resolution reconstruction using a few thick sections and a better resolution reconstruction using many thin sections, generally a tedious chore. This paper describes an approach to 3-D reconstruction which uses stereo images of serial thick sections to reconstruct an object including detail which changes within the depth of an individual thick section.


Author(s):  
J.S. Wall ◽  
V. Maridiyan ◽  
S. Tumminia ◽  
J. Hairifeld ◽  
M. Boublik

The high contrast in the dark-field mode of dedicated STEM, specimen deposition by the wet film technique and low radiation dose (1 e/Å2) at -160°C make it possible to obtain high resolution images of unstained freeze-dried macromolecules with minimal structural distortion. Since the image intensity is directly related to the local projected mass of the specimen it became feasible to determine the molecular mass and mass distribution within individual macromolecules and from these data to calculate the linear density (M/L) and the radii of gyration.2 This parameter (RQ), reflecting the three-dimensional structure of the macromolecular particles in solution, has been applied to monitor the conformational transitions in E. coli 16S and 23S ribosomal RNAs in solutions of various ionic strength.In spite of the differences in mass (550 kD and 1050 kD, respectively), both 16S and 23S RNA appear equally sensitive to changes in buffer conditions. In deionized water or conditions of extremely low ionic strength both appear as filamentous structures (Fig. la and 2a, respectively) possessing a major backbone with protruding branches which are more frequent and more complex in 23S RNA (Fig. 2a).


Author(s):  
M. Boublik ◽  
N. Robakis ◽  
J.S. Wall

The three-dimensional structure and function of biological supramolecular complexes are, in general, determined and stabilized by conformation and interactions of their macromolecular components. In the case of ribosomes, it has been suggested that one of the functions of ribosomal RNAs is to act as a scaffold maintaining the shape of the ribosomal subunits. In order to investigate this question, we have conducted a comparative TEM and STEM study of the structure of the small 30S subunit of E. coli and its 16S RNA.The conventional electron microscopic imaging of nucleic acids is performed by spreading them in the presence of protein or detergent; the particles are contrasted by electron dense solution (uranyl acetate) or by shadowing with metal (tungsten). By using the STEM on freeze-dried specimens we have avoided the shearing forces of the spreading, and minimized both the collapse of rRNA due to air drying and the loss of resolution due to staining or shadowing. Figure 1, is a conventional (TEM) electron micrograph of 30S E. coli subunits contrasted with uranyl acetate.


2021 ◽  
Vol 9 (1) ◽  
Author(s):  
Jerzy Montusiewicz ◽  
Marek Miłosz ◽  
Jacek Kęsik ◽  
Kamil Żyła

AbstractHistorical costumes are part of cultural heritage. Unlike architectural monuments, they are very fragile, which exacerbates the problems of their protection and popularisation. A big help in this can be the digitisation of their appearance, preferably using modern techniques of three-dimensional representation (3D). The article presents the results of the search for examples and methodologies of implementing 3D scanning of exhibited historical clothes as well as the attendant problems. From a review of scientific literature it turns out that so far practically no one in the world has made any methodical attempts at scanning historical clothes using structured-light 3D scanners (SLS) and developing an appropriate methodology. The vast majority of methods for creating 3D models of clothes used photogrammetry and 3D modelling software. Therefore, an innovative approach was proposed to the problem of creating 3D models of exhibited historical clothes through their digitalisation by means of a 3D scanner using structural light technology. A proposal for the methodology of this process and concrete examples of its implementation and results are presented. The problems related to the scanning of 3D historical clothes are also described, as well as a proposal how to solve them or minimise their impact. The implementation of the methodology is presented on the example of scanning elements of the Emir of Bukhara's costume (Uzbekistan) from the end of the nineteenth century, consisting of the gown, turban and shoes. Moreover, the way of using 3D models and information technologies to popularise cultural heritage in the space of digital resources is also discussed.


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