Hydration Stages—Achievements and Obstacles

Author(s):  
V.R. Matricardi ◽  
J. Subjeck ◽  
D.F. Parsons

As we have previously reported our laboratory has developed and perfected a dynamically pumped hydration stage. The stage consists of four collinear apertures: two 100μ and two 200μ. The two 100μ apertures straddle the specimen and define the specimen chamber. The two 200μ apertures define the intermediate chamber which is independently pumped by a normal cryotrapped forepump. The whole stage is translated in order to move the apertures collinear with the beam by the normal specimen translation mechanism, through the bore of the upper polepiece. The specimen may be moved within its 1.2mm gap by an independent translation mechanism thereby allowing the full area of the grid to be examined. Water vapor evaporates from the source and continuously flows out of the apertures thereby providing a very effective anticontamination device.

Author(s):  
Columba de la Parra ◽  
Amanda Ernlund ◽  
Amandine Alard ◽  
Kelly Ruggles ◽  
Beatrix Ueberheide ◽  
...  

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Lian He ◽  
Changfeng Man ◽  
Shouyan Xiang ◽  
Lin Yao ◽  
Xiaoyan Wang ◽  
...  

AbstractCircular RNAs a kind of covalently closed RNA and widely expressed in eukaryotes. CircRNAs are involved in a variety of physiological and pathological processes, but their regulatory mechanisms are not fully understood. Given the development of the RNA deep-sequencing technology and the improvement of algorithms, some CircRNAs are discovered to encode proteins through the cap-independent mechanism and participate in the important process of tumorigenesis and development. Based on an overview of CircRNAs, this paper summarizes its translation mechanism and research methods, and reviews the research progress of CircRNAs translation in the field of oncology in recent years. Moreover, this paper aims to provide new ideas for tumor diagnosis and treatment through CircRNAs translation.


Author(s):  
S. Basu ◽  
D. F. Parsons

We are approaching the invasiveness of cancer cells from the studies of their wet surface morphology which should distinguish them from their normal counterparts. In this report attempts have been made to provide physical basis and background work to a wet replication method with a differentially pumped hydration chamber (Fig. 1) (1,2), to apply this knowledge for obtaining replica of some specimens of known features (e.g. polystyrene latex) and finally to realize more specific problems and to improvize new methods and instrumentation for their rectification. In principle, the evaporant molecules penetrate through a pair of apertures (250, 350μ), through water vapors and is, then, deposited on the specimen. An intermediate chamber between the apertures is pumped independently of the high vacuum system. The size of the apertures is sufficiently small so that full saturated water vapor pressure is maintained near the specimen.


Author(s):  
R. C. Moretz ◽  
G. G. Hausner ◽  
D. F. Parsons

Electron microscopy and diffraction of biological materials in the hydrated state requires the construction of a chamber in which the water vapor pressure can be maintained at saturation for a given specimen temperature, while minimally affecting the normal vacuum of the remainder of the microscope column. Initial studies with chambers closed by thin membrane windows showed that at the film thicknesses required for electron diffraction at 100 KV the window failure rate was too high to give a reliable system. A single stage, differentially pumped specimen hydration chamber was constructed, consisting of two apertures (70-100μ), which eliminated the necessity of thin membrane windows. This system was used to obtain electron diffraction and electron microscopy of water droplets and thin water films. However, a period of dehydration occurred during initial pumping of the microscope column. Although rehydration occurred within five minutes, biological materials were irreversibly damaged. Another limitation of this system was that the specimen grid was clamped between the apertures, thus limiting the yield of view to the aperture opening.


Author(s):  
V. R. Matricardi ◽  
G. G. Hausner ◽  
D. F. Parsons

In order to observe room temperature hydrated specimens in an electron microscope, the following conditions should be satisfied: The specimen should be surrounded by water vapor as close as possible to the equilibrium vapor pressure corresponding to the temperature of the specimen.The specimen grid should be inserted, focused and photo graphed in the shortest possible time in order to minimize dehydration.The full area of the specimen grid should be visible in order to minimize the number of changes of specimen required.There should be no pressure gradient across the grid so that specimens can be straddled across holes.Leakage of water vapor to the column should be minimized.


Author(s):  
K. A. Brookes ◽  
D. Finbow ◽  
Madeleine Samuel

Investigation of the particulate matter contained in the water sample, revealed the presence of a number of different types and certain of these were selected for analysis.An A.E.I. Corinth electron microscope was modified to accept a Kevex Si (Li) detector. To allow for existing instruments to be readily modified, this was kept to a minimum. An additional port is machined in the specimen region to accept the detector, with the liquid nitrogen cooling dewar conveniently housed in the left hand cupboard adjacent to the microscope column. Since background radiation leads to loss in the sensitivity of the instrument, great care has been taken to reduce this effect by screening and manufacturing components that are near the specimen from material of low atomic number. To change from normal transmission imaging to X-ray analysis, the special 4-position specimen rod is inserted through the normal specimen airlock.


Author(s):  
Z.L. Wang

An experimental technique for performing electron holography using a non-FEG, non-biprism transmission electron microscope (TEM) has been introduced by Ru et al. A double stacked specimens, one being a single crystal foil and the other the specimen, are loaded in the normal specimen position in TEM. The single crystal, which is placed onto the specimen, is responsible to produce two beams that are equivalent to two virtual coherent sources illuminating the specimen beneath, thus, permitting electron holography of the specimen. In this paper, the imaging theory of this technique is described. Procedures are introduced for digitally reconstructing the holograms.


Author(s):  
E. L. Vigil ◽  
E. F. Erbe

In cotton seeds the radicle has 12% moisture content which makes it possible to prepare freeze-fracture replicas without fixation or cryoprotection. For this study we have examined replicas of unfixed radicle tissue fractured at room temperature to obtain data on organelle and membrane structure.Excised radicles from seeds of cotton (Gossyplum hirsutum L. M-8) were fractured at room temperature along the longitudinal axis. The fracture was initiated by spliting the basal end of the excised radicle with a razor. This procedure produced a fracture through the tissue along an unknown fracture plane. The warm fractured radicle halves were placed on a thin film of 100% glycerol on a flat brass cap with fracture surface up. The cap was rapidly plunged into liquid nitrogen and transferred to a freeze- etch unit. The sample was etched for 3 min at -95°C to remove any condensed water vapor and then cooled to -150°C for platinum/carbon evaporation.


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