Characterization of frozen hydrated biological specimens by low-loss EELS

Author(s):  
Songquan Sun ◽  
Richard D. Leapman

Analyses of ultrathin cryosections are generally performed after freeze-drying because the presence of water renders the specimens highly susceptible to radiation damage. The water content of a subcellular compartment is an important quantity that must be known, for example, to convert the dry weight concentrations of ions to the physiologically more relevant molar concentrations. Water content can be determined indirectly from dark-field mass measurements provided that there is no differential shrinkage between compartments and that there exists a suitable internal standard. The potential advantage of a more direct method for measuring water has led us to explore the use of electron energy loss spectroscopy (EELS) for characterizing biological specimens in their frozen hydrated state.We have obtained preliminary EELS measurements from pure amorphous ice and from cryosectioned frozen protein solutions. The specimens were cryotransfered into a VG-HB501 field-emission STEM equipped with a 666 Gatan parallel-detection spectrometer and analyzed at approximately −160 C.

Author(s):  
R.D. Leapman ◽  
S.Q. Sun ◽  
S-L. Shi ◽  
R.A. Buchanan ◽  
S.B. Andrews

Recent advances in rapid-freezing and cryosectioning techniques coupled with use of the quantitative signals available in the scanning transmission electron microscope (STEM) can provide us with new methods for determining the water distributions of subcellular compartments. The water content is an important physiological quantity that reflects how fluid and electrolytes are regulated in the cell; it is also required to convert dry weight concentrations of ions obtained from x-ray microanalysis into the more relevant molar ionic concentrations. Here we compare the information about water concentrations from both elastic (annular dark-field) and inelastic (electron energy loss) scattering measurements.In order to utilize the elastic signal it is first necessary to increase contrast by removing the water from the cryosection. After dehydration the tissue can be digitally imaged under low-dose conditions, in the same way that STEM mass mapping of macromolecules is performed. The resulting pixel intensities are then converted into dry mass fractions by using an internal standard, e.g., the mean intensity of the whole image may be taken as representative of the bulk water content of the tissue.


Author(s):  
R.D. Leapman ◽  
R.L. Ornberg

Determination of cellular organelle water content is important in understanding cell volume regulation and also for converting x-ray microanalytical measurements of diffusible ions and elements from dry mass concentration to the biologically more relevant aqueous concentration. It has been proposed that electron energy loss spectroscopy (EELS) can be used to measure mass thickness in frozen hydrated and dehydrated cryosections at low electron dose, and that the method should thus provide a direct estimate of water content. Potentially the EELS inelastic scattering method has a number of advantages over alternative approaches. For example use of the x-ray continuum to measure mass requires much higher doses and cannot be applied at resolutions of 100nm to hydrated samples. Moreover, hydrated cryosections are often too thick to utilize dark field STEM imaging for mass measurement.


Author(s):  
E.G. Bithell ◽  
W.M. Stobbs

It is well known that the microstructural consequences of the ion implantation of semiconductor heterostructures can be severe: amorphisation of the damaged region is possible, and layer intermixing can result both from the original damage process and from the enhancement of the diffusion coefficients for the constituents of the original composition profile. A very large number of variables are involved (the atomic mass of the target, the mass and energy of the implant species, the flux and the total dose, the substrate temperature etc.) so that experimental data are needed despite the existence of relatively well developed models for the implantation process. A major difficulty is that conventional techniques (e.g. electron energy loss spectroscopy) have inadequate resolution for the quantification of any changes in the composition profile of fine scale multilayers. However we have demonstrated that the measurement of 002 dark field intensities in transmission electron microscope images of GaAs / AlxGa1_xAs heterostructures can allow the measurement of the local Al / Ga ratio.


Author(s):  
L. Wan ◽  
R. F. Egerton

INTRODUCTION Recently, a new compound carbon nitride (CNx) has captured the attention of materials scientists, resulting from the prediction of a metastable crystal structure β-C3N4. Calculations showed that the mechanical properties of β-C3N4 are close to those of diamond. Various methods, including high pressure synthesis, ion beam deposition, chemical vapor deposition, plasma enhanced evaporation, and reactive sputtering, have been used in an attempt to make this compound. In this paper, we present the results of electron energy loss spectroscopy (EELS) analysis of composition and bonding structure of CNX films deposited by two different methods.SPECIMEN PREPARATION Specimens were prepared by arc-discharge evaporation and reactive sputtering. The apparatus for evaporation is similar to the traditional setup of vacuum arc-discharge evaporation, but working in a 0.05 torr ambient of nitrogen or ammonia. A bias was applied between the carbon source and the substrate in order to generate more ions and electrons and change their energy. During deposition, this bias causes a secondary discharge between the source and the substrate.


2018 ◽  
Vol 16 (2) ◽  
pp. 219-226
Author(s):  
Sri Rizqi Annisa ◽  
Dewi Larasati ◽  
Endang Bekti K

The aim of this study was to determine the characterization of shredded mureel fish with kluwih substitution on water content, protein content, fiber content and organoleptic (preference for crispness and taste). This study uses a simple Randomized Complete Design (RCD) with the substitution treatment of kluwih and mureel fish, with the following ratio: S1 (240g: 60g), S2 (210g: 90g), S3 (180g: 120g), S4 (150g: 150g), S5 (120g: 180g). Data were analyzed statistically by analysis of variance and if there was a significant effect, further testing was done with BNJ at the level of 5%. The results showed that kluwih substitution in the manufacture of mureel fish shredded had an average: water content of 8.33-10.62%, protein :16.83-22.00%, fiber : 6.79-6.99%, score crispness 2-6.12, taste score 2.6-6.6. Based on the results of the analysis of the variety of kluwih substitutes and mureel fish have a significant effect on water content, protein content and crisp organoleptic test, taste on mureel fish fillet, and no significant effect on fiber content. The best kluwih substitution in S3 treatment with 120 grams of substitute kluwih and 180 grams of mureel fish.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kazunori Matsuda ◽  
Takuya Akiyama ◽  
Satoshi Tsujibe ◽  
Kaihei Oki ◽  
Agata Gawad ◽  
...  

AbstractStool consistency is evaluated mainly in reference to indirect indicators such as water content or the appearance of stool forms using Bristol Stool Form Scale (BSFS). Methods of measurement are limited. We thus aimed to develop a simple protocol for direct measurement of stool consistency using the TA.XTExpress Texture Analyser (Stable Micro Systems Ltd.). We developed a protocol which enables mechanical quantification of the gram-force against a cylindrical probe (ø 6 mm) pushed into the stool surface at 2.0 mm/s to 5 mm depth. The consistency of 252 stools collected from 40 healthy Belgians was evaluated by the direct method and by the indirect indicators (water content and BSFS) for comparison. The log-transformed stool consistency values measured by the texture analyzer had a negative linear correlation with the stool water contents (rrm = − 0.781) with homoscedastic variance, suggesting the appropriateness of the new protocol. They showed a similar correlation with the BSFS, but with a large variance in the consistency values of normal stool forms. This correlation was much smaller for BSFS scored by subjects (rrm = − 0.587) than by experts (rrm = − 0.789), collectively indicating BSFS as a rough indicator of stool consistency susceptible to subjective bias despite its effectiveness in clinical use. The optimized direct method using the texture analyzer enables the accurate quantification of stool consistency, which facilitates understanding of the intestinal environment and function and thus may enhance the value of the stool as a predictor of human health.


2005 ◽  
Vol 37 (5) ◽  
pp. 425-432 ◽  
Author(s):  
Ana PINTADO ◽  
Leopoldo G. SANCHO ◽  
T. G. Allan GREEN ◽  
José Manuel BLANQUER ◽  
Roberto LÁZARO

The Tabernas badlands in semiarid south-east Spain is one of the driest regions in Europe with a mean annual precipitation of c. 240 mm. The landscape is deeply dissected, with canyons, ramblas and sparsely vegetated eroded badland slopes. The vegetation is predominantly a biological soil crust consisting of different types of lichen-rich communities, one of the more conspicuous being dominated by Diploschistes diacapsis (Ach.) Lumbsch. This lichen is mainly restricted to the north- facing slopes, where it forms extensive whitish carpets and probably plays an important role in preventing erosion of the slopes and allowing plant colonization. South-facing slopes are much more eroded and generally lack vegetation. %The photosynthetic performance of north (shade) and south-facing (sun) populations of D. diacapsis was studied to determine if these different populations showed any adaptations to the microclimatic conditions of their individual habitats. The response of CO2 exchange to light intensity, temperature and water content was measured under controlled conditions in the laboratory. Dry weight-based net photosynthetic rates were higher in the southern-exposed population but quantum efficiency, and light compensation points were similar. Thallus weight per unit area (LMA) was considerably higher for shade specimens but maximum water content and optimal water content were very similar and chlorophyll content on a dry weight basis was also similar. Chlorophyll content on an area basis was higher in the northern-exposed population and always much larger than those reported in other studies on the same species (up to 8 times larger) with the result that NP values on a chlorophyll basis were relatively low. The larger LMA meant that shade thalli stored more water per unit area which should ensure longer active periods than sun thalli. The results support a strategy pair of high NP and short active time versus low NP and long active time, both having been reported for other soil crust species. However, the visibly larger biomass of the shade D. diacapsis suggests that the lichen is at the limit of its adaptability in these habitats.


2020 ◽  
Vol 75 (11) ◽  
pp. 913-919
Author(s):  
Frank Krumeich

AbstractSince the 1970s, high-resolution transmission electron microscopy (HRTEM) is well established as the most appropriate method to explore the structural complexity of niobium tungsten oxides. Today, scanning transmission electron microscopy (STEM) represents an important alternative for performing the structural characterization of such oxides. STEM images recorded with a high-angle annular dark field (HAADF) detector provide not only information about the cation positions but also about the distribution of niobium and tungsten as the intensity is directly correlated to the local scattering potential. The applicability of this method is demonstrated here for the characterization of the real structure of Nb7W10O47.5. This sample contains well-ordered domains of Nb8W9O47 and Nb4W7O31 besides little ordered areas according to HRTEM results. Structural models for Nb4W7O31 and twinning occurring in this phase have been derived from the interpretation of HAADF-STEM images. A remarkable grain boundary between well-ordered domains of Nb4W7O31 and Nb8W9O47 has been found that contains one-dimensionally periodic features. Furthermore, short-range order observed in less ordered areas could be attributed to an intimate intergrowth of small sections of different tetragonal tungsten bronze (TTB) based structures.


1944 ◽  
Vol 35 (2) ◽  
pp. 127-139 ◽  
Author(s):  
G. Fraenkel ◽  
M. Blewett

(1) Three insects,Tribolium confusum, Ephestia kuehniellaandDermestes vulpinus, have been grown at several humidities and the following factors have been determined: length of larval period; water content of food and of the freshly formed pupae; wet and dry weight of pupae and wet and dry weight of food consumed during larval development. The “net utilisation” of the food has been calculated as the ratio of dry weight of food eaten per larva to dry weight of pupa.(2) At lower humidities more food is eaten to produce a given unit of body weight. The length of the larval period increases and the weight of the pupae decreases.(3) More food is eaten at low humidities, because part of the food is utilised as water. As a consequence of this, the larva grows more slowly and its final size is smaller. It is shown forDermestesat 30 per cent. andEphestiaat 1 per cent. R.H. that less than 32·9 and 7·6 per cent. of the water in the pupae can be derived from water ingested with the food.


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