A technique for preparing Beauveria spp. for scanning electron microscopy

1980 ◽  
Vol 58 (15) ◽  
pp. 1700-1703 ◽  
Author(s):  
E. C. Quattlebaum ◽  
G. R. Carner

Vapor fixation for 96 h with 1% osmium tetroxide (OsO4) and 3–4 days air drying produced distortion-free specimens of Beauveria spp. for examination with the scanning electron microscope. A combination of 4 h OsO4 vapor fixation and freeze-drying also reduced disruption satisfactorily but specimens were not as well preserved as with the first method. Preparation methods that were ineffective in preventing collapse of hydrophilic structures were Cling Free® sprayed on specimens prior to examination, freeze-drying, critical-point drying (of unfixed material), and vapor fixation with glutaraldehyde.

Author(s):  
Linda M. Sicko ◽  
Thomas E. Jensen

The use of critical point drying is rapidly becoming a popular method of preparing biological samples for scanning electron microscopy. The procedure is rapid, and produces consistent results with a variety of samples. The preservation of surface details is much greater than that of air drying, and the procedure is less complicated than that of freeze drying. This paper will present results comparing conventional air-drying of plant specimens to critical point drying, both of fixed and unfixed material. The preservation of delicate structures which are easily damaged in processing and the use of filter paper as a vehicle for drying will be discussed.


Author(s):  
J R Santos-Mallet ◽  
T D Balthazar ◽  
A A Oliveira ◽  
W A Marques ◽  
A Q Bastos ◽  
...  

Abstract The aim of the present study was to describe the morphology of the eggs of Culex (Culex) saltanensis Dyar that occurs in the Neotropical region. Eggs of the Cx. (Cux.) saltanensis were collected at the Mata Atlântica FIOCRUZ campus, fixed in 1% osmium tetroxide, prepared for mounting on metal supports, observed under a scanning electron microscope, and described morphologically. The eggs had a coniform shape with a length of approximately 0.5 mm (505–510 µm) and a width in the median portion of 117 µm (113–123 µm). Upper portion is lined with tubers of irregular shape and varying sizes (0.64–1.31 µm), located on a cross-linked matrix forming bands observed under optical microscopy. The micropyle is encased in a necklace of approximately 6.6-µm plates arranged in a flower-like shape. Comparing Cx. (Cux.) saltanensis eggs with several species of different genera, important divergent characteristics can be observed. However, this study points to the need for new descriptions of eggs of species belonging to the same subgenus in order to analyze if there will be differences between them. Culex (Cux.) saltanensis eggs have particular characteristics not observed in eggs of other Culicidae genera.


2015 ◽  
Vol 21 (1) ◽  
pp. 17 ◽  
Author(s):  
Poliana Cristina Spricigo ◽  
Jéssica Prada Trento ◽  
Joana Dias Bresolin ◽  
Viviane Faria Soares ◽  
Viviane Faria Soares ◽  
...  

Brazil has great capacity for expansion in the floriculture sector. Studies on postharvest cut flowers contribute to development of the sector, helping to maintain the quality of domestic production. Scanning electron microscopy (SEM) is a powerful tool that allows viewing of flower structures and also microorganisms. The aim of this study was to evaluate methods of preparing flower stem samples for viewing in SEM as a support for studies on postharvest cut flowers. Ways of cutting, fixing, and drying samples were tested. Cutting with a stainless steel blade and through freeze-fracture were tested; fixation was carried out without the use of osmium tetroxide (OsO4); and drying of the samples was performed through freeze-drying and through critical point dryingwithCO2. Cutting with a stainless steel blade proved to be a satisfactory method for stem samples, with low cost and simple application compared to freeze-fracturing. Good fixation and high image contrast were obtained without the use of osmium tetroxide, thus avoiding the use of this toxic compound. Freeze-drying allowed the structure and morphological composition to be viewed, while critical point drying withCO2 preserved the microorganisms present in the samples.


Foods ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 487
Author(s):  
Hana Běhalová ◽  
Bohuslava Tremlová ◽  
Ludmila Kalčáková ◽  
Matej Pospiech ◽  
Dani Dordevic

The aim of the research was to verify the necessity of secondary fixation with osmium tetroxide in various types of meat products and evaluation of structural changes of products using different fixation procedures. The material for the study consisted of 11 types of meat products that were analyzed using a scanning electron microscope (SEM) with two different methods of chemical fixation. The first method included the usual processing of biological samples: glutaraldehyde primary fixation, the use of a buffer, secondary fixation by osmium tetroxide (OsO4), buffer, and dehydration using ethanol of increasing concentrations. The second method comprised the glutaraldehyde primary fixation and dehydration using the ethanol of increasing concentrations only. The results unambiguously suggest that the main difference between these methods is in fixation and visibility of fat. Our analysis principally suggests that fixation of the product with OsO4 allows the tracking of all components (fat droplets, muscle fibers, connective tissue) in meat products. At the same time, our results also support the possibility that the secondary fixation can be skipped during the analysis, where the main objection is an observation of lipid-free structures of the meat products (e.g., connection between muscle and starches or spices) or meat products with an insignificant amount of fat.


Author(s):  
David Joy ◽  
James Pawley

The scanning electron microscope (SEM) builds up an image by sampling contiguous sub-volumes near the surface of the specimen. A fine electron beam selectively excites each sub-volume and then the intensity of some resulting signal is measured. The spatial resolution of images made using such a process is limited by at least three factors. Two of these determine the size of the interaction volume: the size of the electron probe and the extent to which detectable signal is excited from locations remote from the beam impact point. A third limitation emerges from the fact that the probing beam is composed of a finite number of discrete particles and therefore that the accuracy with which any detectable signal can be measured is limited by Poisson statistics applied to this number (or to the number of events actually detected if this is smaller).


Author(s):  
Mamaeva S.N. ◽  
Vinokurov R.R. ◽  
Munkhalova Ya.A. ◽  
Dyakonova D.P. ◽  
Platonova V.A. ◽  
...  

Currently, due to the intensive development of high-tech science-intensive medical and research devices, more and more attention is paid to the development of diagnostics of rare and difficult to diagnose diseases. It is known that among numerous nephropathies, hematuria may be the only symptom of kidney and urinary tract diseases, which complicates their diagnosis and treatment. In order to develop new approaches for the diagnosis of nephropathies, the authors have been studying the morphology of red blood cells in the blood and urine of children and adults using a scanning electron microscope for several years. The paper presents the results of studies of children with various kidney diseases, including IgA-nephropathy, and chronic glomerulonephritis. Scanning electron microscopy was used for the first time to detect nanoparticles on the surface of red blood cells, the size of which is comparable to the size of viruses, which became the basis for one of the authors ' assumptions, namely, the possible transport of certain types of viruses by red blood cells. Thus, some kidney diseases could be considered virus-associated. This paper presents for the first time the results of determining the glomerular filtration rate of both kidneys separately in the study of separate kidney function and of the study of urine smears obtained during catheterization of the ureters in patients with hydronephrosis of one of the kidneys by scanning electron microscopy. As in previous studies, nanoparticles were found on the surface of red blood cells, which leads to the conclusion about the possible viral nature of the disease of the considered patient. In addition, smear images obtained using a microscope showed a significant difference in the elements of the right and left kidneys urine, which did not contradict the data on the study of glomerular filtration rate. According to the authors, the capabilities of the scanning electron microscope can be applied in fundamental research of kidney diseases at the cellular and molecular levels, forming new ideas about their origin, as well as on the basis of which new methods of non-invasive diagnostics can be built.


1972 ◽  
Vol 3 (3) ◽  
pp. 181-188 ◽  
Author(s):  
Christine Dahl

AbstractA method for preparation of alcohol-preserved culicid larvae for Scanning Electron Microscope (SEM) studies is described. It is based on dehydration by ethanol-xylol and fast evaporation of xylol in +8o° C. for ten minutes. For taxonomic studies such as examination of pecten teeth, comb scales and microtrichiae in magnifications up to 6oooX the method is suitable. For studies of receptor structures on hair-tufts and microstructures of the body integument alcohol preserved material is less satisfactory. The microstructure of the comb scales is figured and their function discussed. Differences in the ultrastructure of the abdominal hair-tufts are pointed out.


1988 ◽  
Vol 62 (2) ◽  
pp. 141-147 ◽  
Author(s):  
F. J. Kruger ◽  
V. L. Hamilton-Attwell

ABSTRACTSchistosoma haematobium miracidia were collected from a locality with a high prevalence of human infection with the animal parasite, S. mattheei, which hybridizes with S. haematobium, and from 2 localities with negligible infection rates. The terebratoria of the miracidia from these localities were compared with each other, with laboratory maintained S. haematobium and with four populations of S. mattheei by means of scanning electron microscopy. It was found that the terebratorial membrane of certain of the S. haematobium miracidia from the locality with a high S. mattheei prevalence in humans, resembled the more intricate membrane of S. mattheei. This suggests introgressive hybridization between S. haematobium and S. haematobium x S. mattheei.


1994 ◽  
Vol 40 (134) ◽  
pp. 195-197
Author(s):  
E. W. Wolff ◽  
A. P. Reid

AbstractA snow crystal has been successfully collected on to a scanning electron microscope (SEM) stub in central Greenland. It was preserved at liquid-nitrogen temperature for 5 months, prior to examination in the SEM. This is believed to be the first time a snow crystal has been observed directly in the SEM and offers some new experimental methods for understanding crystals and their chemistry.


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