Electron microscopy of the yeast to mycelial phase conversion of Sporotrichum schenckii

1970 ◽  
Vol 16 (8) ◽  
pp. 747-749 ◽  
Author(s):  
James W. Lane ◽  
Robert G. Garrison

Ultrastructural events occurring during the yeast-like to mycelial phase conversion of the dimorphic fungal pathogen Sporotrichum schenckii were studied by electron microscopy of thin sections. Discrete intracytoplasmic membrane systems were formed as early as 6 h after induction of the conversion stimuli. Within 12 to 18 h, a transitional cell was formed from the parent yeast-like cell. Definite streaming of yeast-like cell cytoplasm and mitochondria was noted before septal formation in the intermediate cell. Although the cell wall of the parent yeast-like phase cell is present in two distinct layers, only the inner layer persists upon transitional cell formation. Yeast-like to mycelial phase reorganization of S. schenckii resembles closely that described for Histoplasma capsulatum, Blastomyces dermatitidis, and Paracoccidioides brasiliensis, and it is suggested that yeast to mold transformation of these dimorphic fungi may proceed via a common conversional mechanism.

1973 ◽  
Vol 19 (10) ◽  
pp. 1305-1309 ◽  
Author(s):  
Robert G. Garrison ◽  
Karen S. Boyd

Aspects of the culturally induced mycelial- to tissue-phase transformation of Penicillium marneffei were studied by electron microscopy of thin sections. The hyphal cell was observed to contain multiple, large lipid bodies scattered throughout the cytoplasm. Five days after induction of conversion, short elements arose which were characterized by the presence of two polar areas of vacuolation containing electron-opaque material thought to be altered lipid. This material could not be demonstrated with permanganate fixation, but reacted strongly with thiocarbohydrazide. The latter reagent is known to enhance the electron density of osmium-stained lipids. After 10 days incubation, the tissue-phase cells appeared to be slightly elongate with one or more septa present as the result of division by fission. The cell wall appeared to be of uniform electron opacity with a slightly roughened appearance to the outer surface. Except for residuals of polar vacuolation, the internal morphology of the tissue phase of P. marneffei appeared similar in many respects to that of the analogous yeast-like phases of certain other of the pathogenic dimorphic fungi.


Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).


Author(s):  
J.R. Walton

In electron microscopy, lead is the metal most widely used for enhancing specimen contrast. Lead citrate requires a pH of 12 to stain thin sections of epoxy-embedded material rapidly and intensively. However, this high alkalinity tends to leach out enzyme reaction products, making lead citrate unsuitable for many cytochemical studies. Substitution of the chelator aspartate for citrate allows staining to be carried out at pH 6 or 7 without apparent effect on cytochemical products. Moreover, due to the low, controlled level of free lead ions, contamination-free staining can be carried out en bloc, prior to dehydration and embedding. En bloc use of lead aspartate permits the grid-staining step to be bypassed, allowing samples to be examined immediately after thin-sectioning.Procedures. To prevent precipitation of lead salts, double- or glass-distilled H20 used in the stain and rinses should be boiled to drive off carbon dioxide and glassware should be carefully rinsed to remove any persisting traces of calcium ion.


Author(s):  
N.C. Lyon ◽  
W. C. Mueller

Schumacher and Halbsguth first demonstrated ectodesmata as pores or channels in the epidermal cell walls in haustoria of Cuscuta odorata L. by light microscopy in tissues fixed in a sublimate fixative (30% ethyl alcohol, 30 ml:glacial acetic acid, 10 ml: 65% nitric acid, 1 ml: 40% formaldehyde, 5 ml: oxalic acid, 2 g: mecuric chloride to saturation 2-3 g). Other workers have published electron micrographs of structures transversing the outer epidermal cell in thin sections of plant leaves that have been interpreted as ectodesmata. Such structures are evident following treatment with Hg++ or Ag+ salts and are only rarely observed by electron microscopy. If ectodesmata exist without such treatment, and are not artefacts, they would afford natural pathways of entry for applied foliar solutions and plant viruses.


Author(s):  
P.J. Lea ◽  
M.J. Hollenberg

Our current understanding of mitochondrial ultrastructure has been derived primarily from thin sections using transmission electron microscopy (TEM). This information has been extrapolated into three dimensions by artist's impressions (1) or serial sectioning techniques in combination with computer processing (2). The resolution of serial reconstruction methods is limited by section thickness whereas artist's impressions have obvious disadvantages.In contrast, the new techniques of HRSEM used in this study (3) offer the opportunity to view simultaneously both the internal and external structure of mitochondria directly in three dimensions and in detail.The tridimensional ultrastructure of mitochondria from rat hepatocytes, retinal (retinal pigment epithelium), renal (proximal convoluted tubule) and adrenal cortex cells were studied by HRSEM. The specimens were prepared by aldehyde-osmium fixation in combination with freeze cleavage followed by partial extraction of cytosol with a weak solution of osmium tetroxide (4). The specimens were examined with a Hitachi S-570 scanning electron microscope, resolution better than 30 nm, where the secondary electron detector is located in the column directly above the specimen inserted within the objective lens.


Author(s):  
T. Baird ◽  
J.R. Fryer ◽  
S.T. Galbraith

Introduction Previously we had suggested (l) that the striations observed in the pod shaped crystals of β FeOOH were an artefact of imaging in the electron microscope. Contrary to this adsorption measurements on bulk material had indicated the presence of some porosity and Gallagher (2) had proposed a model structure - based on the hollandite structure - showing the hollandite rods forming the sides of 30Å pores running the length of the crystal. Low resolution electron microscopy by Watson (3) on sectioned crystals embedded in methylmethacrylate had tended to support the existence of such pores.We have applied modern high resolution techniques to the bulk crystals and thin sections of them without confirming these earlier postulatesExperimental β FeOOH was prepared by room temperature hydrolysis of 0.01M solutions of FeCl3.6H2O, The precipitate was washed, dried in air, and embedded in Scandiplast resin. The sections were out on an LKB III Ultramicrotome to a thickness of about 500Å.


Author(s):  
William H. Massover

Stereoscopic examination of thick sections of fixed and embedded biological tissues by high voltage electron microscopy has been shown to allow direct visualization of three-dimensional fine structure. The present report will consider the occurrence of some new technical problems in specimen preparation and Image interpretation that are not common during lower voltage studies of thin sections.Thick Sectioning and Tissue Coloration - Epon sections of 0.5 μm or more that are cut with glass knives do not have a uniform thickness as Judged by their interference colors; these colors change with time during their flotation on the knife bath, and again when drying onto the specimen support. Quoted thicknesses thus must be considered only as rough estimates unless measured in specific regions by other methods. Chloroform vapors do not always result in good spreading of thick sections; however, they will spread spontaneously to large degrees after resting on the flotation bath for several minutes. Ribbons of thick sections have been almost impossible to obtain.


Author(s):  
J. Cadoz ◽  
J. Castaing ◽  
J. Philibert

Plastic deformation of alumina has been much studied; basal slip occurs and dislocation structures have been investigated by transmission electron microscopy (T.E.M.) (1). Non basal slip has been observed (2); the prismatic glide system <1010> {1210} has been obtained by compression tests between 1400°C and 1800°C (3). Dislocations with <0110> burgers vector were identified using a 100 kV microscope(4).We describe the dislocation structures after prismatic slip, using high voltage T.E.M. which gives much information.Compression tests were performed at constant strainrate (∿10-4s-1); the maximum deformation reached was 0.03. Thin sections were cut from specimens deformed at 1450°C, either parallel to the glide plane or perpendicular to the glide direction. After mechanical thinning, foils were produced by ion bombardment. Details on experimental techniques can be obtained through reference (3).


Author(s):  
Robert M. Fisher

By 1940, a half dozen or so commercial or home-built transmission electron microscopes were in use for studies of the ultrastructure of matter. These operated at 30-60 kV and most pioneering microscopists were preoccupied with their search for electron transparent substrates to support dispersions of particulates or bacteria for TEM examination and did not contemplate studies of bulk materials. Metallurgist H. Mahl and other physical scientists, accustomed to examining etched, deformed or machined specimens by reflected light in the optical microscope, were also highly motivated to capitalize on the superior resolution of the electron microscope. Mahl originated several methods of preparing thin oxide or lacquer impressions of surfaces that were transparent in his 50 kV TEM. The utility of replication was recognized immediately and many variations on the theme, including two-step negative-positive replicas, soon appeared. Intense development of replica techniques slowed after 1955 but important advances still occur. The availability of 100 kV instruments, advent of thin film methods for metals and ceramics and microtoming of thin sections for biological specimens largely eliminated any need to resort to replicas.


Author(s):  
W. T. Donlon ◽  
J. E. Allison ◽  
S. Shinozaki

Light weight materials which possess high strength and durability are being utilized by the automotive industry to increase fuel economy. Rapidly solidified (RS) Al alloys are currently being extensively studied for this purpose. In this investigation the microstructure of an extruded Al-8Fe-2Mo alloy, produced by Pratt & Whitney Aircraft, Goverment Products Div. was examined in a JE0L 2000FX AEM. Both electropolished thin sections, and extraction replicas were examined to characterize this material. The consolidation procedure for producing this material included a 9:1 extrusion at 340°C followed by a 16:1 extrusion at 400°C, utilizing RS powders which have also been characterized utilizing electron microscopy.


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