The significance of SEM in the diagnosis of haematopoietic malignancies

1978 ◽  
Vol 36 (2) ◽  
pp. 314-315
Author(s):  
Etienne de Harven ◽  
Nina Lampen
Keyword(s):  
Room Temperature ◽  
Culture Medium ◽  
Cell Attachment ◽  
Ethanol Dehydration ◽  
Moist Chamber ◽  
Efficient Alternative ◽  
Mononuclear Leucocytes ◽  
Fast Quenching ◽  
Freeze Dryer ◽  
Scanning Electron

Samples of heparinized blood, or bone marrow aspirates, or cell suspensions prepared from biopsied tissues (nodes, spleen, etc. ) are routinely prepared, after Ficoll-Hypaque concentration of the mononuclear leucocytes, for scanning electron microscopy. One drop of the cell suspension is placed in a moist chamber on a poly-l-lysine pretreated plastic coverslip (Mazia et al., J. Cell Biol. 66:198-199, 1975) and fifteen minutes allowed for cell attachment. Fixation, started in 2. 5% glutaraldehyde in culture medium at room temperature for 30 minutes, is continued in the same fixative at 4°C overnight or longer. Ethanol dehydration is immediately followed by drying at the critical point of CO2 or of Freon 13. An efficient alternative method for ethanol dehydrated cells is to dry the cells at low temperature (-75°C) under vacuum (10-2 Torr) for 30 minutes in an Edwards-Pearse freeze-dryer (de Harven et al., SEM/IITRI/1977, 519-524). This is preceded by fast quenching in supercooled ethanol (between -90 and -100°C).

Scanning Electron Microscopy-cuticular Lesions on the Roundworm Ascaris Suum

1970 ◽  
Vol 28 ◽  
pp. 114-115
Author(s):  
P. A. Madden ◽  
W. R. Anderson
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Freeze Drying ◽  
Room Temperature ◽  
Secondary Electron ◽  
Distilled Water ◽  
Freeze Dried ◽  
Silver Paint ◽  
To Come ◽  
Scanning Electron

The intestinal roundworm of swine is pinkish in color and about the diameter of a lead pencil. Adult worms, taken from parasitized swine, frequently were observed with macroscopic lesions on their cuticule. Those possessing such lesions were rinsed in distilled water, and cylindrical segments of the affected areas were removed. Some of the segments were fixed in buffered formalin before freeze-drying; others were freeze-dried immediately. Initially, specimens were quenched in liquid freon followed by immersion in liquid nitrogen. They were then placed in ampuoles in a freezer at −45C and sublimated by vacuum until dry. After the specimens appeared dry, the freezer was allowed to come to room temperature slowly while the vacuum was maintained. The dried specimens were attached to metal pegs with conductive silver paint and placed in a vacuum evaporator on a rotating tilting stage. They were then coated by evaporating an alloy of 20% palladium and 80% gold to a thickness of approximately 300 A°. The specimens were examined by secondary electron emmission in a scanning electron microscope.

Scanning and Transmission Electron Microscopy of Human Red Blood Cells

1969 ◽  
Vol 27 ◽  
pp. 44-45
Author(s):  
A.J. Tousimis ◽  
T.R. Padden
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Blood Cells ◽  
Room Temperature ◽  
Type Specimen ◽  
New Combination ◽  
Human Red Blood Cells ◽  
Transmission Electron ◽  
Microscope Slides ◽  
Scanning Electron

The size, shape and surface morphology of human erythrocytes (RBC) were examined by scanning electron microscopy (SEM), of the fixed material directly and by transmission electron microscopy (TEM) of surface replicas to compare the relative merits of these two observational procedures for this type specimen.A sample of human blood was fixed in glutaraldehyde and washed in distilled water by centrifugation. The washed RBC's were spread on freshly cleaved mica and on aluminum coated microscope slides and then air dried at room temperature. The SEM specimens were rotary coated with 150Å of 60:40- gold:palladium alloy in a vacuum evaporator using a new combination spinning and tilting device. The TEM specimens were preshadowed with platinum and then rotary coated with carbon in the same device. After stripping the RBC-Pt-C composite film, the RBC's were dissolved in 2.5N HNO3 followed by 0.2N NaOH leaving the preshadowed surface replicas showing positive topography.

Scanning Microscopy of Human Intestinal Explants in Organ Culture

1972 ◽  
Vol 30 ◽  
pp. 396-397
Author(s):  
Bruce Wetzel ◽  
Robert Buscho ◽  
Raphael Dolin
Keyword(s):  
Electron Microscopy ◽  
Room Temperature ◽  
Culture Conditions ◽  
Human Fetuses ◽  
Enteric Disease ◽  
Organ Cultures ◽  
Growth Of A Variety ◽  
Normal Human ◽  
Fetal Intestine ◽  
Scanning Electron

It has been reported that explants of human fetal intestine can be maintained in culture for up to 21 days in a viable condition and that these organ cultures support the growth of a variety of known viral agents responsible for enteric disease. Scanning electron microscopy (SEM) has been undertaken on several series of these explants to determine their appearance under routine culture conditions.Fresh specimens of jejunum obtained from normal human fetuses were washed, dissected into l-4mm pieces, and cultured in modified Leibowitz L-15 medium at 34° C as previously described. Serial specimens were fixed each day in 3% glutaraldehyde for 90 minutes at room temperature, rinsed, dehydrated, and dried by the CO2 critical point method in a Denton DCP-1 device. Specimens were attached to aluminum stubs with 3M transfer tape No. 465, and one sample on each stub was carefully rolled along the adhesive such that villi were broken off to expose their interiors.

SEM Observations on the Germination of Euonymous Americanus

1985 ◽  
Vol 43 ◽  
pp. 508-509
Author(s):  
D.R. Hill ◽  
J.R. McCurry ◽  
L.P. Elliott ◽  
G. Howard
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Light Microscopy ◽  
Filter Paper ◽  
Room Temperature ◽  
Food Source ◽  
Environmental Chamber ◽  
Dormant Stage ◽  
Scanning Electron

Germination of Euonymous americanus in the laboratory has previously been unsuccessful. Ability to germinate Euonymous americanus. commonly known as the american strawberry bush, is important in that it represents a valuable food source for the white-tailed deer. Utilizing the knowledge that its seeds spend a period of time in the rumin fluid of deer during their dormant stage, we were successful in initiating germination. After a three month drying period, the seeds were placed in 25 ml of buffered rumin fluid, pH 8 at 40°C for 48 hrs anaerobically. They were then allowed to dry at room temperature for 24 hrs, placed on moistened filter paper and enclosed within an environmental chamber. Approximately four weeks later germination was detected and verified by scanning electron microscopy; light microscopy provided inadequate resolution. An important point to note in this procedure is that scarification, which was thought to be vital for germination, proved to be unnecessary for successful germination to occur. It is believed that germination was propagated by the secretion of enzymes or prescence of acids produced by microorganisms found in the rumin fluid since sterilized rumin failed to bring about germination.

Observations of thick and thin sections in a field-emission SEM

1994 ◽  
Vol 52 ◽  
pp. 1018-1019
Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley
Keyword(s):  
Electron Microscope ◽  
Field Emission ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Chemical Fixation ◽  
Thin Sections ◽  
Transmission Electron ◽  
Conventional Transmission Electron Microscope ◽  
Scanning Electron

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.

Fibrillin molecules aggregate to form an extendible beaded structure which is a major component of “Elastin Associated” microfibrils

1989 ◽  
Vol 47 ◽  
pp. 246-247
Author(s):  
Douglas R. Keene ◽  
B. Kerry Maddox ◽  
Marie B. Spurgin ◽  
Lynn Y. Sakai ◽  
Robert W. Glanville
Keyword(s):  
Molecular Sieve ◽  
Room Temperature ◽  
Culture Medium ◽  
Weight Fraction ◽  
Ammonium Bicarbonate ◽  
High Molecular Weight Fraction ◽  
Bicarbonate Buffer ◽  
Human Amnion ◽  
Gold Conjugate ◽  
Ammonium Bicarbonate Buffer

A mouse monoclonal antibody was used to identify beaded aggregates found in guanidine extracts of human amnion as assemblies of fibrillin molecules. These aggregates were also shown to be a major component of extracellular matrix microfibrils. We further demonstrated that the periodicity of these aggregates can be increased when subjected to mechanical stress.Human amnion was extracted with guanidine and the extracted material purified using ion exchange and molecular sieve chromatography. A high molecular weight fraction was precipitated by dialyzing against dilute acetic acid. Part of the precipitate was suspended in 0.2 M ammonium bicarbonate buffer and rotary shadowed. A second portion was resuspended in culture medium containing antibody which recognizes matrix microfibrils, diluted 1:5 in ammonium bicarbonate and reacted for 120 minutes at room temperature. Antibody labeled precipitate was washed by repeated pelleting and resuspension in buffer and then incubated in Janssen GAM 5 nm gold conjugate for 60 minutes at room temperature.

A new species of Cribraria (Myxomycetes) from Australia

2018 ◽  
Vol 52 (2) ◽  
pp. 379-385 ◽  
Author(s):  
S. L. Stephenson ◽  
Yu. K. Novozhilov ◽  
P. Wellman
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
New Species ◽  
Lemon Yellow ◽  
Moist Chamber ◽  
Light Yellow ◽  
Dark Violet ◽  
The Stability ◽  
A New Species ◽  
Scanning Electron

A new species of Cribraria, described herein as C. bicolor, appeared in moist chamber cultures on samples of the bark of Eucalyptus sp. collected at two localities in Australia. The morphology of representative specimens was examined by light and scanning electron microscopy, and micrographs of relevant morphological details of sporocarps and spores are provided. The species has a number of distinct and unique morphological features, including a glossy bright-violet globose sporotheca and a two-colored long stalk which is bright-red over the lower one-third and light yellow or lemon-yellow over the upper two-thirds. The combination of these characteristics as well as a shallow calyculus which is dark-violet when viewed under a dissecting microscope and bright red in transmitted light when mounted in lactophenol makes C. bicolor a well-defined morphospecies when compared to all other species of Cribraria. The stability of the taxonomic characters of the species was confirmed by an examination of a number of specimens.

scholarly journals Scanning Electron-Assisted Dielectric Microscopy Reveals Autophagosome Formation by LC3 and ATG12 in Cultured Mammalian Cells

2021 ◽  
Vol 22 (4) ◽  
pp. 1834
Author(s):  
Tomoko Okada ◽  
Toshihiko Ogura
Keyword(s):  
Mammalian Cells ◽  
Culture Medium ◽  
Related Gene ◽  
Autophagosome Formation ◽  
Intact Cells ◽  
Microtubule Associated Proteins ◽  
Associated Proteins ◽  
The Difference ◽  
Related Proteins ◽  
Scanning Electron

Autophagy is an intracellular self-devouring system that plays a central role in cellular recycling. The formation of functional autophagosomes depends on several autophagy-related proteins, including the microtubule-associated proteins 1A/1B light chain 3 (LC3) and the conserved autophagy-related gene 12 (Atg12). We have recently developed a novel scanning electron-assisted dielectric microscope (SE-ADM) for nanoscale observations of intact cells. Here, we used the SE-ADM system to observe LC3- and Atg12-containing autophagosomes in cells labelled in the culture medium with antibodies conjugated to colloidal gold particles. We observed that, during autophagosome formation, Atg12 localized along the actin meshwork structure, whereas LC3 formed arcuate or circular alignments. Our system also showed a difference in the distribution of LC3 and Atg12; Atg12 was broadly distributed while LC3 was more localized. The difference in the spatial distribution demonstrated by our system explains the difference in the size of fluorescent spots due to the fluorescently labelled antibodies observed using optical microscopy. The direct SE-ADM observation of cells should thus be effective in analyses of autophagosome formation.

Microstructural Studies of NiCoMnIn Magnetic Shape Memory Ribbons

2013 ◽  
Vol 738-739 ◽  
pp. 436-440 ◽  
Author(s):  
Krystian Prusik ◽  
Katarzyna Bałdys ◽  
Danuta Stróż ◽  
Tomasz Goryczka ◽  
Józef Lelątko
Keyword(s):  
Melt Spinning ◽  
Room Temperature ◽  
Parent Phase ◽  
Magnetic Shape Memory ◽  
Melt Spun ◽  
Columnar Grains ◽  
Ebsd Technique ◽  
Microstructural Studies ◽  
Melt Spinning Technique ◽  
Scanning Electron

In present paper two ribbons of the Ni44Co6Mn36In14 (at.%) were prepared under different melt-spinning technique conditions. Microstructure of the ribbons was studied by scanning electron microscopy (SEM). Depending on the liquid ejection overpressure two types of ribbons microstructures were observed. Ribbon T1 for which ejection overpressure was 1.5 bar showed typical melt-spun ribbon microstructure consisting of a top layer of small equi-axial grains and columnar grains below. For T2 ribbon (ejection overpressure 0.2 bar) only a small fraction of the columnar grains were observed. Structure analysis of the ribbons performed by XRD showed that at room temperature both ribbons have B2 parent phase superstructure. No gamma phase precipitates were observed. In order to determine the orientation of the grains the EBSD technique was applied.

ROOM TEMPERATURE PHOTOLUMINESCENCE OF POLYCRYSTALLINE SIC PREPARED FROM CARBON-SATURATED SI MELT

2002 ◽  
Vol 16 (06n07) ◽  
pp. 1047-1051
Author(s):  
JIANPING MA ◽  
ZHIMING CHEN ◽  
GANG LU ◽  
MINGBIN YU ◽  
LIANMAO HANG ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Photoelectron Spectroscopy ◽  
Room Temperature ◽  
Uv Light ◽  
X Ray Diffraction ◽  
X Ray ◽  
Room Temperature Photoluminescence ◽  
Composition And Structure ◽  
Light Excitation ◽  
Scanning Electron

Intense photoluminescence (PL) has been observed at room temperature from the polycrystalline SiC samples prepared from carbon-saturated Si melt at a temperature ranging from 1500 to 1650°C. Composition and structure of the samples have been confirmed by means of X-ray photoelectron spectroscopy, X-ray diffraction and scanning electron microscopy. PL measurements with 325 nm UV light excitation revealed that the room temperature PL spectrum of the samples consists of 3 luminescent bands, the peak energies of which are 2.38 eV, 2.77 eV and 3.06 eV, respectively. The 2.38 eV band is much stronger than the others. It is suggested that some extrinsic PL mechanisms associated with defect or interface states would be responsible to the intensive PL observed at room temperature.

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