Field Emission Scanning Electron Microscopy Of Mouse Cerebellar Synaptic Contacts

2000 ◽  
Vol 6 (S2) ◽  
pp. 844-845
Author(s):  
O.J. Castejón ◽  
R. P. Apkarian ◽  
H. V. Castejón

Samples of albino mice cerebellar cortex were processed by the cryofracture method for scanning electron microscopy and examined with the field emission scanning electron microscope (FESEM). Albino mouse cerebellar cortex was excised, cut into 1-2 mm slices and inmersed in 4% glutaraldehyde in O. l M phosphate buffer solution, pH 7.4, for 24h at 4°C; and postfixed for 1 h in a similarly buffered 1% osmium tetroxide solution. Specimens were dehydrated in a graded serie of ethanol (30, 50, 70, 80, 90 2x100%) prior to wrapping individual tissue pieces in preformed absolute ethanol filled parafilm cryofracture packets. Rapid freezing of packets was performed by plunging into LN2. First, the packet was transferred from the LN2 storage vessel with LNT chilled forceps in order to avoid themial damage. Secondly, the cooled fracture blade was removed from the LN2, the packet was orientated under the blade, and immediately struck with a heavy tool.

Author(s):  
Yasuaki Hotta ◽  
Masumi Nozaki ◽  
Nobuhiko Honda ◽  
Hiroyuki Kato ◽  
Ying Wei Wang ◽  
...  

Recently, the validity of microwave irradiation (MWI) for electron microscopy has attracted special interest especially in Japan. Recently, we developed a new maceration method for scanning electron microscopy (microwave maceration) and a rapid polymerization method for tissue embedding using epoxy resin (microwave polymerization). Concerning the tissue fixations using MWI (microwave fixation), there are many problems which are not yet clear. In this study, we intend to reveal the effectiveness and mechanism of microwave fixation for scanning and transmission electron mi c roscopy.Liver and trachea taken from Wistar strain male rats were used in this experiment. The fresh livers and tracheae were cut into small pieces and microwave irradiated using a microwave processor (H2500, Bio-Rad) for 1 or 3 min in 2.5% g1utara 1dehyde (GL) bufferized fixatives regulated at pH 7.3 with 0.1M phosphate buffer solution. To prevent the rising of temperature during MWI, specially designed metal case containing water and ice was used (Fig.1).


Author(s):  
Ronald H. Bradley ◽  
R. S. Berk ◽  
L. D. Hazlett

The nude mouse is a hairless mutant (homozygous for the mutation nude, nu/nu), which is born lacking a thymus and possesses a severe defect in cellular immunity. Spontaneous unilateral cataractous lesions were noted (during ocular examination using a stereomicroscope at 40X) in 14 of a series of 60 animals (20%). This transmission and scanning microscopic study characterizes the morphology of this cataract and contrasts these data with normal nude mouse lens.All animals were sacrificed by an ether overdose. Eyes were enucleated and immersed in a mixed fixative (1% osmium tetroxide and 6% glutaraldehyde in Sorenson's phosphate buffer pH 7.4 at 0-4°C) for 3 hours, dehydrated in graded ethanols and embedded in Epon-Araldite for transmission microscopy. Specimens for scanning electron microscopy were fixed similarly, dehydrated in graded ethanols, then to graded changes of Freon 113 and ethanol to 100% Freon 113 and critically point dried in a Bomar critical point dryer using Freon 13 as the transition fluid.


Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).


Author(s):  
Yasushi Kokubo ◽  
Hirotami Koike ◽  
Teruo Someya

One of the advantages of scanning electron microscopy is the capability for processing the image contrast, i.e., the image processing technique. Crewe et al were the first to apply this technique to a field emission scanning microscope and show images of individual atoms. They obtained a contrast which depended exclusively on the atomic numbers of specimen elements (Zcontrast), by displaying the images treated with the intensity ratio of elastically scattered to inelastically scattered electrons. The elastic scattering electrons were extracted by a solid detector and inelastic scattering electrons by an energy analyzer. We noted, however, that there is a possibility of the same contrast being obtained only by using an annular-type solid detector consisting of multiple concentric detector elements.


2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Buzuayehu Abebe ◽  
Enyew Amare Zereffa ◽  
Aschalew Tadesse ◽  
H. C. Ananda Murthy

Abstract Metal oxide nanomaterials are one of the preferences as antibacterial active materials. Due to its distinctive electronic configuration and suitable properties, ZnO is one of the novel antibacterial active materials. Nowadays, researchers are making a serious effort to improve the antibacterial activities of ZnO by forming a composite with the same/different bandgap semiconductor materials and doping of ions. Applying capping agents such as polymers and plant extract that control the morphology and size of the nanomaterials and optimizing different conditions also enhance the antibacterial activity. Forming a nanocomposite and doping reduces the electron/hole recombination, increases the surface area to volume ratio, and also improves the stability towards dissolution and corrosion. The release of antimicrobial ions, electrostatic interaction, reactive oxygen species (ROS) generations are the crucial antibacterial activity mechanism. This review also presents a detailed discussion of the antibacterial activity improvement of ZnO by forming a composite, doping, and optimizing different conditions. The morphological analysis using scanning electron microscopy, field emission-scanning electron microscopy, field-emission transmission electron microscopy, fluorescence microscopy, and confocal microscopy can confirm the antibacterial activity and also supports for developing a satisfactory mechanism. Graphical abstract Graphical abstract showing the metal oxides antibacterial mechanism and the fluorescence and scanning electron microscopic images.


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.


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