scholarly journals Electron microscopic localization of cytoplasmic myosin with ferritin-labeled antibodies.

1981 ◽  
Vol 88 (2) ◽  
pp. 346-351 ◽  
Author(s):  
I M Herman ◽  
T D Pollard
Keyword(s):  
Electron Microscopy ◽  
Light Microscope ◽  
Fluorescent Antibody ◽  
Stress Fibers ◽  
Electron Microscopic ◽  
Cytoplasmic Matrix ◽  
Light Microscope Level ◽  
Antibody Staining ◽  
Terminal Web ◽  
Myosin Filaments

We localized myosin in vertebrate nonmuscle cells by electron microscopy using purified antibodies coupled with ferritin. Native and formaldehyde-fixed filaments of purified platelet myosin filaments each consisting of approximately 30 myosin molecules bound an equivalent number of ferritin-antimyosin conjugates. In preparations of crude platelet actomyosin, the ferritin-antimyosin bound exclusively to similar short, 10-15 nm wide filaments. In both cases, binding of the ferritin-antimyosin to the myosin filaments was blocked by preincubation with unlabeled antimyosin. With indirect fluorescent antibody staining at the light microscope level, we found that the ferritin-antimyosin and unlabeled antimyosin stained HeLa cells identically, with the antibodies concentrated in 0.5-microns spots along stress fibers. By electron microscopy, we found that the concentration of ferritin-antimyosin in the dense regions of stress fibers was five to six times that in the intervening less dense regions, 20 times that in the cytoplasmic matrix, and 100 times that in the nucleus. These concentration differences may account for the light microscope antibody staining pattern of spread interphase cells. Some, but certainly not all, of the ferritin-antimyosin was associated with 10-15-nm filaments. In mouse intestinal epithelial cells, ferritin-antimyosin was located almost exclusively in the terminal web. In isolated brush borders exposed to 5 mM MgCl2, ferritin-antimyosin was also concentrated in the terminal web associated with 10-15-nm filaments.

Fine structure and possible functions of the hermaphrodite duct of some pulmonate snails (Mollusca: Gastropoda)

1990 ◽  
Vol 48 (3) ◽  
pp. 68-69
Author(s):  
Alan N. Hodgson
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Seminal Vesicle ◽  
Reproductive Biology ◽  
Light Microscope ◽  
Light Microscope Level ◽  
Transmission Electron ◽  
Standard Techniques

The hermaphrodite duct of pulmonate snails connects the ovotestis to the fertilization pouch. The duct is typically divided into three zones; aproximal duct which leaves the ovotestis, the middle duct (seminal vesicle) and the distal ovotestis duct. The seminal vesicle forms the major portion of the duct and is thought to store sperm prior to copulation. In addition the duct may also play a role in sperm maturation and degredation. Although the structure of the seminal vesicle has been described for a number of snails at the light microscope level there appear to be only two descriptions of the ultrastructure of this tissue. Clearly if the role of the hermaphrodite duct in the reproductive biology of pulmonatesis to be understood, knowledge of its fine structure is required.Hermaphrodite ducts, both containing and lacking sperm, of species of the terrestrial pulmonate genera Sphincterochila, Levantina, and Helix and the marine pulmonate genus Siphonaria were prepared for transmission electron microscopy by standard techniques.

Bridging the Resolution Gap: Correlated 3D Light and Electron Microscopic Analysis of Large Biological Structures

1999 ◽  
Vol 5 (S2) ◽  
pp. 526-527
Author(s):  
Maryann E. Martone
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Surface Area ◽  
Light Microscope ◽  
Microscopic Analysis ◽  
Electron Microscopic Analysis ◽  
Electron Microscopic ◽  
Transmission Electron ◽  
Biological Structures ◽  
Ganglion Neurons

One class of biological structures that has always presented special difficulties to scientists interested in quantitative analysis is comprised of extended structures that possess fine structural features. Examples of these structures include neuronal spiny dendrites and organelles such as the Golgi apparatus and endoplasmic reticulum. Such structures may extend 10's or even 100's of microns, a size range best visualized with the light microscope, yet possess fine structural detail on the order of nanometers that require the electron microscope to resolve. Quantitative information, such as surface area, volume and the micro-distribution of cellular constituents, is often required for the development of accurate structural models of cells and organelle systems and for assessing and characterizing changes due to experimental manipulation. Performing estimates of such quantities from light microscopic data can result in gross inaccuracies because the contribution to total morphometries of delicate features such as membrane undulations and excrescences can be quite significant. For example, in a recent study by Shoop et al, electron microscopic analysis of cultured chick ciliary ganglion neurons showed that spiny projections from the plasmalemma that were not well resolved in the light microscope effectively doubled the surface area of these neurons.While the resolution provided by the electron microscope has yet to be matched or replaced by light microscopic methods, one drawback of electron microscopic analysis has always been the relatively small sample size and limited 3D information that can be obtained from samples prepared for conventional transmission electron microscopy. Reconstruction from serial electron micrographs has provided one way to circumvent this latter problem, but remains one of the most technically demanding skills in electron microscopy. Another approach to 3D electron microscopic imaging is high voltage electron microscopy (HVEM). The greater accelerating voltages of HVEM's allows for the use of much thicker specimens than conventional transmission electron microscopes.

scholarly journals Physical, immunochemical, and functional properties of Acanthamoeba profilin.

1984 ◽  
Vol 98 (1) ◽  
pp. 214-221 ◽  
Author(s):  
P C Tseng ◽  
M S Runge ◽  
J A Cooper ◽  
R C Williams ◽  
T D Pollard
Keyword(s):  
Fluorescent Antibody ◽  
Deae Cellulose ◽  
Exchange Chromatography ◽  
Sedimentation Equilibrium ◽  
Binding Assays ◽  
Cytoplasmic Matrix ◽  
Unbound Fraction ◽  
Antibody Staining ◽  
High Concentrations ◽  
Equilibrium Ultracentrifugation

Acanthamoebe profilin has a native molecular weight of 11,700 as measured by sedimentation equilibrium ultracentrifugation and an extinction coefficient at 280 nm of 1.4 X 10(4) M-1cm-1. Rabbit antibodies against Acanthamoeba profilin react only with the 11,700 Mr polypeptide among all other ameba polypeptides separated by electrophoresis. These antibodies react with a 11,700 Mr polypeptide in Physarum but not with any proteins of Dictyostelium or Naeglaria. Antibody-binding assays indicate that approximately 2% of the ameba protein is profilin and that the concentration of profilin is approximately 100 mumol/liter cells. During ion exchange chromatography of soluble extracts of Acanthamoeba on DEAE-cellulose, the immunoreactive profilin splits into two fractions: an unbound fraction previously identified by Reichstein and Korn (1979, J. Biol. Chem., 254:6174-6179) and a tightly bound fraction. Purified profilin from the two fractions is identical by all criteria tested. The tightly bound fraction is likely to be attached indirectly to the DEAE, perhaps by association with actin. By fluorescent antibody staining, profilin is distributed uniformly throughout the cytoplasmic matrix of Acanthamoeba. In 50 mM KCl, high concentrations of Acanthamoeba profilin inhibit the elongation rate of muscle actin filaments measured directly by electron microscopy, but the effect is minimal in KCl with 2 MgCl2. By using the fluorescence change of pyrene-labeled Acanthamoeba actin to assay for polymerization, we confirmed our earlier observation (Tseng, P. C.-H., and T. D. Pollard, 1982, J. Cell Biol. 94:213-218) that Acanthamoeba profilin inhibits nucleation much more strongly than elongation under physiological conditions.

Cytology and ultrastructure of Thanatephorus cucumeris and related taxa of the Rhizoctonia complex

1977 ◽  
Vol 55 (18) ◽  
pp. 2419-2436 ◽  
Author(s):  
C. C. Tu ◽  
James W. Kimbrough ◽  
H. C. Aldrich
Keyword(s):  
Electron Microscopy ◽  
Light Microscope ◽  
Light And Electron Microscopy ◽  
Middle Layer ◽  
Ultrastructural Level ◽  
Aniline Blue ◽  
Diploid Nucleus ◽  
Thanatephorus Cucumeris ◽  
Light Microscope Level ◽  
Endoplasmic Reticula

Cytological studies on the vegetative hyphae of members of the Rhizoctonia complex and basidial structures of Thanatephorus cucumeris were performed with light and electron microscopy. Vegetative cells of Thanatephorus and Waitea proved to be multinucleate, whereas those of Uthatobasidium, Ceratobasidium, Athelia. and Botryobasidium are binucleate.Dolipore septa of Thanatephorus, Waitea, Uthatobasidium, and Ceratobasidium are visible with the light microscope when stained with aniline blue in glycerine. Ultrastructurally, pore caps in these genera consisted of two-layered unit membranes, forming cisternae with an electron-dense middle layer. Dolipore septa of Athelia (S. rolfsii) and Botryobasidium are not visible in aniline blue at the light microscope level. At the ultrastructural level, there was an additional cisternal membrane making up a pore cap of three membranes. The fine structure of nuclei, mitochondria, endoplasmic reticula, vacuoles, and other organelles in the basidial structures of T. cucumeris was essentially the same as in other basidiomycetes.Karyogamy of two haploid nuclei occurs in the young basidia of T. cucumeris. The nuclear envelopes of both haploid nuclei break at their adjacent sides and fuse to form a diploid nucleus. After a short interphase, meiosis occurs. No leptotene was observed at prophase I, but a synaptinemal complex was evident and six pairs of chromosomes were observed throughout pachytene, diplotene, and diakinesis. The nuclear envelope disappears at metaphase I and a spindle appears. The second meiotic division is equational. Most of the mature and discharged spores are uninucleate.

scholarly journals ELECTRON MICROSCOPIC LOCALIZATION OF THE NEPHROTOXIC ANTIBODY IN THE GLOMERULI OF THE RAT AFTER INTRAVENOUS APPLICATION OF PURIFIED NEPHRITOGENIC ANTIBODY-FERRITIN CONJUGATES

1968 ◽  
Vol 127 (5) ◽  
pp. 867-878 ◽  
Author(s):  
Arnold Vogt ◽  
Hermann Bockhorn ◽  
Keniti Kozima ◽  
Masamichi Sasaki
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Intravenous Injection ◽  
Fluorescent Antibody ◽  
Fluorescent Antibody Technique ◽  
Electron Microscopic ◽  
Antibody Technique ◽  
Disappearance Time ◽  
Filtration Surface ◽  
Electron Microscopic Localization

Nephritis in rats was induced by intravenous injection of purified ferritin-conjugated rabbit and duck nephrotoxic globulin. Using the fluorescent antibody technique, the same capillary pattern was found as that in glomeruli of rats receiving uncoupled nephrotoxic globulin. Electron microscopy revealed a heavy accumulation of the basement membrane-fixed antibody almost exclusively at the endothelial side. A higher concentration of ferritin was demonstrable in the peripheral basement membrane. The once-fixed antibody remained at the site of reaction though decreasing with time. The half-disappearance time seemed to be shorter than that of the uncoupled nephrotoxic globulin. No difference in localization was observed between rabbit and duck antibody. At least 40 basement membrane-fixed antibody molecules from the rabbit per 3000 mµ2 of filtration surface were needed to cause immediate nephritis. To induce nephritis using duck antibody, a greater number of basement membrane-fixed antibody seemed to be necessary. No evidence of specific reaction with constituents of glomerular cells was obtained.

scholarly journals Comparative Study of Experimental Inclusion Body Hepatitis of Chickens Caused by Two Serotypes of Avian Adenovirus

1978 ◽  
Vol 15 (2) ◽  
pp. 249-263 ◽  
Author(s):  
T. M. Grimes ◽  
O. J. Fletcher ◽  
J. F. Munnell
Keyword(s):  
Inclusion Body ◽  
Hepatitis Virus ◽  
Fluorescent Antibody ◽  
Bursa Of Fabricius ◽  
Electron Microscopic ◽  
Avian Adenovirus ◽  
Antibody Staining ◽  
Inclusion Body Hepatitis ◽  
Significant Depression ◽  
Microscopic Studies

Twenty 1-day-old specific-pathogen-free chickens each were given an intraabdominal inoculation of either a type-8 avian adenovirus, [AMG 5 (2a)], or a type-5 avian adenovirus, inclusion body hepatitis virus (IBHV). The diseases produced were similar. High (60–100%) mortality and statistically significant depression of body weights occurred in both infections. There were necrotizing hepatitis and pancreatitis, lymphoid depletion in the spleen, bursa of Fabricius and thymus, hydropericardium, nephritis and enteritis. Intranuclear inclusions occurred in affected organs. Fluorescent-antibody staining, the Feulgen reaction for deoxyribonucleic acid and electron microscopic studies, as well as studies from the literature, indicated that basophilic inclusions consisted of assembled adenovirions.

scholarly journals Chlamydia pneumoniae in a Free-Ranging Giant Barred Frog (Mixophyes iteratus) from Australia

1999 ◽  
Vol 37 (7) ◽  
pp. 2378-2380 ◽  
Author(s):  
Lee Berger ◽  
Kym Volp ◽  
Sarah Mathews ◽  
Rick Speare ◽  
Peter Timms
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
16S Rrna ◽  
Chlamydia Pneumoniae ◽  
Fluorescent Antibody ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Antibody Staining ◽  
Free Ranging ◽  
Fluorescent Antibody Staining

The koala biovar of Chlamydia pneumoniae was identified in lung tissue from a sick, free-ranging giant barred frog (Mixophyes iteratus) by using electron microscopy, C. pneumoniae-specific fluorescent-antibody staining, cell culture, and sequencing of the ompA, ompB and 16S rRNA genes. This is the first report of a chlamydial strain infecting both a homeotherm and a poikilotherm and only the fourth host (in addition to humans, koalas, and horses) to be naturally infected with this species of Chlamydia. The frog had severe, chronic, mononuclear pneumonia and nonregenerative anemia and pancytopenia.

Selective Staining of Cortical Granules and Golgi Cisternae in Arbacia Punctulata

1969 ◽  
Vol 27 ◽  
pp. 286-287
Author(s):  
Arya K. Bal ◽  
Gilles H. Cousineau
Keyword(s):  
Electron Microscopy ◽  
Light Microscope ◽  
Phosphotungstic Acid ◽  
Dense Material ◽  
Cortical Granules ◽  
Light Microscope Level ◽  
Electron Dense Material ◽  
Selective Staining ◽  
Arbacia Punctulata ◽  
Staining Techniques

Cyto-chemical staining techniques at the light microscope level have revealed the presence of mucopolysaccharides and proteins in the cortical granules of Eichinoderm eggs. In routine electron microscopy preparation the cortical granules appear to have two morphologically distinct components - an electron dense inner component (dark bodies) surrounded by a less-electron dense material. In the present investigation it has been made possible to stain the dense inner material selectively with Phosphotungstic acid (PTA) in non-osmicated aldehyde fixed oocytes and eggs of Arbacia punctulata.

scholarly journals ELECTRON MICROSCOPIC STUDIES OF EXPERIMENTAL NEPHRITIS WITH FERRITIN-CONJUGATED ANTIBODY

1962 ◽  
Vol 115 (5) ◽  
pp. 929-936 ◽  
Author(s):  
Giuseppe A. Andres ◽  
Councilman Morgan ◽  
Konrad C. Hsu ◽  
Richard A. Rifkind ◽  
Beatrice C. Seegal
Keyword(s):  
Electron Microscopy ◽  
Basement Membrane ◽  
Rat Kidney ◽  
Fluorescent Antibody ◽  
Ultrastructural Level ◽  
Fluorescent Antibody Technique ◽  
Electron Microscopic ◽  
Antibody Technique ◽  
Experimental Nephritis ◽  
Microscopic Studies

Ferritin-conjugated antibody has been used to identify by electron microscopy the sites at which nephrotoxic globulins localize in rat kidney during acute experimental glomerulonephritis. Antibody was concentrated in the glomerular basement membrane and in basement membrane-like material contained in distended cisternae of the endoplasmic reticulum. These data confirm and amplify, at the ultrastructural level, the results of studies obtained with the fluorescent antibody technique, and are consistent with the hypothesis that the cisternae and capillary basement membrane possess common proteins.

Harpella leptosa, a new species of Trichomycetes substantiated by electron microscopy

1980 ◽  
Vol 58 (9) ◽  
pp. 1035-1044 ◽  
Author(s):  
Stephen T. Moss ◽  
Robert W. Lichtwardt
Keyword(s):  
Electron Microscopy ◽  
New Species ◽  
Rocky Mountains ◽  
Light Microscope ◽  
Electron Microscopic ◽  
Northern Rocky Mountains ◽  
Microscopic Studies ◽  
Blackfly Larvae ◽  
A New Species

Harpella leptosa (Harpellales, Harpellaceae) from the midget region of blackfly larvae (Simuliidae) is described. With the light microscope it is distinguished from the only known species of the genus, H. melusinae, primarily by its smaller dimensions and less conspicuous holdfast, but electron microscopic studies reveal significant differences in the substructure of the trichospore appendages and holdfast apparatus. Harpella leptosa is presently known only from the northern Rocky Mountains. The two species of Harpella are sympatric, and occasionally live simultaneously in the same host gut.

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