Effect of aldehyde fixatives on the adenosine phosphatases and ultrastructure of Vitreoscilla

1973 ◽  
Vol 19 (9) ◽  
pp. 1059-1064 ◽  
Author(s):  
Jeffrey C. Burnham ◽  
George J. Hageage Jr.
Keyword(s):  
Enzyme Inhibition ◽  
Microscopic Examination ◽  
Adenosine Triphosphatase ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Inhibitory Effect ◽  
Electron Microscopic ◽  
Adenosine Triphosphatase Activity ◽  
Enzymes Inhibition ◽  
Adenosine Phosphate

A variety of aldehydes used in electron cytochemistry including glutaraldehyde, glycidaldehyde, acetaldehyde, and methanol-free formaldehyde were examined for their inhibitory effect on the adenosine phosphate hydrolases of Vitreoscilla species. Enzyme inhibition increased with increasing aldehyde concentration. Of the aldehydes tested glycidaldehyde and acetaldehyde were least inhibitory for both adenosine diphosphatase and adenosine triphosphatase activity. Significantly, a 1% concentration of glutaraldehyde inhibited over 80% of the activity of both enzymes. Inhibition by all fixatives was variably decreased by the addition of 8% sucrose. Electron-microscopic examination of Vitreoscilla species fixed with the various aldehydes revealed that both acetaldehyde and glycidaldehyde followed by osmium tetroxide postfixation gave results comparable with glutaraldehyde–osmium tetroxide fixed cells.

scholarly journals Light and Electron Microscopic Examination of Exocrine Pancreas Using Zinc Iodide-Osmium Tetroxide Technique

1998 ◽  
Vol 75 (5) ◽  
pp. 247-250
Author(s):  
Pergin ATILLA ◽  
Murat AKKUS ◽  
Engin DEVECI ◽  
Yilmaz BILGIN ◽  
Serap INAL^|^Ouml;Z ◽  
...  
Keyword(s):  
Microscopic Examination ◽  
Exocrine Pancreas ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Zinc Iodide

Properties of Isolated Mitochondria of Agaricus Bisporus: Their Relationship to Dormancy and the Germination of Spores

1973 ◽  
Vol 31 ◽  
pp. 458-459
Author(s):  
K. S. McCarty ◽  
R. F. Weave ◽  
L. Kemper ◽  
F. S. Vogel
Keyword(s):  
Mitochondrial Dna ◽  
Ethidium Bromide ◽  
Microscopic Examination ◽  
Agaricus Bisporus ◽  
Osmotic Shock ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Isolated Mitochondria ◽  
Dna Strands ◽  
Cytoplasmic Ribosomes

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.

Electron Microscopic Examination of a Transplantable Rat Mammary Carcinoma

1968 ◽  
Vol 26 ◽  
pp. 20-21
Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher
Keyword(s):  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Undifferentiated Carcinoma ◽  
Uranyl Acetate ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
North West ◽  
X Irradiation ◽  
Tissue Specimens

A transplantable carcinoma was established in North West Sprague Dawley (NWSD) rats by use of X-irradiation by Engle and Spencer. The tumor was passaged through 63 generations over a period of 32 months. The original tumor, an adenocarcinoma, changed into an undifferentiated carcinoma following the 19th transplant. The tumor grew well in NWSD rats of either sex at various ages. It was invariably fatal, causing death of the host within 15 to 35 days following transplantation.Tumor, thymus, spleen, and plasma from 7 rats receiving transplants of tumor at 3 to 9 weeks of age were examined with an electron microscope at intervals of 8, 15, 22 and 30 days after transplantation. Four normal control rats of the same age were also examined. The tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and embedded in Epon. The plasma was separated from heparanized blood and processed as previously described for the tissue specimens. Sections were stained with uranyl acetate followed by lead citrate and examined with an RCA EMU-3G electron microscope.

Artifact-free electron microscopic immunocytochemistry on rat brain tissue

1991 ◽  
Vol 49 ◽  
pp. 272-273
Author(s):  
Veronika Burmeister ◽  
N. Ludvig ◽  
P.C. Jobe
Keyword(s):  
Microscopic Examination ◽  
Free Electron ◽  
Electron Microscopic Examination ◽  
Ultrastructural Localization ◽  
Rabbit Serum ◽  
Electron Microscopic ◽  
Electron Microscopic Immunocytochemistry ◽  
Phosphate Buffered Saline ◽  
Rat Brain Tissue ◽  
Immune Rabbit

Electron microscopic immunocytochemistry provides an important tool to determine the ultrastructural distribution of various molecules in both normal and pathologic tissues. However, the specific immunostaining may be obscured by artifactual immunoreaction product, misleading the investigator. Previous observations show that shortening the incubation period with the primary antibody from the generally used 12-24 hours to 1 hour substantially reduces the artifactual immunostaining. We now extend this finding by the demonstration of artifact-free ultrastructural localization of the Ca2/calmodulindependent cyclic nucleotide phosphodiesterase (CaM-dependent PDE) immunoreactivity in brain.Anesthetized rats were perfused transcardially with phosphate-buffered saline followed by a fixative containing paraformaldehyde (4%) and glutaraldehyde (0.25%) in PBS. The brains were removed, and 40μm sections were cut with a vibratome. The sections were processed for immunocytochemistry as described by Ludvig et al. Both non-immune rabbit serum and specific CaM-dependent PDE antibodies were used. In both experiments incubations were at one hour and overnight. The immunostained sections were processed for electron microscopic examination.

Ultrastructural studies of Chlamydia psittaci 6BC variant strains. I. Ultrastructure of the surface layers of egg-passaged 6BC strain

1973 ◽  
Vol 19 (8) ◽  
pp. 887-894
Author(s):  
Linda Poffenroth ◽  
J. W. Costerton ◽  
Nonna Kordová ◽  
John C. Wilt
Keyword(s):  
Chick Embryo ◽  
Microscopic Examination ◽  
Electron Microscopic Examination ◽  
Chlamydia Psittaci ◽  
Gram Negative Bacteria ◽  
Surface Layers ◽  
Electron Microscopic ◽  
Gram Negative ◽  
Ultrastructural Studies ◽  
First Time

Electron microscopic examination of a semipurified Chlamydia psittaci 6BC strain attenuated in chick embryo yolk sac revealed for the first time two morphologically distinct small elementary bodies which differ both in the ultrastructure of their surface layers and in their buoyant densities in sucrose gradients. Also, the morphology of the surface layers of the larger reticulate forms in cell-free systems is described for the first time. Many points of difference between the surface envelopes and internal structure of chlamydial particles and those of Gram-negative bacteria are discussed.

scholarly journals PROBLEMS ASSOCIATED WITH THE DEMONSTRATION BY LEAD METHODS OF ADENOSINE TRIPHOSPHATASE ACTIVITY IN RESIDENT PERITONEAL MACROPHAGES AND EXUDATE MONOCYTES OF THE GUINEA PIG

1973 ◽  
Vol 21 (5) ◽  
pp. 488-498 ◽  
Author(s):  
R. E. POELMANN ◽  
W. T. DAEMS ◽  
E. J. VAN LOHUIZEN
Keyword(s):  
Plasma Membrane ◽  
Guinea Pig ◽  
Microscopic Study ◽  
Heterogeneous Nucleation ◽  
Adenosine Triphosphatase ◽  
Peritoneal Macrophages ◽  
Electron Microscopic ◽  
Adenosine Triphosphatase Activity ◽  
Eosinophilic Granulocytes ◽  
Membrane Bound

This cytochemical and electron microscopic study on peritoneal macrophages of the guinea pig has raised doubts concerning the validity of lead methods for the demonstration of plasma membrane-bound adenosine triphosphatase activity. The problems encountered are inherent in the use of lead ions as a capture reagent. The nonenzymatically formed precipitates reflect sites of heterogeneous nucleation specific for certain kinds of cells, e.g., resident peritoneal macrophages, eosinophilic granulocytes and, to a lesser degree, exudate monocytes. This type of precipitation is also catalyzed on the surface of nonbiologic matrices such as latex particles. Enzymatic processes may well occur, but they cannot be distinguished from nonenzymatic processes.

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