Molecular hybridization of RNA and DNA in situ: visualization at the electron microscope level

1976 ◽  
Vol 106 (2) ◽  
pp. 185-198 ◽  
Author(s):  
J. Jacob ◽  
M. H. Moar ◽  
Katherine Gillies ◽  
D. Macleod ◽  
K. W. Jones
2000 ◽  
Vol 6 (S2) ◽  
pp. 328-329
Author(s):  
J.M. Robinson ◽  
T. Takizawa ◽  
D.D. Vandré

Immunocytochemistry generally refers to methods directed toward obtaining information on the in situ distribution of antigens in cells and tissues. Immunocytochemical methods can be applied at either the light or electron microscope levels, or both in concert. The detection of antibody recognition of an antigen (i.e., localization of an antigen) relies upon a reporter system. At the light microscope level, enzymes (e.g., horseradish peroxidase) or fluorochromes are the most widely used reporters in immunocytochemistry. At the electron microscope level, particulate probes (e.g., colloidal gold) are the most widely used reporters. However, enzymes and even fluorochromes can be used at the EM level.In this abstract, we discuss our use of gold cluster immunoprobes as the reporter system in both light and electron microscope level immuncytochemistry. These gold cluster immunoprobes, are commercially known as NanogoldTM (NG). These probes are very small with a diameter of 1.4-nm.


1986 ◽  
Vol 102 (5) ◽  
pp. 1646-1653 ◽  
Author(s):  
M Binder ◽  
S Tourmente ◽  
J Roth ◽  
M Renaud ◽  
W J Gehring

A technique has been developed for localizing hybrids formed in situ on semi-thin and ultrathin sections of Lowicryl K4M-embedded tissue. Biotinylated dUTP (Bio-11-dUTP and/or Bio-16-dUTP) was incorporated into mitochondrial rDNA and small nuclear U1 probes by nick-translation. The probes were hybridized to sections of Drosophila ovaries and subsequently detected with an anti-biotin antibody and protein A-gold complex. On semi-thin sections, probe detection was achieved by amplification steps with anti-protein A antibody and protein A-gold with subsequent silver enhancement. At the electron microscope level, specific labeling was obtained over structures known to be the site of expression of the appropriate genes (i.e., either over mitochondria or over nuclei). The labeling pattern at the light microscope level (semi-thin sections) was consistent with that obtained at the electron microscope level. The described nonradioactive procedures for hybrid detection on Lowicryl K4M-embedded tissue sections offer several advantages: rapid signal detection: superior morphological preservation and spatial resolution; and signal-to-noise ratios equivalent to radiolabeling.


IAWA Journal ◽  
1981 ◽  
Vol 2 (4) ◽  
pp. 151-162 ◽  
Author(s):  
A.M. Catesson ◽  
J.C. Roland

Cytochemical techniques and mild extractions were used at the electron microscope level for the study of the cambial zone of several hardwoods and one softwood. The maturation processes of the primary radial and tangential cell walls involve a progressive disappearance of their initial heterogeneity. The buttress-like zone joining these walls appears to be the starting point for a characteristic sequence of changes and intra-wall rearrangement. Topochemical results have suggested an alternative to the 'emboxing concept' of cell wall development.


1968 ◽  
Vol 16 (3) ◽  
pp. 162-171 ◽  
Author(s):  
GUILLERMO JAIM ETCHEVERRY ◽  
LUIS MARIA ZIEHER

Blood platelets obtained from normal rabbits and those isolated from reserpine-treated animals and subsequently incubated in vitro with 5-hydroxytryptamine, norepinephrine and histamine were assayed for amine content or processed for examination under the electron microscope. With the glutaraldehyde-dichromate reaction for unsubstituted catechol- and indoleamines, reactive granules were observed in normal platelets. Formaldehyde fixation prior to the glutaraldehyde-dichromate reaction resulted in a similar image under the electron microscope. In platelets obtained from animals treated with reserpine a decrease of the amine content with a corresponding reduction in the number of dense granules was observed. Following incubation with 5-hydroxytryptamine the concentration of the amine increased markedly and the number of dense granules that reacted with both techniques became practically normal. In norepinephrine-incubated platelets dense granules were demonstrated with the glutaraldehyde-dichromate reaction, but no reactive products were observed using prefixation with formaldehyde. Histamine was also incorporated into depleted platelets but gave no reaction. It is concluded that prefixation with formaldehyde renders negative the reaction with catecholamines, leaving unaffected indoleamine-reactive sites. The previous assumption that the dense granules contain 5-hydroxytryptamine has been confirmed by such a cytochemical approach. The possibility that these organelles constitute a common storage site for different amines is discussed.


2003 ◽  
Vol 81 (7) ◽  
pp. 732-737 ◽  
Author(s):  
Narumol Plaingam ◽  
Sayanh Somrithipol ◽  
E B. Gareth Jones

The genus Infundibulomyces is proposed for coelomycetous fungi with nidulariaceous-like conidiomata and bipolar appendaged conidia. Infundibulomyces cupulata sp. nov., collected from fallen leaves of Lagerstroemia sp. from Thailand, is described as the type species of the genus. This fungus is illustrated at the light microscope and scanning electron microscope level and compared with other morphologically related taxa. Cupulate conidiomata are uncommon and Infundibulomyces is compared with these.Key words: anamorphic fungi, coelomycete, Infundibulomyces, Thailand, tropical.


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