Quantum Dot-Based In Situ Hybridization and Immunohistochemistry to Detect mRNA and Protein at Subcellular Levels, Comparison with Studies Using Electron Microscopy

2015 ◽  
pp. 417-425
Author(s):  
Akira Matsuno ◽  
Yudo Ishii ◽  
Mineko Murakami ◽  
Katsumi Hoya ◽  
Shoko M. Yamada ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Quantum Dot

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

scholarly journals Detection and Confirmation of Reptilian Adenovirus Infection by in Situ Hybridization

2001 ◽  
Vol 13 (4) ◽  
pp. 365-368 ◽  
Author(s):  
Laura E. Leigh Perkins ◽  
Raymond P. Campagnoli ◽  
Barry G. Harmon ◽  
Christopher R. Gregory ◽  
W. L. Steffens ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Adenovirus Infection ◽  
Pathogenic Potential ◽  
Suitable Alternative ◽  
Viral Particles ◽  
Viral Inclusions ◽  
Diagnostic Setting ◽  
Formalin Fixed

Adenovirus infections are documented in at least 12 different species of reptiles. In contrast to their mammalian and avian counterparts reptilian adenoviruses are not well characterized as to their pathogenic potential and their ability to cause primary disease. In the diagnostic setting, fresh tissues are often not available for virus isolation, and the confirmation of reptilian adenovirus infections is dependent largely upon electron microscopy for the identification of intranuclear viral inclusions associated with histopathologic changes. The diagnosis of adenovirus infection in 2 different species of snake was confirmed by the application of DNA in situ hybridization. Using an aviadenovirus specific oligoprobe, adenoviral DNA was observed in the nuclei of hepatocytes, Kupffer cells, endothelial cells, and enterocytes. Electron microscopy of the liver confirmed the presence of intranuclear viral particles morphologically consistent with an adenovirus. DNA in situ hybridization on formalin-fixed tissues can serve as a suitable alternative to electron microscopy in the diagnosis of reptilian adenovirus infections. Both affected snakes had other concurrent diseases, suggesting that the adenovirus may not have been the primary pathogen.

scholarly journals Three-dimensional electron microscopy of ribosomal chromatin in two higher plants: a cytochemical, immunocytochemical, and in situ hybridization approach.

1991 ◽  
Vol 39 (11) ◽  
pp. 1495-1506 ◽  
Author(s):  
P M Motte ◽  
R Loppes ◽  
M Menager ◽  
R Deltour
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Spatial Organization ◽  
Higher Plants ◽  
Three Dimensional ◽  
Spatial Arrangement ◽  
Thin Sections ◽  
Cytoplasmic Dna ◽  
Immunocytochemical Techniques

We report the 3-D arrangement of DNA within the nucleolar subcomponents from two evolutionary distant higher plants, Zea mays and Sinapis alba. These species are particularly convenient to study the spatial organization of plant intranucleolar DNA, since their nucleoli have been previously reconstructed in 3-D from serial ultra-thin sections. We used the osmium ammine-B complex (a specific DNA stain) on thick sections of Lowicryl-embedded root fragments. Immunocytochemical techniques using anti-DNA antibodies and rDNA/rDNA in situ hybridization were also applied on ultra-thin sections. We showed on tilted images that the OA-B stains DNA throughout the whole thickness of the section. In addition, very low quantities of cytoplasmic DNA were stained by this complex, which is now the best DNA stain used in electron microscopy. Within the nucleoli the DNA was localized in the fibrillar centers, where large clumps of dense chromatin were also visible. In the two plant species intranucleolar chromatin forms a complex network with strands partially linked to chromosomal nucleolar-organizing regions identified by in situ hybridization. This study describes for the first time the spatial arrangement of the intranucleolar chromatin in nucleoli of higher plants using high-resolution techniques.

scholarly journals Synthesis of actin in the nonciliated bronchiolar epithelial (Clara) cells of perinatal rats demonstrated by electron microscopy and in situ hybridization.

1990 ◽  
Vol 23 (4) ◽  
pp. 453-465 ◽  
Author(s):  
JUNZO SASAKI ◽  
SADAHIRO WATANABE ◽  
TAKAKO NOMURA ◽  
SHIGETO KANDA ◽  
TOMOICHIRO YAMAAI ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Clara Cells

Ultrastructural localization of the RNA of immunodeficiency viruses using electron microscopy in situ hybridization and in vitroinfected lymphocytes

Micron ◽  
2001 ◽  
Vol 32 (6) ◽  
pp. 579-589 ◽  
Author(s):  
C. Cantó-Nogués ◽  
D. Hockley ◽  
C. Grief ◽  
S. Ranjbar ◽  
J. Bootman ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Ultrastructural Localization

Imaging of multiple mRNA targets using quantum dot based in situ hybridization and spectral deconvolution in clinical biopsies

2006 ◽  
Vol 348 (2) ◽  
pp. 628-636 ◽  
Author(s):  
Eleni Tholouli ◽  
Judith A. Hoyland ◽  
Dolores Di Vizio ◽  
Fionnuala O’Connell ◽  
Sarah A. MacDermott ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Quantum Dot ◽  
Spectral Deconvolution ◽  
Mrna Targets

in situ Hybridization in Electron Microscopy

2001 ◽  
Author(s):  
Gérard Morel ◽  
Annie Cavalier ◽  
Lynda Williams
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization
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