Localization and Anchorage of Maternal mRNAs to Cortical Structures of Ascidian Eggs and Embryos Using High Resolution In Situ Hybridization

2011 ◽  
pp. 49-70 ◽  
Author(s):  
Alexandre Paix ◽  
Janet Chenevert ◽  
Christian Sardet
Keyword(s):  
In Situ Hybridization ◽  
High Resolution ◽  
Maternal Mrnas

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.

An automatic analysis method for in situ hybridization using high-resolution image analysis

1989 ◽  
Vol 281 (5) ◽  
pp. 336-341 ◽  
Author(s):  
W. Stolz ◽  
K. Scharffetter ◽  
W. Abmayr ◽  
W. K�ditz ◽  
T. Krieg
Keyword(s):  
Image Analysis ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Automatic Analysis ◽  
Analysis Method ◽  
Resolution Image ◽  
High Resolution Image

scholarly journals Ultrastructural visualization of cytoskeletal mRNAs and their associated proteins using double-label in situ hybridization.

1989 ◽  
Vol 108 (6) ◽  
pp. 2343-2353 ◽  
Author(s):  
R H Singer ◽  
G L Langevin ◽  
J B Lawrence
Keyword(s):  
In Situ Hybridization ◽  
High Resolution ◽  
Colloidal Gold ◽  
Messenger Rna ◽  
Signal To Noise Ratio ◽  
Simultaneous Detection ◽  
Gold Particles ◽  
Rna Molecules ◽  
Nucleic Acid Molecule

We have been able to visualize cytoskeletal messenger RNA molecules at high resolution using nonisotopic in situ hybridization followed by whole-mount electron microscopy. Biotinated cDNA probes for actin, tubulin, or vimentin mRNAs were hybridized to Triton-extracted chicken embryo fibroblasts and myoblasts. The cells were then exposed to antibodies against biotin followed by colloidal gold-conjugated antibodies and then critical-point dried. Identification of mRNA was possible using a probe fragmented to small sizes such that hybridization of several probe fragments along the mRNA was detected as a string of colloidal gold particles qualitatively and quantitatively distinguishable from nonspecific background. Extensive analysis showed that when eight gold particles were seen in this iterated array, the signal to noise ratio was greater than 30:1. Furthermore, these gold particles were colinear, often spiral, or circular suggesting detection of a single nucleic acid molecule. Antibodies against actin, vimentin, or tubulin proteins were used after in situ hybridization, allowing simultaneous detection of the protein and its cognate message on the same sample. This revealed that cytoskeletal mRNAs are likely to be extremely close to actin protein (5 nm or less) and unlikely to be within 20 nm of vimentin or tubulin filaments. Actin mRNA was found to be more predominant in lamellipodia of motile cells, confirming previous results. These results indicate that this high resolution in situ hybridization approach is a powerful tool by which to investigate the association of mRNA with the cytoskeleton.

High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones

Science ◽  
1990 ◽  
Vol 247 (4938) ◽  
pp. 64-69 ◽  
Author(s):  
P Lichter ◽  
C. Tang ◽  
K Call ◽  
G Hermanson ◽  
G. Evans ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
High Resolution ◽  
Human Chromosome ◽  
Chromosome 11 ◽  
High Resolution Mapping ◽  
Resolution Mapping

High resolution ordering of DNA markers by multi-color fluorescent in situ hybridization of prophase chromosomes

1994 ◽  
Vol 65 (1-2) ◽  
pp. 130-135 ◽  
Author(s):  
J. Inazawa ◽  
T. Ariyama ◽  
T. Tokino ◽  
A. Tanigami ◽  
Y. Nakamura ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
High Resolution ◽  
Dna Markers ◽  
Fluorescent In Situ Hybridization

High-resolution in situ hybridization using DNA halo preparations

1992 ◽  
Vol 1 (8) ◽  
pp. 587-591 ◽  
Author(s):  
J. Wiegant ◽  
W. Kalle ◽  
L. Mullenders ◽  
S. Brookes ◽  
J.M.N. Hoovers ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
High Resolution
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