scholarly journals Sensitive Two-Color Whole-Mount In Situ Hybridizations Using Digoxygenin- and Dinitrophenol-Labeled RNA Probes

BioTechniques ◽  
2002 ◽  
Vol 32 (3) ◽  
pp. 494-500 ◽  
Author(s):  
Sarah Long ◽  
Michael Rebagliati
Keyword(s):  
Rna Probes ◽  
Whole Mount ◽  
Labeled Rna ◽  
In Situ Hybridizations

In Situ Hybridization with Riboprobes: An Overview for Veterinary Pathologists

1998 ◽  
Vol 35 (3) ◽  
pp. 159-167 ◽  
Author(s):  
C. Brown
Keyword(s):  
In Situ Hybridization ◽  
Molecular Biology ◽  
Infectious Agents ◽  
Probe Target ◽  
Cellular Genes ◽  
Rna Probes ◽  
Pcr Technology ◽  
Labeled Rna ◽  
Target Combination

In situ hybridization using nonradioactively-labeled RNA probes is a technique that combines understanding of basic molecular biology and histopathologic interpretation. Recombinant or PCR technology can be used to produce probes that hybridize with a wide variety of cellular genes and infectious agents. Hybridization conditions can be optimized for each probe/target combination.

scholarly journals A fixative suitable for in situ hybridization histochemistry.

1993 ◽  
Vol 41 (6) ◽  
pp. 947-953 ◽  
Author(s):  
F Uehara ◽  
N Ohba ◽  
Y Nakashima ◽  
T Yanagita ◽  
M Ozawa ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Proteinase K ◽  
Optimal Time ◽  
Hybridization Histochemistry ◽  
Tissue Sections ◽  
In Situ Hybridization Histochemistry ◽  
Rna Probes ◽  
Labeled Rna ◽  
The Stability

We compared the morphology and stability of hybridization signals between paraffin sections of rat retina fixed with commonly used 4% paraformaldehyde/PBS and those fixed with a fixative containing glutaraldehyde in in situ hybridization histochemistry, using a digoxigenin-labeled RNA probe complementary for beta-galactoside alpha 2,6-sialyltransferase mRNA. Retinal detachment was frequently observed in the sections fixed with 4% paraformaldehyde-PBS, whereas the morphology was satisfactorily preserved in those fixed with either 0.5% glutaraldehyde, 4% paraformaldehyde-PBS, or 2.5% glutaraldehyde-PBS. Without glutaraldehyde, it was difficult to determine the most appropriate length of proteinase K digestion of tissue sections for facilitating probe penetration, since the optimal time for definite hybridization was variable among the retinal cells in heterogeneous layers. By addition of glutaraldehyde to paraformaldehyde or with glutaraldehyde alone, it was easy to establish the appropriate time for the unmasking procedure, since intense mRNA signals were constant throughout the retina by proteinase K digestion for more than 30-40 min. Using a fixative that causes stronger cross-linking (e.g., glutaraldehyde) is recommended to improve not only the morphology but also the stability of hybridization signals in in situ hybridization histochemistry with paraffin embedding and digoxigenin-labeled RNA probes.

scholarly journals 2. Histocytochemistry Series In Situ Hybridization Using Non-radioisotope labeled RNA Probes (4)

2010 ◽  
Vol 6 (1) ◽  
pp. 23-29
Author(s):  
Takuya Mishima ◽  
Shan-shun Luo ◽  
Toshiyuki Takeshita ◽  
Toshihiro Takizawa
Keyword(s):  
In Situ Hybridization ◽  
Rna Probes ◽  
Labeled Rna
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