In Situ Hybridization of Mouse Embryo and Tissue Sections with Radiolabeled RNA Probes

2007 ◽  
Vol 2007 (11) ◽  
pp. pdb.prot4821 ◽  
Author(s):  
Andras Nagy ◽  
Marina Gertsenstein ◽  
Kristina Vintersten ◽  
Richard Behringer
Keyword(s):  
In Situ Hybridization ◽  
Mouse Embryo ◽  
Tissue Sections ◽  
Rna Probes

In situ hybridization methods for the detection of somatostatin mRNA in tissue sections using antisense RNA probes

1986 ◽  
Vol 18 (11-12) ◽  
pp. 597-604 ◽  
Author(s):  
Heinz Hoefler ◽  
Henry Childers ◽  
Marc R. Montminy ◽  
Ronald M. Lechan ◽  
Richard H. Goodman ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Antisense Rna ◽  
Tissue Sections ◽  
Rna Probes

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 In situ hybridization using PEG-embedded tissue and riboprobes: increased cellular detail coupled with high sensitivity.

1989 ◽  
Vol 37 (3) ◽  
pp. 389-393 ◽  
Author(s):  
D F Clayton ◽  
A Alvarez-Buylla
Keyword(s):  
In Situ Hybridization ◽  
High Sensitivity ◽  
Cresyl Violet ◽  
High Signal ◽  
Tissue Sections ◽  
Glass Slides ◽  
Cresyl Violet Staining ◽  
Rna Probes ◽  
Hybridization Protocol

We describe a procedure for preparing tissue sections by embedding in polyethylene glycol for subsequent in situ hybridization analysis using single-stranded RNA probes. Improved tissue morphology is obtained as compared to frozen sections, and the embedding procedure is milder and faster than paraffin embedding. Sections as thin as 2 microns are readily cut from PEG-embedded brain tissue. A simplified hybridization protocol (Clayton et al.: Neuron 1:249, 1988) supports the detection of even low-abundance brain mRNAs (less than or equal to 10(-4) fractional mRNA mass). By employing high stringency washes in place of ribonuclease treatment after hybridization, cell RNA is retained for cresyl violet staining, and high signal:noise ratios are achieved. Solutions to problems with section mounting and adherence to glass slides are presented. The combination of improved morphology, high signal levels, and relative simplicity should make this procedure useful in a variety of applications.

A gene with sequence similarity to Drosophila engrailed is expressed during the development of the neural tube and vertebrae in the mouse

Development ◽  
1988 ◽  
Vol 104 (2) ◽  
pp. 305-316 ◽  
Author(s):  
D. Davidson ◽  
E. Graham ◽  
C. Sime ◽  
R. Hill
Keyword(s):  
In Situ Hybridization ◽  
Neural Tube ◽  
Mouse Embryo ◽  
Sequence Similarity ◽  
Anterior Part ◽  
Limb Bud ◽  
Tissue Type ◽  
Restricted Domains ◽  
Restricted Region

The mouse genes En-1 and En-2 display sequence similarity, in and around the homeobox region, to the engrailed family in Drosophila. This paper describes their pattern of expression in the 12.5-day mouse embryo as determined by in situ hybridization. En-2 is expressed in a subset of cells expressing En-1. Both genes are expressed in the developing midbrain and its junction with the hindbrain. In addition, En-1 is expressed in the floor of the hindbrain, a restricted ventrolateral segment of the neural tube throughout the trunk and anterior part of the tail, the dermatome of tail somites, the centrum and costal processes in developing vertebrae, a restricted region of facial mesenchyme and the limb-bud ectoderm. Supplementary studies of 9.5-day and 10.5-day embryos showed that the same pattern of expression pertained in the neural tube, but that expression in the somites is at first confined to the dermatome and later found at a low level in restricted sclerotomal regions. Both genes are expressed in restricted domains which do not cross tissue-type boundaries. In several instances, however, boundaries of expression lie within morphologically undifferentiated tissue. These results suggest that En-1 and En-2 may be involved in the establishment or maintenance of the spatial integrity of specific domains within developing tissues.

Application of in situ hybridization with a novel phenytoin-labeled probe to conventional formalin-fixed, paraffin-embedded tissue sections

1995 ◽  
Vol 52 (3) ◽  
pp. 309-316 ◽  
Author(s):  
Yoshito Eizuru ◽  
Yoichi Minamishima ◽  
Tadashi Matsumoto ◽  
Toshinari Hamakado ◽  
Mikio Mizukoshi ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Labeled Probe ◽  
Formalin Fixed
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