scholarly journals Sensitive Nonradioactive Detection of mRNA in Tissue Sections: Novel Application of the Whole-mount In Situ Hybridization Protocol

2001 ◽  
Vol 49 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Antoon F. M. Moorman ◽  
Arjan C. Houweling ◽  
Piet A. J. de Boer ◽  
Vincent M. Christoffels
Keyword(s):  
In Situ Hybridization ◽  
Tissue Sections ◽  
Whole Mount ◽  
Hybridization Protocol

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.

An efficient in situ hybridization protocol for multiple tissue sections and probes on miniaturized slides

2002 ◽  
Vol 212 (8) ◽  
pp. 403-406 ◽  
Author(s):  
Gunnar Weisheit ◽  
Kirsten Mertz ◽  
Karl Schilling ◽  
Christoph Viebahn
Keyword(s):  
In Situ Hybridization ◽  
Tissue Sections ◽  
Hybridization Protocol

Whole-mount in situ hybridization: minimizing the folding problem of thin-sheet tissue-like crayfish haematopoietic tissue

Crustaceana ◽  
2018 ◽  
Vol 91 (1) ◽  
pp. 1-15
Author(s):  
Aleksandra Zečić ◽  
Chadanat Noonin
Keyword(s):  
In Situ Hybridization ◽  
Histological Study ◽  
Thin Sheet ◽  
Hybridization Technique ◽  
Tissue Sections ◽  
Specific Transcript ◽  
Optimized Protocol ◽  
Whole Mount ◽  
Haematopoietic Tissue

Crayfish haematopoietic tissue (HPT) has a thin-sheet-like structure with a thickness of 100-160 μm and a width of approximately 1-2 cm. This structure makes HPT extremely easy to fold after removal from the animal. Therefore, it is difficult to handle the tissue without folding when processing for sectioning and histological study. The degree of tissue folding reflects the size of the tissue sections obtained, how complicated it is to interpret the location of each tissue section, and the accuracy of the interpretation of the location of a specific transcript. To facilitate the interpretation of a specific transcript location in the HPT, we optimized a whole-mount in situ hybridization technique to minimize tissue folding. This optimized protocol effectively reduced the tissue folding. Therefore, the location of a specific transcript in the HPT was easily and accurately defined. This protocol will be useful for whole-mount staining of other tissues with similar structure.

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