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.

Localization of Classical Swine Fever Virus from Chronically Infected Pigs by In Situ Hybridization and Immunohistochemistry

2003 ◽  
Vol 40 (1) ◽  
pp. 107-113 ◽  
Author(s):  
C. Choi ◽  
C. Chae
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Mononuclear Cells ◽  
Classical Swine Fever ◽  
Lymphoid Tissues ◽  
Hybridization Technique ◽  
Tissue Sections ◽  
Background Staining ◽  
The Central Nervous System

Classical swine fever (CSF) virus (CSFV) nucleic acid and antigen were detected in 15 pigs with naturally occurring chronic CSF by in situ hybridization and immunohistochemistry. The most consistent and prominent microscopic lesions were perivascular mononuclear cell infiltration and gliosis in the central nervous system of pigs with chronic CSF. Positive cells typically exhibited a dark brown (in situ hybridization) or red (immunohistochemistry) reaction product in the cytoplasm without background staining. A positive signal for both in situ hybridization and immunohistochemistry was detected in mononuclear cells and lymphocytes of lymphoid tissues. Viral nucleic acid was detected in some tissue sections in the absence of viral antigen. The in situ hybridization technique developed in this study was useful for the detection of CSFV RNA in tissues taken from chronically infected pigs and may be a valuable technique for studying the pathogenesis of chronic CSFV infection.

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

In situ hybridization technique for mRNA detection in whole mount Arabidopsis samples

2006 ◽  
Vol 1 (4) ◽  
pp. 1939-1946 ◽  
Author(s):  
Jan Hejátko ◽  
Ikram Blilou ◽  
Philip B Brewer ◽  
Jiří Friml ◽  
Ben Scheres ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Hybridization Technique ◽  
Mrna Detection ◽  
Whole Mount

scholarly journals Combination of Direct Viable Count and Fluorescent In Situ Hybridization (DVC-FISH) as a Potential Method for Identifying Viable Vibrio parahaemolyticus in Oysters and Mussels

Foods ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1502
Author(s):  
Jorge García-Hernández ◽  
Manuel Hernández ◽  
Yolanda Moreno
Keyword(s):  
In Situ Hybridization ◽  
Vibrio Parahaemolyticus ◽  
Fluorescent In Situ Hybridization ◽  
Potential Method ◽  
Viable Count ◽  
Hybridization Technique ◽  
Fish Technique ◽  
Viable Cells

Vibrio parahaemolyticus is a human food-borne pathogen with the ability to enter the food chain. It is able to acquire a viable, non-cultivable state (VBNC), which is not detected by traditional methods. The combination of the direct viable count method and a fluorescent in situ hybridization technique (DVC-FISH) makes it possible to detect microorganisms that can present VBNC forms in complex samples The optimization of the in vitro DVC-FISH technique for V. parahaemolyticus was carried out. The selected antibiotic was ciprofloxacin at a concentration of 0.75 μg/mL with an incubation time in DVC broth of 5 h. The DVC-FISH technique and the traditional plate culture were applied to detect and quantify the viable cells of the affected pathogen in artificially contaminated food matrices at different temperatures. The results obtained showed that low temperatures produced an important logarithmic decrease of V. parahaemolyticus, while at 22 °C, it proliferated rapidly. The DVC-FISH technique proved to be a useful tool for the detection and quantification of V. parahaemolyticus in the two seafood matrices of oysters and mussels. This is the first study in which this technique has been developed to detect viable cells for this microorganism.

scholarly journals FISHing for chick genes: Triple-label whole-mount fluorescence in situ hybridization detects simultaneous and overlapping gene expression in avian embryos

2004 ◽  
Vol 229 (3) ◽  
pp. 651-657 ◽  
Author(s):  
Nathaniel Denkers ◽  
Pilar García-Villalba ◽  
Christopher K. Rodesch ◽  
Kandice R. Nielson ◽  
Teri Jo Mauch
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Avian Embryos ◽  
Overlapping Gene ◽  
Whole Mount
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