scholarly journals Locked Nucleic Acid In situ Hybridization Analysis of miR-21 Expression during Colorectal Cancer Development

2009 ◽  
Vol 15 (12) ◽  
pp. 4009-4016 ◽  
Author(s):  
Nobutake Yamamichi ◽  
Ryoichi Shimomura ◽  
Ken-ichi Inada ◽  
Kouhei Sakurai ◽  
Takeshi Haraguchi ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Hybridization Analysis ◽  
Locked Nucleic Acid ◽  
Cancer Development

scholarly journals Improved In Situ Hybridization Efficiency with Locked-Nucleic-Acid-Incorporated DNA Probes

2006 ◽  
Vol 72 (8) ◽  
pp. 5311-5317 ◽  
Author(s):  
Kengo Kubota ◽  
Akiyoshi Ohashi ◽  
Hiroyuki Imachi ◽  
Hideki Harada
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Fluorescence Intensity ◽  
Dna Probes ◽  
Locked Nucleic Acid ◽  
Factors Affecting ◽  
Hybridization Efficiency ◽  
Probe Hybridization ◽  
Major Factors

ABSTRACT Low signal intensity due to poor probe hybridization efficiency is one of the major drawbacks of rRNA-targeted in situ hybridization. There are two major factors affecting the hybridization efficiency: probe accessibility and affinity to the targeted rRNA molecules. In this study, we demonstrate remarkable improvement in in situ hybridization efficiency by applying locked-nucleic-acid (LNA)-incorporated oligodeoxynucleotide probes (LNA/DNA probes) without compromising specificity. Fluorescently labeled LNA/DNA probes with two to four LNA substitutions exhibited strong fluorescence intensities equal to or greater than that of probe Eub338, although these probes did not show bright signals when they were synthesized as DNA probes; for example, the fluorescence intensity of probe Eco468 increased by 22-fold after three LNA bases were substituted for DNA bases. Dissociation profiles of the probes revealed that the dissociation temperature was directly related to the number of LNA substitutions and the fluorescence intensity. These results suggest that the introduction of LNA residues in DNA probes will be a useful approach for effectively enhancing probe hybridization efficiency.

scholarly journals Optimizing locked nucleic acid/2’-O-methyl-RNA fluorescence in situ hybridization (LNA/2’OMe-FISH) procedure for bacterial detection

PLoS ONE ◽  
2019 ◽  
Vol 14 (5) ◽  
pp. e0217689 ◽  
Author(s):  
Andreia S. Azevedo ◽  
Inês M. Sousa ◽  
Ricardo M. Fernandes ◽  
Nuno F. Azevedo ◽  
Carina Almeida
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Fluorescence In Situ Hybridization ◽  
Locked Nucleic Acid ◽  
Bacterial Detection

scholarly journals Discrimination of bacteriophage infected cells using locked nucleic acid fluorescent in situ hybridization (LNA-FISH)

Biofouling ◽  
2016 ◽  
Vol 32 (2) ◽  
pp. 179-190 ◽  
Author(s):  
Diana Vilas Boas ◽  
Carina Almeida ◽  
Sanna Sillankorva ◽  
Ana Nicolau ◽  
Joana Azeredo ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Fluorescent In Situ Hybridization ◽  
Locked Nucleic Acid ◽  
Infected Cells

scholarly journals Detection and discrimination of biofilm populations using locked nucleic acid/2′-O-methyl-RNA fluorescence in situ hybridization (LNA/2′OMe-FISH)

2015 ◽  
Vol 104 ◽  
pp. 64-73 ◽  
Author(s):  
Andreia S. Azevedo ◽  
Carina Almeida ◽  
Bruno Pereira ◽  
Pedro Madureira ◽  
Jesper Wengel ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Fluorescence In Situ Hybridization ◽  
Locked Nucleic Acid

Application of locked nucleic acid-based probes in fluorescence in situ hybridization

2016 ◽  
Vol 100 (13) ◽  
pp. 5897-5906 ◽  
Author(s):  
Sílvia Fontenete ◽  
Daniel Carvalho ◽  
Nuno Guimarães ◽  
Pedro Madureira ◽  
Céu Figueiredo ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Fluorescence In Situ Hybridization ◽  
Locked Nucleic Acid

scholarly journals A novel approach for microRNA in situ hybridization using locked nucleic acid probes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Isabella W. Paulsen ◽  
Michael Bzorek ◽  
Jesper Olsen ◽  
Birgitte Grum-Schwensen ◽  
Jesper T. Troelsen ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
High Sensitivity ◽  
Staining Intensity ◽  
Locked Nucleic Acid ◽  
Proteinase K ◽  
Fixation Time ◽  
Target Tissue ◽  
Messenger Ribonucleic Acid

AbstractIdentification of target tissue microRNAs (miR) using in situ hybridization (ISH), with digoxigenin-labeled locked nucleic acid (LNA) probes, is influenced by preanalytic parameters. To determine the best retrieval method for common microRNAs, a multiblock composed of paraffin-embedded tonsil, cervix, placenta, and hyperplastic prostate tissue were included. Tissue were fixed in 10% formalin in a range of 5–144 hours (h). Cut sections (5 μm) from the multiblock were subjected to combinations of pretreatment procedures: variable periods of proteinase K (PK) digestion or Heat-induced microRNA Retrieval (HmiRR) using target retrieval solution (TRS) pH 6.1 or 9, with or without enzymatic treatment (pepsin). Results for the overall categories: TRS pH 9 versus PK; p = 2.9e−23, TRS pH 9 versus TRS pH 6.1; p = 1.1e−14, TRS pH 6.1 versus PK; p = 2.9e−03. A long fixation time, resulted in the best microRNA preservation and staining intensity (long vs. short: p = 3.5e−47, long vs. moderate: p = 1.6e−44, moderate vs. short: p = 4.3e−16), was enhanced using HmiRR TRS pH 9 with or without pepsin providing high sensitivity and specificity. These observations conflict with other ISH techniques (e.g., messenger ribonucleic acid), which typically require shorter fixation periods, and therefore, further studies are warranted.

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