High-Resolution Whole-Mount In Situ Hybridization Using Quantum Dot Nanocrystals
Joint Authors
Lubatti, Andrea
Skourides, Paris A.
Charalambous, Anna
Ioannou, Andriani
Eleftheriou, Iro
Source
Issue
Vol. 2012, Issue 2012 (31 Dec. 2012), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2012-01-12
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
The photostability and narrow emission spectra of nanometer-scale semiconductor crystallites (QDs) make them desirable candidates for whole-mount fluorescent in situ hybridization to detect mRNA transcripts in morphologically preserved intact embryos.
We describe a method for direct QD labeling of modified oligonucleotide probes through streptavidin-biotin and antibody-mediated interactions (anti-FITC and anti-digoxigenin).
To overcome permeability issues and allow QD conjugate penetration, embryos were treated with proteinase K.
The use of QDs dramatically increased sensitivity of whole-mount in situ hybridization (WISH) in comparison with organic fluorophores and enabled fluorescent detection of specific transcripts within cells without the use of enzymatic amplification.
Therefore, this method offers significant advantages both in terms of sensitivity, as well as resolution.
Specifically, the use of QDs alleviates issues of photostability and limited brightness plaguing organic fluorophores and allows fluorescent imaging of cleared embryos.
It also offers new imaging possibilities, including intracellular localization of mRNAs, simultaneous multiple-transcript detection, and visualization of mRNA expression patterns in 3D.
American Psychological Association (APA)
Ioannou, Andriani& Eleftheriou, Iro& Lubatti, Andrea& Charalambous, Anna& Skourides, Paris A.. 2012. High-Resolution Whole-Mount In Situ Hybridization Using Quantum Dot Nanocrystals. BioMed Research International،Vol. 2012, no. 2012, pp.1-9.
https://search.emarefa.net/detail/BIM-991816
Modern Language Association (MLA)
Ioannou, Andriani…[et al.]. High-Resolution Whole-Mount In Situ Hybridization Using Quantum Dot Nanocrystals. BioMed Research International No. 2012 (2012), pp.1-9.
https://search.emarefa.net/detail/BIM-991816
American Medical Association (AMA)
Ioannou, Andriani& Eleftheriou, Iro& Lubatti, Andrea& Charalambous, Anna& Skourides, Paris A.. High-Resolution Whole-Mount In Situ Hybridization Using Quantum Dot Nanocrystals. BioMed Research International. 2012. Vol. 2012, no. 2012, pp.1-9.
https://search.emarefa.net/detail/BIM-991816
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-991816