Abstract
BackgroundLuminescent semiconductor nanocrystals, also known as quantum dots (QD), possess highly desirable optical properties that account for development of a variety of exciting biomedical techniques. These properties include long-term stability, brightness, narrow emission spectra, size tunable properties and resistance to photobleaching. QD have many promising applications in biology and the list is constantly growing. These applications include DNA or protein tagging for in vitro assays, deep-tissue imaging, fluorescence resonance energy transfer (FRET), and studying dynamics of cell surface receptors, among others. Here we explored the potential of QD-mediated labeling for the purpose of tracking an intracellular protein inside live cells.ResultsWe manufactured dihydrolipoic acid (DHLA)-capped CdSe-ZnS core-shell QD, not available commercially, and coupled them to the cell cycle regulatory protein Cyclin E. We then utilized the QD fluorescence capabilities for visualization of Cyclin E trafficking within cells of Xenopus laevis embryos in real time.ConclusionsThese studies provide “proof-of-concept” for this approach by tracking QD-tagged Cyclin E within cells of developing embryos, before and during an important developmental period, the midblastula transition. Importantly, we show that the attachment of QD to Cyclin E did not disrupt its proper intracellular distribution prior to and during the midblastula transition. The fate of the QD after cyclin E degradation following the midblastula transition remains unknown.Electronic supplementary materialThe online version of this article (doi:10.1186/s12951-015-0092-6) contains supplementary material, which is available to authorized users.
Highlights
Luminescent semiconductor nanocrystals, known as quantum dots (QD), possess highly desirable optical properties that account for development of a variety of exciting biomedical techniques
We successfully synthesized functional dihydrolipoic acid (DHLA)-capped CdSe-ZnS QD tailored to our application of attachment of recombinant (His6)-Cyclin E protein to its surface
We followed our QD-tagged Cyclin E complexes inside the cells of developing Xenopus embryos in real time, which allowed us to discriminate between cytoplasmic Cyclin E localization before the midblastula transition (MBT) (4 hpf ) and its accumulation in the nucleus at the MBT (6 hpf )
Summary
Luminescent semiconductor nanocrystals, known as quantum dots (QD), possess highly desirable optical properties that account for development of a variety of exciting biomedical techniques. These properties include long-term stability, brightness, narrow emission spectra, size tunable properties and resistance to photobleaching. QD have many promising applications in biology and the list is constantly growing These applications include DNA or protein tagging for in vitro assays, deep-tissue imaging, fluorescence resonance energy transfer (FRET), and studying dynamics of cell surface receptors, among others. CdSe nanocrystals are prepared by reacting organometallic precursors at high temperatures in a coordinating solvent mixture This results in capping of the inorganic core with an organic layer of trioctylphosphine/trioctylphosphineoxide mixture (TOP/TOPO) [3]. The shell protects the QD from oxidation and prevents
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