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- Title
Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes.
- Authors
Feugang, Jean M.; Youngblood, Ramey C.; Greene, Jonathan M.; Willard, Scott T.; Ryan, Peter L.
- Abstract
Background: The fertility performance of animals is still a mystery and the full comprehension of mammalian gametes maturation and early embryonic development remains to be elucidated. The recent development in nanotechnology ofers a new opportunity for real-time study of reproductive cells in their physiological environments. As a irst step toward that goal, we evaluated the efectiveness of a luorescent and luminescent nanoparticle for in vitro and ex vivo imaging of porcine gametes. Methods: Freshly harvested boar sperm were labeled with red-shifted (655 nm) quantum dot nanoparticles conjugated (QD+) or not (QD-) with plasminogen antibody and evaluated. Subsets of labeled spermatozoa were loaded into straws and placed within the lumen of gilt reproductive tracts for ex vivo intra-uterine imaging. Porcine cumulusoocyte complexes (COCs) were matured in the presence of QD- or QD+. Ovarian follicles were microinjected with QD- or QD+ and placed in culture for up to 4 days. After labeling, all samples were supplemented with coelenterazine, the luciferase substrate, and immediately submitted to bioluminescence analysis, followed by luorescence and hyperspectral imaging. Data were analyzed with ANOVA and P < 0.05 indicated signiicant diferences. Results: All labeled-samples revealed bioluminescence emission that was conirmed by luorescence and hyperspectral imaging of the QD localization within the cells and tissues. Over 76% of spermatozoa and both immature and mature COCs were successfully labeled with QD- or QD+. The QD- luorescence appeared homogenously distributed in the oocytes, while found in the entire sperm length with a higher accumulation within the mid-piece. Labeled-follicles exhibited a progressive migration of QD nanoparticles within the follicle wall during culture. In contrast, QD+ luorescence signals appeared condensed and stronger in the follicle cells, sperm head, and sub-plasma membrane area of mature oocytes. Weaker QD+ signals were detected in the cumulus cells. Fluorescence and hyperspectral microscope imaging showed comparable intracellular QD localization. Ex-vivo intra-uterine bioluminescence imaging of labeled spermatozoa revealed stronger signals captured over the oviducts, with uterine body allowing the lowest signal detection. Conclusion: Findings indicate that conjugated and non-conjugated luorescent nanoparticles can be used for efective labeling of mammalian gametes for in vitro monitoring and potential in vivo targeted-imaging.
- Subjects
QUANTUM dots; QUANTUM electronics; BIOLUMINESCENCE; GAMETES; NANOPARTICLES
- Publication
Journal of Nanobiotechnology, 2015, Vol 13, Issue 1, p1
- ISSN
1477-3155
- Publication type
Article
- DOI
10.1186/s12951-015-0097-1