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ARS Home » Southeast Area » Stoneville, Mississippi » Warmwater Aquaculture Research Unit » Research » Publications at this Location » Publication #330349
Title: Bioluminescent luciferase-modified magnetic nanoparticles as potential imaging agents for mammalian spermatozoa detection and tracking

Author
item VASQUEZ, ERICK - Mississippi State University
item FEUGANG, JEAN - Mississippi State University
item WILLARD, SCOTT - Mississippi State University
item RYAN, PETER - Mississippi State University
item WALTERS, KEISHA - Mississippi State University

Submitted to: Journal of Nanobiotechnology (Biomed Central Open Access)
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/17/2016
Publication Date: 3/17/2016
Citation: Vasquez, E.S., Feugang, J.M., Willard, S.T., Ryan, P.L., Walters, K.B. 2016. Bioluminescent luciferase-modified magnetic nanoparticles as potential imaging agents for mammalian spermatozoa detection and tracking. Journal of Nanobiotechnology (Biomed Central Open Access). 14:20-29.

Interpretive Summary: Magnetic nanoparticles were synthesized and complexed with the firefly luciferase enzyme to produce a multifunctional nanocomposite (Luc + MNP). The bioluminescence generated in the presence of mammalian spermatozoa was examined, showing Luc + MNP as a promising candidate to enhance or replace some current bioimaging technologies. By utilizing FDA-approved iron oxide magnetic nanoparticles and a natural enzyme, such as firefly luciferase, this nanocomposite has potential for a lower toxicity than some other methods, as well as the ability to magnetically manipulate cells and track them. The results presented here demonstrate the possibility of using this light-emitting magnetic nanoparticle for cellular binding and imaging. Additional studies on the optimum concentrations and solution composition for cell binding and viability, cell tracking and magnetic manipulation, and time-dependence of the labeling and bioluminescence will allow for a better understanding of the parameters for implementing this imaging method for assisted reproductive technologies.

Technical Abstract: Background: Nanoparticles have emerged as key materials for developing applications in nanomedicine, nanobiotechnology, bioimaging and theranostics. Existing bioimaging technologies include bioluminescent resonance energy transfer-conjugated quantum dots (BRET-QDs). Despite the current use of BRET-QDs for bioimaging, there are strong concerns about QD nanocomposites containing cadmium which exhibits potential cellular toxicity. Results: In this study, bioluminescent composites comprised of magnetic nanoparticles and firefly luciferase (Photinus pyralis) are examined as potential light-emitting agents for imaging, detection, and tracking mammalian spermatozoa. Characterization was carried out using infrared spectroscopy, TEM and cryo-TEM imaging, and '-(Zeta) potential measurements to demonstrate the successful preparation of these nanocomposites. Binding interactions between the synthesized nanoparticles and spermatozoon were characterized using confocal and atomic/magnetic force microscopy. Bioluminescence imaging and UV–visible-NIR microscopy results showed light emission from sperm samples incubated with the firefly luciferase-modified nanoparticles. Therefore, these newly synthesized luciferasemodified magnetic nanoparticles show promise as substitutes for QD labeling, and can potentially also be used for in vivo manipulation and tracking, as well as MRI techniques. Conclusions: These preliminary data indicate that luciferase-magnetic nanoparticle composites can potentially be used for spermatozoa detection and imaging. Their magnetic properties add additional functionality to allow for manipulation, sorting, or tracking of cells using magnetic techniques.