Abstract
Tailless T4 nanoparticles (NPs) have large surface areas consisting of more than 105 diverse surface reactive groups and offer great flexibility in chemical modification for tailoring the desired functionality. Dye-conjugated T4 NPs exhibiting bright fluorescence are biocompatible and can be internalized by various eukaryotic cells which land themselves as excellent cellular imaging agents. Here, we describe the preparation of engineered T4 NPs including dye-conjugation and characterization, and the procedure for cellular uptake and confocal microscopy.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Manchester M, Singh P (2006) Virus-based nanoparticles (VNPs): Platform technologies for diagnostic imaging. Adv Drug Delivery Rev 58:1505–1522
Li K, Nguyen HG, Lu X, Wang Q (2010) Viruses and their potential in bioimaging and biosensing applications. Analyst 135:21–27
Steinmetz NF (2010) Viral nanoparticles as platforms for next-generation therapeutics and imaging devices. Nanomedicine 6:634–641
Archer MJ, Liu JL (2009) Bacteriophage T4 nanoparticles as materials in sensor applications: variables that influence their organization and assembly on surfaces. Sensors 9:6298–6311
Robertson KR, Soto CM, Archer MJ, Odoemene O, Liu JL (2011) Engineered T4 viral nanoparticles for cellular imaging and flow cytometry. Bioconj Chem 22(4): 595–604
Selick HE, Kreuzert KN, Alberts BM (1988) The bacteriophage T4 insertion/substitution vector system: a method for introducing site-specific mutations into the virus chromosome. J Biol Chem 263(23):11336–11347
Soto CM, Blum AS, Vora GJ, Lebedev N, Meador CE, Won AP, Chatterji A, Johnson JE, Ratna BR (2006) Fluorescent signal amplification of carbocyanine dyes using engineered viral nanoparticles. J Am Chem Soc 128: 5184–5189
Soto CM, Blaney KM, Dar M, Khan M, Lin B, Malanoski AP, Tidd C, Rios MV, Lopez DM, Ratna BR (2009) Cowpea mosaic virus nanoscaffold as signal enhancement for DNA microarrays. Biosens Bioelectron 25:48–54
Berlier JE, Rothe A, Buller G, Bradford J, Gray DR, Filanoski BJ, Telford WG, Yue S, Liu J, Cheung C-Y, Chang W, Hirsch JD, Beechem JM, Haugland RP, Haugland RP (2003) Quantitative comparison of long-wavelength Alexa Fluor dyes to Cy dyes: fluorescence of the dyes and their bioconjugates. J Histochem Cytochem 51:1699–1712
Chen H, Ahsan SS, Santiago-Berrios MEB, Abruna HD, Webb WW (2010) Mechanisms of quenching of Alexa fluorophores by natural amino acids. J Am Chem Soc 132: 7244–7245
Gruber HJ, Hahn CD, Kada G, Riener CK, Harms GS, Ahrer W, Dax TG, Knaus H-G (2000) Anomalous fluorescence enhancement of Cy3 and Cy3.5 versus anomalous fluorescence loss of Cy5 and Cy7 upon covalent linking to IgG and noncovalent binding to avidin. Bioconjugate Chem 11:696–704
West W, Pearce S (1965) The dimeric state of cyanine dyes. J Phys Chem 69:1894–1903
Acknowledgements
We would like to thank Dr. Carissa Soto for her technical assistance on the measurement of fluorescent intensity and Dr. Lindsay W. Black from University of Maryland at Baltimore for his kind gift of T4 K10. This work was supported by the Office of Naval Research. The opinions and assertions contained herein are the private ones of the authors and are not to be construed as official or reflecting the views of the Department of the Navy or the military at large.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Liu, J.L., Robertson, K.L. (2014). Engineered Bacteriophage T4 Nanoparticles for Cellular Imaging. In: Lin, B., Ratna, B. (eds) Virus Hybrids as Nanomaterials. Methods in Molecular Biology, vol 1108. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-751-8_14
Download citation
DOI: https://doi.org/10.1007/978-1-62703-751-8_14
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-750-1
Online ISBN: 978-1-62703-751-8
eBook Packages: Springer Protocols