Light Image Patterned Molecular Delivery into Live Cells Using Gold Particle Coated Substrate

Department of Electrical Engineering, University of California, Los Angeles (UCLA), 420 Westwood Plaza, 48-121 Engineering IV, Los Angeles, CA 90095-1597, USA.
Optics Express (Impact Factor: 3.49). 01/2010; 18(2):938-46. DOI: 10.1109/LEOSST.2008.4590556
Source: PubMed

ABSTRACT An image-patterned molecular delivery system for mammalian cells is demonstrated by pulsed laser irradiation of gold particles immobilized on a substrate below a cell monolayer. Patterned cavitation bubble nucleation was captured using a time-resolved imaging system and molecular delivery verified by observing the uptake of a membrane-impermeable fluorescent dye, calcein. Delivery efficiency as high as 90% was observed and multiplexed, patterned dye delivery was demonstrated. (C) 2009 Optical Society of America

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Available from: Pei-Yu Chiou, Sep 26, 2015
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    • "Since no ablation of the AuNP from the cell surface was observed under GNOME laser transfection conditions (Fig. 4), the appearance of vapour or cavitation bubbles seems to be unlikely as those should lead to particle detachment. Explosive boiling or the generation of plasmonic nanobubbles, as described by Wu et al. [45] and Lukianova-Hleb et al. [16], [46], [47], respectively, therefore is most likely not involved in the perforation mechanism. To gain complete understanding of the mechanism and to distinguish which process is dominant, further investigations are needed. "
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    ABSTRACT: Laser based transfection methods have proven to be an efficient and gentle alternative to established molecule delivery methods like lipofection or electroporation. Among the laser based methods, gold nanoparticle mediated laser transfection bears the major advantage of high throughput and easy usability. This approach uses plasmon resonances on gold nanoparticles unspecifically attached to the cell membrane to evoke transient and spatially defined cell membrane permeabilization. In this study, we explore the parameter regime for gold nanoparticle mediated laser transfection for the delivery of molecules into cell lines and prove its suitability for siRNA mediated gene knock down. The developed setup allows easy usage and safe laser operation in a normal lab environment. We applied a 532 nm Nd:YAG microchip laser emitting 850 ps pulses at a repetition rate of 20.25 kHz. Scanning velocities of the laser spot over the sample of up to 200 mm/s were tested without a decline in perforation efficiency. This velocity leads to a process speed of ∼8 s per well of a 96 well plate. The optimal particle density was determined to be ∼6 particles per cell using environmental scanning electron microscopy. Applying the optimized parameters transfection efficiencies of 88% were achieved in canine pleomorphic adenoma ZMTH3 cells using a fluorescent labeled siRNA while maintaining a high cell viability of >90%. Gene knock down of d2-EGFP was demonstrated and validated by fluorescence repression and western blot analysis. On basis of our findings and established mathematical models we suppose a mixed transfection mechanism consisting of thermal and multiphoton near field effects. Our findings emphasize that gold nanoparticle mediated laser transfection provides an excellent tool for molecular delivery for both, high throughput purposes and the transfection of sensitive cells types.
    PLoS ONE 03/2013; 8(3):e58604. DOI:10.1371/journal.pone.0058604 · 3.23 Impact Factor
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    • "The biomedical applications of plasmonic nanoparticles (NPs) use their optical scattering properties for imaging and diagnostics 1-4, and their photothermal properties for various types of therapies through the generation of heat 1,5,6, bubbles 6-12 and acoustic waves 13,14. The specificity and sensitivity of imaging and diagnostics, as well as the selectivity and efficacy of the therapeutic methods, depend upon the selectivity of NP delivery and activation within their targets, typically diseased cells or tissues. "
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    ABSTRACT: Theranostic applications require coupling of diagnosis and therapy, a high degree of specificity and adaptability to delivery methods compatible with clinical practice. The tunable physical and biological effects of selective targeting and activation of plasmonic nanobubbles (PNB) were studied in a heterogeneous biological microenvironment of prostate cancer and stromal cells. All cells were targeted with conjugates of gold nanoparticles (NPs) through an antibody-receptor-endocytosis-nanocluster mechanism that produced NP clusters. The simultaneous pulsed optical activation of intracellular NP clusters at several wavelengths resulted in higher optical contrast and therapeutic selectivity of PNBs compared with those of gold NPs alone. The developed mechanism was termed "rainbow plasmonic nanobubbles." The cellular effect of rainbow PNBs was tuned in situ in target cells, thus supporting a theranostic algorithm of prostate cancer cell detection and follow-up guided destruction without damage to collateral cells. The specificity and tunability of PNBs is promising for theranostic applications and we discuss a fiber optic platform that will capitalize on these features to bring theranostic tools to the clinic.
    Theranostics 01/2011; 1:3-17. · 8.02 Impact Factor
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    ABSTRACT: We have used short laser pulses to generate transient vapor nanobubbles around plasmonic nanoparticles. The photothermal, mechanical, and optical properties of such bubbles were found to be different from those of plasmonic nanoparticle and vapor bubbles, as well. This phenomenon was considered as a new complex nanosystem-plasmonic nanobubble (PNB). Mechanical and optical scattering properties of PNB depended upon the nanoparticle surface and heat capacity, clusterization state, and the optical pulse length. The generation of the PNB required much higher laser pulse fluence thresholds than the explosive boiling level and was characterized by the relatively high lower threshold of the minimal size (lifetime) of PNB. Optical scattering by PNB and its diameter (measured as the lifetime) has been varied with the fluence of laser pulse, and this has demonstrated the tunable nature of PNB.
    ACS Nano 03/2010; 4(4):2109-23. DOI:10.1021/nn1000222 · 12.88 Impact Factor
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