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Saklayen, Nabiha; Huber, Marinus; Madrid, Marinna; Nuzzo, Valeria; Vulis, Daryl I.; Shen, Weilu; Nelson, Jeffery; McClelland, Arthur A.; Heisterkamp, Alexander und Mazur, Eric (2017): Intracellular Delivery Using Nanosecond-Laser Excitation of Large-Area Plasmonic Substrates. In: Acs Nano, Bd. 11, Nr. 4: S. 3671-3680

Volltext auf 'Open Access LMU' nicht verfügbar.

Abstract

Efficiently delivering functional cargo to millions of cells on the time scale of minutes will revolutionize gene therapy, drug discovery, and high throughput screening. Recent studies of intracellular delivery with thermoplasmonic structured surfaces show promising results but in most cases require time- or cost-intensive fabrication or lead to unreproducible surfaces. We designed and fabricated large-area (14 X 14 mm), photolithography based, template-stripped plasmonic substrates that are nanosecond laser-activated to form transient pores in cells for cargo entry. We optimized fabrication to produce plasmonic structures that are ultrasmooth and precisely patterned over large areas. We used flow cytometry to characterize the delivery efficiency of cargos ranging in size from 0.6 to 2000 kDa to cells (up to 95% for the smallest molecule) and viability of cells (up to 98%). This technique offers a throughput of 50000 cells/min, which can be scaled up as necessary. This technique is also cost-effective as each large-area photolithography substrate can be used to deliver cargo to millions of cells, and switching to a nanosecond laser makes the setup cheaper and easier to use. The approach we present offers additional desirable features: spatial selectivity, reproducibility, minimal residual fragments, and costeffective fabrication. This research supports the development of safer genetic and viral disease therapies as well as research tools for fundamental biological research that rely on effectively delivering molecules to millions of living cells.

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