Photothermal nanoblade for large cargo delivery into mammalian cells

Abstract

We report a photothermal nanoblade that utilizes a metallic nanostructure to harvest laser pulse energy and convert it into a localized explosive vapor bubble, which rapidly punctures a lightly-contacting cell membrane via high-speed fluidic flows and induced transient shear stress. Integrating the metallic nanostructure with a micropipette, the nanoblade generates a micron-sized membrane access port for delivering concentrated cargo (5×10 8 bacteria/ml) with high efficiency (46%) and cell viability (>90%) into mammalian cells. Additional biologic and inanimate cargo over 3-orders of magnitude in size including DNA, 200 nm polystyrene beads to 2 μm bacteria have been delivered into multiple cell types.

Dissection of the Burkholderia intracellular life cycle using a photothermal nanoblade

“…To accomplish this, we exploited the capabilities of our recently developed photothermal nanoblade to place bacteria directly into the cytoplasm of host cells ( Fig. 2A) (16). Because a nanoblade has not yet been customized and approved for use with Bp in a select agent BSL-3 facility, we focused our analysis on Bt, which can be safely manipulated under BSL-2 conditions.…”

“…T3SS Bsa has been proposed to be required for late events associated with cell-cell spread, but direct investigation has been difficult since mutants are defective in earlier steps in the intracellular life cycle. To address this conundrum, we have used a photothermal nanoblade to deliver live bacteria directly into the cytoplasm of mammalian cells (16). The photothermal nanoblade device uses a laser pulse to excite a thin titanium coating on the tip of a glass capillary pipette.…”

Electrical Impedance Monitoring of Photothermal Porated Mammalian Cells

“…Among current delivery approaches, the photothermal nanoblade has shown its effectiveness in delivering large cargo into live cells with high viability and high efficiency. 11,12,14 The photothermal nanoblade uses a metallic nanostructure to harvest energy from short optical pulses, typically at a laser fluence of ~180 mJ/cm 2 , and converts this energy into localized and patterned nanocavitation bubbles. When a mechanically soft and fragile cell membrane is near or in contact with the metallic nanostructure, an explosive cavitation bubble rapidly punctures the adjacent lipid bilayer via high-speed and localized fluid flows.…”

Intracellular Delivery by Shape Anisotropic Magnetic Particle–Induced Cell Membrane Cuts

“…[14][15][16] However, cell viability, efficiency, setup cost, conjugation of molecules onto magnetic nanoparticles, and cytotoxicity from iron oxide nanoparticles are current obstacles. [17][18][19][20] A low-cost, high-efficiency, and high-throughput method for intracellular delivery remains highly desirable.…”