Influence of laser parameters on nanoparticle-induced membrane permeabilization

Abstract

Light-absorbing nanoparticles that are heated by short laser pulses can transiently increase membrane permeability. We evaluate the membrane permeability by flow cytometry assaying of propidium iodide and fluorescein isothiocyanate dextran (FITC-D) using different laser sources. The dependence of the transfection efficiency on laser parameters such as pulse duration, irradiant exposure, and irradiation mode is investigated. For nano- and also picosecond irradiation, we show a parameter range where a reliable membrane permeabilization is achieved for 10-kDa FITC-D. Fluorescent labeled antibodies are able to penetrate living cells that are permeabilized using these parameters. More than 50% of the cells are stained positive for a 150-kDa IgG antibody. These results suggest that the laser-induced permeabilization approach constitutes a promising tool for targeted delivery of larger exogenous molecules into living cells.

In vitro and ex vivo strategies for intracellular delivery

“…Recent efforts indicate that tuning the laser pulse parameters and properties of the absorbing particles can bias the mechanism towards a particular mode 95,96 . Although parameters are still being explored, a proof-of-concept study demonstrated conditions under which irradiation of gold nanoparticles was proposed to trigger localized thermal damage that permitted more than half of the treated cells to take up labelled antibodies 97 .…”

In vitro perforation of human epithelial carcinoma cell with antibody-conjugated biodegradable microspheres illuminated by a single 80 femtosecond near-infrared laser pulse

“…17,18 The use of a femtosecond laser with gold nanoparticles for the cell membrane perforation has also been reported. [19][20][21] In this method, collective free electron oscillation (surface plasmon) inside the gold particle generates a highly enhanced near field, resulting in the generation of cavitation bubbles and shock waves in the close vicinity of the cell membrane.…”

Immobilization of gold nanoparticles on cell culture surfaces for safe and enhanced gold nanoparticle-mediated laser transfection

“…Antibody conjugated 30-nm nanospheres were used in their study. 3 Heinemann et al 1 presented a fully automated setup employing 850-ps pulses at 532 nm. The delivery of siRNA, Morpholinos, and dextrans of sizes up to 2000 kDa was demonstrated using this setup.…”

“…© A versatile alternative to common transfection techniques, like electroporation and lipofection, is based on the interaction of laser irradiation and gold nanoparticles. [1][2][3] The routine procedure consists of the incubation of cells with gold nanoparticles with sizes of 80 to 200 nm for 3 to 6 h, which is sufficient for sedimentation for these large particles. 1,2,4 Alternatively, smaller antibody-conjugated particles are used.…”

“…1,2,4 Alternatively, smaller antibody-conjugated particles are used. 3 Irradiation of the gold nanoparticles can be performed at various pulse lengths and laser wavelengths. Yao et al applied nanosecond and picosecond laser pulses at 532 nm to deliver 10-kDa dextrans and antibodies into cells.…”