Single-cell optoporation using nanosecond laser irradiation on grid-marked Au nanostars layers

Abstract

Recently we developed Au nanoparticle (AuNP) layers as photothermal substrates for cell optoporation using nanosecond laser irradiation [10.1002/jbio.201800166]. Among several types of AuNP with varying geometry and optical properties, the Au nanostars layers have been shown as most preferable platforms for effective cell optoporation and transfection. The typical optoporation procedure include several steps: the assembling of Au nanostars directly on the culture plastic, followed by harvesting the cells on its surface, and subjection to single laser exposure. In the present study we demonstrate the advantageous features of our system for precise controllable optoporation with single cell performance. For this purpose, we prepared Au nanostar layers on the bottoms of 24-well culture plates and gridded them using nanosecond laser marking with 100x100 micron mesh. Thus we obtained the plasmonic surfaces for cell harvesting and optoporation that facilitate the tracing of individual cells. We tested the optoporation performance for propidium iodide delivery on the CHO (Chinese hamster ovarian) cells. These CHO cells are commonly used for in vitro single-cell investigations due to their ability to form confluent monolayer from one cell. We have adjusted the irradiation regimes for effective optoporation of the rarely seeded individual CHO cells on the grid-marked Au nanostars layers. The grid-marked substrates sufficiently facilitated the long-term monitoring of the same cells after optoporation during several days using standard fluorescent microscopy. Thus, we developed a simple approach which have further perspectives for practical commercialization as novel tool for precisely controlled laser transfection with single-cell performance.

File with abstract

Speaker

Elena Avdeeva
Institute of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences
Russia

Discussion

Ask question