Targeted microinjection into cells and retina using optoporation.

Area of Science:

• Biomedical Engineering
• Cell Biology
• Neuroscience

Background:

• Traditional methods struggle with precise, non-damaging delivery of exogenous materials into cells.
• Laser microbeams offer a promising noncontact approach for targeted cellular delivery.

Purpose of the Study:

• To report targeted delivery of impermeable substances into mammalian cells and goldfish retinal explants using ultrafast laser microbeam assisted injection.
• To demonstrate the capability of laser-assisted optoporation for precise transfection in three-dimensional neuronal tissue.

Main Methods:

• Utilized ultrafast laser microbeams for localized pore formation and direct injection into cells.
• Investigated indirect optoporation via external bubble formation.
• Optimized laser parameters (intensity, exposure, spatial pattern) for efficient transfection.
• Confirmed delivery and diffusion patterns using fluorescent dyes and gene expression (channelrhodopsin-2).

Main Results:

• Successfully introduced impermeable dye into cells, observing spatiotemporal diffusion patterns.
• Demonstrated successful spatial transfection in goldfish retina explants, confirmed by fluorescent protein expression.
• Characterized differences in injection dynamics between direct and indirect optoporation.

Conclusions:

• Ultrafast laser microbeam assisted injection is a highly precise method for delivering exogenous materials, including impermeable substances, into cells and complex tissues.
• This technique holds significant potential for advancing the understanding of neuronal circuitry in both healthy and diseased retinas.