High-Efficiency Targeted Gene Transfection of Cells Using Temporal and Spatial Shaping Femtosecond Laser Irradiation

Area of Science:

• Biotechnology
• Optics
• Cell Biology

Background:

• Laser-assisted cell gene transfection offers sterile and non-toxic delivery but suffers from low efficiency.
• Existing methods struggle to meet the demands of practical applications due to suboptimal transfection rates.

Purpose of the Study:

• To enhance the efficiency of femtosecond laser-assisted cell gene transfection.
• To develop a method for improving transfection rates while preserving cell viability.

Main Methods:

• A temporal and spatial shaping method for femtosecond laser pulses was developed.
• Temporal shaping involved segmenting pulses into variable-energy subpulses with controlled delays.
• Spatial shaping extended the working distance and reduced focal position sensitivity.

Main Results:

• The proposed method significantly improved transfection efficiency in HEK-293T cells.
• A maximum transfection efficiency of 45.1% was achieved.
• Cell viability remained high, with a survival rate of 93.6%.

Conclusions:

• Femtosecond laser pulse shaping effectively enhances gene transfection efficiency and cell viability.
• This technique offers a promising solution for advancing laser-based gene delivery.
• The method provides crucial technical support for broader clinical adoption of femtosecond laser transfection.