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Infrared Laser-Based Single Cell Permeabilization by Plasma Membrane Temperature Gradients.

Allen L Garner1,2,3, Bogdan Neculaes4, Dmitry V Dylov4

  • 1School of Nuclear Engineering, Purdue University, West Lafayette, IN 47906, USA.

Membranes
|June 23, 2022
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Summary

Laser microinjection offers a simpler, more cost-effective alternative to traditional methods for precise cell delivery. This study demonstrates its potential for selective cell permeabilization using a 1550 nm femtosecond laser.

Keywords:
membrane permeabilizationmicroinjectionoptoinjectionoptoporationtemperature gradientstransfection

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Laser Physics

Background:

  • Single cell microinjection is precise but complex and requires skilled operators.
  • Laser-based microinjection offers a standardized, accessible alternative.

Purpose of the Study:

  • To investigate laser-based microinjection using a 1550 nm femtosecond laser.
  • To elucidate the physical mechanism behind laser-induced cell permeabilization.

Main Methods:

  • Utilized a 1550 nm, 100 fs pulse duration laser at a 20 ns repetition rate for microinjection.
  • Exposed Chinese Hamster Ovarian (CHO) cells to the laser and assessed propidium iodide uptake.
  • Calculated transmembrane potential changes due to laser-induced temperature gradients.

Main Results:

  • Demonstrated selective cell permeabilization in CHO cells via propidium iodide uptake.
  • Observed agreement between experimental conditions and electropermeabilization thresholds.
  • Hypothesized a mechanism involving laser-induced plasma membrane temperature gradients.

Conclusions:

  • 1550 nm femtosecond lasers show potential for cost-effective, operator-agnostic microinjection.
  • This method may induce fewer deleterious effects compared to traditional lasers.
  • Laser-induced temperature gradients are a likely mechanism for cell permeabilization.