Application of dynamic diffractive optics for enhanced femtosecond laser based cell transfection.
We demonstrate the advantages of a dynamic diffractive optical element, namely a spatial light modulator (SLM) for the controlled and enhanced optoinjection and phototransfection of mammalian cells with a femtosecond light source. The SLM provides full control over the lateral and axial positioning of the beam with sub-micron precision. Fast beam translation enables time-sequenced irradiation, which is shown to enhance the optoinjection efficiency and alleviate the problem of exact beam positioning on the cell membrane. We show that irradiation in three axial positions doubles the number of viably optoinjected cells when compared with a single dose. The presented system also enables untargeted raster scan irradiation which provides a higher throughput transfection of adherent cells at the rate of 1 cell per second. Additionally, fluorescent imaging is used to demonstrate cell selective two-step gene therapy.
Antkowiak
Maciej
M
SULSA, School of Biology, University of St Andrews, St Andrews KY169TS, UK. ma81@st-andrews.ac.uk
Torres-Mapa
Maria Leilani
ML
Gunn-Moore
Frank
F
Dholakia
Kishan
K
D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion; D006224:Cricetinae; D003412:Cricetulus; D007834:Lasers; D008027:Light; D055620:Optical Phenomena; D011829:Radiation Dosage; D012542:Scattering, Radiation; D013997:Time Factors; D014162:Transfection / Q000295:instrumentation / Q000379:methods*