TY - GEN
T1 - Femtosecond optical transfection as a tool for genetic manipulation of human embryonic stem cells
AU - Torres-Mapa, M. L.
AU - Gardner, J.
AU - Bradburn, H.
AU - King, J.
AU - Dholakia, K.
AU - Gunn-Moore, F.
PY - 2013
Y1 - 2013
N2 - We demonstrate the use of femtosecond optical transfection for the genetic manipulation of human embryonic stem cells. Using a system with an SLM combined with a scanning mirror allows poration of both single-cell and colony-formed human embryonic stem cells in a rapid and targeted manner. In this work, we show successful transfection of plasmid DNA tagged with fluorescent reporters into human embryonic stem cells using three doses of focused femtosecond laser. A significant number of transfected cells retained their undifferentiated morphological feature of large nucleus with high nucleus to cytoplasmic ratio, 48h after photoporation. Furthermore, DNA constructs driven by different types of promoters were also successfully transfected into human embryonic stem cells using this technique.
AB - We demonstrate the use of femtosecond optical transfection for the genetic manipulation of human embryonic stem cells. Using a system with an SLM combined with a scanning mirror allows poration of both single-cell and colony-formed human embryonic stem cells in a rapid and targeted manner. In this work, we show successful transfection of plasmid DNA tagged with fluorescent reporters into human embryonic stem cells using three doses of focused femtosecond laser. A significant number of transfected cells retained their undifferentiated morphological feature of large nucleus with high nucleus to cytoplasmic ratio, 48h after photoporation. Furthermore, DNA constructs driven by different types of promoters were also successfully transfected into human embryonic stem cells using this technique.
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U2 - 10.1117/12.2003739
DO - 10.1117/12.2003739
M3 - Conference contribution
AN - SCOPUS:84878128792
SN - 9780819493804
T3 - Proceedings of SPIE - The International Society for Optical Engineering
BT - Frontiers in Ultrafast Optics
T2 - Frontiers in Ultrafast Optics: Biomedical, Scientific, and Industrial Applications XIII
Y2 - 3 February 2013 through 5 February 2013
ER -