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23 August 2011 Enhancement of the efficiency of femtosecond optical transfection
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Proceedings Volume 8173, Photonics 2010: Tenth International Conference on Fiber Optics and Photonics; 817307 (2011) https://doi.org/10.1117/12.899117
Event: International Conference on Fiber Optics and Photonics, 2010, Guwahati, India
Abstract
Cell transfection is the process in which extra cellular nucleic acids such as DNA, RNA, Si-RNA can be deliberately injected into the cytoplasm of the cell. This technique of cell transfection forms a central tool in the hands of a cell biologist to explore the mechanism within the cell. In optical transfection a well focused laser spot alters the permeability of the cell membrane so as to allow the entry of extra-nuclear materials into the cell. Femto-second optical transfection have proved to be better than other laser based cell transfection, owing to the three dimensionally confined multi-photon effects on the cell membrane thereby leaving the rest of the cell unaffected. Even though the femto-second optical transfection has proved to be sterile, non-invasive and highly selective, it has to improve in terms of efficiency, and throughput to address real life problems. We report here a method to achieve significant enhancement in the efficiency of femto-second optical transfection. The protocol of the transfection procedure is modified by adding a suitable biochemical reagent - Nupherin-neuron - into the cell medium during the transfection, which can assist the delivery of DNA into the nucleus once the DNA gets injected into the cytoplasm of the cell. We achieved a 3 fold enhancement in the transfection efficiency with this modified protocol. Also we report for the first time the transfection of recently trypsinised cells with a very high transfection efficiency, which would pave way to the development of high throughput microfluidic optical transfection devices.
© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Bavishna B. Praveen, David Stevenson, Maciej Antkowiak, Frank J. Gunn-Moore, and Kishan Dholakia "Enhancement of the efficiency of femtosecond optical transfection", Proc. SPIE 8173, Photonics 2010: Tenth International Conference on Fiber Optics and Photonics, 817307 (23 August 2011); https://doi.org/10.1117/12.899117
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