Enhancement of the efficiency of femtosecond optical transfection

Bavishna B. Praveen; David Stevenson; Maciej Antkowiak; Frank J. Gunn-Moore; Kishan Dholakia

doi:10.1117/12.899117

23 August 2011 Enhancement of the efficiency of femtosecond optical transfection

Bavishna B. Praveen, David Stevenson, Maciej Antkowiak, Frank J. Gunn-Moore, Kishan Dholakia

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.

Citation Download Citation

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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KEYWORDS

Femtosecond phenomena

Ovary

Microfluidics

Tissue optics

Biology

Fluorescent proteins

Laser optics

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