Application of optical tweezers and excimer laser to study protoplast fusion

Titirat Kantawang; Sompid Samipak; Jumras Limtrakul; Nattaporn Chattham

doi:10.1117/12.2195700

29 July 2015 Application of optical tweezers and excimer laser to study protoplast fusion

Titirat Kantawang, Sompid Samipak, Jumras Limtrakul, Nattaporn Chattham

Proceedings Volume 9659, International Conference on Photonics Solutions 2015; 96590F (2015) https://doi.org/10.1117/12.2195700
Event: International Conference on Photonics Solutions 2015, 2015, Hua Hin, Thailand

Abstract

Protoplast fusion is a physical phenomenon that two protoplasts come in contact and fuse together. Doing so, it is possible to combine specific genes from one protoplast to another during fusion such as drought resistance and disease resistance. There are a few possible methods to induce protoplast fusion, for example, electrofusion and chemical fusion. In this study, chemical fusion was performed with laser applied as an external force to enhance rate of fusion and observed under a microscope. Optical tweezers (1064 nm with 100X objective N.A. 1.3) and excimer laser (308 nm LMU-40X-UVB objective) were set with a Nikon Ti-U inverted microscope. Samples were prepared by soaking in hypertonic solution in order to induce cell plasmolysis. Elodea Canadensis and Allium cepa plasmolysed leaves were cut and observed under microscope. Concentration of solution was varied to induce difference turgor pressures on protoplasts pushing at cell wall. Free protoplasts in solution were trapped by optical tweezers to study the effect of Polyethylene glycol (PEG) solution. PEG was diluted by Ca⁺ solution during the process to induced protoplast cell contact and fusion. Possibility of protoplast fusion by excimer laser was investigated and found possible. Here we report a novel tool for plant cell fusion using excimer laser. Plant growth after cell fusion is currently conducted.

Citation Download Citation

Titirat Kantawang, Sompid Samipak, Jumras Limtrakul, and Nattaporn Chattham "Application of optical tweezers and excimer laser to study protoplast fusion", Proc. SPIE 9659, International Conference on Photonics Solutions 2015, 96590F (29 July 2015); https://doi.org/10.1117/12.2195700

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KEYWORDS

Excimer lasers

Microscopes

Optical tweezers

Laser welding

Objectives

Resistance

Spherical lenses

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