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Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses

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dc.contributor.author Thobakgale, Setumo L
dc.contributor.author Manoto, Sello L
dc.contributor.author Ombinda-Lemboumba, Saturnin
dc.contributor.author Maaza, M
dc.contributor.author Mthunzi-Kufa, Patience
dc.date.accessioned 2017-11-06T12:50:04Z
dc.date.available 2017-11-06T12:50:04Z
dc.date.issued 2017-02
dc.identifier.citation Thobakgale, S.L. et al. 2017. Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses. Proceedings Volume 10062, Optical Interactions with Tissue and Cells XXVIII; 1006206 (2017), San Francisco, California, United States, 28 January- 2 February 2017 en_US
dc.identifier.isbn 9781510605657
dc.identifier.issn 1605-7422
dc.identifier.uri https://www.spiedigitallibrary.org/conference-proceedings-of-spie/10062/1/Phototransfection-of-mouse-embryonic-stem-cells-with-plasmid-DNA-using/10.1117/12.2252188.full?SSO=1
dc.identifier.uri DOI: 10.1117/12.2252188
dc.identifier.uri http://hdl.handle.net/10204/9724
dc.description Copyright: 2017 SPIE. Due to copyright restrictions, the attached PDF file only contains the abstract of the full text item. For access to the full text item, kindly consult the publisher's website. en_US
dc.description.abstract Cellular manipulation by delivery of molecules into cells has been applied extensively in tissue engineering research for medical applications. The different molecular delivery techniques used range from viral and chemical agents to physical and electrical methods. Although successful in most studies, these techniques have inherent difficulties such as toxicity, unwanted genetic mutations and low reproducibility respectively. Literature recognizes pulsed lasers at femtosecond level to be most efficient in photonic interactions with biological material. As of late, laser pulses have been used for drug and DNA delivery into cells via transient optical perforation of the cellular membrane. Thus in this study, we design and construct an optical system coupled to a femtosecond laser for the purpose of phototransfection or insertion ofplasmid DNA (pDNA) into cells using lasers. We used fluorescent green protein (pGFP) to transfect mouse embryonic stem cells as our model. Secondly, we applied fluorescence imaging to view the extent of DNA delivery using this method. We also assessed the biocompatibility of our system by performing molecular assays of the cells post irradiation using adenosine triphosphate (ATP) and lactate dehydrogenase (LDH). en_US
dc.language.iso en en_US
dc.publisher SPIE en_US
dc.relation.ispartofseries Worklist;18602
dc.subject Phototransfection en_US
dc.subject Femtosecond en_US
dc.subject Green protein en_US
dc.subject pGFP en_US
dc.title Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses en_US
dc.type Conference Presentation en_US
dc.identifier.apacitation Thobakgale, S. L., Manoto, S. L., Ombinda-Lemboumba, S., Maaza, M., & Mthunzi-Kufa, P. (2017). Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses. SPIE. http://hdl.handle.net/10204/9724 en_ZA
dc.identifier.chicagocitation Thobakgale, Setumo L, Sello L Manoto, Saturnin Ombinda-Lemboumba, M Maaza, and Patience Mthunzi-Kufa. "Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses." (2017): http://hdl.handle.net/10204/9724 en_ZA
dc.identifier.vancouvercitation Thobakgale SL, Manoto SL, Ombinda-Lemboumba S, Maaza M, Mthunzi-Kufa P, Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses; SPIE; 2017. http://hdl.handle.net/10204/9724 . en_ZA
dc.identifier.ris TY - Conference Presentation AU - Thobakgale, Setumo L AU - Manoto, Sello L AU - Ombinda-Lemboumba, Saturnin AU - Maaza, M AU - Mthunzi-Kufa, Patience AB - Cellular manipulation by delivery of molecules into cells has been applied extensively in tissue engineering research for medical applications. The different molecular delivery techniques used range from viral and chemical agents to physical and electrical methods. Although successful in most studies, these techniques have inherent difficulties such as toxicity, unwanted genetic mutations and low reproducibility respectively. Literature recognizes pulsed lasers at femtosecond level to be most efficient in photonic interactions with biological material. As of late, laser pulses have been used for drug and DNA delivery into cells via transient optical perforation of the cellular membrane. Thus in this study, we design and construct an optical system coupled to a femtosecond laser for the purpose of phototransfection or insertion ofplasmid DNA (pDNA) into cells using lasers. We used fluorescent green protein (pGFP) to transfect mouse embryonic stem cells as our model. Secondly, we applied fluorescence imaging to view the extent of DNA delivery using this method. We also assessed the biocompatibility of our system by performing molecular assays of the cells post irradiation using adenosine triphosphate (ATP) and lactate dehydrogenase (LDH). DA - 2017-02 DB - ResearchSpace DP - CSIR KW - Phototransfection KW - Femtosecond KW - Green protein KW - pGFP LK - https://researchspace.csir.co.za PY - 2017 SM - 9781510605657 SM - 1605-7422 T1 - Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses TI - Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses UR - http://hdl.handle.net/10204/9724 ER - en_ZA


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