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

Lebogang Thobakgale; Sello Lebohang Manoto; Saturnin Ombinda Lemboumba; Malik Maaza; Patience Mthunzi-Kufa

doi:10.1117/12.2252188

15 February 2017 Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses

Lebogang Thobakgale, Sello Lebohang Manoto, Saturnin Ombinda Lemboumba, Malik Maaza, Patience Mthunzi-Kufa

Proceedings Volume 10062, Optical Interactions with Tissue and Cells XXVIII; 1006206 (2017) https://doi.org/10.1117/12.2252188
Event: SPIE BiOS, 2017, San Francisco, California, United States

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 of plasmid 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).

Conference Presentation

Citation Download Citation

Lebogang Thobakgale, Sello Lebohang Manoto, Saturnin Ombinda Lemboumba, Malik Maaza, and Patience Mthunzi-Kufa "Phototransfection of mouse embryonic stem cells with plasmid DNA using femtosecond laser pulses", Proc. SPIE 10062, Optical Interactions with Tissue and Cells XXVIII, 1006206 (15 February 2017); https://doi.org/10.1117/12.2252188

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available