Please use this identifier to cite or link to this item:https://hdl.handle.net/20.500.12259/58393
Type of publication: Straipsnis Clarivate Analytics Web of Science ar/ir Scopus / Article in Clarivate Analytics Web of Science or / and Scopus (S1)
Field of Science: Biologija / Biology (N010)
Author(s): Kadikis, Roberts;Saknite, Inga;Kilikevičius, Audrius;Lihachev, Alexey;Petrovska, Ramona;Jakovels, Dainis;Tamošiūnas, Mindaugas;Baltušnikas, Juozas;Šatkauskas, Saulius
Title: Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporation efficiencies
Is part of: Journal of biomedical optics. Bellingham, WA : SPIE, 2016, Vol. 21, iss. 4
Extent: p. 1-12
Date: 2016
Note: E-ISSN: 1560-2281. Impact Factor: 2.859
Keywords: Sonoporation;Electroporation;Green fluorescent proteins;Multispectral imaging;Fluorescence spectroscopy
Abstract: We highlight the options available for noninvasive optical diagnostics of reporter gene expression in mouse tibialis cranialis muscle. An in vivo multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) expression, providing information on location and duration of EGFP expression and allowing quantification of EGFP expression levels. For EGFP coding plasmid (pEGFP-Nuc Vector, 10  μg/50  ml10  μg/50  ml) transfection, we used electroporation or ultrasound enhanced microbubble cavitation [sonoporation (SP)]. The transcutaneous EGFP fluorescence in live mice was monitored over a period of one year using the described parameters: area of EGFP positive fibers, integral intensity, and mean intensity of EGFP fluorescence. The most efficient transfection of EGFP coding plasmid was achieved, when one high voltage and four low voltage electric pulses were applied. This protocol resulted in the highest short-term and long-term EGFP expression. Other electric pulse protocols as well as SP resulted in lower fluorescence intensities of EGFP in the transfected area. We conclude that noninvasive multispectral imaging technique combined with fluorescence spectroscopy point measurements is a suitable method to estimate the dynamics and efficiency of reporter gene transfection in vivo
Internet: https://doi.org/10.1117/1.JBO.21.4.045003
https://doi.org/10.1117/1.JBO.21.4.045003
Affiliation(s): Biochemijos katedra
Biologijos katedra
Gamtos mokslų fakultetas
Lietuvos sporto universitetas
Vytauto Didžiojo universitetas
Appears in Collections:Universiteto mokslo publikacijos / University Research Publications

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