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Journal of Biomedical Optics

Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporation efficiencies
Author(s): Mindaugas Tamošiūnas; Roberts Kadikis; Inga Saknīte; Juozas Baltušnikas; Audrius Kilikevičius; Alexey Lihachev; Ramona Petrovska; Dainis Jakovels; Saulius Šatkauskas
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Paper 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  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.

Paper Details

Date Published: 29 April 2016
PDF: 12 pages
J. Biomed. Opt. 21(4) 045003 doi: 10.1117/1.JBO.21.4.045003
Published in: Journal of Biomedical Optics Volume 21, Issue 4
Show Author Affiliations
Mindaugas Tamošiūnas, Vytautas Magnus Univ. (Lithuania)
Roberts Kadikis, Institute of Electronics & Computer Science (Latvia)
Inga Saknīte, Univ. of Latvia (Latvia)
Juozas Baltušnikas, Lietuvos Sporto Univ. (Lithuania)
Audrius Kilikevičius, Lietuvos Sporto Univ. (Lithuania)
Alexey Lihachev, Univ. of Latvia (Latvia)
Ramona Petrovska, Latvian Biomedical Research & Study Ctr. (Latvia)
Dainis Jakovels, Univ. of Latvia (Latvia)
Saulius Šatkauskas, Vytautas Magnus Univ. (Lithuania)

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