Research Papers: General

Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporation efficiencies

[+] Author Affiliations
Mindaugas Tamošiūnas

Vytautas Magnus University, Biophysical Research Group, Faculty of Natural Sciences, Vileikos 8, Kaunas LT-44404, Lithuania

Vytautas Magnus University, Department of Biochemistry, Faculty of Natural Sciences, Vileikos 8, Kaunas LT-44404, Lithuania

Roberts Kadikis

Institute of Electronics and Computer Science, 14 Dzerbenes Street, Riga LV-1006, Latvia

Inga Saknīte, Alexey Lihachev, Dainis Jakovels

University of Latvia, Institute of Atomic Physics and Spectroscopy, 19 Rainis Boulevard, Riga LV-1586, Latvia

Juozas Baltušnikas, Audrius Kilikevičius

Lithuanian Sports University, Institute of Sports Sciences and Innovation, Sporto 6, LT-44221 Kaunas, Lithuania

Ramona Petrovska

Latvian Biomedical Research and Study Centre, Ratsupites iela 1, Riga LV-1067, Latvia

Saulius Šatkauskas

Vytautas Magnus University, Biophysical Research Group, Faculty of Natural Sciences, Vileikos 8, Kaunas LT-44404, Lithuania

J. Biomed. Opt. 21(4), 045003 (Apr 29, 2016). doi:10.1117/1.JBO.21.4.045003
History: Received August 31, 2015; Accepted April 12, 2016
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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.

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© 2016 Society of Photo-Optical Instrumentation Engineers

Citation

Mindaugas Tamošiūnas ; Roberts Kadikis ; Inga Saknīte ; Juozas Baltušnikas ; Audrius Kilikevičius, et al.
"Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporation efficiencies", J. Biomed. Opt. 21(4), 045003 (Apr 29, 2016). ; http://dx.doi.org/10.1117/1.JBO.21.4.045003


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