0000000000376819
AUTHOR
Roberts Kadikis
Benign — A typical nevi discrimination using diffuse reflectance and fluorescence multispectral imaging system
Early diagnostics of skin cancer is of interest for dermatologists. Atypical nevi are not considered to be malignant, but are suspects that should be detected and monitored over time. The multispectral imaging system Nuance operating in spectral range 450–950 nm was adapted for clinical in vivo measurements in diffuse reflectance and fluorescence mode. Mean and standard deviation values of optical density and fluorescence intensity were extracted from segmented pigmented lesions (21 benign and 26 atypical nevi) and used for further analysis. It was possible to achieve 62% sensitivity and 67% specificity for discrimination between atypical and benign lesions using averaged fluorescence mean …
Application of fluorescence spectroscopy and multispectral imaging for non-invasive estimation of GFP transfection efficiency
Electroporation and ultrasound induced sonoporation has been showed to induce plasmid DNA transfection to the mice tibialis cranialis muscle. It offers new prospects for gene therapy and cancer treatment. However, numerous experimental data are still needed to deliver the plausible explanation of the mechanisms governing DNA electro- or sono-transfection, as well as to provide the updates on transfection protocols for transfection efficiency increase. In this study we aimed to apply non-invasive optical diagnostic methods for the real time evaluation of GFP transfection levels at the reduced costs for experimental apparatus and animal consumption. Our experimental set-up allowed monitoring …
pEGFP transfection into murine skeletal muscle by electrosonoporation
In this study, we aimed to determine whether the combination of electroporation (EP) and ultrasound (US) waves (sonoporation) can affect the plasmid DNA transfection to mice tibialis cranialis muscle. Multispectral imaging technique combined with fluorescence spectroscopy point measurements has been used for the transcutaneous detection of enhanced green fluorescent protein (EGFP) fluorescence, providing information on location and duration of EGFP expression. We found that electrosonoporation, commonly enhancing pDNA transfection in vitro , had no positive effect on EGFP transfection efficiency increase in vivo with respect to electroporation alone. We presume that this may be associated w…
Noninvasive optical diagnostics of enhanced green fluorescent protein expression in skeletal muscle for comparison of electroporation and sonoporation efficiencies.
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 10 μg/50 ml ) transfection, we used electroporation or ultrasound enhanced microbubble cavitation [sonoporation (SP)]. The transcutaneous EGFP fluorescence in live mice was monit…