The recent advancements of three laser-based diagnostic technologies developed at the Riga group are briefly reviewed: (i) RGB imaging of cw-laser excited skin autofluorescence intensity and photobleaching rate distributions, (ii) ps-laser excited skin autofluorescence and diffuse reflectance kinetics analysis, (iii) snapshot RGB skin chromophore mapping under triple-laser illumination. These techniques have passed preliminary laboratory and clinical tests which have demonstrated a promising potential for further implementation in portable devices for routine clinical applications. Operation principles, set-up schemes and some clinical results obtained by the above-mentioned techniques are discussed in this paper.
It is widely known that not all of the treated cells survive after introduction of exogenous molecules via any physical method. Therefore, it is important to develop methods that would allow simultaneous evaluation of both molecular delivery efficiency and cell viability. This study presents Förster resonance energy transfer (FRET)-based method that allows molecular transfer and cell viability evaluation in a single measurement by employing two common fluorescent dyes, namely, ethidium bromide and trypan blue. The method has been validated using cell sonoporation. The FRET-based method allows the efficiency evaluation of both reversible and irreversible sonoporation in a single experiment. Therefore, this method could be used to reduce time, labor, and material cost while improving the accuracy of evaluations.
The feasibility of smartphones for in vivo skin autofluorescence imaging has been investigated. Filtered autofluorescence images from the same tissue area were periodically captured by a smartphone RGB camera with subsequent detection of fluorescence intensity decreasing at each image pixel for further imaging the planar distribution of those values. The proposed methodology was tested clinically with 13 basal cell carcinoma and 1 atypical nevus. Several clinical cases and potential future applications of the smartphone-based technique are discussed.
Experimental methodology for parallel measurements of in-vivo skin autofluorescence (AF) lifetimes and photobleaching dynamic has been developed and tested. The AF lifetime decay distributions were periodically collected from fixed tissue area with subsequent detection of the fluorescence intensity decrease dynamic at different time gates after the pulse excitation. Temporal distributions of human <i>in-vivo </i>skin AF lifetimes and bleaching kinetics were collected and analyzed by means of commercial time-correlated single photon counting system.
The autofluorescence lifetime of healthy human skin was measured using excitation provided by a picosecond diode laser operating at a wavelength of 405 nm and with fluorescence emission collected at 475 and 560 nm. In addition, spectral and temporal responses of healthy human skin and intradermal nevus in the spectral range 460 to 610 nm were studied before and after photobleaching. A decrease in the autofluorescences lifetimes changes was observed after photobleaching of human skin. A three-exponential model was used to fit the signals, and under this model, the most significant photoinduced changes were observed for the slowest lifetime component in healthy skin at the spectral range 520 to 610 nm and intradermal nevus at the spectral range 460 to 610 nm.
The impact of visible cwlaser irradiation on skin autofluorescence lifetimes was investigated in spectral range from 450 nm to 600 nm. Skin optical provocations were performed during 1 min by 405 nm low power cw laser with power density up to 20 mW/cm<sup>2</sup>. Autofluorescence lifetimes were measured before and immediately after the optical provocation.
The effect of ultrasound exposure on bleomycin fluorescence and pharmacological properties is studied. Bleomycin was treated by ultrasound for 7 min. Bleomycin fluorescence was measured during ultrasound exposure by means of fiber-optic spectrometry. Cell colony test was used to evaluate blemycin cytotoxity before and after ultrasound exposure.
Influence of low power laser irradiance on healthy skin using diffuse reflectance spectroscopy and multispectral
imaging was studied. Changes of diffuse reflectance spectra in spectral range from 500 to 600 nm were observed after
405 nm, 473 nm and 532 nm laser provocation, leading to conclusion that the content of skin hemoglobin has changed.
Peaks in spectral absorbance (optical density) curves corresponded to well-known oxy-hemoglobin absorbance peaks at
542 and 577 nm.