Multispectral and autofluorescence RGB imaging for skin cancer diagnostics
This paper presents the results of statistical clinical data, combining two diagnostic methods. A combination of two skin imaging methods – diffuse reflectance and autofluorescence – has been applied for skin cancer diagnostics. Autofluorescence (AF) and multispectral diffuse reflectance images were acquired by custom made prototype with 405 nm, 526 nm, 663 nm and 964 nm LEDs and RGB CMOS camera. Parameter p’ was calculated from diffuse reflectance images under green, red and infrared illumination, AF intensity (I’) was calculated from AF images exited at 405nm wavelength. Obtained results show that criterion p` > 1 gives possibility to discriminate melanomas and different kind of keratosis…
Lasers for in-vivo skin diagnostics: some recent developments
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 …
Towards combined multispectral, FLIM and Raman imaging for skin diagnostics
To explore challenges for further improvement of diagnostic performance, a project aimed at development of technology for tri-modal skin imaging by combining multispectral, fluorescence lifetime and Raman band imaging was initiated. In this study, each of the mentioned imaging modalities has been preliminary tested and updated. Four different multispectral imaging devices were tested on color standards. Picosecond laser-excited fluorescence lifetime imaging equipment was examined on ex-vivo skin samples. Finally, a new Raman spectroscopy setup with 785 nm laser was launched and tested on cell cultures and ex-vivo skin. Advantages and specific features of the tri-modal skin imaging are discu…
FRET-based method for evaluation of the efficiency of reversible and irreversible sonoporation.
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 Forster 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. …
Riga Group’s recent results on laser applications for skin diagnostics
Abstract The laser-related activities are reviewed of the Biophotonics Laboratory at UL Institute of Atomic Physics and Spectroscopy following the previous ICSQE-2018 conference. Four recent research projects are considered, including one EC Horizon-2020 project, two European Regional Development Fund (ERDF) projects and one project funded by the Latvian Council of Science (LCS). The projects are generally aimed at developing new optical methods and technologies for non-invasive in-vivo skin assessment to facilitate the early diagnostics of skin malformations (including cancers). Most of the projects explore novel approaches of camera-based biomedical imaging to the clinical diagnostics and…
Agar-based phantoms for skin diagnostic imaging
Agar-based skin phantoms with different thicknesses and hemoglobin concentration were evaluated for diagnostics of skin lesions by RGB imaging. Scattering properties of the phantoms were simulated using intralipid, absorption properties – using lyophilized powder of human hemoglobin. RGB images of phantoms were captured by self-developed laboratory made devices. The algorithm for calculation of chromophore concentrations are based on Beer-Lambert law and includes the photon path length evaluated from the measured photon-time-of-flight signals. Optical properties and chromophore concentration maps of phantoms obtained from RGB images were analyzed. The influence of chromophore concentration …
Skin-remitted photon path lengths: experimental study
Skin-remitted picosecond laser pulses were detected at four input-output fiber distances in the spectral range 560-800 nm. After deconvolution procedures, distributions and mean values of the remitted photon path lengths in forearm skin were analyzed.
Towards direct measurements of remitted photon path lengths in skin: kinetic studies in the range 520-800 nm
Skin-remitted picosecond laser pulses have been detected at variable input-output fiber distances (8 … 20 mm) in the spectral range 520-800 nm, with subsequent analysis of the pulse shape changes. Transfer functions representing the temporal responses of remitted photons to infinitely narrow δ-pulse excitation have been calculated. Parameters related to the photon path length in skin – input-output pulse peak delays, pulse FWHM, travel times of the “initial” photons and distributions of the remitted photon path lengths – are presented and analyzed. The measurement results are in general agreement with the photon propagation model expectations
Remitted photon path lengths in human skin: in-vivo measurement data
The remitted photon path lengths in human skin can be estimated by modelling; however, there are very few experimental data available to validate the simulations. This study exploited the photon time of flight method where picosecond laser pulses at seven wavelength bands in the spectral range 560-800 nm were launched into in-vivo forearm skin of 10 volunteers via an optical fiber. The pulses of back-scattered light were detected via another optical fiber placed at variable distance (1, 8, 12, 16 or 20 mm) from the input fiber, with subsequent analysis of their shapes for all 35 spectral-spatial combinations. Using a deconvolution algorithm, the distribution functions of remitted photon arr…
Remitted photon path length in human skin, skin phantoms and cell cultures
An experimental method for remitted photon path length measurements in scattering media has been developed and tested on human skin and skin neoplasms, skin phantoms and cell cultures. The photon time-of-flight (PTOF) measurement method was used in this study, where the photon travel time was converted into path length. Remitted light signals were obtained using a picosecond broadband laser and a set of narrowband interference filters in spectral rang 520 – 760 nm. Five different distances of 1, 8, 12, 16 and 20 mm between the source and detector fibers were used. Measurements were performed at different wavelengths and distance combinations; they were taken from human skin and skin malform…
Skin chromophore mapping from multi-spectral laser line images
Skin chromophore maps can be used for assessment of various skin malformations and early cancer diagnostics. Commercially available devices are bulky and expensive. We present two portable proof-of-concept device prototypes for multi-spectral laser line imaging with three (448 nm, 532 nm and 659 nm) and four (450 nm, 523 nm, 638 nm and 850 nm) wavelength laser illumination. Laser modules and special optics that ensure uniform light distribution over the region of interest have been exploited. Skin chromophore maps were calculated using Beer-Lambert law, considering light scattering properties in the skin and including photon path length evaluated from the directly measured photon-time-of-fl…