Modern trends in optical bioimaging require novel nanoproducts combining high image contrast with efficient treatment capabilities. Silicon nanoparticles are a wide class of nanoobjects with tunable optical properties, which has potential as contrasting agents for fluorescence imaging and optical coherence tomography. In this paper we report on developing a novel technique for fabricating silicon nanoparticles by means of picosecond laser ablation of porous silicon films and silicon nanowire arrays in water and ethanol. Structural and optical properties of these particles were studied using scanning electron and atomic force microscopy, Raman scattering, spectrophotometry, fluorescence, and optical coherence tomography measurements. The essential features of the fabricated silicon nanoparticles are sizes smaller than 100 nm and crystalline phase presence. Effective fluorescence and light scattering of the laser-ablated silicon nanoparticles in the visible and near infrared ranges opens new prospects of their employment as contrasting agents in biophotonics, which was confirmed by pilot experiments on optical imaging.

Photodynamic therapy (PDT) is a modern minimally invasive treatment technique that has demonstrated efficiency for a wide range of clinical applications. PDT is based on the cytotoxic effect of singlet oxygen produced in biotissue as a result of irradiation with light of a specific spectral range of a photosensitizer (PS) delivered to the target area prior to the procedure.^1,2 Being highly reactive with a short radius of action, the produced singlet oxygen causes damage in cells contacting with PS, thus providing a local impact with minimal effects on the surrounding normal tissue. Currently, PDT is actively employed for antitumor treatment^3,4 and is being involved into anti-inflammatory treatment due to antimicrobial action, including cases of microbes resistant to traditional drug treatment.⁵ PDT also demonstrates positive aesthetic effects, such as skin rejuvenation, that have been well documented in a number of clinical trials.⁶ Both anti-inflammatory treatment and aesthetic medicine applications typically employ low-dose PDT regimes⁷ with total light doses not exceeding $150{\mathrm{J}/\mathrm{cm}}^2$.

Fluorescence imaging modalities are currently a routine tool for the assessment of marker distribution within biological tissues, including monitoring of fluorescent photosensitizers (PSs) in photodynamic therapy (PDT). Conventional fluorescence imaging techniques provide en-face two-dimensional images, while depth-resolved techniques require complicated tomographic modalities. In this paper, we report on a cost-effective approach for the estimation of fluorophore localization depth based on dual-wavelength probing. Owing to significant difference in optical properties of superficial biotissues for red and blue ranges of optical spectra, simultaneous detection of fluorescence excited at different wavelengths provides complementary information from different measurement volumes. Here, we report analytical and numerical models of the dual-wavelength fluorescence imaging of PS-containing biotissues considering topical and intravenous PS administration, and demonstrate the feasibility of this approach for evaluation of the PS localization depth based on the fluorescence signal ratio. The results of analytical and numerical simulations, as well as phantom experiments, were translated to the in vivo imaging to interpret experimental observations in animal experiments, human volunteers, and clinical studies. The proposed approach allowed us to estimate typical accumulation depths of PS localization which are consistent with the morphologically expected values for both topical PS administration and intravenous injection.

Plants are affected by numerous environmental factors that influence their physiological processes and productivity. Early revealing of their action based on measuring spectra of reflected light and calculating reflectance indices is an important stage in the protection of agricultural plants. Photochemical reflectance index (PRI) is a widely used parameter related to photosynthetic changes in plants under action of stressors. We developed a new system for proximal imaging of PRI based on using short pulses of measuring light detected simultaneously in green (530 nm) and yellow (570 nm) spectral bands. The system has several advances compared to those reported in literature. Active light illumination and subtraction of the ambient light allow for PRI measurements without periodic calibrations. Short duration of measuring pulses (18 ms) minimizes their influence on plants. Measurements in two spectral bands operated by separate cameras with aligned fields of visualization allow one to exclude mechanically switchable parts like filter wheels thus minimizing acquisition time and increasing durability of the setup. Absolute values of PRI and light-induced changes in PRI (ΔPRI) in pea leaves and changes of these parameters under action of light with different intensities, water shortage, and heating have been investigated using the developed setup. Changes in ΔPRI are shown to be more robust than the changes in the absolute value of PRI which is in a good agreement with our previous studies. Values of PRI and, especially, ΔPRI are strongly linearly related to the energy-dependent component of the non-photochemical quenching and can be potentially used for estimation of this component. Additionally, we demonstrate that the developed system can also measure fast changes in PRI (hundreds of milliseconds and seconds) under leaf illumination by the pulsed green-yellow measuring light. Thus, the developed system of proximal PRI imaging can be used for PRI measurements (including fast changes in PRI) and estimation of stressors-induced photosynthetic changes.