Vladislav A. Kamensky

Complementary use of cross-polarization and standard OCT for differential diagnosis of pathological tissues

An experimental standard optical coherence tomography (OCT) setup that can be easily modified for cross-polarization OCT (CP OCT) operation has been developed to perform differential diagnosis of pathological tissues. The complementary use of CP OCT, a technique that provides a map of cross-polarization backscattering properties of an object being studied by means of low-coherence interferometry, and standard OCT imaging improves the specificity of diagnostics of pathological changes occurring in tissues. It is shown that healthy, neoplastic and scar tissues of the esophagus have different cross-polarization backscattering properties. A comparative analysis of CP OCT, OCT and histological images from one and the same tissue area has been made. A close correlation between the location of collagen fibers in biological tissue and signal intensity in CP OCT images is found.

Contrasting properties of gold nanoparticles for optical coherence tomography: phantom, in vivo studies and Monte Carlo simulation.

The possibility of using silica-gold nanoshells with 150 nm silica core size and 25 nm thick gold shell as contrasting agents for optical coherence tomography (OCT) is analyzed. Experiments on agar biotissue phantoms showed that the penetration of nanoshells into the phantoms increases the intensity of the optical coherence tomography (OCT) signal and the brightness of the corresponding areas of the OCT image. In vivo experiments on rabbit skin demonstrated that the application of nanoshells onto the skin provides significant contrasting of the borders between the areas containing nanoshells and those without. This effect of nanoshells on skin in vivo is manifested by the increase in intensity of the OCT signal in superficial parts of the skin, boundary contrast between superficial and deep dermis and contrast of hair follicles and glands. The presence of nanoshells in the skin was confirmed by electron microscopy. Monte Carlo simulations of OCT images confirmed the possibility of contrasting skin-layer borders and structures by the application of gold nanoshells. The Monte Carlo simulations were performed for two skin models and exhibit effects of nanoparticles similar to those obtained in the experimental part of the study, thus proving that the effects originate exactly from the presence of nanoparticles.

Novel algorithm of processing optical coherence tomography images for differentiation of biological tissue pathologies

A numerical algorithm based on a small-angle approximation of the radiative transfer equation (RTE) is developed to reconstruct scattering characteristics of biological tissues from optical coherence tomography (OCT) images. According to the algorithm, biological tissue is considered to be a layered random medium with a set of scattering parameters in each layer: total scattering coefficient, variance of a small-angle scattering phase function, and probability of backscattering, which fully describe the OCT signal behavior versus probing depth. The reconstruction of the scattering parameters is performed by their variation to fit the experimental OCT signal by the theoretical one using a time-saving genetic algorithm. The proposed reconstruction procedure is tested on model media with known scattering parameters. The possibility to estimate scattering parameters from OCT images is studied for various regimes of OCT signal decay. The developed algorithm is applied to reconstruct optical characteristics of epithelium and stroma for normal cervical tissue and its pathologies, and the potential to distinguish between the types of pathological changes in epithelial tissue by its OCT images is demonstrated.

Phototoxic effects of fluorescent protein KillerRed on tumor cells in mice

KillerRed is known to be a unique red fluorescent protein displaying strong phototoxic properties. Its effectiveness has been shown previously for killing bacterial and cancer cells in vitro. Here, we investigated the photototoxicity of the protein on tumor xenografts in mice. HeLa Kyoto cell line stably expressing KillerRed in mitochondria and in fusion with histone H2B was used. Irradiation of the tumors with 593 nm laser led to photobleaching of KillerRed indicating photosensitization reaction and caused significant destruction of the cells and activation of apoptosis. The portion of the dystrophically changed cells increased from 9.9% to 63.7%, and the cells with apoptosis hallmarks from 6.3% to 14%. The results of this study suggest KillerRed as a potential genetically encoded photosensitizer for photodynamic therapy of cancer.

In vivo study of the effect of mechanical compression on formation of OCT images of human skin.

Modern optical diagnostic techniques often require deformations of the studied bio-tissues for image acquisition. This paper discusses the effect of mechanical compression on the formation of OCT images of human skin. The study was performed in vivo on human volunteers of different age. We show that application of compression to human skin induces changes in optical properties of the sample associated with elasticity of different skin layers. These changes induce an increase in the contrast of interlayer boundaries. Further application of compression causes the appearance of dark areas in the OCT images obtained, likely associated with interstitial or intracellular water inflow to the observed region. The effects studied are of importance for proper interpretation of obtained OCT images in diagnosis of skin pathologies.

In situ monitoring of laser modification process in human cataractous lens and porcine cornea using coherence tomography

We demonstrate that optical coherence tomography (OCT) is a convenient diagnostic tool to monitor pulse-to-pulse kinetics in laser interactions with biological tissue. In experiments on laser modification and ablation of the cataractous human lens and the porcine cornea we have applied this technique in situ to investigate different modes of preablation tissue swelling, crater formation and thermally affected zone development. The cataractous lens is an example of highly scattering media whereas the cornea is initially low scattering. The radiation with different wavelengths has been employed including that of a YAG:Er laser (λ=2.94 μm), a glass:Er laser (λ=1.54 μm), YAG:Nd lasers (λ=1.32 μm and λ=1.44 μm), as well as of the fifth harmonic of a Nd:YAP laser (λ=0.216 μm). Pulse-to-pulse OCT monitoring has been accompanied by the probe beam shielding diagnostics to provide the time-resolved observation of the interaction dynamics.

Compression as a method for increasing the informativity of optical coherence tomography of biotissues

The efficiency of the mechanical compression of biotissues for improving the differentiation between pathological changes in the structure of a biotissue observed by the method of optical coherence tomography (OCT) is investigated. The effect of the compression in the OCT-images of samples of the human rectum affected by inflammation and carcinoma is studied ex vivo. It is shown that the use of compression makes it possible to differentiate between these pathological changes. To interpret experimental data, images of an inflamed part of rectum are modeled by the Monte Carlo method for different degrees of compression. The results of modeling agree qualitatively with the experimental data.

Fluorescence diffuse tomography for detection of red fluorescent protein expressed tumors in small animals.

A fluorescence diffuse tomography (FDT) setup for monitoring tumor growth in small animals has been created. In this setup an animal is scanned in the transilluminative configuration by a single source and detector pair. To remove stray light in the detection system, we used a combination of interferometric and absorption filters. To reduce the scanning time, an experimental animal was scanned using the following algorithm: (1) large-step scanning to obtain a general view of the animal (source and detector move synchronously); (2) selection of the fluorescing region; and (3) small-step scanning of the selected region and different relative shifts between the source and detector to obtain sufficient information for 3D reconstruction. We created a reconstruction algorithm based on the Holder norm to estimate the fluorophore distribution. This algorithm converges to the solution with a minimum number of fluorescing zones. The use of tumor cell lines transfected with fluorescent proteins allowed us to conduct intravital monitoring studies. Cell lines of human melanomas Mel-P, Mel-Ibr, Mel-Kor, and human embryonic kidney HEK293 Phoenix were transfected with DsRed-Express and Turbo-RFP genes. The emission of red fluorescent proteins (RFPs) in the long-wave optical range permits detection of deep-seated tumors. In vivo experiments were conducted immediately after subcutaneous injection of fluorescing cells into small animals.

Simultaneous photoacoustic and optically mediated ultrasound microscopy: phantom study.

An experimental setup for combined photoacoustic (PA) and optically mediated ultrasound (US) microscopy is presented. A spherically focused 35 MHz polyvinylidene fluoride (PVDF) ultrasonic detector with a numerical aperture of 0.28, a focal distance of 9 mm, and a bandwidth (-6 dB level) of 24 MHz was used to obtain PA and US data with a 3 mm imaging depth. A fiber-optic system was employed to deliver laser excitation pulses from a tunable laser to the studied medium. A single optical pulse was used to form both PA and US A-scans. The probing US pulses were generated thermoelastically due to absorption of backscattered laser radiation by the metalized surface of a PVDF film.

OCT‐guided laser hyperthermia with passively tumor‐targeted gold nanoparticles

The goal of this study is the development of a method of local laser hyperthermia with gold nanoparticles under noninvasive optical monitoring of nanoparticle accumulation in tumor tissue in vivo. Bifunctional plasmon resonant nanoparticles that are optimal for OCT diagnostics and laser heating at the wavelength of 810 nm were used in the study. The OCT examination showed that the accumulation of gold nanoparticles in the tumor invading into skin was maximal 4-5 h after intravenous injection. It was demonstrated that nanoparticle accumulation in tumor allowed more local heating and enhanced thermal sensitivity of tumor tissue. Laser hyperthermia that heated tumor up to 44-45 °C at maximum nanoparticle accumulation induced apoptotic death of tumor cells and inhibited tumor growth by 104% on the 5th day after treatment.