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Carotenoids play multiple roles in insects, including coloration and protection. Most insects can obtain carotenoids only from their diet. Therefore, carotenoids are proposed to reflect trophic chains and lifestyles of insects. We investigated the mini-ecosystem of a gall on a hawkweed Hieracium × robustum induced by the gall wasp Aulacidea hieracii (Hymenoptera: Cynipidae) that is attacked by parasitoid wasp Eurytoma cynipsea (Hymenoptera: Eurytomidae). The parasitoid larvae consume the gall wasp larvae that consume the gall tissues. We employed resonance Raman spectroscopy to trace the fate of carotenoids in living larvae and pupae of these insects. We showed that carotenoid composition in the parasitoid closely corresponds to that of its diet—the gall wasp. On the contrary, carotenoid composition in the gall wasp was independent of that in the gall tissues, and the carotenoid concentration increases as non-feeding larvae mature. Thus, A. hieracii is suggested as a candidate among insects to have the ability to synthesize and modify carotenoids. Our findings give rise to the question of the relevance of using carotenoids as markers of trophic flow in the gall community.

The results of the study of the morphology and surface topography of chitosan films of the salt (S-) and basic (B-) chemical form by atomic force microscopy are presented. The films were cast from polymer solutions in acetic, lactic, citric and succinic acid. NaOH and triethanolamine were used for the salt → chitosan base reaction. Surface tomograms were obtained; the main morphological characteristics and roughness parameters of the film samples were estimated. It was found that the morphology, the degree of order, root-mean-square roughness and the height of the surface roughness were determined by the polymer chemical form, the nature of the acid used and the reagent of the polymer-like conversion reaction. The surface of the S-form chitosan films is characterized by fibrillar structural ordering (also dendritic for chitosan succinate), and that of the B-form is globular. The smallest size of surface supramolecular elements was observed for the S-form chitosan films, while the greatest roughness was for the B-form ones. Changing the reagent of the chitosan S → B reaction did not affect the morphological characteristics of the films; however, it affected the microrelief roughness. A more uniform basic chitosan film is formed in an organic base environment. It was suggested that the formation of fibrillar supramolecular structures was due to the unfolding and straightening of macrochains because of the repulsion of the same charged monomer units, while the globular ones were formed due to the folding and densification of macrocoils after neutralizing the total charge of the macrochain.

Label-free characterization of cell subpopulations is a very promising biomedical approach. Nowadays, there are several label-free methods based on different physical properties such as size, density, stiffness, etc. allowing the characterization of biological objects. However, fluorescence properties are the most suitable feature for the label-free study of tissue and cells. Understanding the autofluorescence level peculiarities of normal and pathological / live and dead cells can become a helpful tool for cells' metabolic activity, viability evaluation, and diagnostics of a number of diseases. In this study, we applied a series of mouse cell lines (RAW 264.7 - macrophages, L929 - fibroblasts, C2C12 - myoblasts, and B16-F10 - melanoma) to compare cell autofluorescence of live and dead cells under 488 nm laser excitation and found the difference between their autofluorescence depending on a cell state and type.

Surface-enhanced Raman scattering (SERS) is widely used for in vitro and in vivo bioimaging applications. However, reproducible and controllable fabrication of SERS tags with a high density of elec...

Surface-enhanced Raman scattering (SERS) has proven to be a promising technique for different types of imaging including preoperative and intraoperative in vivo tumor visualization. However, the strong scattering of the turbid tissue limits its use in subcutaneous areas. In this paper, we used an optical clearing technique to improve the SERS signal from a subcutaneous tumor phantom. The phantom is a 2 mm sphere of calcium alginate with incorporated petal-like gap-enhanced Raman tags. The use of optical clearing increases the SERS signal target-to-background ratio for 5 times and allow to decrease the total imaging time for at least 10 times. In addition, SERS imaging assisted with optical clearing made it possible to more precisely determine the shape and boundaries of the implanted phantom. The combination of optical clearing and SERS is a promising strategy for the clinical imaging of subcutaneous objects that are usually shielded by dermal tissue. This article is protected by copyright. All rights reserved.

Photoswitchable fluorescent proteins (PFPs) that can change fluorescence color upon excitation have revolutionized many applications of light such as tracking protein movement, super-resolution imaging, identification of circulating cells, and optical data storage. Nevertheless, the relatively weak fluorescence of PFPs limits their applications in biomedical imaging due to strong tissue autofluorecence background. Conversely, plasmonic nanolasers, also called spasers, have demonstrated potential to generate super-bright stimulated emissions even inside single cells. Nevertheless, the development of photoswitchable spasers that can shift their stimulated emission color in response to light is challenging. Here, we introduce the novel concept of spasers using a PFP layer as the active medium surrounding a plasmonic core. The proof of principle was demonstrated by synthesizing a multilayer nanostructure on the surface of a spherical gold core, with a non-absorbing thin polymer shell and the PFP Dendra2 dispersed in the matrix of a biodegradable polymer. We have demonstrated photoswitching of spontaneous and stimulated emission in these spasers below and above the spasing threshold, respectively, at different spectral ranges. The plasmonic core of the spasers serves also as a photothermal (and potentially photoacoustic) contrast agent, allowing for photothermal imaging of the spasers. These results suggest that multimodal photoswitchable spasers could extend the traditional applications of spasers and PFPs in laser spectroscopy, multicolor cytometry, and theranostics with the potential to track, identify, and kill abnormal cells in circulation.

Graphene and graphene oxide (GO) both being two-dimensional materials are gaining popularity among researchers as a promising nanomaterial for various medical and biological applications. The aim of this study is to elucidate the influence of nanostructured GO sheets on viability of a model species of gram-negative E. coli bacteria transformed with pRSET-emGFP plasmid in in vitro experiments. It was shown that GO at concentrations between 0.0025 and 2.5 g/l in growth medium inhibits growth of bacterial colonies, while in physiological saline solution (PS) this effect decreases dramatically to the point of complete disappearance. It was shown that in order to obtain a pronounced antibacterial effect one needs to introduce high concentrations of GO into the media (up to 2.5 g/l), which can be important for development of antibacterial materials for biomedical applications. Some of the obtained data provide clear evidence to electrostatic nature of interaction between bacterial and GO sheets. A number of previous papers suggested the process of biofilms formation by bacteria as the primary reason for aggregation between graphene-like materials and bacterial cells. However, formation of flocculent structures consisting of GO and dead bacteria and accompanied with decrease in zeta-potential of particles in the suspension to 18 mV proves that electrostatic interactions play the major role in aggregation. The obtained data can be used for employing GO and similar materials in new systems for water-purification from biological contaminants. Besides, our results stress the importance of accounting for the conditions in which goods and coatings containing graphene-like materials as an antibacterial agent are used, as well as unification of the experimental conditions.

Surface-enhanced Raman scattering (SERS) tags have proven to be excellent labels for tissue bioimaging because of their low interference from biological matrices, high photostability, and possibility for using as theranostic agents. Although SERS tags are widely used for the imaging of tumors in vivo, in practice, the low contrast of the tag accumulation in the tissue and strong light scattering can significantly affect their detectability. In this work, we studied these effects by using a phantom of tumor tissue with incorporated SERS tags. The phantom is a 2 mm sphere of calcium alginate with incorporated SERS tags at a concentration of 0.625 × 108–2 × 109 cm−3. To simulate the surrounding medium with differing turbidities, the phantom was placed in a 4 mm thick agarose gel containing intralipid at a concentration of 0–1%. SERS bioimaging was carried out using standard backscattering geometry with different light focusing conditions. We found that shielding the phantom with a turbid medium led not only to a decrease in detectability but also to a decrease in the apparent size of the imaging object. Our results can help develop more accurate algorithms for processing SERS data for bioimaging.

Polyelectrolyte microcapsules and other targeted drug delivery systems could substantially reduce the side effects of drug and overall toxicity. At the same time, the cardiovascular system is a unique transport avenue that can deliver drug carriers to any tissue and organ. However, one of the most important potential problems of drug carrier systemic administration in clinical practice is that the carriers might cause circulatory disorders, the development of pulmonary embolism, ischemia, and tissue necrosis due to the blockage of small capillaries. Thus, the presented work aims to find out the processes occurring in the bloodstream after the systemic injection of polyelectrolyte capsules that are 5 μm in size. It was shown that 1 min after injection, the number of circulating capsules decreases several times, and after 15 min less than 1% of the injected dose is registered in the blood. By this time, most capsules accumulate in the lungs, liver, and kidneys. However, magnetic field action could slightly increase the accumulation of capsules in the region-of-interest. For the first time, we have investigated the real-time blood flow changes in vital organs in vivo after intravenous injection of microcapsules using a laser speckle contrast imaging system. We have demonstrated that the organism can adapt to the emergence of drug carriers in the blood and their accumulation in the vessels of vital organs. Additionally, we have evaluated the safety of the intravenous administration of various doses of microcapsules.

In vivo liquid biopsy, especially using the photoacoustic (PA) method, demonstrated high clinical potential for early diagnosis of deadly diseases such as cancer, infections, and cardiovascular disorders through the detection of rare circulating tumor cells (CTCs), bacteria, and clots in the blood background. However, little progress has been made in terms of standardization of these techniques, which is crucial to validate their high sensitivity, accuracy, and reproducibility. In the present study, we addressed this important demand by introducing a dynamic blood vessel phantom with flowing mimic normal and abnormal cells. The light transparent silica microspheres were used as white blood cells and platelets phantoms, while hollow polymeric capsules, filled with hemoglobin and melanin, reproduced red blood cells and melanoma CTCs, respectively. These phantoms were successfully used for calibration of the PA flow cytometry platform with high-speed signal processing. The results suggest that these dynamic cell flow phantoms with appropriate biochemical, optical, thermal, and acoustic properties can be promising for the establishment of standardization tool for calibration of PA, fluorescent, Raman, and other detection methods of in vivo flow cytometry and liquid biopsy.

Detection and extraction of circulating tumor cells and other rare objects in the bloodstream are of great interest for modern diagnostics, but devices that can solve this problem for the whole blood volume of laboratory animals are still rare. Here we have developed SPIM-based lightsheet flow cytometer for the detection of fluorescently-labeled objects in whole blood. The bypass channel between two blood vessels connected with the external flow cell was used to visualize, detect, and magnetically separate fluorescently-labeled objects without hydrodynamic focusing. Carriers for targeted drug delivery were used as model objects to test the device performance. They were injected into the bloodstream of the rat, detected fluorescently, and then captured from the bloodstream by a magnetic separator prior to filtration in organs. Carriers extracted from the whole blood were studied by a number of in vitro methods.

Targeting drug delivery systems is crucial to reducing the side effects of therapy. However, many of them are lacking effectiveness for kidney targeting, due to systemic dispersion and accumulation in the lungs and liver after intravenous administration. Renal artery administration of carriers provides their effective local accumulation but may cause irreversible vessel blockage. Therefore, the combination of the correct administration procedure, suitable drug delivery system, selection of effective and safe dosage is the key to sparing local therapy. Here, we propose the 3-μm sized fluorescent capsules based on poly-L-arginine and dextran sulfate for targeting the kidney via a mice renal artery. Hemodynamic study of the target kidney in combination with the histological analysis reveals a safe dose of microcapsules (20 × 106), which has not lead to irreversible pathological changes in blood flow and kidney tissue, and provides retention of 20.5 ± 3% of the introduced capsules in the renal cortex glomeruli. Efficacy of fluorescent dye localization in the target kidney after intra-arterial administration is 9 times higher than in the opposite kidney and after intravenous injection. After 24 h microcapsules are not observed in the target kidney when the safe dose of carriers is being used but a high level of fluorescent signal persists for 48 h indicating that fluorescent cargo accumulation in tissues. Injection of non-safe microcapsule dose leads to carriers staying in glomeruli for at least 48 h which has consequences of blood flow not being restored and tissue damage being observed in histology.

There has been growing interest in recent years in developing multifunctional materials for studying the structure interface in biological systems. In this regard, the multimodal systems, which possess activity in the near-infrared (NIR) region, become even more critical for the possibility of improving examined biotissue depth and, eventually, data analysis. Herein, we engineered bi-modal contrast agents by integrating carbon nanotubes (CNT) and gold nanoparticles (AuNP) around silica microspheres using the Layer-by-Layer self-assembly method. The experimental studies revealed that microspheres with CNT sandwiched between AuNP exhibit strong absorption in the visible and NIR regions and high optoacoustic contrast (OA, also called photoacoustics) and Raman scattering when illuminated with 532 nm and 785 nm lasers, respectively. The developed microspheres demonstrated amplification of the signal in the OA flow cytometry at the laser wavelength of 1064 nm. This finding was further validated with ex vivo brain tissue using a portable Raman spectrometer and imaging with the Raster-scanning OA mesoscopy technique. The obtained data suggest that the developed contrast agents can be promising in applications of localization OA tomography (LOT), OA flow cytometry, and multiplex SERS detection.

Macroscopic fluorescence spectroscopy parameters of colon benign and malignant tissues were obtained with spectrofluorimeter FluoroLog 3 (HORIBA Jobin Yvon, France). Investigated biological tissue samples were excised during standard surgical procedure for tumour removal, with cancerous and healthy tissue and the measurements are performed as soon as possible after the excision. The tissue samples were stored in safe-keeping perfusion solution and isothermal conditions and transported from the hospital to the spectroscopy laboratory. The fluorescence spectroscopy measurements were performed for excitation wavelengths in the range of 280-440 nm, with 10 nm increment step, and emission was detected in the range of 300-800 nm for the healthy and cancerous areas of the sample with a step of 1 nm. Micro-spectroscopy was performed with confocal laser scanning microscopy system using 405 nm excitation for fluorescence images of tissue pathologies. We had used a Leica TCS SP system (Leica Inc.). Images were acquired using a 40x magnification objective with a numerical aperture of 0.75. In Lambda-scan regime the spectral data about the selected areas of normal and abnormal tissue sites were obtained and compared. A spectral and image detection in the range of 420-680 nm had place. Fixed tissue samples are unstained, deparaffinized and rehydrated histology tissue slides. Comparison between the evaluated optical macro and micro spectroscopy parameters and their value for clinical application of autofluorescence spectroscopy in cancer diagnosis are presented and discussed.

Flow cytometry nowadays is among the main working instruments in modern biology paving the way for clinics to provide early, quick, and reliable diagnostics of many blood-related diseases. The major problem for clinical applications is the detection of rare pathogenic objects in patient blood. These objects can be circulating tumor cells, very rare during the early stages of cancer development, various microorganisms and parasites in the blood during acute blood infections. All of these rare diagnostic objects can be detected and identified very rapidly to save a patient’s life. This review outlines the main techniques of visualization of rare objects in the blood flow, methods for extraction of such objects from the blood flow for further investigations and new approaches to identify the objects automatically with the modern deep learning methods.

We demonstrate a novel approach to the controlled loading of inorganic nanoparticles and proteins into submicron- and micron-sized porous particles. The approach is based on freezing/thawing cycles, which lead to high loading densities. The process was tested for the inclusion of Au, magnetite nanoparticles, and bovine serum albumin in biocompatible vaterite carriers of micron and submicron sizes. The amounts of loaded nanoparticles or substances were adjusted by the number of freezing/thawing cycles. Our method afforded at least a three times higher loading of magnetite nanoparticles and a four times higher loading of protein for micron vaterite particles, in comparison with conventional methods such as adsorption and coprecipitation. The capsules loaded with magnetite nanoparticles by the freezing-induced loading method moved faster in a magnetic field gradient than did the capsules loaded by adsorption or coprecipitation. Our approach allows the preparation of multicomponent nanocomposite materials with designed properties such as remote control (e.g. via the application of an electromagnetic or acoustic field) and cargo unloading. Such materials could be used as multimodal contrast agents, drug delivery systems, and sensors.

The lateral flow immunoassay (LFIA) has emerged as a powerful tool for rapid screening owing to its simplicity and flexibility for detection of various biomarkers. However, conventional LFIA strips have several disadvantages, including limits in quantitative analysis and low sensitivity. Here we developed a novel surface-enhanced Raman scattering LFIA based on nonspherical gap-enhanced Raman tags (GERTs), with Raman molecules (RMs) embedded in a 1-nm gap between Au nanorod core and Au shell. Such tags have a strong and uniform SERS response, an order of magnitude higher than that of other common SERS tags such as Au nanorods, nanostars, Au nanoshells with surface-adsorbed RMs, or spherical GERTs with embedded RMs. The feasibility of the tags was demonstrated by the semiquantitative and sensitive detection of the heart disease biomarker cardiac troponin I (cTnI). GERTs were conjugated with monoclonal antibodies and used for LFIA in the same way as ordinary functionalized colloidal gold. The presence of the target antigen, cTnI, was identified by Raman microscopy mapping of the test zone. With the SERS-based LFIA, the limit of cTnI detection was about 0.1 ng/mL. This value is within the diagnostic range of cTnI in the blood serum of patients with heart infarction and is 30 times lower than that of the colorimetric LFIA test using the same antibodies and either GERTs or colloidal gold as labels.

A process has been described for obtaining polymer (polyelectrolyte) microcapsules and polyelectrolyte/magnetite composite microcapsules, as well as silica-based hybrid capsules via silane-type precursor hydrolysis, which gives rise to the formation of silica nanoparticles in the structures of shells containing different numbers of polymer layers. The polymer and nanocomposite microcapsules have been formed by successive adsorption of oppositely charged polyelectrolytes and/or nanoparticles on the surface of microparticles of vaterite, which is a polymorphous modification of calcium carbonate. The obtained shells of microcapsules of all types have been studied by atomic force microscopy, confocal fluorescence microscopy, and scanning electron microscopy before and after sonication. The effect of high-intensity focused ultrasound on the obtained samples of microcapsules has been investigated and the sonication parameters that ensure efficient disruption of their shells in an aqueous medium have been determined. It has been found that composite microcapsules containing magnetite nanoparticles in their shells are most sensitive to sonication. The obtained composite microcapsules are promising for developing new systems for delivery of biologically active substances and multifunctional coatings.

Photoluminescent macroporous silicon is studied for tumor imaging application. Gamma-radiation is used for modification of morphology of pores and their size. It is shown by Raman and photoluminescence study that after the irradiation of gamma rays new nanocrystallites of smaller diameter in comparison to the initial porous silicon appeared.

Surface-enhanced Raman scattering (SERS) performance of Au nanorods (AuNRs) can be enhanced by the tip adsorption of Raman molecules (RMs) through anisotropic polymer stabilization or through the embedding of RMs between AuNR cores and Ag shells of AuNR@RM@Ag composite particles. We propose a new strategy to design ultrabright SERS probes composed of high-aspect-ratio AuNRs with anisotropic Ag coatings, with preferred adsorption of RMs to open AuNR tips. Specifically, for 4-nitrobenzenethiol (NBT) concentrations above a threshold value c > ctr, the fabricated Au@NBT@Ag particles had NBT-functionalized open Au tips, as well as anisotropic Ag shells grown on the AuNR sides. The SERS response of these probes with an optimal Ag shell was highest in the off-resonance mode, when the excitation wavelength was far from the plasmon resonance of the Au@NBT@Ag composites. Growing the Ag shell further to completely cover the AuNRs decreased the SERS enhancement. For biocompatibility and stability, the probes were add...

Surface-enhanced Raman scattering (SERS) has emerged as an excellent analytical tool for the effective detection and fingerprint identification of various chemicals. Recently, significant progress has been made in the fabrication of SERS-active substrates using simple, inexpensive, and affordable methods. The full potential of universal SERS diagnostics will likely be realized with the development of approaches and devices capable of effectively detecting analytes on various surfaces as well as in multicomponent media. In addition, the combination of implantable or wearable SERS-active substrates and remote portable devices enables real-time diagnostics that ideally fit the concept of personalized medicine. In this paper, we summarize recent achievements in fabricating flexible SERS substrates made of cellulose paper, polymer membranes, and textile fibrous films. Emphasis is placed on the in-situ extraction and detection of various chemicals in real-world surfaces and complex media using flexible nanofibrous SERS platforms. The potential SERS applications and future perspectives in on-site diagnostics are also discussed.

The formation of lead arachidate (Pb(AA)(2)) coatings produced by Langmuir tensiometry (LT) was studied in dependence of lead nitrate (Pb(NO3)(2)) concentration in the aqueous subphase at pH 5.5. The stable Pb(AA)(2) monolayers were transferred to Si wafers and TEM grids at 35 mN/m in the liquid condensed phase of the AA monolayer applying the Langmuir-Blodgett (LB) (ten monolayers) or the Langmuir-Schaefer (LS) technique, respectively. The most homogeneous monolayer was obtained at a Pb(NO3)(2) concentration of 0.1 mM by using the LS method. Most homogeneous films (five bilayers) were obtained for a Pb(NO3)(2) concentration of 0.01 mM by using the LB method. The formed films are investigated by transmission electron microscopy (monolayer produced by LS), X-ray reflectivity (XRR), and atomic force microscopy (films produced by LB). These methods revealed the formation of homogeneously distributed lead inclusions in the formed monolayers and films. Pb aggregates increase in number whereas their average size stays constant at ae 50 nm(2) with increasing Pb(NO3)(2) concentration. Thickness parameters were determined by XRR. With increasing Pb(NO3)(2) concentration, the bilayer thickness was found to increase. Furthermore, the patterns (111) and (200) for a cubic lead structure were found. The experimental results are compared with simulations obtained from geometry optimization on a semi-empirical quantum level to discuss the lead-ion binding to the AA molecules at pH 5.5.

Toxicity of reduced graphene oxide (rGO) has been a topic of multiple studies and was shown to depend on a variety of characteristics of rGO and biological objects of interest. In this paper, we demonstrate that when studying the same dispersions of rGO and fluorescent Escherichia coli (E. coli) bacteria, the outcome of nanotoxicity experiments also depends on the type of culture medium. We show that rGO inhibits the growth of bacteria in a nutrition medium but shows little effect on the behavior of E. coli in a physiological saline solution. The observed effects of rGO on E. coli in different media could be at least partially rationalized through the adsorption of bacteria and nutrients on the dispersed rGO sheets, which is likely mediated via hydrogen bonding. We also found that the interaction between rGO and E. coli is medium-dependent, and in physiological saline solutions they form stable flocculate structures that were not observed in nutrition media. Furthermore, the aggregation of rGO and E. coli in saline media was observed regardless of whether the bacteria were alive or dead. Filtration of the aggregate suspensions led to nearly complete removal of bacteria from filtered liquids, which highlights the potential of rGO for the filtration and separation of biological contaminants, regardless of whether they include live or dead microorganisms.

One of the most effective ways of targeted drug delivery is the intravenous injection of carriers. However, to prevent undesirable side effects it is necessary to develop microencapsulation drug delivery systems that satisfy following requirements: biocompatibility, non-cytotoxity, biodegradability, colloidal stability in different medium solutions. In case of polymeric microcapsules, aggregation is a crucial and challenging question for biomedical applications that has not been studied well yet. We investigated polymeric microcapsules behavior in PBS, human plasma, and human blood in time. Aggregation degree was measured after 1, 10, 30, and 60 minutes of incubation in a stationary state. As a result, dynamics of capsules aggregation was shown for each medium.

A new type of bimodal contrast agent was made that is based on the self-quenching of indocyanine green (ICG) encapsulated in a biocompatible and biodegradable polymer shell. The increasing of a local ICG concentration that is necessary for the obtaining of self-quenching effect was achieved by freezing-induced loading and layer-by-layer assembly. As a result, an efficient photoacoustic(optoacoustic)/fluorescent contrast agent based on composite indocyanine green/polymer particles was successfully prepared and was characterized by fluorescence and photoacoustic(optoacoustic) tomography in vitro. This type of contrast agent holds good promise for clinical application owing to its high efficiency and biosafety.

Fluorescence spectroscopy is a sensitive, fast and non-invasive tool for a diagnostics of cancerous gastrointestinal lesions. It could be applied for in situ detection of tumours during primary endoscopic observations or as add-on measurement modality during microscopic observations of tissue histology slides for their initial or retrospective diagnosis. Therefore, we are looking for diagnostically important features of normal and cancerous tissue areas in a broad spectral range for gastrointestinal tissues ex vivo using two steady-state macroscopic fluorescent spectroscopic modalities and by confocal fluorescent microscopic detection. Results obtained from autofluorescence spectroscopy of benign and malignant lower part gastrointestinal tract (GIT) lesions from freshly excised tissues during surgical removal of the lesions in 18 patients (22 lesions), were compared with the spectral measurements obtained during confocal fluorescent microscopy observations of unstained tissue slides using 405 nm excitation. Excitation–emission matrices (EEMs) were used for ex vivo measurements with applied excitation in 280-440 nm spectral region and emission observed between 300 and 700 nm. Synchronous fluorescence spectroscopy (SFS) approach was also applied to improve the spectral resolution of the observed complex emission spectra. Specific fluorescent features observed, related to presence of structural proteins, co-enzymes and endogenous porphyrins in the tissues investigated, allow discriminating normal mucosa from benign polyps and malignant carcinoma lesions with diagnostic accuracy up to 94.4%.

At the present time a new direction of theranostics associated with using of nanostructured multifunctional carriers is developed rapidly. Appreciative examples of such carriers are core-shell nanoparticles, liposomes, polymer micelles, capsules produced by Layer by Layer assembly (LbL) method. Every type of nanostructured carriers has its own advantages and disadvantages. Undoubted advantages of LbL capsules are high loading capacity, wide range of possible internal payloads and wide possibilities of controlling physical and chemical properties of nanostructured carriers by variation of shell structure, thickness and its chemical composition [М.Delcea, et al, Adv. drug delivery reviews, 2011, 63(9), 730, A.S.Timin, et al, Adv. Mater. Interfaces,2016, 4, 1600338]. Application of nanostructured carriers is limited by absence of methods for its in vivo visualization with sufficient spatial resolution and significant tissue penetration depth [A.S.Timin, et al, Adv. Mater. Interfaces, 2016, 4,1600338]. One of the method that allows to visualize and to detect the nanostructured carriers efficiently is a photoacoustic (PA) method. PA approach allows to use the photoacoustic cytometry in vivo [E.I. Galanzha et al, Cancer Res. 2009, 69, 7926]. It is also very important for detection and killing of free circulating cancer cells required for early cancer diagnostics and decreasing the metastases probability [E.I. Galanzha, et al, J. Biophotonics, 2009, 2, 725]. Unique combination of photoacoustic cytometry and nanostructured carriers prepared by LbL assembly approach gives clinicians and researchers significant advantages in the developing of new therapy method based on delivery of nanostructured carriers loaded by bioactive substances into lesion focus and also their visualization and remote controlled release of encapsulated bioactive substances using PA approach. Visualization of two types of nanostructured carriers produced by LbL assembly method was demonstrated recently in diluted and undiluted blood [E.I. Galanzha, et al, J. Biophotonics, 2009, 2, 725]. It was established that hollow microcapsules exhibited greater photoacoustic signal comparing to core-shell type of microparticles with the same composition of polymeric shell [E.I. Galanzha, et al, J. Biophotonics, 2009, 2, 725]. Nevertheless, there are no known examples of in vivo photoacoustic detection of LbL nanostructured carriers, therefore the main goal of present study is formation of biocompatible nanostructured carriers (BNCs) that will exhibit giant photoacoustic signal allowing in vivo detection of such structures. It was demonstrated for the first time, the super (giant) PA contrast of BNCs both in vitro and in vivo. It was verified the unique PA characteristics of these advanced contrast agents using PA flow cytometry diagnostic platform. The obtained data suggest the high PA contrast of BNCs that can be associated with synergistic plasmonic, thermal and acoustic effects, especially in nonlinear mode with nanobubble formation in overheated absorbing layers, in particular gold nanoclusters between two light transparent shells. This work was partly supported by RFBR (project №18-29-08046), the Government of the Russian Federation (grant no. 14.Z50.31.0044 to support scientific research projects implemented under the supervision of leading scientists at Russian institutions and Russian institutions of higher education) and Saratov State University.

Increasing antimicrobial resistance requires the development of novel materials and approaches for treatment of various infections. Utilization of photodynamic therapy represents an advanced alternative to antibiotics and metal-based agents. Here, we report the fabrication of electrospun material that possesses benefits of both topical antimicrobial and photodynamic therapies. This material combines chitosan, as a biocompatible polymer, and a second generation photosensitizer. The incorporation of photosensitizer doesn't affect the material morphology and its nearly uniform distribution in fibers structure was observed by confocal Raman microscopy. Owing to photosensitizer the prepared material exhibits the light-induced and spatially limited antimicrobial activity that was demonstrated against Staphylococcus aureus, an important etiological infectious agent. Such material can be potentially used in antibacterial therapy of chronic wounds, infections of diabetic ulcers, and burns, as well as rapidly spreading and intractable soft-tissue infections caused by resistant bacteria.

Dot immunoassay has been proposed as a simple and fast method for detection of various analytes. However, this qualitative method has a narrow working range and is limited to multiplex detection. To overcome these drawbacks, we suggest a highly specific, quantitative, and multiplex dot immunoassay using plasmonic gap-enhanced Raman tags (GERTs) and a nitrocellulose membrane as a substrate. The assay principle is based on reading surface-enhanced Raman scattering (SERS) spectra from analyte drops on the membrane strip after incubation with GERTs conjugated to biospecific probes. Three types of GERTs were synthesised by embedding nitrobenzenethiol (NBT), naphthalenethiol (NT), and acetamidothiophenol (AcTP) Raman molecules inside Au core/shell nanoparticles. For proof-of-concept experiments, the NBT, NT, and AcTP GERTS were further functionalized with rabbit anti-rat, anti-human, and anti-chicken antibodies, respectively. For all three corresponding antigens, the detected Raman signals linearly depended on the analyte amount within the range from 10 to 300 ng. The multiplex capability of the assay is illustrated by simultaneous one-step determination of rat, human, and chicken IgGs with a mixture of functionalized GERTs by recording Raman maps for whole membrane to avoid the point-to-point repeatability problem. Thus, GERTs are promising SERS nanotags for advanced versions of immunoassays instead of common plasmonic SERS labels with Raman reporters excited by the external near field.

Carbon dots (CDs) are usually used as an alternative to other fluorescent nanoparticles. Apart from fluorescence, CDs also have other important properties for use in composite materials, first of all their ability to absorb light energy and convert it into heat. In our work, for the first time, CDs have been proposed as an alternative to gold nanostructures for harvesting light energy, which results in the opening of polymer-based containers with biologically active compounds. In this paper, we propose a method for the synthesis of polylactic acid microchamber arrays with embedded CDs. A comparative analysis was made of the damage to microchambers functionalized with gold nanorods and with CD aggregates, depending on the wavelength and power of the laser used. The release of fluorescent cargo from the microchamber arrays with CD aggregates under laser exposure was demonstrated.

Synthesis of hydrophobic luminescent carbon nanostructures in non-aqueous solution has been performed. The system consisted of hexane as nonpolar “oil” phase, nonionic polyoxyethylene (4) lauryl ether (Brij L4) as a surfactant, and either water or dextran sulfate aqueous solution as a polar phase. Carbon nanodots (CND) with a particle size of 5.0 ± 0.7 nm were formed by solvothermal treatment at 180 °C of two- (hexane–Brij L4) and three-component (hexane–Brij L4–aqua solutions) systems. It was found that CND with blue-green-yellow photoluminescence (PL) with linear dependence of emission peak position on excitation wavelength and stability of PL over at least 3 weeks were formed from Brij L4. We found that using small volume of water as the third component helps to reorganize Brij L4 molecules which results in CND PL intensity increase by a factor of 1.5 and quantum yield increase from 2.0 to 2.6% compared to CND from the two-component micellar system. Replacing water with dextran sulfate solution resulted in a lower quantum yield to 2.2%. Thus, addition of small amount of water clearly shows an effect of molecular level organization on properties of CND and results in enhanced quantum yield.

Progress in understanding the cell biology and diseases depends on advanced imaging and labeling techniques. Here, we address this demand by exploring novel multilayered nanocomposites (MNCs) with plasmonic nanoparticles and absorbing dyes in thin nonabsorbing shells as supercontrast multimodal photoacoustic (PA) and fluorescent agents in the near-infrared range. The proof of concept was performed with gold nanorods (GNRs) and indocyanine green (ICG) dispersed in a matrix of biodegradable polymers. We demonstrated synergetic PA effects in MNCs with the gold-ICG interface that could not be achieved with ICG and GNRs alone. We also observed ultrasharp PA and emission peaks that could be associated with nonlinear PA and spaser effects, respectively. Low-toxicity multimodal MNCs with unique plasmonic, thermal and acoustic properties have the potential to make a break-through in PA flow cytometry and near-infrared spasers in vivo by using the synergetic interaction of plasmonic modes with a nearby absorbing medium.

Gold and composite Au@Ag layered nanoparticles, in which Raman molecules (RMs) are embedded in a nanometer-sized gap between metal layers, have great potential in biomedical applications as highly efficient imaging SERS probes. Compared to common tags with outer RMs exciting by plasmonic near field, the embedded RMs of new probes are protected from environmental conditions and subjected to a strongly enhanced internal field in the gap. Another type of efficient SERS tags are the tip functionalized Au(core)@Ag(shell) nanorods (NRs) operating in off-resonance mode. In this talk, we summarize our recent efforts in fabrication, electromagnetic simulation, and bioimaging application of gold based Au@RM@Au and hybrid AuNR@RM@Ag SERS tags.

A sensor based on microstructured waveguides (MWGs) with a hollow core inner surface covered with polyelectrolyte-layer-stabilized gold nanostars was developed for the SERS sensing of dissolved analytes. A polyelectrolyte-layer coating over the inner surface of glass cladding served as a spacer, reducing nonlinear optical effects in the glass near plasmonic hotspots of nanoparticles, as a stabilizing agent for thermodynamically unstable gold nanostars and as an optical coating for the fine-tuning of MWG bandgaps. This approach can be used to construct different kinds of SERS sensors for dissolved analytes, providing conservation, the prevention of coagulation, and the drying of a liquid sample for the time required to record the signal.

Photonic crystal fibers with hollow core (HC PCFs) are a specific class of optical fibers characterized by microstructure with periodic holes oriented along fiber. The combination of HC PCF with Raman spectroscopy for biosensors creation is attractive in the terms of the low sample volume, the possibility to increase the integration time without sample degradation and maintaining constant focus during experiments. Here we propose layer-by-layer polyelectrolyte coating of HC PCF inner surface in order to obtain charge-selective absorption of analyte, stabilization of Surface-Enhanced Raman scattering (SERS)-active gold nanoparticles. Distance between SERS hotspots and glass reduces nonlinear signals from glass, and increases signal-to-noise ratio of SERS spectra.

Microcapsules, made of biodegradable polymers, containing magnetite nanoparticles with tunable contrast in both the T1 and T2 MRI modes, were successfully prepared using a layer-by-layer approach. The MRI contrast of the microcapsules was shown to depend on the distance between magnetite nanoparticles in the polymeric layers, which is controlled by their concentration in the microcapsule shell. A fivefold increase in the average distance between the nanoparticles in the microcapsule shell led to a change in the intensity of the MR signal of 100% for both the T1 and T2 modes. Enzyme treatment of biodegradable shells resulted in a change of the microcapsules' MRI contrast. In vivo degradation of nanocomposite microcapsules concentrated in the liver after intravenous injection was demonstrated by MRI. This method can be used for the creation of a new generation of drug delivery systems, including drug depot, with combined navigation, visualization and remote activated release of bioactive substances in vivo.

The impact of high-frequency (1.2 MHz) ultrasound with a power density of 0.33 W cm(-2) on microcapsule nanocomposite shells with embedded zinc oxide nanoparticles was investigated by exploring modeling simulations and direct visualization. For the first time the sonication effect has been monitored in situ on individual microcapsules upon exposure of their aqueous suspension to ultrasound. The stress distribution on the microcapsule shell for the impact of ultrasound with high (1.2 MHz) and low (20 kHz) frequency at two fixed intensities (0.33 and 30 W cm(-2)) has been modeled. As shown in silico and experimentally the nanocomposite microcapsules were destroyed more effectively by the action of high-frequency (1.2 MHz) ultrasound in comparison to the low frequency (20 kHz) one with the same power density.

A wet-chemical approach is used to fabricate centimeter-scale gold nanoisland films (NIFs) with tunable morphology of islands and with strong electromagnetic coupling between them. The approach consists in a uniform seeding of small gold nanoparticles on a glass or silicon substrate, followed by controllable growth of the seeds into small nanoislands. A special technique for TEM sampling was developed to follow the gradual formation of larger-sized isolated nanoparticles, nanoislands of sintered overgrown seeds, and a complete gold layer with nanoscale cracks. The electromagnetic field distribution inside the fabricated NIFs was calculated by FDTD simulations applied to actual TEM images of the fabricated samples rather than to artificial models commonly used. SERS measurements with 1,4-aminothiophenol (ATP) molecules demonstrated the analytical enhancement factor about of 10(7) and the fundamental enhancement factor about of 10(8) for optimized substrates. These values were at least 1 order of magnitude higher than that for self-assembled arrays of gold nanostars and silver nanocubes. SERS spectra of independent samples demonstrated good sample-to-sample reproducibility in terms of the relative standard deviation (RSD) of the main peaks less than 20%. Additionally, Raman maps with 1 μm increment in X-Y directions of NIFs (800 spectral spots) demonstrated good point-to-point repeatability in the intensity of the main Raman vibration modes (RSD varied from 5% to 15% for 50 randomly selected points). A real-life application of the fabricated SERS substrates is exemplified by the detection of the thiram fungicide in apple peels within the 5-250 ppb linear detection range. Specifically, the NIF-based SERS technology detected thiram on apple peel down to level of 5 ng/cm(2).