Олейников Владимир Александрович

Личная информация

2007-н. вр.: Учёный секретарь ИБХ РАН

2011-н.вр.: Заведующий лабораторией Молекулярной биофизики ИБХ РАН

Избранные публикации

  1. Mochalov K.E., Chistyakov A.A., Solovyeva D.O., Mezin A.V., Oleinikov V.A., Vaskan I.S., Molinari M., Agapov I.I., Nabiev I., Efimov A.E. (2017). An instrumental approach to combining confocal microspectroscopy and 3D scanning probe nanotomography. Ultramicroscopy 182, 118–123 [+]

    In the past decade correlative microscopy, which combines the potentials of different types of high-resolution microscopies with a variety of optical microspectroscopy techniques, has been attracting increasing attention in material science and biological research. One of outstanding solutions in this area is the combination of scanning probe microscopy (SPM), which provides data on not only the topography, but also the spatial distribution of a wide range of physical properties (elasticity, conductivity, etc.), with ultramicrotomy, allowing 3D multiparametric examination of materials. The combination of SPM and ultramicrotomy (scanning probe nanotomography) is very appropriate for characterization of soft multicompound nanostructurized materials, such as polymer matrices and microstructures doped with different types of nanoparticles (magnetic nanoparticles, quantum dots, nanotubes, etc.), and biological materials. A serious problem of this technique is a lack of chemical and optical characterization tools, which may be solved by using optical microspectroscopy. Here, we report the development of an instrumental approach to combining confocal microspectroscopy and 3D scanning probe nanotomography in a single apparatus. This approach retains all the advantages of SPM and upright optical microspectroscopy and allows 3D multiparametric characterization using both techniques. As the first test of the system developed, we have performed correlative characterization of the morphology and the magnetic and fluorescent properties of fluorescent magnetic microspheres doped with a fluorescent dye and magnetic nanoparticles. The results of this study can be used to obtain 3D volume images of a specimen for most high-resolution near-field scanning probe microscopies: SNOM, TERS, AFM-IR, etc. This approach will result in development of unique techniques combining the advantages of SPM (nanoscale morphology and a wide range of physical parameters) and high-resolution optical microspectroscopy (nanoscale chemical mapping and optical properties) and allowing simultaneous 3D measurements.

  2. Efimov A.E., Agapov I.I., Agapova O.I., Oleinikov V.A., Mezin A.V., Molinari M., Nabiev I., Mochalov K.E. (2017). A novel design of a scanning probe microscope integrated with an ultramicrotome for serial block-face nanotomography. Rev Sci Instrum 88 (2), 023701 [+]

    We present a new concept of a combined scanning probe microscope (SPM)/ultramicrotome apparatus. It enables "slice-and-view" scanning probe nanotomography measurements and 3D reconstruction of the bulk sample nanostructure from series of SPM images after consecutive ultrathin sections. The sample is fixed on a flat XYZ scanning piezostage mounted on the ultramicrotome arm. The SPM measuring head with a cantilever tip and a laser-photodiode tip detection system approaches the sample for SPM measurements of the block-face surface immediately after the ultramicrotome sectioning is performed. The SPM head is moved along guides that are also fixed on the ultramicrotome arm. Thereby, relative dysfunctional displacements of the tip, the sample, and the ultramicrotome knife are minimized. The design of the SPM head enables open frontal optical access to the sample block-face adapted for high-resolution optical lenses for correlative SPM/optical microscopy applications. The new system can be used in a wide range of applications for the study of 3D nanostructures of biological objects, biomaterials, polymer nanocomposites, and nanohybrid materials in various SPM and optical microscopy measuring modes.

  3. Bobrovsky A., Mochalov K., Oleinikov V., Solovyeva D., Shibaev V., Bogdanova Y., Hamplová V., Kašpar M., Bubnov A. (2016). Photoinduced Changes of Surface Topography in Amorphous, Liquid-Crystalline, and Crystalline Films of Bent-Core Azobenzene-Containing Substance. The journal of physical chemistry. B 120 (22), 5073–82 [+]

    Recently, photofluidization and mass-transfer effects have gained substantial interest because of their unique abilities of photocontrolled manipulation with material structure and physicochemical properties. In this work, the surface topographies of amorphous, nematic, and crystalline films of an azobenzene-containing bent-core (banana-shaped) compound were studied using a special experimental setup combining polarizing optical microscopy and atomic force microscopy. Spin-coating or rapid cooling of the samples enabled the formation of glassy amorphous or nematic films of the substance. The effects of UV and visible-light irradiation on the surface roughness of the films were investigated. It was found that UV irradiation leads to the fast isothermal transition of nematic and crystalline phases into the isotropic phase. This effect is associated with E-Z photoisomerization of the compound accompanied by a decrease of the anisometry of the bent-core molecules. Focused polarized visible-light irradiation (457.9 nm) results in mass-transfer phenomena and induces the formation of so-called "craters" in amorphous and crystalline films of the substance. The observed photofluidization and mass-transfer processes allow glass-forming bent-core azobenzene-containing substances to be considered for the creation of promising materials with photocontrollable surface topographies. Such compounds are of principal importance for the solution of a broad range of problems related to the investigation of surface phenomena in colloid and physical chemistry, such as surface modification for chemical and catalytic reactions, predetermined morphology of surfaces and interfaces in soft matter, and chemical and biochemical sensing.

  4. Ivanov A.A., Koval V.S., Susova O.Y., Salyanov V.I., Oleinikov V.A., Stomakhin A.A., Shalginskikh N.A., Kvasha M.A., Kirsanova O.V., Gromova E.S., Zhuze A.L. (2015). DNA specific fluorescent symmetric dimeric bisbenzimidazoles DBP(n): the synthesis, spectral properties, and biological activity. Bioorg. Med. Chem. Lett. 25 (13), 2634–8 [+]

    A series of new fluorescent symmetric dimeric bisbenzimidazoles DBP(n) bearing bisbenzimidazole fragments joined by oligomethylene linkers with a central 1,4-piperazine residue were synthesized. The complex formation of DBP(n) in the DNA minor groove was demonstrated. The DBP(n) at micromolar concentrations inhibit in vitro eukaryotic DNA topoisomerase I and prokaryotic DNA methyltransferase (MTase) M.SssI. The DBP(n) were soluble well in aqueous solutions and could penetrate cell and nuclear membranes and stain DNA in live cells. The DBP(n) displayed a moderate effect on the reactivation of gene expression.

  5. Zaitsev S.Y., Lukashev E.P., Solovyeva D.O., Chistyakov A.A., Oleinikov V.A. (2014). Controlled influence of quantum dots on purple membranes at interfaces. Colloids Surf B Biointerfaces 117, 248–51 [+]

    The development of bio-sensitized nanofilms engineered from biomembrane components and inorganic nanoparticles is a promising field of colloid and interface science and technologies. Recent nano-bioengineering approaches employing quantum dots (QDs) permit the enhancement of the purple membrane (PM) "light-harvesting capacity" compared to native PMs. The influence of QDs on the PM properties, especially the bacteriorhodopsin (bR) photocycle, has been found that has both fundamental (mechanisms of photoreception) and applied implications (including the fabrication of hybrid bionanomaterials). Samples of PM-QD complexes capable of energy transfer and characterized by increased rates of M-intermediate formation and decay have been obtained. The modified bR photocycle kinetic parameters may be explained by changes in the PM interface upon QD adsorption. The increase and decrease in absorption at 410 nm (or photopotential) for PM-QD complexes are, on average, several times more rapid than for PM suspensions or PM dry films. These results provide a strong impetus for the development of nanomaterials with advanced properties.

  6. Rakovich A., Donegan J.F., Oleinikov V., Molinari M., Sukhanova A., Nabiev I., Rakovich Y.P. (2014). Linear and nonlinear optical effects induced by energy transfer from semiconductor nanoparticles to photosynthetic biological systems. J. Photochem. Photobiol., С: Photocgem. Rev. 20, 17–32 [+]

    The development of new hybrid materials that can be integrated into current technologies is one of the most important challenges facing material scientists today. The purpose of this work is to review recent studies in one largely unexplored area of nanobiotechnology: the development of nano-bio hybrid materials that exploit Fцrster Resonance Energy Transfer (FRET) to enhance the functionalities of technologically promising photosynthetic biomaterials. One of very promising approaches is to employ semiconductor quantum dots having a broad absorption spectrum as nanoantennae coupled with the natural lightharvesting complexes of photosynthetic reaction centers. This system reveals great potential for the utilization of quantum dots in artificial photosynthetic devices. The second very useful functionality, which is discussed in this review, is the possibility to enhance the efficiency of the main biological function (proton pumping) of the protein bacteriorhodopsin using nonradiative energy transfer from quantum dots. Also recent studies revealed that FRET-based improvement of the biological function of bacteriorhodopsin in the presence of quantum dots allows for strong wavelength-dependent enhancement of the nonlinear refractive index of bacteriorhodopsin. These new hybrid bio-nanomaterials with exceptional light-harvesting and nonlinear properties will have numerous photonic applications employing their photochromic, energy transfer, and energy conversion properties.

  7. Bobrovsky A., Mochalov K., Chistyakov A., Oleinikov V., Shibaev V. (2013). AFM study of laser-induced crater formation in films ofazobenzene-containing photochromic nematic polymer andcholesteric mixture. J. Photochem. Photobiol., A 275, 30–36 [+]

    Впервые выполнены исследования методом атомно-силовой микроскопии индуцированной лазерным излучением деформации поверхности и образования дефектов (кратеров) в жидкокристаллических, содержащих азобензол фотохромных полимерах. В исследовании использовали сочетание методов поляризационной микроскопии, микроскопии атомных сил и возможности облучения хорошо сфокусированным лазерным пучком с длиной волны 532 нм. Были исследованы пленки нематического азобензольного полимера, содержащего полиакрилат и холестерические смеси, включающие хиральные примеси. Облучение сфокусированным лазерным лучом  приводит к образованию кратера в результате в массопереноса за пределами центра луча. Глубина кратеров составляет от десятков нанометров и увеличивается с увеличением времени облучения. Обнаружено, что явление наблюдается только в «толстых» пленках (5 – 10) мкм, в отличие от тонких, приготовленных методом нанесения на вращающуюся подложку (spin-cotting), в которых подобного не происходит. Скорость образования кратера, глубина и диаметр кратера не зависят от хиральности системы и одинаковы для нематических и холестерических пленок. Показано, что для ориентированных вдоль оси нематических полимерных пленок массоперенос наблюдается только вдоль ЖК оси и не зависит от направления вектора поляризации лазерного излучения

  8. Mochalov K.E., Efimov A.E., Bobrovsky A., Agapov I.I., Chistyakov A.A., Oleinikov V., Sukhanova A., Nabiev I. (2013). Combined scanning probe nanotomography and optical microspectroscopy: a correlative technique for 3D characterization of nanomaterials. ACS Nano 7 (10), 8953–62 [+]

    Combination of 3D structural analysis with optical characterization of the same sample area on the nanoscale is a highly demanded approach in nanophotonics, materials science, and quality control of nanomaterial. We have developed a correlative microscopy technique where the 3D structure of the sample is reconstructed on the nanoscale by means of a "slice-and-view" combination of ultramicrotomy and scanning probe microscopy (scanning probe nanotomography, SPNT), and its optical characteristics are analyzed using microspectroscopy. This approach has been used to determine the direct quantitative relationship of the 3D structural characteristics of nanovolumes of materials with their microscopic optical properties. This technique has been applied to 3D structural and optical characterization of a hybrid material consisting of cholesteric liquid crystals doped with fluorescent quantum dots (QDs) that can be used for photochemical patterning and image recording through the changes in the dissymmetry factor of the circular polarization of QD emission. The differences in the polarization images and fluorescent spectra of this hybrid material have proved to be correlated with the arrangement of the areas of homogeneous distribution and heterogeneous clustering of QDs. The reconstruction of the 3D nanostructure of the liquid crystal matrix in the areas of homogeneous QDs distribution has shown that QDs do not perturb the periodic planar texture of the cholesteric liquid crystal matrix, whereas QD clusters do perturb it. The combined microspectroscopy-nanotomography technique will be important for evaluating the effects of nanoparticles on the structural organization of organic and liquid crystal matrices and biomedical materials, as well as quality control of nanotechnology fabrication processes and products.

  9. Melnikau D., Savateeva D., Lesnyak V., Gaponik N., Fernández Y.N., Vasilevskiy M.I., Costa M.F., Mochalov K.E., Oleinikov V., Rakovich Y.P. (2013). Resonance energy transfer in self-organized organic/inorganic dendrite structures. Nanoscale 5 (19), 9317–23 [+]

    Hybrid materials formed by semiconductor quantum dots and J-aggregates of cyanine dyes provide a unique combination of enhanced absorption in inorganic constituents with large oscillator strength and extremely narrow exciton bands of the organic component. The optical properties of dendrite structures with fractal dimension 1.7-1.8, formed from J-aggregates integrated with CdTe quantum dots (QDs), have been investigated by photoluminescence spectroscopy and fluorescence lifetime imaging microscopy. Our results demonstrate that (i) J-aggregates are coupled to QDs by Förster-type resonant energy transfer and (ii) there are energy fluxes from the periphery to the centre of the structure, where the QD density is higher than in the periphery of the dendrite. Such an anisotropic energy transport can be only observed when dendrites are formed from QDs integrated with J-aggregates. These QD/J-aggregate hybrid systems can have applications in light harvesting systems and optical sensors with extended absorption spectra.

  10. Generalova A.N., Oleinikov V.A., Sukhanova A., Artemyev M.V., Zubov V.P., Nabiev I. (2013). Quantum dot-containing polymer particles with thermosensitive fluorescence. Biosens Bioelectron 39 (1), 187–93 [+]

    Создан новый тип термонаносенсора, основанный на эффекте дистанционно зависсимого тушения флуоресцентных полупроводниковых нанокристаллов включенных в матрицу из термочувствительного полимера (поливинил-N-капролактам). Термосенсор чувствителен к изменению температуры в области 30-40 С. Показаны примеры использования нанотермометра для решения прикладных задач. 

  11. Bobrovsky A., Mochalov K., Oleinikov V., Sukhanova A., Prudnikau A., Artemyev M., Shibaev V., Nabiev I. (2012). Optically and electrically controlled circularly polarized emission from cholesteric liquid crystal materials doped with semiconductor quantum dots. Adv. Mater. Weinheim 24 (46), 6216–22 [+]

    Создан новый электро- и фотоконтролируемый материал на основе холестерической жидкокристаллической матрицы и внедренных в нее полупроводниковых флуоресцентных нанокристаллов (квантовых точек). УФ облучение и/или приложение электрического поля позволяет изменять степень круговой поляризации нового материала и значение эмиссии флуоресценции квантовых точек внедренных в ЖК матрицу

  12. Sukhanova A., EvenDesrumeaux K., Kisserli A., Tabary T., Reveil B., Millot J.M., Chames P., Baty D., Artemyev M., Oleinikov V., Pluot M., Cohen J.H., Nabiev I. (2012). Oriented conjugates of single-domain antibodies and quantum dots: toward a new generation of ultrasmall diagnostic nanoprobes. Nanomedicine 8 (4), 516–25 [+]

    Common strategy for diagnostics with quantum dots (QDs) utilizes the specificity of monoclonal antibodies (mAbs) for targeting. However QD-mAbs conjugates are not always well-suited for this purpose because of their large size. Here, we engineered ultrasmall nanoprobes through oriented conjugation of QDs with 13-kDa single-domain antibodies (sdAbs) derived from llama IgG. Monomeric sdAbs are 12 times smaller than mAbs and demonstrate excellent capacity for refolding. sdAbs were tagged with QDs through an additional cysteine residue integrated within the C terminal of the sdAb. This approach allowed us to develop sdAbs-QD nanoprobes comprising four copies of sdAbs coupled with a QD in a highly oriented manner. sdAbs-QD conjugates specific to carcinoembryonic antigen (CEA) demonstrated excellent specificity of flow cytometry quantitative discrimination of CEA-positive and CEA-negative tumor cells. Moreover, the immunohistochemical labeling of biopsy samples was found to be comparable or even superior to the quality obtained with gold standard protocols of anatomopathology practice. sdAbs-QD-oriented conjugates as developed represent a new generation of ultrasmall diagnostic probes for applications in high-throughput diagnostic platforms.

  13. Mahmoud W., Rousserie G., Reveil B., Tabary T., Millot J.M., Artemyev M., Oleinikov V.A., Cohen J.H., Nabiev I., Sukhanova A. (2011). Advanced procedures for labeling of antibodies with quantum dots. Anal. Biochem. 416 (2), 180–5 [+]

    Semiconductor quantum dots (QDs) are proved to be unique fluorescent labels providing excellent possibilities for high-throughput detection and diagnostics. To explore in full QDs' advantages in brightness, photostability, large Stokes shift, and tunability by size fluorescence emission, they should be rendered stable in biological fluids and tagged with the target-specific capture molecules. Ideal QD-based nanoprobes should not exceed 15nm in diameter and should contain on their surface multiple copies of homogeneously oriented highly active affinity molecules, for example, antibodies (Abs). Direct conjugation of QDs with the Abs through cross-linking of QDs' amines with the sulfhydryl groups issued from the reduced Abs' disulfide bonds is the common technique. However, this procedure often generates conjugates in which the number of functionally active Abs on the surface of QDs does not always conform to expectations and is often low. Here we have developed an advanced procedure with the optimized critical steps of Ab reduction, affinity purification, and QD-Ab conjugation. We succeeded in reducing the Abs in such a way that the reduction reaction yields highly functional, partially cleaved, 75-kDa heavy-light Ab fragments. Affinity purification of these Ab fragments followed by their tagging with the QDs generates QD-Ab conjugates with largely improved functionality compared with those produced according to the standard procedures. The developed approach can be extended to conjugation of any type of Ab with different semiconductor, noble metal, or magnetic nanocrystals.

  14. Generalova A.N., Oleinikov V.A., Zarifullina M.M., Lankina E.V., Sizova S.V., Artemyev M.V., Zubov V.P. (2011). Optical sensing quantum dot-labeled polyacrolein particles prepared by layer-by-layer deposition technique. J Colloid Interface Sci 357 (2), 265–72 [+]

    Optical sensing polymer particles with tailored semiconductor nanocrystal (QD) loading are prepared by layer-by-layer deposition technique (LbL). Polyacrolein particles of 1.2 μm diameter are used as solid support for deposition of hydrophilic CdSe/ZnS nanocrystal/polyelectrolyte multilayers formed by electrostatic interactions. The pH-dependent fluorescence of QDs and pH-dependent conformations of polyelectrolytes, which likely passivate the surface state of nanocrystals, allow a creation of both mono- and multiplex coded polymer particles with pH-dependent fluorescence intensity. Bovine serum albumin (BSA) as outermost layer makes it possible to design the optical sensing polymer particles with reversibly responded fluorescence at pH variations. The fluorescence of such polymer particles with BSA outer layer is sensitive to copper(II) ion while the fluorescence of these particles is practically insensitive to the other divalent cations (Zn(2+), Ca(2+), Ba(2+), Co(2+), Mg(2+)). The detection limit of Cu(2+) is about 15 nM. Adaptation of LbL method to prepare QD-labeled polymer particles with enhanced complexity (e.g. several types of QDs, multiple biofunctionality) is expected to open new opportunities in biotechnological applications.

  15. Generalova A.N., Sizova S.V., Zdobnova T.A., Zarifullina M.M., Artemyev M.V., Baranov A.V., Oleinikov V.A., Zubov V.P., Deyev S.M. (2011). Submicron polymer particles containing fluorescent semiconductor nanocrystals CdSe/ZnS for bioassays. Nanomedicine (Lond) 6 (2), 195–209 [+]

    This study aimed to design a panel of uniform particulate biochemical reagents and to test them in specific bioassays. These reagents are polymer particles of different sizes doped with semiconductor nanocrystals and conjugated with either full-size antibodies or recombinant mini-antibodies (4D5 scFv fragment) designed by genetic engineering approaches.

  16. Kholodenko I.V., Kholodenko R.V., Vodovozova E.L., Oleinikov V.A., Polyakov N.B., Molotkovskaya I.M., Petrov R.V. (2009). Ganglioside GM1-binding sites in interleukin-4: a photoaffinity labeling study. Dokl. Biochem. Biophys. 418, 31–5 ID:1119
  17. Oleinikov V.A. (2009). [Semiconductor fluorescent nanocrystals (quantum dots) in biological biochips]. Bioorg. Khim. 37 (2), 171–89 [+]

    Comprehension of biological processes in cells, tissues and organisms requires identification and analysis of numerous biological objects, mechanisms of their action and regulation. Microarray (biochips) technology is a rare tool to solve this problem. It is based on high-throughput recognition of a target to the probe and has the potential to measure simultaneously the presence of numerous molecules in multiplexed testes, all contained in a small drop of test fluid. Biochips allow the parallel analysis of genomic or proteomic content in healthy versus disease-affected or altered tissues or cells. The signals read-out from the biochips is done with organic dyes which often suffer from photobleaching, low brightness and background fluorescence. Recent data show that the use of fluorescent nanocrystals "quantum dots" (QDs) allows push away these restrictions. The QDs are sufficiently bright to be detected as individual particles, extremely resistant to photobleaching and provide unique possibilities for multiplexing thus supplying the microarray technology with the novel read-out option enabling the sensitivity of detection reaching the single molecule level. This paper is aimed at the development of the approaches to the QDs application in microarray-based detection. Possibilities of QDs application both in solid state (planar) biochips as well as intensively developing technique of suspension biochips (bead-based assays or liquid biochips) are demonstrated. The latter are more and more applied for simultaneous identification of very large numbers of molecules in proteomics, genomics, drug screening and clinical diagnostics. This assays base on spectral encoded elements (as a rule polymer microbeads). The benefits of using optically encoded microbeads (instead of the solid-state two-dimensional arrays) are derived from the freedom of bead to move in three dimensions. Polymeric beads optically encoded with organic dyes allow for a limited number of unique codes, whereas the use of semiconductor nanocrystals as fluorescent tags improves the beads multiplexed imaging capabilities, photostability and sensitivity of the biological objects detection. Additionally, an employment in suspension biochips of Frster resonance energy transfer (FRET) allows improving detection specificity. The absence of fluorescent background from non-interacting with the beads dye-labelled antibodies additionally increases the sensitivity of detection and further facilitates the multiplexing capabilities of nanocrystals-based detection and diagnostics. So the combination of the biochips and QDs techniques allow increasing detection sensitivity and significantly raising the number of detected objects (multiplexing capacities). Such combination should provide the breakthrough in proteomics, particularly in new drugs development, clinical diagnostics, new disease markers identification, better understanding of intracellular mechanisms.

  18. Tuzikov A.B., Chinarev A.A., Gambaryan A.S., Oleinikov V.A., Klinov D.V., Matsko N.B., Kadykov V.A., Ermishov M.A., Demin I.V., Demin V.V., Rye P.D., Bovin N.V. (2003). Polyglycine II nanosheets: supramolecular antivirals? Chembiochem 4 (2-3), 147–54 [+]

    Tetraantennary peptides [glycine(n)-NHCH(2)](4)C can form stable noncovalent structures by self-assembly through intermolecular hydrogen bonding. The oligopeptide chains assemble as polyglycine II to yield submicron-sized, flat, one-molecule-thick sheets. Attachment of alpha-N-acetylneuraminic acid (Neu5Acalpha) to the terminal glycine residues gives rise to water-soluble assembled glycopeptides that are able to bind influenza virus multivalently and inhibit adhesion of the virus to cells 10(3)-fold more effectively than a monomeric glycoside of Neu5Acalpha. Another antiviral strategy based on virus-promoted assembly of the glycopeptides was also demonstrated. Consequently, the self-assembly principle offers new perspectives on the design of multivalent antivirals.