Предложен метод 3D-TIRF-микроскопии, основанный на объединении методик ультрамикротомии и флуоресцентной микроскопии полного внутреннего отражения поверхности образца после среза и позволяющий реконструировать трехмерную ультраструктуру объектов.

A system of a low-profile SPM-deflectometer (SPM - scanning probe microscopy) which makes it possible to increase the aperture of the supplied objectives to the currently record value NA = 0.75 has been developed, manufactured and tested. The introduction of such a system will significantly improve the performance of optical techniques for combined SPM / optical microspectroscopy systems.

Competitive schemes for constructing an optical system for combining a scanning probe microscope and optical microspectrometer, allowing the study of opaque samples by the method of tip enhancement of the Raman scattering intensity, are considered. Optimal objectives for the implementation of each scheme, taking into account the presence of a scanning microscope probe, are selected. The efficiency of usage of each optical scheme is estimated quantitatively for both excitation of a Raman scattering signal and collecting secondary radiation. As a result, the most effective optical system in terms of "excitation / collection“ parameter is revealed.

A unified analytical technology based on the integration of scanning probe nanotomography and optical microscopy (SPNT–OM) is presented, and the potential of the technology is demonstrated by the example of a multiparameter analysis of encoded microspheres. The presented SPNT–OM technology can become a powerful tool for multimodal nanocharacterization of a wide range of microparticles, composites, and hybrid polymers.

The principal limitation of the use of the Tip Enhanced Raman Scattering spectroscopy (TERS) is due to the lack of reliable scanning probes. The paper presents a simple technique for the fabrication of cantilever-based TERS-probes with highly reproducible characteristics, high performance, reliable and giving sufficient enhancement. The technique is based on the modification of standard scanning probe microscope contilevers by plasma etching and the subsequent formation of a TERS-amplifying region on the tip of probe by deposition of colloidal nanoparticles by dielectrophoresis.

A united analytical technology based on the integration of scanning probe nanotomography and optical microscopy (SPNT-OM) is presented and the potential of the technology is demonstrated by the example of multi-parameter analysis of encoded microspheres. The presented SPNT-OM technology can become a powerful tool for multimodal nanocharacterization of a wide range of microparticles, composite and hybrid polymers.

AbstractA new experimental method for nanoscale measurements of the absorption spectra of single nanoobjects has been developed based on scanning near-field optical microspectroscopy (SNOM) and nanospectrophotometry (NSP). The main distinctive feature of the proposed SNOM-NSP technique consists in depositing a sample onto a coverglass followed by its probing in the total internal reflection spectroscopy mode. This approach allows the number of analyzed samples to be significantly increased and provides the possibility of combining measurements with other optical techniques. The proposed SNOM-NSP method has been successfully used for studying single plasmonic nanoparticles and their complexes with Rhodamine 6G dye.

Histone proteins are promising carriers for delivery of recombinant nucleic acids in various cell cultures. Conjugates of proteins with fluorescent dyes are modern tools in the study of cellular transport and intracellular distribution of important biopolymers. The aim of this work was to optimize the method of recombinant histone H1.3 conjugation with photoactivatable fluorescent dye. Also we studied intracellular penetration and subcellular localization of the resulting conjugate. Resulting conjugate was capable of intensive fluorescence in the red region of the spectrum after irradiation with violet light. The degree of conjugation was determined under different reaction conditions. A comparison of the cytotoxicity of histone H1.3 and its conjugate with photoactivated fluorescent dye in HeLa cells and its intracellular localization was characterized. We found that conjugates mainly localize in recycling endosomes and to a lesser extent in peroxisomes. Thus, the resulting conjugate of recombinant histone H1.3 with photoactivatable fluorescent dye can be used for further studies its anticancer activity and as carrier for drugs and nucleic acids delivery into human and animal cells.

Particles size of lipase from porcine pancreas, polystyrenesulfonate-Na and their complex PSS-Lipase were investigated for the first time. The obtained data suggest the formation of stable complex PSS-Lipase with average particle size about 23.4 nm.

The influence of polyelectrolyte and polymer environment on the lipase activity was studied. The models of supramolecular systems based on lipases and polyelectrolytes were proposed. The prospects of using the polymer-enzyme systems in practice were showed.