A method of extraction of quercetin from a plant cell based on the combined action of ultrasound and a metastable fraction of an aqueous solution is proposed. This treatment leads to more efficient release of the cytoplasmic component due to etching and/or mechanical destruction of the plant cell envelope. The oxidized fraction of the solution has the most pronounced extractive properties; however, it acts on quercetin by oxidizing the chromophore part of the molecule. According to the criterion of pigment preservation, the best extraction medium is the reduced fraction of water. Analytical methods were used to analyze the extract samples: UV-Vis spectroscopy, gel electrophoresis of proteins, 1H NMR spectroscopy, and QCM weighing, as well as scanning electron microscopy.

The determination of changes in the properties of polysaccharides in electrochemically activated water solution were carried out by means of UV-Vis spectrophotometry, time-of-flight secondary ion mass spectrometry and scanning electron microscopy. It was shown how the properties of samples analyzed (agar, modified starch, edible starch) changed depending on a fraction of electrochemically activated water solution (catholyte/anolyte). Our findings demonstrate that the determined changes can be valuable information in designing novel approaches to managing the properties of biological raw material used for food manufacturing process. Furthermore, the technology of electrochemically activated water solutions is a suitable green chemical process that can be translated to industrial level.

Changes in the properties of polysaccharides in an electrochemically activated aqueous solution have been studied using UV-Vis spectrometry, time-of-flight secondary ion mass spectrometry and scanning electron microscopy. It is shown that the studied solutions of polysaccharides of plant origin (agar, soluble starch, and food starch) are subject to modification, the direction of which is determined by the fraction of electrochemically activated solution used (catholyte or anolyte). The identified features may be in demand when developing new ways to control the properties of biological raw materials in food production technology. It was noted that the technology of an electrochemically activated aqueous solution, which has been classified as “green,” can be reproduced on an industrial scale.

A possibility of nonenzymatic hydrolysis of glucose polymers in a cell under variation in the cytoplasm redox balance is studied. Experiments have been carried out in vitro in aqueous solutions using water-soluble oligosaccharides and insoluble polysaccharides. To simulate different redox balance conditions, fractions of electrochemically reduced and oxidized distilled water are taken. Comparative investigations are performed using UV-vis spectrometry, time-of-flight secondary ion mass spectrometry (TOF-SIMS), quartz crystal microbalance (QCM) weighing, and scanning electron microscopy (SEM). It is shown that a change in the aqueous solution redox balance causes an amylolytic effect. The revealed features allow estimating the role played by nonenzymatic hydrolysis of polysaccharides in maintaining the intracellular level of glucose, a substrate needed for synthesis of adenosine triphosphate (ATF).

A nanocarrier was obtained by coating the natural clinoptilolite particle surface with a phosphatidylcholine layer. The shell effective size does not exceed the thickness of the phospholipid molecular layer, which was confirmed by UV spectrometry and molecular mass spectrometry. The hydrodynamic diameter of the formulated nanocarrier, which was determined by dynamic light scattering, is smaller than the clinoptilolite core size. This effect is assumed to be caused by the phospholipid shell, which reduces the aqueous medium friction. The nanosize of the formulated nanocarrier, the natural clinoptilolite core, and the phospholipid shell together allow a combination of fruitful features that can be used for the fabrication of multifunctional platforms for the delivery of biologically active substances, bioimaging, or as a basis for biosensors.

For in vitro fertilization technology, the quality of oocytes has a direct impact on the egg fertilization and developmental competence of early embryo. The morphological criteria are used for the estimation of oocyte quality before its fertilization in vitro. To date, only one method is known to determine the maturity of oocyte. This is the routine observation with a light microscope. The aim of this article was to adapt the noninvasive quantitative laser scanning microtomography (QLSM) for the investigation of morphological features of a human oocyte in vitro. This approach was used to accumulate the Z-stack gallery of optical sections of IVF oocyte. The layer-by-layer acquisition allows the fine cytoplasmic structure imaging. Applying the QLSM Z-stack of optical sections, the cellular volume was calculated with quantitative 3D reconstruction of a human oocyte. The volume value and intracellular structure were used as novel criteria to assess the oocyte state after the stress evoked by cryopreservation procedure.

A technology for obtaining nanoparticles of natural clinoptilolite, a mineral from the zeolite family, under laboratory conditions has been developed. The size of zeolite particles was determined by scanning electron microscopy, and the size of nanocapsules was determined by dynamic light scattering. A method for applying a layer of alcohol-soluble phosphatidylcholine (lecithin) onto the surface of a nanoparticle has been proposed. The adsorption of lecithin by clinoptilolite particles was studied by ultraviolet spectrophotometry. It has been shown that the formation of the complex of phospholipid and zeolite in ethyl alcohol had a two-phase kinetics: in the initial phase, the adsorption of lecithin was recorded; it then desorbed into the solvent.

В данной работе изучали тонкое строение бактериальной плёнки, сформированной на внутренней поверхности проточного реактора. Используя подходы сканирующей электронной микроскопии (SEM), исследовали детали рельефа биоплёнки. Анализировали действие электрохимически восстановленного водного (ERW) раствора на ультраструктуру биоплёнки, созданную планктонной формой E.coli и/или лактобактериями. Показано, что обработка посредством ERW-раствора разрушает полимерный матрикс биоплёнки и её клеточную компоненту.

The action of electrochemically activated water on the fine structure of biofilms formed by the plankton forms of lactic acid bacteria and E. coli was investigated. Bacterial biofilms were grown on the inner surface of the tube of flow reactor, imitating a pipeline. The ultrastructure of the relief of biofilms was visualized using scanning electron microscopy. Sequential treatment with anolyte and catholyte destroyed both the organic polymer matrix of biofilms and bacterial cells embedded in the matrix.

The aim of this work was to study the fine structure of bacterial films grown on the inner tube surface of a flow reactor. Using the scanning electron microscopy (SEM) approaches, the detailed biofilm relief was visualized. The action of electrochemically reduced water (ERW) on the biofilm ultrastructure generated by the plankton form of E. coli and/or lacto bacteria was investigated. The treatment with an ERW solution destroyed the biofilm organic polymer matrix and bacterial cells embedded in the matrix.

The technique for the preparation of biological tissue sections developed for Electron Probe Microanalysis has been adapted for ToF-SIMS analysis of mouse GV stage oocytes. GV-oocyte sample preparation involves the following steps: plunge freezing, freeze drying, impregnation in an embedding medium, and section cutting. Molecule-specific images of the distribution of molecules in a single oocyte have been obtained with the described technique and ToF-SIMS analysis. The ToF-SIMS analysis data show that the efficient lateral image resolution is approximately 1 μm. Hence, ToF-SIMS enables us to study the distribution of chemical substances in relation to the morphological data obtained by scanning electron microscopy or conventional light microscopy.

The structural organization of the biological material in the nucleoli (Nucleolus-Like Bodies (NLBs)) in mouse GV oocytes has been studied with atomic force microscopy (AFM), scanning electron microscopy (SEM), and time-of-flight secondary ion mass spectrometry (ToF-SIMS). Experimental samples are prepared by quick plunge freezing, followed by freeze-drying, embedding in Epon epoxy resin, and sectioning with a microtome. The biological material in the NLBs is found to be organized as a domain structure of densely packed granules with sizes from tenths of a micron to a micron.

Методами атомно-силовой микроскопии, электронной сканирующей микроскопии и времяпролетной масс-спектрометрии вторичных ионов исследована структурная организация биологического материала в ядрышках (ядрышко подобных тельцах — nucleolus-like bodies, NLBs) преовуля- торных ооцитов мыши на стадии зародышевого пузырька. Экспериментальные образцы получены методом быстрой криофиксации с последующей мягкой лиофилизацией, заливкой эпоксидной смолой Epon и нарезкой срезов с помощью микротома. Установлено, что биологический материал в NLBs организован в виде гранул с размером от долей микрона до микрона, плотно упакованных в доменную структуру, которые проявляются в изображениях, полученных методами атомно-силовой микроскопии, электронной сканирующей микроскопии и времяпролетной масс- спектрометрии вторичных ионов.

This paper updates our knowledge on quantitative laser scanning microscopy and summarizes the capabilities of this method as applied to cytometry and analysis of cell structure of the mouse early embryo and the oocyte. This method requires a stack of optical sections obtained as Z-series with subsequent 3D reconstruction. This approach was used for visualization of the 3D cell model, measurement of the cell volume and surface area, as well as a study of the cell interior via optical sections. To maintain the dimensional characteristics of embryos or oocytes the method of sample preparation involved the following consequent steps: rapid cryofixation, low-temperature dehydration, infiltration by optically transparent mounting media, and laser-scanning microscopy. This strategy enables volume measurement, even in the case of a single cell within the multicellular system of the mouse early embryo.

Regulatory volume decrease in response to hypotonic stress is typical of the oocytes and early mouse embryos. Changes in the kinetics of osmotic reaction can be used as a marker of the modulating effect of the incubation medium on transmembrane transport in embryonic cells. Quantitative laser scanning microtomography (QLSM) was used to measure oocyte volume. In this paper, it is shown that addition of 5 μM glycine, taurine, or GABA, as well as ATP to Dulbecco’s medium abolished the regulatory volume decrease in mature mouse oocytes.