rus
DOI: 10.22184/1993-8578.2020.13.6.360.363
The digital bionanoscopy platform can be used to solve a wide range of problems in biology and medicine. Its capabilities are applicable to study actual problems in biology and medicine, such as early detection of viruses, bacterial resistance to antibiotics and topological issues of neuronal networks.
The digital bionanoscopy platform can be used to solve a wide range of problems in biology and medicine. Its capabilities are applicable to study actual problems in biology and medicine, such as early detection of viruses, bacterial resistance to antibiotics and topological issues of neuronal networks.
Теги: digital bionanoscopy platform scanning probe microscopy сканирующая зондовая микроскопия цифровая платформа бионаноскопии
Our research group develops an experimental bionanoscopy platform on the base of a scanning probe microscope and microlens technology to provide the following:
optical observations with a resolution up to 40–50 nm and a time interval of a fraction of a millisecond;
registration of the biology objects topography and their characteristic movement in a liquid and air with sub-nanometer resolution and a time resolution of a fraction of a millisecond;
local exposure to chemical reagents in the nanometer-sized area with control over the dose of the agent, while the volume of the delivered dose can reach the level of 10–15 microlitres and less;
detection of the electrical potential distribution pattern over the surface of the object;
observation of electric signals flow along the selected tracks.
The bionanoscopy platform makes it possible to study biological objects, detect viruses and the antibiotic resistance of bacteria. Nowadays, there exist either very sensitive (enzyme-linked immunosorbent assay, ELISA or polymerase chain reaction, PCR) or rapid detection methods (immunochromatographic strips). The gap between the highly sensitive methods and express techniques is 100–10,000 times the difference in the virus detection limit [1–4].
Our experiments in detection of the influenza A virus we use a sensor surface with biospecific interaction based on an optimal probe (antibodies, aptamers, synthetic receptors) intended for use in a flow-through liquid cell. When choosing a probe, the following parameters are taken into account: sensitivity, specificity, analysis time, binding constant, false positive and false negative results, durability, and a possibility of regeneration. The experiments are controlled with the aid of enzyme immunoassay and colorimetric analyses and PCR diagnostics. In the experiments, a possibility of using Rayleigh light scattering by viral particles both in the flow and when attached to the sensor surface of the biochip is considered.
In experiments with bacteria, a method of cells delicate immobilization on a sensor surface or a surface modified with reagents (polylysine, antibodies) is used. In this case the task of optical and probe microscopy is to monitor submicron vibrations of the living bacterial cell membrane (or a single bacterium) and their changes (attenuation) under the influence of antibiotics.
Determining the nature of the antibiotic effect on a bacterial cell by a significant decrease in the nature of oscillations of the cell membrane can significantly reduce the time of the antibiotic test compared to a conventional test used for observing cell division and colony growth. The method to record cell oscillations can reduce the total testing time to 20 minutes or less.
The bionanoscopy platform aims to study the detailed structure of living neurons networks. Here we propose the following approaches:
the first is the geometric alignment of the microlens array [5] with the area of neurons;
the second is to provide conditions for the full functioning and growth of the neural network;
the third is the initiation of signal transmission using scanning capillary microscopy [6] as a result of local chemical action by a mediator and / or an external impulse (electrical, electrochemical, mechanical).
Particular attention is paid to the formation pattern of the contacts between dendrites, the interaction between synapses and the formation and passage of nerve impulses through the neural network. Large-scale observation of the neural network is carried out using an array of microlenses while a detailed characterization of the topography of dendrites and axons the picture of signal passage will be carried out using atomic force, capillary and electrochemical microscopy. In the case of capillary microscopy, multichannel capillary probes are used to simultaneous recording of topography, measurement of the electric potential and local delivery of reagents.
The developed bionanoscopy platform makes it possible to determine:
the effectiveness of the designed receptor surfaces to a specific strain of the virus. Demonstration of the possibilities will be carried out on the influenza A virus and various model plant viruses (tobacco mosaic virus, potato virus, etc.);
bacterial resistance to antibiotics during the test duration of no more than 20 minutes;
the role of the dendritic processes of the neuron in the formation of nerve impulses of the neural network and the relationship of their topology with the functions of neural networks. ■
This work was supported by the Russian Science Foundation, project No. 20-12-00389, and the Russian Foundation for Basic Research, project No. 20-32-90036.
optical observations with a resolution up to 40–50 nm and a time interval of a fraction of a millisecond;
registration of the biology objects topography and their characteristic movement in a liquid and air with sub-nanometer resolution and a time resolution of a fraction of a millisecond;
local exposure to chemical reagents in the nanometer-sized area with control over the dose of the agent, while the volume of the delivered dose can reach the level of 10–15 microlitres and less;
detection of the electrical potential distribution pattern over the surface of the object;
observation of electric signals flow along the selected tracks.
The bionanoscopy platform makes it possible to study biological objects, detect viruses and the antibiotic resistance of bacteria. Nowadays, there exist either very sensitive (enzyme-linked immunosorbent assay, ELISA or polymerase chain reaction, PCR) or rapid detection methods (immunochromatographic strips). The gap between the highly sensitive methods and express techniques is 100–10,000 times the difference in the virus detection limit [1–4].
Our experiments in detection of the influenza A virus we use a sensor surface with biospecific interaction based on an optimal probe (antibodies, aptamers, synthetic receptors) intended for use in a flow-through liquid cell. When choosing a probe, the following parameters are taken into account: sensitivity, specificity, analysis time, binding constant, false positive and false negative results, durability, and a possibility of regeneration. The experiments are controlled with the aid of enzyme immunoassay and colorimetric analyses and PCR diagnostics. In the experiments, a possibility of using Rayleigh light scattering by viral particles both in the flow and when attached to the sensor surface of the biochip is considered.
In experiments with bacteria, a method of cells delicate immobilization on a sensor surface or a surface modified with reagents (polylysine, antibodies) is used. In this case the task of optical and probe microscopy is to monitor submicron vibrations of the living bacterial cell membrane (or a single bacterium) and their changes (attenuation) under the influence of antibiotics.
Determining the nature of the antibiotic effect on a bacterial cell by a significant decrease in the nature of oscillations of the cell membrane can significantly reduce the time of the antibiotic test compared to a conventional test used for observing cell division and colony growth. The method to record cell oscillations can reduce the total testing time to 20 minutes or less.
The bionanoscopy platform aims to study the detailed structure of living neurons networks. Here we propose the following approaches:
the first is the geometric alignment of the microlens array [5] with the area of neurons;
the second is to provide conditions for the full functioning and growth of the neural network;
the third is the initiation of signal transmission using scanning capillary microscopy [6] as a result of local chemical action by a mediator and / or an external impulse (electrical, electrochemical, mechanical).
Particular attention is paid to the formation pattern of the contacts between dendrites, the interaction between synapses and the formation and passage of nerve impulses through the neural network. Large-scale observation of the neural network is carried out using an array of microlenses while a detailed characterization of the topography of dendrites and axons the picture of signal passage will be carried out using atomic force, capillary and electrochemical microscopy. In the case of capillary microscopy, multichannel capillary probes are used to simultaneous recording of topography, measurement of the electric potential and local delivery of reagents.
The developed bionanoscopy platform makes it possible to determine:
the effectiveness of the designed receptor surfaces to a specific strain of the virus. Demonstration of the possibilities will be carried out on the influenza A virus and various model plant viruses (tobacco mosaic virus, potato virus, etc.);
bacterial resistance to antibiotics during the test duration of no more than 20 minutes;
the role of the dendritic processes of the neuron in the formation of nerve impulses of the neural network and the relationship of their topology with the functions of neural networks. ■
This work was supported by the Russian Science Foundation, project No. 20-12-00389, and the Russian Foundation for Basic Research, project No. 20-32-90036.
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