RU2348694C1 - Device for bioculture cell irradiation - Google Patents

Abstract

FIELD: chemistry; biochemistry.

SUBSTANCE: invention concerns biotechnology, biology, medicine and biophysics. Device includes panoramic gauge of standing tension and relaxation wave factor, including generator and indicator, reflectometre with one arm connected to generator and another arm connected to indicator, vessel with bioculture, and additional thermostat. Electromagnetic generator is used, with third arm connected to adjusted load and fourth arm connected to thermostat. Mentioned connections are implemented in the form of quazioptic routes. End route part of fourth arm is inserted into thermostat through a sealed hole in its wall. Vessel with bioculture is positioned inside thermostat in radiation zone and made of material transparent for radiation. Procreation microbe cells cultivated to logarithmic phase of bioculture growth are used. Device can include container for reference bioculture, positioned inside thermostat and screened from radiation. End route part of fourth arm can feature horn. Generator with air oxygen absorption and irradiation frequency can be used. Vessel for bioculture can also feature teflon film lid.

EFFECT: significant increase of cell numbers in growing bioculture population.

5 cl, 2 dwg, 1 ex

Description

The invention relates to the field of biotechnology, biology, medicine and biophysics and can be used in biotechnological industries and for scientific purposes to remotely stimulate the growth rate and study the biological characteristics of developing cell cultures of prokaryotic and eocaryotic cells.

It is known to grow cells in thermostats, while growth is stimulated by selecting temperatures and a nutrient medium.

The disadvantage of this device is the restrictions in obtaining high concentrations of cells.

Known circular rocking chair UVMT-12-250, which is a holder for flasks with culture, placed in a thermostatically controlled volume, providing the optimum temperature for the development of cells. The flasks are constantly shaken, which provides aeration (saturation of the medium with oxygen) of the grown culture.

In this case, the number of cells in the population increases sharply.

The closest in technical essence to the proposed solution is the installation consisting of a panoramic meter of the standing voltage wave coefficient (VSWR) and attenuation, consisting of a oscillating frequency generator and indicator, directional reflectometer, MSW cell, transmitted wave sensor, electromagnet, ammeter, stabilized current source Hall sensor; magnetic induction meter, test tubes with bioculture (see patent for the invention of the Russian Federation No. 2287014, IPC C12N13 / 00).

A disadvantage of the known device is the complexity of the installation, the lack of effectiveness of the influence of magnetic fields on the biological activity of microorganisms. In addition, the device cannot be used to affect developing cells, i.e. cells in a state of biological activity.

An object of the present invention is to provide a device for influencing developing cells.

The technical result is a significant increase in the number of cells in a developing population.

This object is achieved in that the device for irradiating bioculture cells, including a panoramic meter of the coefficient of the standing wave of voltage and attenuation, containing a generator and an indicator, a reflectometer, one arm of which is connected to the generator, the other to the indicator, a vessel with bioculture, according to the solution further comprises a thermostat, the generator of electromagnetic radiation is selected as the generator, while the third arm is connected to the matched load, and the fourth is connected to the thermostat, the mentioned connections filled in the form of quasi-optical paths, the end of the fourth shoulder tract is placed inside the thermostat through a sealed hole in its wall, the bioculture vessel is placed inside the thermostat in the radiation zone and is made of a material transparent to radiation, prokriatic microorganism cells grown to a logarithmic are selected as bioculture phases of bioculture development.

The device contains a container for control bioculture, placed inside the thermostat, shielded from radiation.

The end of the fourth shoulder tract is equipped with a speaker.

A generator with a frequency of absorption and radiation of atmospheric oxygen was selected as a generator.

The bioculture vessel is equipped with a Teflon film lid.

The solution is illustrated by drawings: in Fig. 1 is a block diagram of the inventive device, in Fig. 2 - growth curves of cultures of E.Coli K-12, where

1) a panoramic meter of the coefficient of the standing voltage wave (VSWR) and attenuation;

2) oscillating frequency generator;

3) indicator;

4) the first quasi-optical path;

5). reflectometer;

6) the second quasi-optical path;

7) the third quasi-optical path;

8) the fourth quasi-optical path;

9) thermostat;

10) a vessel with irradiated bioculture;

11) containers with a control bioculture;

12) agreed load;

13) transmitted wave sensor.

It consists of: a panoramic meter of the coefficient of the standing wave of voltage (VSWR) and attenuation 1; as a part of a quasi-optical oscillator of oscillating frequency 2, for example, a quasi-optical programmable EHF generator, and indicator 3 (Ya2R - 67); quasi-optical paths 4, 6-8; reflectometer 5; dry electric thermostat 9 (for example, brand TS-80M-2); a vessel 10, for example an Ermleiner flask with irradiated culture; load agreed 12; shielded container 11 with three flasks (irradiated, aerated, control culture); transmitted wave sensor 13.

The installation works as follows. The signal from the oscillator of the oscillating frequency 2 using a quasi-optical path 4 is fed to a directional reflectometer 5, where part of the incident high-frequency power is branched into the quasi-optical path 8 and detected by the transmitted wave sensor 13. The detected signal is fed to the input of the indicator 3 through the quasi-optical path 6. The main part incident high-frequency power is fed to the Ermleiner flask with the irradiated culture 10, which is at a stabilized temperature set in thermostat 9. Using the coordinated load 12 minutes, a quasi-optical path 7, the transmitted wave multipath excluded. The signal reflected from the bulb along path 8 through an OTDR enters path 6, where it is detected and fed to the input of indicator 3. Both detected signals in indicator 3 are compared. As a result of processing, on the screen of the cathode ray tube of indicator 3 information is displayed on the magnitude of the introduced attenuation by the irradiated medium. The flask 10 is made of a material transparent to electromagnetic radiation. Technical parameters of the thermostat ensure that a constant temperature is maintained with a spread of up to 0.25 ° C in the range from + 28 ° C to + 55 ° C. To study non-thermal effects associated with the influence of electromagnetic radiation and the exclusion, in this regard, the influence of the temperature factor on the experimental data, the accuracy of maintaining the temperature in the working chamber of the thermostat was increased to 0.05 ° C. The working chamber of the thermostat was a parallelepiped with an internal size of 395x400x500 mm with conductive walls. A quasi-optical programmable generator provides irradiation of objects of research at frequencies of the terahertz range, in particular at a frequency of a molecular absorption spectrum of atmospheric oxygen, 129 ± 2 GHz. The end part of the quasi-optical path 8 is mounted in the working chamber of the thermostat using heat-insulating material. For flask 10, a cap was made of a Teflon film 30 μm thick (transparent for terahertz waves), which made it possible to irradiate the culture without contact with the atmosphere. The latter allowed for research in hypoxia. Using the principle of separate separation and detection of incident and transmitted wave signals, it was possible to carry out controlled irradiation of the culture at various frequencies in the operating range of the apparatus and measure the absorption parameters of the medium with the culture in the flask 10.

An example of a specific implementation. The daily agar culture of E. coli K-12 Escherichia coli test strain was washed with physiological saline and 0.1 ml of this suspension was inoculated into meat-peptone broth (MPB), poured 100 ml into 3 300 ml Ermleiner flasks. The optical density of the crops on a spectrophotometer was adjusted to 0.31. All three flasks were placed in a metal container 11 inside the thermostat. To ensure the same development conditions for 24 hours, the temperature inside the container and inside the thermostat's working chamber was kept constant and equal to 37 ° С. At the 4th hour of culture development (logarithmic phase of development), the first flask was removed from the container and installed for 30 minutes under the quasi-optical path 8; the second was installed for thirty minutes in a thermostatic circular rocking chair UVMT-12-250, the temperature in which was previously brought to a value of 37 ° C; the third flask remained in the container and was not exposed to radiation and aeration. After 30 minutes, the flask, irradiated and aerated, was returned to the metal container 11.

To build three growth curves: irradiated, aerated and control samples, samples of 5 ml volume were taken at different times (0, 2, 4, 6, 8, 12, 24 hours), the optical density of which was measured on an SF-26 spectrophotometer at a wavelength λ = 560 nm.

As can be seen from the data presented, up to 4 hours of development, the optical density indices of all three samples are almost identical and amount to 0.33-0.4 relative units D. After 30 minutes of exposure to electromagnetic radiation and aeration, by the sixth hour of cultivation, the optical density of the irradiated and aerated culture sharply increases and amounts to 2.2 D. In the control sample, this indicator was 0.6 D. Subsequently, the development of populations of irradiated and aerated cultures occurred with the same intensity. The optical density at 8, 12, 24 hours of development was 2.9, 3.3, 4.3, and 3, 3.5, 4.5 D., respectively. The corresponding indices of the control sample were 1.1, 2.1, and 2.3 D. Also, the values of biomass growth after the 4th hour of development are given. The calculation of biomass was carried out by finding the particular values of optical density, for one of the crops, at the corresponding monitoring hour (0, 2, 4, 6, 8, 12, 24) to the previous one. Thus, after a thirty-minute exposure to the 4-hour population of Escherichia coli radiation, at the frequency of the molecular absorption spectrum of atmospheric oxygen, the biomass increase was 6.3 and was 1.4 times higher than with thirty-minute aeration (for which the indicator was 4.4). In the control culture, this indicator was 1.6. The obtained data confirmed the established fact, which indicates that irradiation of the E. coli culture in the logarithmic phase of the population’s development in its biological effect is similar to the effect observed during the enrichment of the culture medium with oxygen (aeration).

Thus, the thirty-minute effect of electromagnetic radiation on the molecular absorption frequency of atmospheric oxygen on a four-hour E. coli K-12 Escherichia coli culture leads to increased growth and increased biomass accumulation and is comparable in its severity with the effect of thirty-minute aeration. It was also found that after exposure, in the next two hours, the growth rate of the biomass of the irradiated culture is 1.4 times higher than that of the aerated one. When irradiated before the lag phase of the effect, and also during the stationary phase of the development of the population, stimulation of its development is not observed.

Claims (5)

1. A device for irradiating bioculture cells, including a panoramic meter of the coefficient of the standing wave of voltage and attenuation, containing a generator and an indicator, a reflectometer, one arm of which is connected to the generator, the second to the indicator, a vessel with bioculture, characterized in that it further comprises a thermostat, The generator of electromagnetic radiation was chosen as the generator, while the third arm is connected to a matched load, and the fourth is connected to a thermostat, the mentioned connections are made in the form of quasi-optical tracts, the end of the fourth shoulder tract is placed inside the thermostat through a sealed hole in its wall, the bioculture vessel is placed inside the thermostat in the radiation zone and is made of a material transparent to radiation, prokriatic microorganism cells grown to the logarithmic phase of bioculture development were selected as bioculture. 2. The device according to claim 1, characterized in that it contains a container for a control bioculture, located inside the thermostat, shielded from radiation. 3. The device according to claim 1, characterized in that the end part of the path of the fourth shoulder is equipped with a speaker. 4. The device according to claim 1, characterized in that a generator with a frequency of absorption and radiation of atmospheric oxygen is selected as a generator. 5. The device according to claim 1, characterized in that the vessel for bioculture is equipped with a lid of Teflon film.

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