Method for detection and identification of biological microobjects and their nanocomponents and related device for implementation thereof

FIELD: medicine.

SUBSTANCE: detection and identification of biological objects and their nanocomponents are enabled by exposure to radiation, monochromatic or nonmonochromatic radiation, including laser, sensing of one or more samples containing microobjects and their nanokcomponents with the use of a set of sample response measurement and record devices. The responses characteristics of each radiation conversion event are measured separately or in the aggregate, transferred and reduce in a diagnostically linear form. It is followed normalisation, correction and creation of a base of the reference and diagnosed parametres of microobjects and/or their nanocomponents. Further, the reference, diagnosed and identified microobjects and/or their nanocomponents are recognised and compared with experience data of required parametres on the basis of measurement with the use of a matrix.

EFFECT: use of the declared method allows precise qualitative and quantitative analysis of detected, identified, diagnosed parametres of the microobjects and their nanocomponents on the basis of optical measurement.

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The invention relates to the field of medicine, Microbiology, food and industrial biotechnology, namely to the study of biological materials by determining their physical and chemical properties using optical means, for systems in which the material probing or excited by optical means and it fluorescent.

There is a method of detection and identification of microbes from mixtures of microbes, which is based on a two-dimensional centrifugation according to the degree of sedimentation of bands equal to the density and on the detection of the associated particles on the properties of reflected light and fluorescence specific fluorescent labels (United States Patent 7070739).

The method allows to distinguish infection, to identify microbes, to study and characterize new germs. Also the way you confidently identify the microbes and promotes more effective treatment of the respective disease.

However, the use of the method and the appropriate equipment with consumables requires a significant financial cost. Also the method does not possess the noninvasive diagnosis is not clinical, and laboratory, which complicates its use by the physician, as required by the transportation of the material under investigation in the laboratory and transfer of research results back to the clinic, which additionally complicates the monitoring of the treatment process and evaluate its effectiveness.

The closest in technical essence of the present invention is a method of selective optical diagnostics of microbes and device for its implementation, which is based on the Desk as absorption and fluorescence characteristics of the studied substance, placed in the adjusted cell. At the same time as significant diagnostic data using the spectral characteristics for all pre-selected wavelengths of absorption and fluorescence, which is treated with computers and software) based on statistical regression methods to obtain information about the species of microorganisms in the substrate (Metalicana. Laser clinical biophotometer. M, 2008).

This method allows real-time to determine the concentration of microorganisms in biological substrates.

However, it is not possible to diagnose effluorescence biological substances and does not contain features intracorporeal survey. Us, he was chosen as a prototype.

The technical result of the invention is the possibility of qualitative and quantitative content analysis of organic substances on the basis of optical measurements for the purposes of industry, medicine, ecology and the food of those who of ology.

This technical result is achieved in that in the method applied technical solutions, which consists in the fact that, as a phenomenon, having a valid factor, for example therapeutic, and characterized by physical quantities, bearing diagnostic information, use the conversion rate of the laser (S) or other light source and/or other electromagnetic radiation with such aspects relevant characteristics of quantitative measures individually and/or collectively, which is measured by means of appropriate devices, such as reflection, scattering, absorption, fluorescence, Raman and nonlinear scattering, photoacoustic effects, thermo-optical and other, in the samples of objects of animate and inanimate nature of medical, industrial, environmental, food, biosphere, space and other purposes, with the subsequent porting of the received information, and diagnostic and analytical processing it in accordance with the based on a statistical regression model of component concentrations hardware-adapted algorithm of the method on a computer, which is that the characteristics remain on the device for storing information on computers and are linear in the diagnostic parameter form, and then their obrabecim the Ute according to hardware adapted the algorithm of the way so that first normalized all measured characteristics, for example, with the use of standards for the transmission, reflection, fluorescence and other phenomena of conversion, WHETHER in testing facilities, which are appropriate hardware functions for normalization parameters by means of multiplication by the latter, and/or, for example, in medical diagnosis in the case ectocarpales survey, for example, using the condition of continuity of the integral intensity of the reflected in the opposite direction of the exciting radiation, and/or using the condition of continuity of the integral intensity of the Raman bands of water at a selected wavelength of the exciting radiation, and/or using the condition of continuity of the integral intensity of the band fluorescence added to the samples in a standard concentration of quantum dots and/or fluorophores with a narrow-band fluorescence, and when andcorporate survey, using the condition of constancy of any performance conversion, WHETHER in a biological object (BO) or linear characteristics of homeostatic biological parameter of a person, and/or use of the multilevel, the normalization that takes into account the local values on the background obscheorganizmenny, according to the formula

Xpi=Xai/(1-αi·Xe),

where Xpi- Ravenna characteristic conversion, Xai- ectocarpaceae absolute characteristic conversion, Xe- andcorporate relative characteristics of conversion, αi- the coefficient at the i-th feature in the expression for homeostatic parameter P=Σαk·Ikand/or and/or other types of diagnostics restore the correct fluorescence spectra, using the previously measured spectral characteristics of the extinction using the measuring path of the complex spectra of actual fluorescence by the formula

Xi corr=Xi/(a1+a2·ei+a3·ei2),

where Xi corr- is corrected spectral characteristics of the i-th channel of the device, Xi- the value of the corresponding spectral characteristics without adjustment, a1, a2and a3- correction factors that depend on the geometry of the samples and the method of collecting light, eispatial damping rate due to extinction in the i-th wavelength, on the part of the stored spectral information for a sufficiently large number of samples of organic substances with known analytical content is used to create the model concentrations by implementing, in accordance hardware adapted the algorithm of the method, procedures, regression analytic in the information on the statistical principal components of the spectral characteristics of these samples, and for the rest of the samples of unknown content is in the calculation according to hardware adapted the algorithm of the method of using the resulting model, and calculated quantitative and/or qualitative characteristics that may characterize a biological or molecular composition, use for industry, medicine, ecology, food, biosphere, aerospace and other technologies, while producing the control treatment until full recovery in medicine and follow the cycles of transformations BO in production, and in the case of completion of the production cycle or during convalescence the patient's diagnosis is terminated, and medical diagnostic setup is used for radiation therapy patients also before each series of diagnostics to measure the characteristics of the transfer standards and make them into a computer to calculate the response function, and the samples are prepared, for example, by taking a certain number of investigated liquid, solid or gaseous object animate and/or inanimate nature and location in standardized cell for measuring the absorption spectra and laser-induced fluorescence and other characteristics of the radiation conversion laser or other light source, which can be as large a volume of approximately NESCO who are milliliters, and a small, approximately in the proportion of microlitres and collected on a tablet-based, containing also flow cell, this also could include integrated with plansyou device for photoacoustic, opto-thermal, and other diagnostics, which measure the relevant characteristics of the conversion of radiation is placed in the tablet of the cell samples, and for dilute solutions with the analyzed contents of the pre-increase the concentration of content filtering substrate, with subsequent determination of the concentration and type of substances, for example on the filter, and in the case of non-shared objects with spatially extended surface of the measurements associated with the light response run directly by holding the light-harvesting device to the surface of organic matter, and that in the global implementation of optical diagnostics using remote methods on measured characteristics of the conversion of the laser or other source to determine the quantitative and qualitative parameters close or distant samples, while the diagnostic complex is of single unit analysis of information processing fluorescence spectra of several blocks measuring these spectra and transmitting the information for processing the operations of analysis on a local or global network using standard protocols, including encryption protocols, and/or provide the implementation of individually normalized diagnostic clinic, which measure spectral and signal characteristics of the conversion of optical radiation in the living substance of each individual of the organism at different stages of its development and entered into the database of the parent computer, and then used for analysis, according to the above principle, and compare its results with the analysis made in the current time, and in case of deviations of the current diagnostic parameters from the individual and collective normal ranges make the diagnosis on the corresponding computed parameter pathology

and the fact that the device for its implementation, containing the cell to which the supply of the exciting radiation and the removal of the light response is complex fiber optic cable from the first end of the completed stranded, with the second end for transmitting excitation radiation is made solid, vein which enters the Central wire of the first end, the third end to transmit light response in the spectrum analyzer performed stranded, the strands which pass into the peripheral veins of the first end, a second end connected with the output of laser radiation, and the cell is made in the form, dimensions and materials that are specic to the task of the applications, the registration part is made of a matrix of semiconductor photodetectors, which are placed and/or not placed in thermostat is cooled to low temperatures, and/or multi-channel photomultiplier tube made according to the design principle of the photomultiplier tube louver type with common cathode and many of the anodes corresponding to the multiple channels, the number of approximately 150-300, and measurements of the transmission spectra is performed on the spectrum analyzer, which is connected with the single-core fiber, which is supplied from the side of the cell opposite side of the light source of polychromatic light, in addition and/or separately measured spectral fluorescence picture of the sample using a fluorescent microscope equipped with projecting optical part, a laser source for excitation of the sample, a filter, a polarizer, a dispersing element eyepiece for visual inspection and/or without eyepiece, high-sensitivity camera and digitizer device signal and interfacing with a computer (PC), and the system of collecting fluorescent radiation emitted diagnosed by the sample, can be implemented in several ways, for example, represents a simple end of the fiber optic catheter for ectocarpales and andcorporate applied what I or, for example, represents a lens or mirror collimator system, which allows you to collect and focus rays from a relatively small sample at different angles, the end face of the receiving light guide, or, for example, represents a dielectric antenna comprising two glued together dielectric plates, one of which is on the outside of the first side plate has the form of a slow wave structure, and the boundary of dielectrics on one edge of the plate, where the maximum focusing of the excited surface waves, enters the optical fiber, as well as to increase the collective power of the antenna, the volume of the second dielectric have a network of fine fiber lived that allow you to effectively translate the surface wave into the receiving light guide, or, for example, representing the optical resonator, with the capacity of holding it samples in cuvettes, special shapes, such as ellipsoidal with the receiving window of the light guide near the top of the semimajor axis, and the cuvette for measuring characteristics of fluorescence and extinction of the content in them is made in the form of containers hole forms with transparent walls and volumes, for example in the tenth, a hundredth of a milliliter to visualize objects animate and/or inanimate nature, and measured the I characteristics of conversion, and the distance between the walls of the ditch lie in the range greater than the transverse dimensions of microbes by 5-10% and/or a few thousandths of a milliliter for very turbid objects, and what to automate and speed up the measurement of large quantities of samples of the objects of animate and/or inanimate nature of the cell equal and/or different amounts of 0.001 to 1 ml are collected in tablets and cell are performed with a larger volume and/or additional built-in tablets, including running version, for measuring available in large quantities of sample substrates, this also includes integrated plansyou device for photoacoustic, opto-thermal and other diagnostics, with elements of interfacing with computers, and devices for collection of integrated and/or reintegrating light response, mating with a fluorescent microscope and a mechanism for horizontal and vertical movement tablets, part of the ditch in the tablet can be filled with substances that serve as benchmarks, positive, negative, etc.

Description of the method

The method of optical diagnostics of living matter is carried out by means of devices, the first of which is shown in figure 1. Fluorescent device for diagnostics has an interface for connecting computers. It is implemented using multi-channel light-sensitive items analog-to-digital converters, transmitting the spectral signal of each channel, which is the analogue of the spectral intensity at a certain part of the spectrum at the input channel of the computer (interface). The instrument is used in the proposed method is by using the appropriate device-adaptive algorithm of the method for installation of laser fluorescent diagnostics (figure 1), consisting of a laser 17 with suitable wavelengths generation of complex optical cable with fibers for the supply of the excitation radiation 11 to analyze the sample and removal of the fluorescent response and supply it to the spectrum analyzer 9, spectrum analyzer 6, consisting of an optical filter wavelength excitation 11, the dispersion element, and multi-channel optoelectronic detector 3, amplifier electric signal, an analog-to-digital Converter (2), computer (1) with the appropriate hardware-adapted algorithm method for processing and storing spectral information. Measure the spectrum of the extinction of the substrate used source nemonokhromaticheskogo (multi-frequency) radiation 7. When applying a tablet ditch the laser beam 17 passes through the selected cell tablets, and the light response of the cell 15 is assembled on the end of fiber 12 and is directed to the input of the spectrophotometer 6, d is more spectrum is digitized and transmitted to a computer (1). Also, figure 1 shows the following items: 4 - analog-to-digital Converter measuring transmission spectra, 5 - matrix of semiconductor detectors for measuring transmission spectra, 8 - investigated sample, 13 of the endoscope optical path of the probe transmission spectra, 14 - tablet, 16 - flow cuvette tablets. The tablet is made of optically transparent material and contains on the front surface of several (2-500) recesses, a ditch, with a small amount of about 0.1-0.001 milliliters, and some of them may be of the same size, and some different, and may also contain flow cell, the input and output ends of which are connected to the hoses that communicates with the tank inlet liquid and the tank to drain the fluid from the cell and/or a reference cuvette containing water for calibration of the Raman spectra. For measuring other characteristics of the conversion of laser and/or optical radiation use appropriate devices to register 19, the signal which is digitized by the ADC 20 and is input to the computer 1. For collecting fluorescent radiation and transmission by fiber optics uses light-harvesting device 18, schemes for the implementation of which is shown in figure 2. In the first embodiment, the light-harvesting device is a simple end of the fiber optic catheter is 7, which contains a Central core, intended for the supply of the excitation radiation to the sample 1 and the peripheral veins for collection and removal of fluorescent response 2. In the second embodiment, the light-harvesting device consists of a lens or mirror collimator system to collect and focus rays of the relatively small size of the sample 1 at different angles, the end face of the receiving light conductor 2. This system consists of a mirror 3 and a lens 4 or of the two lenses 4 and/or without a mirror 3 collected so that is the focus of the fluorescence radiation of the sample 1 in the output window of the light guide 2. In the third embodiment, the light-harvesting device is a dielectric antenna 5, collected from two glued together dielectric plates with different refractive indices n1 - 1 and n2 - 2, one of which is on the outside of the first side plate has the form of a slow wave structure (figure 3). The boundary of dielectrics on one edge of the plate, where the maximum focusing of the excited surface waves, enters the optical fiber. Also to increase the collective power of the antenna, the volume of the second dielectric has a network of thin fiber lived 3 that allow you to effectively translate the surface wave into the receiving optical fiber 4 (figure 3). Using t the one device you can sbirt fluorescent radiation from objects with a large surface (table, wall, surface tools and skin), and with small samples of fluorescent liquids. In the fourth embodiment, the light-harvesting device (figure 2) represents the optical resonator 6 with the possibility to accommodate in examples 1 half pans adapted for this form 1. The resonator is made of a special form, which provides a more efficient collection and transmission of radiation in the output window of the light guide 2, for example, in the form of an ellipsoid 6 with a mirrored inner surface, to which the fiber is fed to the top of the semimajor axis of the ellipsoid.

Multi-channel optoelectronic detector is performed in several different ways. In the first embodiment, this multi-channel matrix of semiconductor photodetectors. In the second embodiment, a multi-matrix of semiconductor photodetectors placed in a cooled thermostat low temperature. This will allow you to achieve greater resistance to its own thermal noise. In the third embodiment, this multi-channel photomultiplier tube, made on the principle of the photomultiplier tube louver type (figure 4). It contains one extended photocathode 1 and many of the anodes 2, the number corresponding to the number of recording channels, approximately 150-300. Between the anode and cathode is the system dinodon 3, consisting of metal plates inclined at primulinum angle to the plane of the cathode, moreover, plate facing directly one over another dinodon rejected in opposite directions. Working on dinode served cascade voltage multiplying the photocurrent of each channel. The number plates on dinetah one more than the number of anodes.

For diagnostics it is necessary to perform the correction of fluorescence spectra using the extinction spectra as due to absorption and scattering of light in the substrate, the fluorescence intensity decreases and the spectrum while still changes shape. Correction of the spectra is carried out by the formula (1).

where Xi corr- is corrected spectral characteristics of the i-th channel of the device, Xi- the value of the corresponding spectral characteristics without adjustment, and1and2and a3- correction factors that depend on the geometry of the samples and the method of collecting light, eispatial damping rate due to extinction in the i-th wavelength.

The cell can be made relatively large, and a small amount of and executed prisoners in protective casing, which is opaque, marred with the inner side and has only a cover for placing and removal of samples, the holes for the inlet and outlet hose flow-through cuvette and connectors for the Union with the light guide and/or the projection path of the transmission of light information. For example, to detect cells of microbes in low concentrations apply a thin cuvette with a volume of a few hundredths of a milliliter. And for substrates with large concentrations of use of the cell volume to a thousandth of a milliliter. These small, equal or unequal in volume of the cell can be collected on one tablet basis, placing the cell in a matrix manner. The distance between the transparent walls of the cuvette can be no more than 5-10% larger than the studied bacteria and/or microbes, which further increases the accuracy of measurement of the concentrations and species of microbes, and in addition allows you to get a true characteristic fluorescence of microbes without the contribution of the spectrum of the scattering environment, especially in substrates with low concentration, such as 101-103CFU/ml, where a CFU is the number of colony forming units of microbes in their seeding in the nutrient medium at 1 ml test substrate and/or suspension of microbes.

You can apply special tablet technology (Fig), implemented in the form of holes 3...4 cut quartz plate 2, which are produced with equal and unequal sizes, such as different and the same cross-sectional area or the same and different height, and covering the top plate 1. This technology has many what rikusentai compared with similar technologies. For example, the simplicity in the preparation of the measured substrates on it and when cleaning, when the content is easily washed off with water or alcohol. Also at the same transverse area of the holes is easy to implement standardization of the measurement results. For example, when measuring the response of spatially integrating devices such as a microscope, the measured value normalized to the amount of holes:

Xn=X/V,

where Xnthe normalized value, X is the value without normalization, V is the volume of the hole. But, for example, when measuring fiber-optic catheter up to the surface of the plate 1, and with a uniform light holes by laser, the measured value normalized so:

Xn=X·h/V,

where h is the thickness of the plate 1. When measuring fiber-optic catheter up to the surface of the plate 1, and the light holes of the exit window of the catheter measured value normalized so:

Xn=X·h3/V.

Measuring the fluorescence of advanced and/or separately performed on the fluorescent microscope, the scheme of which is shown in figure 5. Here polarized radiation, for example, excimer laser 8 is directed in cell 7 with a test specimen 6. The emission of fluorescence from single bacteria through the polarizer 5 and the spectral filter 4 enters the microscope 3, provided with, for example, the camera 2. RGB camera signal is surgical the characteristics of the sample, filter and wavelength of excitation. The computer input 1 which is applied the signal from the CCD camera 2, compares this signal with the existing library and determines the type of bacteria. Knowing the volume of the cell and counting apiece bacteria, calculate their concentration. In some cases, the degree of depolarization can clarify and complement the spectral signal.

For measuring the spectral characteristics of fluorescence and extinction of aquatic environments with low concentrations of microbes used pre-enrichment of the substrate diagnosed by produced by driving a large volume of aqueous medium through the filter. While the filter remains microbes that are either removed and dissolved in water to the desired concentration and placed in a cuvette for measurement, or all diagnostic measurements performed directly on the filter, and knowing, for example, the volume passed through it liquid, say water, it is easy to recalculate the results diagnostic concentrations of species of microbe or microbes per unit surveyed total volume of liquid such as water.

A second device, which is used as an optional source of diagnostic information in the expected way, is a hardware node to measure photoacoustics effect, consisting of a standard set of components for its devices. the composition includes a laser and a photoacoustic cell. The cell with the test substance is placed in a cell, include the laser pulse and measure the photoinduced acoustic response. The received signal locations are digitized and entered into the database on the mainframe. If the signal due to technical or other reasons is nonlinear distorted, his pre-linearized using pre-measured response function.

The third device, which is used as an optional source of diagnostic information in the expected way, is a hardware node for measuring opto-thermal effect. This device is in the standard version. Remove the signal is translated by transforming the computer into a linear data format and is saved in the database.

The fourth device, which is used as an optional source of diagnostic information in the expected way, is a hardware node for measuring lines of Raman scattering of laser radiation. This device also runs the standard version. Remove the signal is translated by transforming the computer into a linear data format and is saved in the database.

To obtain diagnostic information about the examined object can be additionally used devices, the principles and methods of work which is s based on measurement of one or more of the other effects of the conversion of electromagnetic radiation into other forms of energy within the object when the corresponding object. The resulting signal is digitized and fed to the input port/output systems. On the computer using device-adaptive algorithm of the way the signal is converted into a linear quantitative internal parameters of the object form.

The method allows to normalize the measured spectral characteristics without the use of additional devices normalization, as the object and means of normalization is itself water environment test substrate and/or the insertion into it of the quantum dot, when applying the proposed methods of normalization. It is also possible at the beginning of each series of measurements to calculate the hardware functions for normalization of all of the readings. For this measure relevant characteristics of the conversion of WHETHER the standards and using the obtained characteristics, calculate corrections for the response function, using the criteria necessary constancy of these characteristics in time for standards.

For normalization the absolute and relative values of the measurement results of the characteristics of conversion DOES BO and NC use two types of calibrator: andcoronary and ectocarpales. In the first case, all measured characteristics are normalized with regard to the relevant correction factors for any individually expressed and recorded internal characteristic or d is esteticheski parameter conversion IF (opto-thermal, photoacoustic, fluorescence, Raman and others), which is proportional to the intensity of WHETHER each local area BO as in norm and in pathology (and also taking into account the prehistory of these parameters before and during pathology), direct features are normalized, are linear in the diagnostic parameter mind:

Xe=Ii/P, P=Σαk·Ik,

where Xenormalized value characteristics, Ii, Ik- direct readings, converted to linear diagnostic parameter mind, R - normalizing factor defined in General as the sum of the measured values of Ikmultiplied by the weighting coefficients αkfound by the algorithm described below, as a result of statistical studies to determine, for example, homeostatic parameter species or another do not change from individuals const parameter, i, k is the sequence number of characteristics.

In the second case, all measured characteristics during in vitro examination is standardized and adjusted absolute values using the characteristics of the conversion, WHETHER in the studied object as an internal source of amplified molecules, cells), and an external source of converse the local signal (fluorescent additives, quantum dots, different standards and so on), and subtracting the background effects from the apparatus, tubes), which is determined by the formula:

Xai=(Ii-If)/P,

where Xai- the absolute value of the characteristics, If- readings in the background.

You can use multilevel quantitative value obtained taking into account both the above amendments, to consider local changes of characteristics in the background obscheorganizmenny:

Xpi=Xai/(1-αi·Xe),

where Xpithe multilevel value characteristics.

For the objective diagnosis of the type and concentration of chemical component or microbes with the purpose of normalization of the spectra using several methods, depending on the specific substrate. Use normalization to the intensity of the reflected excitation radiation, passed through the measuring path and gave their contribution to the overall spectrum of a light response of the substrate. Normalization factor equal to the inverse of the integral intensity of the reflected excitation radiation. Just use the normalized intensity of the bands of Raman scattering by water molecules of the laser radiation. This method of normalization gives the most accurate normalization factor, since the intensity of the Raman bands of property nalina only the intensity of the laser radiation in the substrate. Normalization factor equal to the inverse of the integral intensity of the Raman bands of water molecules. For normalization of the signal instead of the Raman spectra to use a range of Rayleigh scattering of water molecules, but it is much more difficult to eliminate the contribution of the laser radiation (glare). Still using the normalization using fluorescent or luminescent additives with a standard concentration directly in the substrate. This method of normalization is also quite accurate. This requires the use of quantum dots or fluorophores with a narrow spectral bands of fluorescence. Normalization factor equal to the inverse of the integral intensity of the band fluorescence additives.

Hardware-adapted algorithm of the method is based on the method of concentration, allowing on the basis of comparison of the database and the obtained spectra to assess the qualitative and quantitative composition of biological and chemical component content diagnosable substance in the form of concentration of the microorganism and its species. This algorithm allows specific quantitative analysis microbacterial biological substrate and dissolved elements by their spectral information through the implementation of linear regression on the main com is ananti (ha) spectral characteristics. Ledger and the regression coefficients are stored in a database and are static source of information during the process of diagnosis. The signal resulting from fluorescent diagnostics, is the spectrum at a given wavelength and a given discretization step. The signal for the other methods used in the method can also be present in the form of a set of data as a multidimensional vector magnitude (dimension equal to the number of discrete values in the data) or range.

The method is based concentrations was based on a phenomenological model of the characteristics of the conversion, WHETHER in the sample as the sum of the corresponding characteristics of molecular or other component. According to her, in samples of biological substrate are a few common for all samples types of such components. Upon excitation by radiation of the red range of these components, such as fluorophores or chromophores are usually porphyrins. We believe that the characteristics captured in the sample, in the first approximation is the sum of the characteristics of the contained components. The analysis of fluorescence spectra or other characteristics of the conversion IF we get a discrete set of intensities of fluorescence or other measurable phenomena emitted by the sample at different wavelengths. We assume that any analysera the range of spectrum is a convolution of several spectra, generated by various components in the sample. Then any intensity value for each sample from this set according to the model is the sum of spectral intensities of fluorescent or other component, multiplied by their concentration coefficients

where xsjdiscrete spectral intensity of the s-th sample of the j-th wavelength, ykjdiscrete intensity of the k-th component in the j-th wavelength, csk- concentration factor, proportional quantitative measure of the presence of the k-th component in S-th sample.

The component may be a separate molecule, a biological cell, for example cell of the germ. In terms of linear algebra spectral intensity of the samples represent the coordinates corresponding to these sample vectors in a multidimensional space, whose size is determined by the number of steps of sampling data in the original spectra. In this representation, the spectral information of the discrete spectral intensity component ykjbaseline vector, on which are arranged the vectors of characteristics of samples. This circumstance basicity is due to the fact that the set of vectors of samples included in the linear envelope of the component vectors ykjbecause of the vector y kjhave the same dimension as xsjand they completely exhaust the possibilities of building total fluorescence spectra, assuming that the components are present in all samples and only these components are present.

For the stability of the model, as well as its universality, these basis vectors are not for the original space of the spectral characteristics, the dimension of which is relatively large, and for the parameter space of reduced dimensionality, which is using statistical factor analysis or principal component analysis of the spectral intensities of fluorescence of a phenomenon a large number of samples with known content. This preliminary data conversion is necessary because due to the large similarity of the fluorescence spectra of such objects the problem of nding the basis is complicated by computing the area close to the singularity. This behavior of the spectral characteristics is not possible to find the directions of the basis vectors that are resistant to noise, caused by deposits of hardware noise of the spectrometer in a single cell of the opto-electronic detector. But the transition to a statistically stable parameters of fluorescence, which is, for example, the main factors characteristics, instead of the space is nsta noisy individual spectral intensities, allows you to bypass this danger. The partitioning of the feature vector for factor and random components is the end result of cyclic iterative algorithm consistently engaged in each cycle calculations decomposition of the data matrix on the structural part and noise:

Here X is the data matrix, F is the matrix of the factors of interest in the basis of the principal component, the Q - matrix of size (J×P), which is the transposed matrix of loadings, F is the matrix of residuals. This algorithm assumes an initial set of signs should be centered, i.e. with the zero of the coordinate system of the feature space in the centre of the statistical scatter. It is assumed that the vector of residuals e=(e1e2,...,ek) obeys p-dimensional normal distribution with zero vector mean and diagonal covariance matrix Ve.

To conduct factor analysis to model concentrations of pre-coordinate system center by moving the origin to the point defined by the feature vector, averaged over all samples, and conduct a principal component analysis. Then found the factors used to find the regression coefficients at scale concentrations when implementing regression using the least squares method for experimental the x points, corresponding samples of the reference sample. This means that in the model

where r0iand rpiregression coefficients for the i - th biological parts (germ), known csiand fspsearched coefficients r01and rpithe best approximation of this model for all samples from the reference sample using the method of least squares.

The result of regression on principal components is the calibration matrix, which can be used to hold the projection of the vector of characteristics (a discrete set of spectral intensities of fluorescence of the sample) on a scale of concentrations fluorophore component. Matrix are determined by the regression procedure of the main factors on matrix component concentrations of these samples, i.e. microbes.

where W is the calibration matrix of size J×I matrix of concentrations of dimensions S×I whose elements are the concentrations fluorophore component i in the samples from the reference sample with non-s. The regression procedure of the main factors on the scale of concentrations allows you to find the calibration matrix and the basis to calculate (predict) the concentration of individual species of microbes by the following formula

Here ci- oncentrate the i-th component (germ), woi- zero values of the regression coefficients, mj- value component of the vector that translates the values of the feature vector xjdiagnosed sample centered in the coordinate system of signs. The mathematical solution of the problem of recognition as the implementation of regression on principal components is solved in two stages: search for the space of linear characteristics (factors) source characteristics (spectra), with the greatest level of statistical significance, and within this space are vector, corresponding to the independent contributions of the analytical component in these characteristics. Calculate quantitative measures of the contribution from each analytical components (concentrations of components) are the projection of the vector linear statistical characteristics of fluorescence or other phenomena on the vector of this analytical components in the space of reduced dimension.

On Fig presents the General scheme of diagnostics and monitoring of the proposed method that allows to determine the pathology and to monitor treatment until full recovery in medicine, and to monitor the cycles of transformations BO in production, as well as in other applications. The scheme in total shows the main stages of hardware adapted algorithm method. The General scheme R is the implementation of the algorithm. Legend: 1 - in vitro measurement of conversion DOES BO (sample and tablet technologies), 2 - andcorporate measurement of conversion, WHETHER in BO using different signalisierung devices, 3 - normalization of absolute values, 4 - diagnosis and accumulation of data on monitoring of the treatment process or transforming the object in the process of manufacturing cycles, monitoring, ecology, etc., 5 - end 6 - archive database.

The calculated quantitative and qualitative characteristics of the samples used are, for example, when medical differential diagnosis of microbial diseases of nature, when the component is a cell pathogen in samples taken from patients. In industry, this method can be used for the optical method of monitoring the production of biological and chemical substances associated with biotechnology. In ecology is to control, for example, clean drinking water, contamination of natural and artificial reservoirs, disease diagnosis flora. In the food industry, the method is used for production control associated with the operation of the cultures of microorganisms, or for quality control of food products. Here the component is a cell of a microorganism, may be a molecule and t is xing.

Based on the above hardware and methodological elements in the implementation of optical diagnostics of living matter can be arranged following technical solution allowing to investigate living matter for the needs of medical and other diagnostics. This system of global diagnostics network consisting of a single unit analysis of information processing the set of spectra of several blocks measuring these spectra and transmitting the information for processing the unit of analysis in a local or global network using standard protocols, including encryption protocols. Cm. the scheme of the network MultiTerminal computerized system of Express-diagnostics and expert assessment of effectiveness of treatment and rehabilitation of patients with diseases and microbial processes of nature and for the systematic study of the person individually (Fig. Network MultiTerminal computerized system of individual rapid diagnosis and peer review processes. Legend: 1 - the branch medical applications, 2 - branch industrial applications, 3 - sectoral food applications, 4 - head center expert system for diagnosis and control processes, 5 - branch sites of ecological applications, 6 - field is passed the object space assignment 7 - branch objects biosphere destination), (Fig.9. Network MultiTerminal computerized system of individual rapid diagnosis and expert assessment of effectiveness of treatment of diseases and microbial processes of nature, and for the systematic study of man individually. Legend: 1 - the basic center of obstetrics, 2 - basic clinical gynecology center, 3 - basic clinical center of gastroenterology, 4 - basic clinical Pediatrics, 5 - basic, clinical center of dentistry, 6 - basic clinical therapy centre, 7 points lifetime individual medical examination of the population on the way and collect information on individual diseases, 8 - computer unit 9 measuring diagnostic node apparatus for optical studies of medical, industrial, food and environmental interest, 10 - implementation for intracorporeal survey, 11, 14 - a device for fluorescence diagnosis, 12, 15 is a device for FA diagnosis, 13 - implementation for in-vitro examination, 16 - gastroenterology, 17 - surgery, 18 - therapy, 19 - Pediatrics, 20 - dentistry, 21 - remote local clinical facility, 22 - mobile personal tool diagnostic tool porting head with computers, 23 - library microbes (Museum strains), library of microbes (CL the clinical strains). Mixture, differential diagnosis, data Bank on the preferred antimicrobial agent) is also diagnostic for the industry (figure 10. Network MultiTerminal computerized individual Express diagnostics industry, food technology, ecology, and other assignments. Legend: 1 - main centre of industrial Microbiology, 2 - basic centre of industrial biotechnology, 3 - core center of agriculture, 4 - base center chemical production, 5, 6 - computer unit 7 measuring diagnostic node apparatus for optical studies of medical, industrial, food and environmental interest, 8 - device for fluorescence diagnosis, 9 - device for FA diagnosis, 10 - remote local industrial Microbiology, 11 - remote local object agriculture, 12 - remote local object chemical production, 13 - database on industrial objects), environment (11. Network MultiTerminal computerized system of individual rapid diagnosis for industry, food technology, ecology, and other assignments. Legend: 1 - basic center for urban ecology, 2 - basic technical ecology, 3 - core center of ecology of the sea, the 4 - base control center management, 5, 6 - computer BC is to, 7 - measuring diagnostic node apparatus for optical studies of medical, industrial, food and environmental interest, 8 - device for fluorescence diagnosis, 9 - device for FA diagnosis, 10 - remote local urban ecology, 11 - remote local industrial ecology, 12 - remote local object ecology of the sea and other, 13 - database of industrial facilities), food technology and other assignments.

The system consists of a single server local and remote objects (Fig 8). The server contains a systematic library of microbes Museum and clinical strains, also a database of industrial, food and environmental objects, and hardware adapted algorithm of the method of conducting a diagnosis and gives treatment recommendations. Clients include diagnostic systems on remote local clinical, targeted industrial (environmental and others) and mobile objects. Some clients and acts as a consumer information server capabilities and supplier of additional information on variable kinds of microorganisms (basic).

In a centralized system is based on the interoperability of the medical diagnostic facilities in different institutions located in remote geographically the areas with the main computer system via the global Internet network. For this purpose, connected to a global computer network diagnostic and treatment facilities supplied hardware adapted algorithm method, which produces the connection with the server of the computer according to standard protocols. Computer remote diagnostic units have an interface for managing the complex and to control the interaction with the main computer for processing diagnostic information. As a result of transfer of processed data to the querying about handling the remote client on the computer screen diagnostic displays the result of diagnosis and basic treatment recommendations for a patient, the sample was investigated. Further, for example, monitoring the treatment process operator diagnostic complex enters and directs to the main computer records about the treatment. Based on this additional information, the server of the computer adjusts and sending to the remote client updated recommendations that will improve the conditions of treatment and to implement the expert evaluation of the effectiveness of rehabilitation and treatment of patients with diseases and microbial processes of nature.

Head of the computer network system of personal rapid diagnosis is a base for clicks the processing and storage of diagnostic information, coming from diagnostic systems in remote geographical areas. Also head of the computer contains diagnostic library microbes Museum and clinical strains, as well as mixtures of microbes and background options common biological substrates, both pure and contaminated with germs. In addition, it contains a database of the others listed in this proposed invention. Such libraries hardware-adapted algorithm of the method of the parent computer for examination and calculation of preliminary static information that is also used in the diagnostic process. Additionally it stores the data Bank on the preferred antimicrobial agent for each type strains and preferred tailored to the individual specificity of the object. Library of microbes can be supplemented by information about new types of microbes and by measuring clinical microbes on some remote clinical centers. Hardware-adapted algorithm of the method of the parent computer consists of parts that are responsible for the transmission and reception of information from remote clients, from parts, service the library of microbes and data Bank on antimicrobial agents and part of the host computing diagnostic information, the choice of antimicrobial preparation the TA and expert assessment of effectiveness of treatment and rehabilitation, as well as carrying out the adjustment of the diagnostic criteria. The process head computer is adjusted by the operators at the right time.

Part of clinical interest as a basic clinical centers, where special monitoring clinical diagnosis of microbes and processes of rehabilitation and treatment of patients with diseases and microbial processes of nature. These basic centers are in the process of checking the resistance of clinical strains to antimicrobial drugs and their effectiveness last, check the actions of these drugs on microbes inside the body of the patient and identify the germ that caused the disease. This system will provide timely detection of pathogens and assessment of the distribution of the germ. At a basic center for the study of industrial, food and environmental objects by a primary special monitoring samples of these objects with the purpose of expanding the database for these objects.

Based on the above system can be implemented individually normalized diagnostic clinic. The idea of such a center is in use in the diagnosis, individual quantitative and qualitative criteria for each person. The content of certain substances, the population of macroorganizational, the status and structure of tissues in health and disease are different, which gives an incentive to create a new support paradigm "normal-normal" and methods of implementation of its recommendations. For this measure the spectral-signal characteristics of the conversion of optical radiation in the living substance of each individual organism in norm, i.e. in the absence of disease at different stages of its development. The measured data are entered into the database of the parent computer, and then used for analysis, according to the above principle, and compare its results with the analysis made in the current time. In case of deviations of the current diagnostic parameters from the individual and collective (on the principle of nationality, geography, and of mankind in General) normal ranges make the diagnosis on the corresponding computed parameter pathology. Also on the basis of a statistical analysis is performed, the prediction of the probability of occurrence of certain diseases in the future for each organism.

A positive effect.

This technical solution has the following advantages (opportunities and prospects):

- screening for rapid assessment of infectious diseases (tuberculosis, AIDS, syphilis, cholera, and others),

- prophylactic medical examination in order to identify diseases and microbial processes of nature and Regis the radio background (control) characteristics of different biological fluids (urine, saliva, blood and other) to create passport health,

the population gets a modern environmentally friendly technology without the use of consumables and economic benefit (cost per analysis 10-100 times cheaper than the standard applied analogues),

Express-diagnostics and evaluation of the effectiveness of treatment of a dysbacteriosis (suffers 90% of the population)

- learning system with obtaining new informative opportunities, taking into account changes of species of microorganisms in medical institutions located in different climatic, ecological and geographical conditions, economic viability and a significant reduction of the costs of diagnosis due to tsentralizovannomy system,

- the feasibility of a system of individual monitoring, diagnosis and prediction of each of the human organism from the moment of birth to the current based on the technical solutions and support paradigm "norm-norm"

- in the industrial, food and environmental industries implemented the principle of cumulatively diagnostic data.

In the system structure includes universal optical fluorescence device for determination of species Express the indicating and differential diagnosis of microbes under different n is sologig. This device is a fluorescent microscope containing a dispersion element and the receiving CCD camera, as well as lasers for excitation of fluorescence in the samples, including excimer lasers. The object table of the microscope has a mount for standardized containers intended for diagnosable biological substrate. Additionally, the microscope has an eyepiece for visual monitoring of the process of removing data. Reception CCD camera microscope passes through the cable fluorescent image on port computer diagnostic system, where the hardware-adapted algorithm of the method is further maintenance of this information.

Examples of the application

Example 1.

An example illustrates one of the directions of a wide range of possible applications of the proposed method in medical, industrial and food biotechnology. To obtain reliable data about the studied microbacterium the facility must have a clear understanding of its characteristics absorption, scattering, and on the likely concentration in it of those or other fluorescent substances. Therefore, to improve the accuracy of microbes and their concentrations in mixtures by this method were measured transmission spectra of the samples with which the correction is Lee (restored) spectra of laser-induced fluorescence of the same samples. Based on the reconstructed spectra were calculated concentration in the samples treated with detergent.

Table of concentrations for the two samples of mixtures of mycobacteria, calculated taking into account dispersion and absorption.

12345
6321.8-129.62287.45602.29-130.64
7260.35301.79184.65460.69-17.75

(Legend: 1 - M. Avium (Mb.a), 2 - M. Vaccae (Mb.v), 3 - Tuberculosis(H37) (Mb.h), 4 - M. Intracellulare (Mb.i), 5 - M. Kansasii (Mb.k). Above the horizontal shows the types computed mycobacteria, on the right, the vertical axis shows the samples of mixtures of bacteria (6 (Mix 1): Mb.a. (1)+Mb.h. (3)+Mb.I. (4), 7 (Mix 2): Mb.a. (1)+Mb.b. (2)+Mb.I. (4))).

These microbiological descriptions give the following results on the composition of the mixtures (characters to the right of the colon shows the number of mixtures, to the left is present in the mixture mycobacteria, short symbols are separated from the zoom):

Mix 1: Mb.a. and Mb.h. and Mb.I.

Mix 2: Mb.a. and Mb.b. and Mb.I.

The table shows that the calculated concentration of mycobacteria in mixtures Mix1 and Mix2 have agreed with microbiological data. It is seen that in the first mixture is greater than zero the value of the concentration of mycobacteria have Mb.a, Mb.h and Mb.I, and the second mixture are observed mycobacteria Mb.a, Mb.b and Mb.I. that is in full accord with the above list. Such a good computation of quantitative characteristics occurs due to the optical properties of the substrate for bacteria.

Example 2.

There were also samples and mixed systems of microbes grown on non-selective medium. Wash with medium diluted in saline solution and placed in the tablet of the cell, having a volume of 0.01 ml. Next, the cuvette was placed in the mount fluorescent microscope, in which fluorescence is excited by a radiation of a KrF-excimer laser with a wavelength of 248 nm radiation. The emission spectrum of the laser cut standard test glass plate of the microscope, as their transmission starts at about 310 nm. Using the optical system and the chromatic dispersion system and CCD camera microscope were kept and digitized fluorescent image of the local area of the cuvette with the sample, which are shown in Fig.7 (Fluorescent image of microbes in the mixture, the implementation is given in three different spectral intervals: 1-400 nm, 2-500 nm and 3-600 nm. These images visible colony of Mycobacterium tuberculosis in the bottom left, since it is different from other microbes in intensity and color fluorescent images).

At the bottom of each picture shows the wavelengths that fall within the scope of the spectral ranges of the fixed CCD camera radiation. Next, the measurement results were processed using device-adaptive algorithm of the method for the differential diagnosis, resulting in the identified microbial species .Tuberculosis H37. In the pictures they form a colony in the lower left part of the screen. These experiments are preliminary.

Example 3.

The object of study called the surgery has a database of germs of surgical diseases in the field of pulmonology. While it includes the features of microbes that is registered with the device for measuring and conversion of laser radiation. These components included in the software product, and they are simultaneously and objects of comparison in the diagnosis and identification of possible etiological factors encountered in this disease. In particular, when the bedside A. with lung diseases with suspected pneumonia (bilateral lobar pneumonia) examine and register characteristics are appropriate to ejstvujuschij devices. These features are then processed by the diagnostic complex containing the appropriate device, and when receiving positive results reveal pathogenic microbes. In the subsequent action is determined by the hardware-adapted algorithm of this method in combination with the analysis of time series alive or not these microbes, then the information is passed to the doctor (graphs, tables), from which the doctor can definitely put the diagnosis. What this technology identifies the characteristics of the pathology and determines the biological state of germs that can achieve the goal of determining with a high degree of probability, whether it is an etiological factor.

1. The method of detection and identification of biological micro-objects and their nanocomponents, including radiation sensing monochromatic or nemonokhromaticheskogo radiation, including laser, one or more samples containing measurable objects, measurement and recording of the response parameters of samples for irradiation in the form of fluorescence characteristics or extinction, the data processing computer, wherein using the set of devices for measuring and recording one or more of the following responses samples in the form of characteristics of the radiation conversion, including by placing the sample in the cuvette and/or Sandero the radiation objects of animate or inanimate nature, including through the application of the light-harvesting device to the surface of the object, including measure and record simultaneously, sequentially these responses in the form of conversion of radiation, including from one or more sources of radiation, measure and record the responses, including reflection, scattering, fluorescence, extinction of the probing radiation, photoacoustic, optoterminal feedback, nonlinear, including Raman scattering, the spatial distribution of scattered and/or fluorescent radiation, commonly used to measure characteristics of the responses from each type of the phenomenon of conversion of radiation separately or together in any combination of these phenomena as at the place, and for deleted samples, while pre-measure and record the characteristics of the responses to the effects of radiation on the reference sample and/or objects of comparison with a known diagnosed own parameters and/or well-known custom response, and then measure the performance of response samples/objects diagnosed with unknown parameters and/or unknown physical parameters of response to radiation exposure, while these responses are passed, including using optical fibers, including a ditch, from the tablet on the cell with the corresponding matrix elements of the photodetectors, if necessary, placed in a low-temperature thermostat, and/or through a fluorescent microscope, and/or photoacoustic, optoterminal and/or cavitation sensors, including integrated on the walls of the ditch, including measuring the spectral characteristics of the response is performed using blocks analyzer spectra and after transfer in the computer measured and digitized signals remain in computer memory, result in linear diagnosed parameter form, thus producing a linear normalization of the measured characteristics for each of the diagnosed parameter according to the formula
Xpi=Xai/(1-αi·Xe),
where Xpithe multilevel characteristic conversion, Xai- ectocarpaceae absolute characteristic conversion, Xe- andcorporate relative characteristics of conversion, αi- the coefficient at the i-th feature in the expression for homeostatic parameter P=Σαk·Ikwhere k is the number of measured characteristics, then this data is normalized to the previously measured and/or known values of the spectral characteristics and/or other homeostatic parameter reference samples/objects, in doing so, a data correction and hardware features by the formula
Xi corr=Xi/(a1+a2·ei+a3·ei2),
where x icorr- is corrected spectral characteristics of the i-th channel of the device (on the i-th wavelength), Xi- the value of the corresponding spectral characteristics without adjustment, and1and2and a3- correction factors that depend on the geometry of the samples and the method of collecting light, eispatial damping rate due to extinction in the i-th wavelength corresponding to the spectral characteristics, then to create a statistical model of the diagnosed parameters complex spectral characteristics representing a set of corresponding eigenvectors reference and diagnostic parameters of micro-objects and their nanocomponents share on factor and random components sequentially performed in each cycle calculations decomposition of the data matrix on the structural part and the noise by the formula
X=FQ+E,
where X is the data matrix, F is the matrix of the factors of interest in the basis of the principal component, the Q - matrix of size (J×P), which is the transposed matrix of loadings, F is the matrix of residuals (noise), then according to the selected structural portion of each of the required characteristics of its models specify the appropriate search option diagnosed micro-objects and their nanocomponents, while the measurement results, calculation of modelirovaniya computer characteristics of the responses of the micro-objects and their nanocomponents with known and relevant diagnosed simulated parameters remain on the computer and create a database using them on the computer using the algorithm described above conduct is detected and compared with that obtained on the basis of the measurements, the experimental data of the desired parameters other identifiable micro-objects and their nanocomponents.

2. The method according to claim 1, characterized in that the measurement results of the conversion response remote diagnosed samples and/or objects and/or their respective designated parameter to diagnose the micro-objects and their nanocomponents passed with the help of remote sensing data for local, global, wired and/or wireless communication networks of friends and/or remote sampling and/or objects in the diagnostic unit of analysis information, which provides processing of the response characteristics of the samples and/or objects from several blocks measuring these characteristics, and are used in the diagnosis of processes involving identifiable objects and their nanocomponent.

3. The method according to claim 1, characterized in that diagnosed the parameters of the micro-objects and their nanocomponents in samples and/or sites in vivo determines the status of each individual organism in norm - as a reference object at different stages of its development and findings diagnosed parameters and/or characteristics of an organism are entered in the database DB of the specified computer.

4. Tablet cuvette for implementing the method according to claim 1, containing a tightly folded together two plane-parallel plates, the first plate from the side adjacent to the second plate of optically transparent material, one or more recesses, wherein the tablet contains on the front surface of the multiple recesses ditch (2-500) with a volume of about 0,1-0,001 ml, the distance between the transparent walls of the ditch lie in the range greater than the transverse dimensions of microbes by 5-10%, and the proportion of these cells may be the same size or volume, and some different, while the cell can be performed in flowing embodiment, the input and output ends are connected to the hoses for inlet and drain the fluid, and the cell can be equipped with sensors to detect conversion response of the sample to the sensing optical radiation.



 

Same patents:

FIELD: physics.

SUBSTANCE: X-ray optical endoscope has a housing in which there are optically interfaced X-ray and viewing optical channels for viewing object images, the X-ray channel has an X-ray sensitive CCD matrix of size B×B, the optical channel has a first semitransparent mirror mounted in front and on the longitudinal axis of the X-ray sensitive CCD matrix at an angle of 45°, a lens installed on the axis which passes through the centre of the first semitransparent mirror perpendicular to the axis of the X-ray sensitive CCD matrix, a monitor and/or a computer with an image display, wherein a second colour CCD matrix of size A×A is mounted in the focal plane of the lens with focal distance F outside the zone of propagation of the X-ray beam, near the lens there is a light-emitting diode with angle of radiation W>=arctg (A/2F) for illuminating the object. A matrix of N>8 microlasers is placed symmetrically around the X-ray sensitive CCD matrix.

EFFECT: possibility of matching considerably different characteristics of X-ray and optical channels.

2 dwg

FIELD: physics.

SUBSTANCE: X-ray optical endoscope has a housing inside of which there are optically interfaced X-ray and viewing optical channels for viewing the image of an object, the X-ray channel has a first focon with an X-ray fluorescent transducer at the input end, a second focon identical to the first whose output end is mated with the input fibre-optic washer of an image-intensifier tube, a first regular fibre-optic bundle which interfaces butt-ends of focons by smaller diametres, a first collimating lens whose focal plane coincides with the plane of the output fibre-optic washer of the image intensifier, and the optical axis is superimposed with its axis of symmetry and a first semitransparent mirror mounted on that axis at an angle of 45°, wherein the endoscope also includes an annular matrix of N>=8 microlasers in front of the X-ray fluorescent transducer concentrically with the transducer outside the zone of propagation of the information-bearing part of the stream of X-rays, optical axes of the microlasers are parallel each other and axis of the first focon and they lie symmetrically on a circle of diametre Dl, through which an annular structure of laser spots forms on the object, where the diametre Dl remains constant with variation of distance from the object to the endoscope and is equal to the diametre of the zone of the object illuminated by X-rays.

EFFECT: possibility of matching considerably different characteristics of X-ray and optical channels.

1 dwg

FIELD: physics.

SUBSTANCE: X-ray optical endoscope has a housing inside of which there are optically interfaced X-ray and viewing optical channels for projecting images of an object onto a CCD matrix of a television system which forms an image on a monitor, wherein the X-ray channel has a focon with an X-ray phosphor at its input end, a first fibre-optic regular bundle, a first collimating lens with focal distance F1, whose focal plane coincides with the output end of this bundle and a first mirror installed on the optical axis of the first collimating lens at an angle of 45°, wherein the endoscope also includes an annular matrix of N>8 microlasers placed symmetrically about the axis of the focon on a circle having diametre D equal to the diametre of the input end of the focon. Optical axes of the microlasers are parallel each other and the axis of the focon, owing to which they form an annular structure of laser spots of diametre D on the object, where the value D remains constant with variation of distance from the object to the endoscope.

EFFECT: possibility of matching considerably different characteristics of X-ray and optical channels, high image quality, reduced weight and size of the device.

3 dwg

FIELD: physics.

SUBSTANCE: X-ray optical endoscope has a housing in which there is a focon with an X-ray fluorescent transducer at the input end, a CCD matrix in whose A×A bit map the image of the output end of the focon is completely inscribed and a television monitor for viewing X-ray images formed on said matrix, wherein the endoscope also has a colour television camera having a lens with focal distance F and a CCD matrix with an A×A bit map and an annular structure of N>8 microlasers whose axes are parallel to each other and axis of the focon, lying symmetrically about the input end of the focon on a circle of diametre D. The colour television camera is mounted at the centre of the input end of the focon or lies outside its limits. The focal distance of the lens is such that the image of the annular structure of laser spots can be completely inscribed in the bit map of that camera. The object is illuminated with a light-emitting diode with controlled brightness and colour index of radiation and in a certain way, with given angle of radiation, dimensions of defects are measured using a standard metric scale with a certain division value on the screen of the monitor.

EFFECT: higher matching of characteristics of X-ray and optical channels, reduced wearing of the semitransparent mirror.

2 dwg

FIELD: medicine.

SUBSTANCE: invention contains body which consists of base and cover, made from fluoroplast, and device for imitation of optical properties of various types of live biological tissues. Device includes set of layer-by-layer placed elements which possess light-dispersing, fluorescent and light-absorbing optical properties. Layer-by-layer placed elements represent polymer optical films, whose spectral linear coefficients of absorption and dispersion correspond to spectral linear coefficients of light absorption by biological tissues with superficial and epidermal melanin, blood saturated with oxygen on 5-100%, spectral linear coefficient of light dispersion by collagen fibres and dense tissue structure without blood. Wavelengths of fluorescence of polymer optic films correspond to wavelengths of fluorescence of respiratory cell enzymes of biological tissue.

EFFECT: application of invention extends functional possibilities of device and ensures calibration and check-up of vast range of devices and apparatuses of non-invasive medical spectrophotometry.

3 cl, 1 dwg, 1 tbl

FIELD: measurement technique.

SUBSTANCE: using: for nondestructive control of products and materials. X-ray optical endoscope contains body with X-ray and optical channels located in it. Herewith X-ray optical endoscope has the second colour CCD-matrix of B*B size additionally introduced in it which matrix is installed on the axis of optical channel lens in the plane of its image. Focal length of this lens f0 is selected with regard to relation f0=L*B/D, where L is minimum distance from inlet butt end of focon to the lens, D is diametre of this butt end, and radiation angle of optical channel illuminator is selected from condition W=arctg(B/2f0). Herewith video data from both CCD-matrices enters computer with colour display with possibility of simultaneous or sequential viewing X-ray or optical images of object in various modes of their digital processing and superimposing on display screen.

EFFECT: providing possibility to reconcile significantly different characteristics of X-ray and optical channels using single CCD-matrix.

3 dwg

FIELD: physics.

SUBSTANCE: radiooptical endoscope comprises body with X-ray and optical channels arranged in it, besides radiooptical endoscope additionally contains the second colour CCD-matrix with B*B size installed on axis of optical channel lens in plane of its image, in front of optical channel lens symmetrically to it there are the following components installed coaxially - circular matrix with diametre D from N microlasers, optical axes of which are parallel to each other and axis of optical channel lens, which os used to generate image of laser spots circular structure on object surface, and circular structure diametre equals inlet parametre of focon and remains constant as distance from object to focon changes, and circular matrix from M light diodes with diametre Dc>D, angle of radiation of which is selected based on the conditon W=arctg(B/2f), and length of their radiation wave Y2 is selected with account of provision of maximum contrast of spots images from microlasers with radiation wave length Y1, besides video information from both CCD-matrices arrives to inlet of computer with colour display, with the possibility of simultaneous or serial view of X-ray and optical images of object in various modes of their digital processing and matching on display screen.

EFFECT: invention provides for the possibility to match substantially different characteristics of X-ray and optical channels with the help of single CCD-matrix.

3 dwg

FIELD: physics.

SUBSTANCE: radiooptical endoscope comprises body with X-ray and optical channels arranged in it, at the same time additionally the second colour CCD matrix is added to radiooptical endoscope, being installed on axis of optical channel lens in the plane of its image, besides video information from both CCD-matrices arrives to inlet of computer with colour display, with the possibility of simultaneous or serial view of X-ray and optical images of object, between the object and mirror from plexiglass there is the second rectangular mirror installed as arranged at the angle β to axis parallel to longitudinal axis of focon, and arranged at the distance H from it, centre of the second rectangular mirror is located at the distance Δ from plane of object, in front of the second rectangular mirror on axis passing through its centre and parallel to longitudinal axis of focon there is a collimator arranged, in point of back focus of collimator there is a light diode installed, at the outlet of collimator with the help of rectangular diaphragm a beam of light is generated, having rectangular shape with size dxD, where d is lowest size of beam, D is inlet diametre of focon, which after reflection from the second rectangular mirror sends a beam of light to object at the angle to its surface, which makes it possible to control defects in the mode of dark field.

EFFECT: invention provides for the possibility to match substantially different characteristics of X-ray and optical channels with the help of single CCD-matrix, provides for the possibility to detect defects having apparent relief.

2 dwg

FIELD: physics; optics.

SUBSTANCE: invention relates to detecting flaws in objects in difficult to access cavities. A device for complex X-ray and visual inspection of objects inside difficult to access cavities has a housing inside of which there are optically interfaced X-ray-optical and visual-optical channels for projecting the image of the object onto a CCD matrix of a television system, which forms an image on a monitor. Inside the housing there is also a fourth lens, the optical axis of which is parallel to the optical axis of the focusing cone and lies at a distance R from it. The focusing cone and the fourth lens are fitted with possibility of mutual displacement without violating parallel alignment of their optical axes within the limits of value R and successive fixation in positions for which the centre of the input face of the focusing cone and the centre of image of the fourth lens are superposed with the centre of the regular optical fibre bundle of the X-ray-optical channel, and in front of a broadband light source of the visual-optical channel, which emits in the visible and ultraviolet spectra, a filter is placed with possibility of input-output from the radiation flux, where the filter selects from the radiation spectrum of this source waves in the UV range, which excite photoluminescence in the penetrant used in luminescent defectoscopy.

EFFECT: possibility of inspecting objects in dark conditions.

5 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to methods of laboratory diagnostics. According to the method of blood test and automated technical blood tester. Blood sample is mixed with diluent and/or solvent reagent. Blood sample flow is formed and exposed to coherent polarised radiation of wavelength 330-680 nm directed along the flow axis. Named blood sample is analysed for polyangular light dispersion with blood cells detected and counted by polyangular light dispersion. Blood analyser applied for method implementation contains flow-type chamber, blood sample flow former in specified flow-type chamber, coherent radiation source, photodetector, and data control and processing unit connected to specified photodetector. Flou-type chamber represents scanning dish with the case made of optically transparent material and internal straight canal, and spherical mirror axis of which matches with that of internal channel of scanning dish. Coherent radiation source is polarised coherent radiation source of wavelength 330-680 nm mounted so that emitting radiation is directed along the axis of specified internal channel of scanning dish. The internal surface of spherical mirror of scanning dish is optically connected with specified photodetector.

EFFECT: possibility receive complex data referring to qualitative and quantitative blood structure providing minimum modification of measured blood corpuscles (cells) and high accuracy, besides, complete express blood analysis within no more than two stages.

26 cl, 11 dwg

FIELD: medicine.

SUBSTANCE: cultivated microbiological objects count is ensured by the measurements of their morphological compositions by determining the size distribution of microorganism cells in a nutrient fluid by variation of scattered light intensity. The fluid flow is sounded with monochromatic coherent light; interaction signal of probe radiation and analysed microbiological objects are recorded by the measurement of amplitude and duration of scattered light pulses by analysed particles; and functions derived from the measurements are plotted in the form of two-dimensional distribution of specified amplitudes and durations expressing statistic parameters of light scattering intensity by particles. After said functions, the size distribution of analysed cultivated microbiological objects and decay products of the nutrient fluid is derived.

EFFECT: higher measurement accuracy due to eliminated error caused by foreign particles that are decay products of the nutrient fluid in cultivation of the analysed microbiological objects.

9 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: invention concerns medicine, particularly method of cardiomyocyte selection from cardiomyocyte-containing cell mix without genetic alteration incardiomyocytes. Method of cardiomyocyte content boost in cardiomyocyte-containing cell mix without genetic alteration. Method of cardiomyocyte obtainment without genetic alteration to cardiomyocytes. Method of cardiomyocyte content assessment in cardiomyocyte-containing cell mix.

EFFECT: possible efficient selection of cardiomyocytes from cardiomyocyte-containing cell mix without genetic alterations.

20 cl, 16 dwg, 14 ex

FIELD: chemistry, biochemistry.

SUBSTANCE: method can be used in microbiology, food industry for estimation of viability of unicellular organisms (yeasts, etc.), which demonstrate difference of dielectric properties. Per cent content in mixture of live and dead unicellular microorganisms is determined by deviation of measured value of dielectric permeability from dielectric permeability of mixtures, consisting only of live and only of dead unicellular microorganisms, or by experimental dependence of second derivative of dielectric permeability on humidity.

EFFECT: elaboration of distant methods of assessment in continuous flow, in elaboration of industrial methods of control over live microorganism production.

2 dwg

FIELD: medicine.

SUBSTANCE: invention concerns medical microbiology. The method of chronic urogenital gonococcal infection course forecast involves seeding of accompanying fungi of Candida genus in case of gonococcus detection, and persistence factors of accompanying microorganisms is evaluated. If titration of fungi of Candida genus gives not less than 102 colony-forming cells per millilitre and antilysozyme activity evolves simultaneously in the quantity not less than 1.3 mcg/ml per optical density unit, and anticomplementary activity not less than 1.5·106 antilytic complement units, then chronic character of urogenital infection is confirmed diagnosed.

EFFECT: increased accuracy of chronic urogenital gonococcal infection course forecast.

2 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: method involves carrying out bacteriological study of esophageal mucous membrane biopsy samples. No microorganism growth or predominant Streptococcus spp., Peptostreptococcus spp., Staphylococcus spp. in monoculture or culture association in the amount of equal to or greater than 103-104 CFU/g (colony formation units), and no Escherichia coli, Bacteroides spp., Enterococcus faecalis, Enterococcus faecium, Candida spp. in monoculture or culture association in the amount of equal to or greater than 102-107 CFU/g, and optionally increased total microorganisms quantity to 104-107 CFU/g being observed, alkaline ingredient availability in refluxate in gastroesophageal reflux cases is declared to take place.

EFFECT: enabled alkaline ingredient availability and microbiocenosis disorders intensity evaluation.

FIELD: medicine.

SUBSTANCE: method involves determining duodenal juice acidity, duodenum bulbary and postbulbary department insemination degree with H.pylori. Ulcer edge and pyloric canal area bioptates are subjected to immunological and histological examination. Duodenal juice acidity being equal to or higher than 6.5 mmole/h and duodenal mucous membrane insemination degree being equal to or higher than 100 bacteria in vision field, IgG antibodies to H.pylori diluted in 1:160 proportion and higher, gastric metaplasia being detected in pyloric canal bioptates of ulcer edges among hypertrophied smooth muscle cells of separate groups of atrophied and deformed smooth muscle cells divided by layers of loose connecting tissue having blood vessels, fibroblasts, lymphocytes and macrophages, and anisochromia being detected when staining hypertrophied smooth muscle cells, pylorostenosis development is to be predicted.

EFFECT: high accuracy in predicting pylorostenosis development clinical course.

FIELD: medicine.

SUBSTANCE: method involves separating pure microorganism cultures from nasal mucous membrane and/or rhinopharynx microflora and identifying them. Anti-lysozyme activity is determined in pure culture and microbial insemination share in the general microbial insemination index is calculated for biotope under study. The first value being equal to or greater than 3 mcg/ml and the second one greater than 45%, rhinotubal microorganism migration into middle ear tympanic cavity is to be predicted.

EFFECT: high accuracy in predicting clinical course of inflammation in middle ear.

2 tbl

FIELD: biotechnology, microbiology.

SUBSTANCE: method involves sowing out samples of mixed cultures in liquid selective media, determination of cells number accumulating in media during microorganisms growth by bioluminescent method and mathematical treatment of kinetic data of the growth of individual bacterial cultures, determination of the parent concentration of microorganisms relating to different taxonomic groups. Invention allows carrying out the simultaneous identification of bacterial cells relating to different taxonomic groups and presenting in mixed cultures simultaneously, and to enhance precision in determination of cells number in the broad concentration range and to reduce the total analysis time significantly. In industry mixed cultures are used widely in different branches of food industry (dairy, meat, brewing and others) and in other biotechnological processes (biological treatment of sewage waters, bioremediation of soils, producing methane from waste in different manufactures and others). Invention can be used for differentiated determination of microorganisms number in mixed cultures that are widely distributed in nature: in air, soil, ponds, as components of natural microflora in higher organisms and among contaminants causing injury of different objects.

EFFECT: improved method for determination.

1 tbl, 3 dwg, 5 ex

FIELD: microbiology, optics.

SUBSTANCE: invention relates to investigations of materials by assay of their physical or chemical properties using optical devices and to systems wherein material is excited by optical agents resulting to it luminescence. Invention proposes a test carrier as a centrifugal tube. Carrier is separated for upper and bottom cavities by partition. Volume of lower cavity is 0.1 of tube volume. A hole is made in partition near a wall. The constructive decision of partition provides efflux of sample from lower cavity with minimal overcoming the combined forces of wetting and surface tension. Also, invention proposes methods/variants for rapid measurement of absolute concentration of microorganisms in biosubstrate by their photoluminescence. Methods involve using fluorescent or phosphorescent measuring device and above said test carrier. Methods provides increasing rate and precision of assay, to use serial measuring devices and to carry out measurement of the concentration of particles in substrate with another specific gravity value as compared with that of liquid in substrate. Invention can be used in food and biotechnological industry for determination of absolute concentration of microorganisms in different substrates.

EFFECT: improved method for assay, valuable properties of carrier.

2 tbl, 1 dwg

The invention relates to food industry, in particular to the baking industry, and can be used to determine the number of fungi on the surface of bakery products

FIELD: microbiology, optics.

SUBSTANCE: invention relates to investigations of materials by assay of their physical or chemical properties using optical devices and to systems wherein material is excited by optical agents resulting to it luminescence. Invention proposes a test carrier as a centrifugal tube. Carrier is separated for upper and bottom cavities by partition. Volume of lower cavity is 0.1 of tube volume. A hole is made in partition near a wall. The constructive decision of partition provides efflux of sample from lower cavity with minimal overcoming the combined forces of wetting and surface tension. Also, invention proposes methods/variants for rapid measurement of absolute concentration of microorganisms in biosubstrate by their photoluminescence. Methods involve using fluorescent or phosphorescent measuring device and above said test carrier. Methods provides increasing rate and precision of assay, to use serial measuring devices and to carry out measurement of the concentration of particles in substrate with another specific gravity value as compared with that of liquid in substrate. Invention can be used in food and biotechnological industry for determination of absolute concentration of microorganisms in different substrates.

EFFECT: improved method for assay, valuable properties of carrier.

2 tbl, 1 dwg

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