KR20210041157A - Real-Time Biopsy Analysis System and Method Thereof - Google Patents

Abstract

Disclosed are a real-time biological tissue analysis system capable of determining types of biological tissues in real time by analyzing a fluorescence ratio of a fluorescence signal excited from a biological tissue, and a method thereof. It is possible to determine the types of biological tissues in real time by spectroscopy of a fluorescence signal of the biological tissue generated in the form of complex light to measure the same by wavelength, and by analyzing the fluorescence signal ratio using the overlapping wavelengths of the measured signals. Accordingly, it is possible to solve problems appertaining to the conventional biopsy, which include time delay, burden on the patient's body, and additional cost.

Description

Real-Time Biopsy Analysis System and Method Thereof}

The present invention relates to a real-time biological tissue analysis system and method, and more particularly, to a real-time biological tissue analysis system and method capable of determining the type of biological tissue in real time by analyzing the fluorescence ratio of the fluorescence signal excited in the biological tissue. About.

Cancer, the number one cause of death in modern people, is very important to diagnose the disease early because the cure rate and survival rate drop sharply as the disease persists.

In order to reliably diagnose the onset of cancer, a biopsy must be performed. The most frequently performed biopsy method to determine the presence of abnormalities in the current tissue cells is a physical biopsy (biopsy), which is endoscopy through the oral cavity or anus. The camera and forceps are inserted into the body to physically collect the suspected part of the abnormal tissue, and finally determine whether the tissue cells are abnormal through processes such as incision, staining, and culture.

This biopsy method is highly dependent on the experience and skill of the specialist performing the test, and if the abnormal tissue is of a small size that cannot be identified with the naked eye, or if the abnormal tissue is not protruding to the surface and is buried and cannot be detected, the biopsy itself is performed. There is a problem that it cannot be achieved.

In addition, even if a biopsy is performed, a time delay occurs due to a complicated examination process in order to confirm the final tissue cell abnormality. In addition, although it is a fatal disadvantage that is a test method to check the health status of a patient, it has several problems, such as a burden on the patient's health through physical tissue collection and a secondary infection problem.

The most representative non-invasive test method suggested as an alternative to such a physical biopsy is an optical biopsy.

When implementing an optical biopsy, unlike a conventional physical biopsy, it is possible to check a small size that is difficult to check with the naked eye or an abnormal tissue that is buried, and it is possible to solve the temporal delay problem by simplifying a complex inspection process. In addition, it is free from the physical burden of the patient due to physical tissue collection, which is the most fatal problem, or from secondary infection, and additionally simplifies the examination process and reduces the need for specialists, thereby reducing costs. There are many advantages that can solve most of the problems arising from biopsy).

Because of these advantages, attempts and prior art developments have been made to analyze biological tissues using various optical phenomena (reflection, diffraction, fluorescence, etc.) that occur when light is irradiated on biological tissues. Due to its unique molecular structure, it is attracting attention as a material analysis technique not only in the medical field but also in various fields.

In addition, unlike phosphorescence, since fluorescence has ^{a characteristic that a natural wavelength is generated within a short time (10 -9} ~ 10 ^-7 seconds) after light absorption according to its physical principle, medical examinations such as biopsy that require rapid and immediate identification are required. As a technology with many advantages in the field, it is more demanded.

However, since fluorescence analysis can most clearly derive the material analysis result according to the principle that the difference in fluorescence signal occurs due to the unique molecular structure of the material, it is still being rapidly applied to research on material analysis. It has not been commercialized in such medical fields.

This is because there is a lack of research on fluorescence signals generated in living tissues, and optical biopsy technology using fluorescence is merely irradiating a light source on living tissues and collecting fluorescence signals generated therefrom in real time. Since the fluorescence signal generated in the living tissue appears in the form of complex light rather than single light, real-time biopsy cannot be implemented simply by collecting the fluorescence signal in real time.

Korean Patent Registration 10-1172745

The technical problem to be achieved by the present invention is to measure the fluorescence signal by wavelength by spectroscopic measurement of a unique fluorescence signal according to the type of biological tissue, and by using the fluorescence signal ratio information for the overlapping wavelength point of the fluorescence signals measured by wavelength, the biological tissue can be measured in real time. It is to provide a real-time biological tissue analysis system and method capable of discriminating the type.

In the real-time biological tissue analysis system of the present invention for solving the above problem, the unique fluorescence signal according to the type of biological tissue is measured for each wavelength, and each peak of the fluorescence signals measured for each wavelength is overlapped. A fluorescence information collection unit that stores ratio information data of a fluorescence signal using a wavelength point, and a biological tissue analysis unit that determines the type of biological tissue in real time by filtering the wavelength of the fluorescence signal based on the information stored in the fluorescence information collection unit. Includes.

The overlapping wavelength point may include a reference overlapping point corresponding to the first overlapping peak among the overlapping peaks, and partial overlapping points corresponding to peaks overlapping after the reference overlapping point.

The fluorescence signal information includes an entire fluorescence signal collected by blocking light based on the reference overlapping point and a partial fluorescence signal collected by blocking light based on the partial overlapping point, and the It may be information stored by extracting the difference in the ratio of the partial fluorescence signal.

The fluorescence information collection unit includes: a first light source unit for irradiating light to the living tissue, a reaction unit for blocking external interference light to detect the fluorescence signal, a detection unit for detecting a fluorescence signal excited from the living tissue, and the It may include a control unit for storing unique fluorescence information according to the biological tissue.

The sensing unit may include a spectroscopic unit that adjusts a wavelength of the fluorescent signal incident on the sensing unit, and a sensor unit that senses a fluorescent signal whose wavelength is adjusted by the spectroscopic unit.

The spectroscope may include a monochromator or an optical system.

The first light source unit and the sensing unit may be connected to the reaction unit using an optical fiber unit.

The optical fiber unit may include a first optical fiber connecting the first light source unit and the reaction unit, and a second optical fiber connecting the sensing unit and the reaction unit, and arranged to surround the first optical fiber.

The biological tissue analysis unit may include a body, a second light source unit disposed at the center of the body and transmitting light for generating the fluorescent signal to the biological tissue, disposed around the second light source unit, and detecting the fluorescent signal. , A detection unit for detecting a blocked fluorescence signal based on the overlapping wavelength point, and a circuit unit located below the second light source unit and the detection unit, and outputting a difference in conductivity of the fluorescent signal detected by the detection unit as a ratio Can include.

The sensing unit includes a filter disposed above the body and filtering a wavelength corresponding to the overlapping wavelength point, and a fluorescence sensor disposed below the filter and sensing the fluorescence signal incident through the filter. can do.

The filter may be formed to have different amounts of light of the fluorescent signal incident on the fluorescent sensor.

The fluorescence information collection unit or the biological tissue analysis unit may be arranged to be in contact with the biological tissue when measuring the biological tissue.

The number of sensing units may have the same number as the number of overlapping wavelength points.

Each of the sensing units may be arranged such that the same distance is spaced apart from the second light source unit.

The fluorescent sensor may include a substrate, an electrode formed on the substrate and formed in an IDT structure, and a zinc oxide nanorods layer formed on the electrode and formed in a zigzag shape.

The circuit unit has a number equal to the number of detection units, and includes a fluorescent sensor resistor connected in parallel with each other and a load resistor connected in series with the fluorescent sensor resistor so as to correspond to the fluorescent sensor resistance, and the fluorescent sensor resistor and The load resistor may have a shape of a Wheatstone Bridge in order to output a difference in conductivity of the fluorescent signal detected by the sensing unit, respectively.

The analysis method of the real-time biological tissue analysis system of the present invention for solving the above problem includes the steps of measuring a unique fluorescence signal according to the type of biological tissue by wavelength after spectroscopy using a fluorescence information collection unit, and the fluorescence signal measured for each wavelength. Setting an overlapping wavelength point at which each peak of the field overlaps, storing the fluorescence signal ratio information for the overlapping wavelength point, and the fluorescence signal through a biological tissue analysis unit based on the stored fluorescence signal ratio information. And determining the type of biological tissue in real time by filtering the wavelength.

The setting of the overlapping wavelength point may include checking an overlapping peak for the fluorescence signals measured for each wavelength, setting a reference overlapping point corresponding to the first overlapping peak among the overlapping peaks, and the The step of setting partial overlapping points corresponding to the overlapping peaks after the reference overlapping point may be further included.

The storing of the fluorescence signal information includes: collecting a full fluorescence signal by blocking light based on the reference overlapping point, collecting a partial fluorescence signal by blocking light based on the partial overlapping points, and It may further include the step of extracting and storing the difference in the ratio of the partial fluorescence signal to the fluorescence signal.

The determining of the type of biological tissue in real time includes: detecting a fluorescent signal excited from the biological tissue, filtering the detected fluorescent signal based on the overlapping wavelength point and sensing the sensed fluorescent signal. It may further include the step of determining the type of biological tissue by comparing the stored fluorescence signal information.

According to the present invention, it is possible to determine the type of biological tissue in real time by spectroscopically measuring a fluorescence signal of a biological tissue generated in the form of a complex light and measuring the fluorescence signal ratio by using the overlapping wavelength of the measured signals. . Accordingly, it is possible to solve problems such as a time delay, a physical burden of a patient, and an additional cost according to a conventional biopsy.

In addition, it is not limited to the biopsy module in the form of an endoscope, and the identification of tissue cells in real time, such as the identification of tissue cells that are difficult to identify with the naked eye in an incision surgery for confirming and removing tumors, etc. It can be used wherever it is necessary to check whether or not there is an abnormality.

Furthermore, if it is based on general material analysis methods such as incision, dyeing, cultivation, and observation after the conventional physical collection, and the fluorescence wavelength signal generated from each material is converted into data and used for real-time analysis, not only in the medical field, but also in agriculture and space. It has a wide range of applications such as aviation and environmental analysis.

The technical effects of the present invention are not limited to those mentioned above, and other technical effects that are not mentioned will be clearly understood by those skilled in the art from the following description.

1 is an image showing cell specimens of various biological tissue cells.
FIG. 2 is a graph showing intrinsic fluorescence wavelengths for each of the biological tissue cells shown in FIG. 1.
3 is a block diagram showing the real-time biological tissue analysis system of the present invention.
4 is a block diagram showing a fluorescence information collection unit of the present invention.
5 is a diagram showing a connection configuration of a fluorescence information collection unit of the present invention.
6 is a view showing an embodiment of measuring a biological tissue using the fluorescence information collection unit of the present invention.
7 is a graph showing an embodiment of analyzing the fluorescence wavelength of a biological tissue using the fluorescence information collection unit of the present invention.
8 is a view showing a biological tissue analysis unit of the present invention.
9 is a diagram showing an arrangement structure of the sensing unit of the present invention.
10 is a diagram showing a fluorescence sensor of the present invention.
11 is a circuit diagram showing a circuit part of the present invention.
12 is a flow chart showing an analysis method using the real-time biological tissue analysis system of the present invention.

Since the present invention can apply various transformations and have various embodiments, specific embodiments will be illustrated in the drawings and will be described in detail in the detailed description. However, this is not intended to limit the present invention to a specific embodiment, it should be understood to include all conversions, equivalents, and substitutes included in the spirit and scope of the present invention. In describing the present invention, when it is determined that a detailed description of a related known technology may obscure the subject matter of the present invention, a detailed description thereof will be omitted.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, and in the description with reference to the accompanying drawings, the same or corresponding components are assigned the same reference numbers, and redundant descriptions thereof will be omitted. It should be.

1 is an image showing cell specimens of various biological tissue cells.

FIG. 2 is a graph showing intrinsic fluorescence wavelengths for each of the biological tissue cells shown in FIG. 1.

Here, the cell specimen of Fig. 1(a) represents the epidermal side cell specimen, the cell specimen of Fig. 1(b) represents the inside of the epidermis, that is, the Pia fat cell specimen, and the cell specimen of Fig. 1(c) is used for cancer cells. Shows the cell specimen for In addition, FIGS. 2(a), (b), and (c) are graphs measuring intrinsic fluorescence wavelengths excited in each cell specimen shown in FIG. 1.

1 and 2, when comparing the fluorescence wavelengths of the normal cells shown in FIGS. 2(a) and 2(b) with the fluorescence wavelengths of the cancer cells shown in 2(c), the fluorescence wavelengths of the normal cells and the cancer cells It can be seen that the silver fluorescence peak or the shape difference does not occur significantly.

This is because the fluorescence signal generated in the biological tissue of the polymer unit is in the form of a complex light rather than a single light, so even if the tissue is mutated, such as a cancer cell, the peak or shape difference of the complex light wavelength does not occur. In other words, among the constituents of mammals including humans, the material that accounts for the largest proportion except for water is generally a protein, and a protein is a linkage in which about 20 amino acids form a sequence through numerous peptide bonds. However, even if cells are mutated like cancer, the kind, sequence, or ratio of amino acids does not change completely.

From the result of this fluorescence wavelength, it can be confirmed that it is essential to perform spectroscopy in order to discriminate it because the fluorescence signal of the living body has a complex light form. In addition, it can be seen that the spectral resolution must also be high because there is no significant difference such that the entire wavelength of the fluorescent signal changes.

Since fluorescence is generated naturally according to the scanning of the light source, the detection itself can be performed in real time. However, measuring and recording a fluorescent signal in the form of a complex light by wavelength requires a wavelength transition time from at least tens of seconds to up to tens of minutes depending on measurement conditions such as measurement range and resolution, and the measured fluorescent signal Actual real-time biopsy is impossible because additional time is generated depending on the algorithm used for the analysis. In other words, it takes several tens of minutes to obtain fluorescence information of a local area, and it may take several hours or more to perform a biopsy by analyzing the entire fluorescence signal of a living tissue that is required to be measured.

Accordingly, in order to implement a biopsy on a living tissue in real time, a method capable of spectral processing a fluorescent signal in the form of a complex light to be collected and physical equipment for implementing the same are required.

3 is a block diagram showing the real-time biological tissue analysis system of the present invention.

Referring to FIG. 3, the real-time biological tissue analysis system according to the present invention determines the type of biological tissue in real time based on the fluorescence information collection unit 100 and the fluorescence information collection unit 100 for storing fluorescence signal information with data. It includes a biological tissue analysis unit 200 to determine.

The fluorescence information collection unit 100 spectroscopes a unique fluorescence signal according to the type of biological tissue and measures it by wavelength, and collects fluorescence signal information on an overlapping wavelength point at which each peak of the fluorescence signals measured by wavelength overlaps. Save it.

In addition, the fluorescence information collection unit 100 may include a first light source unit 110, a reaction unit 120, a detection unit 130, and a control unit 140.

4 is a block diagram showing a fluorescence information collection unit of the present invention.

Referring to FIG. 4, the first light source unit 110 of the fluorescence information collecting unit 100 according to the present invention emits excitation light that excites fluorescence from a light source to excite a fluorescence signal from a living tissue.

Here, when the light source to be irradiated is fixed to a specific wavelength, the system can be miniaturized by using an LED or laser having a unique wavelength, and it can be utilized for various advantages such as reducing cost and increasing convenience. In addition, if the wavelength of the irradiation light source cannot be specified because the fluorescence wavelength range of the biological tissue to be measured cannot be identified, a white light source such as an Xe lamp is used to prevent detection because the wavelength of the irradiation light source overlaps with the fluorescence signal wavelength. By providing a monochromator, the wavelength of the irradiation light source can be flexibly adjusted.

The reaction unit 120 serves to block external interference light in order to detect a fluorescent signal excited from a living tissue.

That is, the reaction unit 120 is manufactured in the form of a dark box, and may be manufactured in various ways according to the type and size of a biological tissue or specimen to be measured. In addition, the reaction unit 120 may be formed in an environment that thoroughly blocks external interference light that interferes with detecting a fluorescent signal having a weak luminous intensity of less than 1% compared to the luminous intensity of the irradiated light source.

The detection unit 130 serves to detect a fluorescence signal excited from a living tissue. In addition, the sensing unit 130 may include a spectroscopic unit 131 and a sensor unit 132.

The spectroscopic unit 131 serves to adjust a wavelength of an optical signal incident on the sensing unit 130. To this end, the spectroscope 131 may be a spectroscope (Monochromator) or an optical system (Optical System).

However, since the spectrometer takes a lot of time from a few seconds to several tens of minutes depending on the measurement conditions such as the range of the wavelength, the interval, and the delay time when measuring the fluorescence signal of a living tissue. In an experimental environment where the state of living tissue can change rapidly, continuous measurement is limited. In addition, when a spectroscope is used for fluorescence analysis, the amount of measurable fluorescence signal is reduced compared to the total amount of fluorescence signal generated by attenuation of the signal generated while passing through the spectrometer, which is disadvantageous in measuring weak fluorescence signals.

In order to compensate for these shortcomings, it is preferable to apply an optical system to the spectroscope 131 of the present invention rather than a spectrometer.

The optical system may include an optical filter, a condensing lens, and the like. Currently, low-cost optical filters (Long Pass Filter (LPF), Short Pass Filter (SPF), etc.) of tens of thousands of won have high performance such as OD (Optical Density) 5 or higher and Transmittance 90% or higher. (131) can be configured at low cost, and unlike the use of a spectrometer, signal attenuation is very small (transmittance of 90% or more), and fluorescence signals can be concentrated by configuring an optical system through a condensing lens. It is very advantageous for detecting the signal. However, if the measurement range is set excessively wide, the error in signal intensity generated according to the SR (Spectral Response) of the sensor (PMT, etc.) to be used may increase. Therefore, the SR must be taken into account when analyzing the fluorescence signal.

The sensor unit 132 serves to detect a substantial fluorescence signal. In addition, the sensor unit 132 may be a high-performance PMT (Photo Multiplier Tube) capable of detecting optical signals in a wide wavelength range, or a high-sensitivity fluorescence sensor capable of detecting optical signals in a specific wavelength range according to a measurement object. .

As an example, the optical signal incident to the sensing unit 130 is a composite signal having various wavelengths such as light source, fluorescence, noise, etc., and since the fluorescence signal desired to be measured also has various wavelength distributions, the intensity information for each wavelength of the input signal is stored. In order to obtain it, it is essential to spectral the optical signal incident on the sensing unit 130 and transmit it to the sensor unit 132 for each wavelength. Accordingly, the sensor unit 132 may be configured as an optical sensor having a wide measurement range and excellent sensitivity, such as a PMT, so as to detect the intensity of each wavelength in a wide range.

However, PMT has the advantage of having a high sensitivity and a wide measurement area, but has a disadvantage in that it requires a vacuum tube and a power controller capable of applying a high voltage for electron acceleration, resulting in an increase in volume. Therefore, it is desirable to use a highly sensitive fluorescent sensor that is capable of high-efficiency and real-time measurement, and is miniaturized.

The controller 140 serves to store unique fluorescence information according to a biological tissue.

In addition, the control unit 140 may be composed of a data acquisition unit (DAQ) for collecting data, a source measure unit (SMU), and a PC for controlling equipment.

5 is a diagram showing a connection configuration of a fluorescence information collection unit of the present invention.

6 is a view showing an embodiment of measuring a biological tissue using the fluorescence information collection unit of the present invention.

5 and 6, the first light source unit 110 and the detection unit 130 may be connected to the reaction unit 120 to each other using an optical fiber unit. In addition, the optical fiber unit 150 includes a first optical fiber 151 connecting the first light source unit 110 and the reaction unit 120 to each other, and a second optical fiber connecting the sensing unit 130 and the reaction unit 120 to each other. 152). That is, the light irradiated from the light source of the first light source unit 110 is transmitted to the biological tissue disposed in the reaction unit 120 through the first optical fiber 151, and the fluorescence signal excited by the biological tissue by the light is transmitted to the second It may be transmitted from the reaction unit 120 to the sensing unit 130 through the optical fiber 152.

Here, the first optical fiber 151 may be disposed at the center of the optical fiber unit 150, and a plurality of second optical fibers 152 may be disposed to surround the first optical fiber 151.

This is because the fluorescent signal generated when the light source is irradiated to the living tissue in the reaction unit 120 is emitted in all directions, so the arrangement of the optical fiber unit 150 can improve the light receiving efficiency of the fluorescent signal.

In addition, when the contact measurement with the living tissue is performed using the fluorescence information collecting unit 100 of the present invention as shown in FIG. 6(b), the collection rate of the reflected light is reduced compared to the non-contact measurement of FIG. 6(a). It is possible to generate more fluorescence by penetrating more light into living tissues while lowering. Accordingly, by attenuating unnecessary signals such as reflected light in the fluorescence signal measurement, the ratio of the fluorescence signal to the input signal and the signal to noise ratio (SNR) are improved.

In the process of measuring the fluorescence signal of a living body using the fluorescence information collecting unit 100, the light source irradiated from the light source of the first light source unit 110 generates monochromatic light having a shorter wavelength than the fluorescence wavelength region generated in the living tissue. The fluorescence wavelength is transmitted to the reaction unit 120, and in the reaction unit 120, the excited fluorescence wavelength due to the light irradiated to the living tissue is transmitted to the sensing unit 130, and in the sensing unit 130, through the spectroscopic unit 131. By shifting the wavelength and receiving the fluorescence signal, the controller 140 may finally store unique fluorescence information for each biological tissue.

Here, the unique fluorescence information for each biological tissue may be information on a ratio of a fluorescence signal for each biological tissue using an overlapping wavelength at which peaks of fluorescence signals measured for each wavelength overlap.

7 is a graph showing an embodiment of analyzing the fluorescence wavelength of a biological tissue using the fluorescence information collection unit of the present invention.

That is, FIG. 7 is a graph showing fluorescence wavelengths measured using the fluorescence information collection unit 100 for each of the cell specimens shown in FIG. 1 as a fluorescence intensity ratio.

As shown in FIG. 7, it can be seen that the wavelengths for the fluorescence signals of normal cells and cancer cells are not clearly distinguished. That is, all of the peaks of the fluorescence wavelength except for the reflected light source are formed in the mid-300nm, and the difference in wavelength is not large. This is because tissue cells of a high molecular unit are not composed of a single protein, and even if mutation occurs in tissue cells, the composition of all proteins in the tissue does not change, so the fluorescence signal generated from tissue cells can generally have a single wavelength. There is no change, and the fluorescence wavelength does not change significantly.

In addition, even in the same living tissue, the amount of fluorescence signal may vary depending on the shape, density, humidity, etc. of the tissue, and the difference may be greater than the difference in the absolute value of the fluorescence signal according to the type of tissue cell. Depending on the contact distance, the absolute amount of fluorescence generated and actually collected may show a large difference, so the method of determining the type of tissue cell using the absolute intensity difference of the conventional fluorescence signal has a disadvantage of inferior accuracy.

Therefore, the biological tissue analysis system of the present invention is a fluorescence signal using an overlapping wavelength point 10 at which each peak of the fluorescence signals measured for each wavelength is superimposed by spectroscopically measuring a unique fluorescence signal according to the type of biological tissue. By storing the information data on the ratio, the biological tissue cells are classified in real time.

Here, the overlapping wavelength point 10 is a reference overlapping point 11 corresponding to the first overlapping peak among the overlapping peaks and peaks overlapping after the reference overlapping point 11 in the measured fluorescence signal. It may include partial overlapping points 12. The overlapping wavelength point 10 then functions as a cut-off wavelength of the optical filter.

A method of measuring the fluorescence signal ratio using the overlapping wavelength point 10 will be described using the fluorescence signal shown in FIG. 7.

Since tissue cells of a high molecular unit are not composed of a single protein, the fluorescence signal also does not have a single peak. As an example, when light of 280 nm is irradiated from a light source to the living tissue shown in FIG. 1, the fluorescent signal excited by the living tissue generates peaks at around 310 nm and 340 nm, respectively, as shown in FIG. 7. In addition, the overlapping wavelength point 10 is set as a point where each peak of the fluorescence signal generated according to the type of biological tissue overlaps.

Here, in order to analyze the fluorescence signal ratio, at least two overlapping wavelength points 10 are required, and as the overlapping wavelength points 10 increase, the analysis accuracy of the fluorescence signal may be improved.

As an example, when two overlapping wavelength points 10 are designated in order to analyze the fluorescence signal ratio in the fluorescence wavelength shown in FIG. 7, the reference overlapping point 11 among the overlapping wavelengths corresponds to a 300 nm position, and a partial overlapping point (12) corresponds to the 320nm position. Accordingly, the entire fluorescence signal is collected by blocking the light at 300 nm corresponding to the reference overlapping point 11, and the partial fluorescence signal is collected by blocking the light at 320 nm corresponding to the partial overlapping point 12 as a comparison point. Here, the method of collecting the fluorescence signal may be a form in which the collected fluorescence signal wavelengths are integrated.

When the total fluorescence signal and the partial fluorescence signal are collected, the ratio of the partial fluorescence signal to the total fluorescence signal is extracted. In addition, in an exemplary embodiment, the fluorescence signal ratio is extracted by setting one reference overlapping point 11 and one partial overlapping point 12, but two or more partial overlapping points 12 may be set. That is, as the overlapping wavelength point 10 increases, the analysis accuracy of the fluorescence signal may be improved.

The collected information on the ratio of fluorescence signals for each biological tissue is converted into a database and used in the biological tissue analysis unit 200 to determine the type of biological tissue in real time.

Subsequently, referring to FIG. 3, the biological tissue analysis unit 200 determines the type of biological tissue in real time by filtering the wavelength of the fluorescence signal measured based on the ratio information of the fluorescence signal stored in the fluorescence information collection unit 100. do.

8 is a view showing a biological tissue analysis unit of the present invention.

Referring to FIG. 8, the biological tissue analysis unit 200 according to the present invention may include a body 210, a second light source unit 220, a sensing unit 230, and a circuit unit 240.

The second light source 220 performs a function of transmitting light for generating a fluorescent signal to a living body tissue.

The second light source unit 220 is preferably disposed at the center of the body 210 in order to increase the light-receiving efficiency of the fluorescent signal because the fluorescent signal generated when light is irradiated to the biological tissue is emitted in all directions. In addition, when performing contact measurement with a living body tissue, it is possible to increase the generation of fluorescence excited in the living body tissue because the collection rate of the reflected light source can be lowered and more light can penetrate into the measurement object. Therefore, in measuring a fluorescence signal, it is possible to attenuate an unnecessary signal such as reflected light and increase a necessary signal such as a fluorescence signal, thereby improving a signal to noise ratio (SNR).

Here, it is preferable to use light having a shorter wavelength than that of the measurement area of the living body tissue as the wavelength of light irradiated from the second light source unit 220. For example, the second light source unit 220 may directly insert a miniaturized monochromatic LED or laser into the body 210 together with an optical lens, or place a light source outside when a change in the wavelength of the light source is required to be transmitted to the optical fiber.

The detection unit 230 detects a fluorescence signal excited from a biological tissue, but detects a blocked fluorescence signal based on the overlapping wavelength point 10.

The detection unit 230 is disposed so as to surround the second light source unit 220 around the second light source unit 220, and may be disposed to be spaced apart from the second light source unit 220 by a predetermined distance. Here, it is preferable to arrange the number of detection units 230 to be the same as the number of overlapping wavelength points 10 designated by the fluorescence information collection unit 100. As an example, although four detection units 230 are shown in FIG. 8, when the number of overlapping wavelength points 10 is two, two detection units 230 may be disposed.

In this case, it is preferable that the detection units 230 are arranged so that the same distance as the second light source unit 220 is spaced apart from each other.

9 is a diagram showing an arrangement structure of the sensing unit of the present invention.

Referring to FIG. 9, as an example, when the number of overlapping wavelength points 10 is two, the detection unit 230 is disposed to surround the second light source unit 220, but the same distance as the second light source unit 220 is spaced apart. It may be arranged in a pair so as to be possible. That is, they may be arranged to be symmetrical to each other around the second light source unit 220. This is to minimize the difference in the fluorescence signal received from the biological tissue sample 201 to be measured by allowing each paired sensing unit 230 to be spaced apart from the light source by the same distance.

In addition, the detection unit 230 may include a filter 231 and a fluorescence sensor 232.

The filter 231 is disposed on the body 210 and filters a wavelength corresponding to the overlapping wavelength point 10. Accordingly, each filter 231 used in the sensing unit 230 may filter different bands according to the corresponding overlapping wavelength point 10. That is, the filter 231 may be formed so that the amount of light of the fluorescent signal incident on the fluorescent sensor 232 is different.

As the filter 231, an LPF (Long Pass Filter) optical filter may be used, and it is disposed on the body 210 to protect the fluorescence sensor 232 disposed under the filter 231, and performs a spectral function.

The fluorescence sensor 232 is disposed under the filter 231 and senses a fluorescence signal incident through the filter 231. That is, the amount of light of the fluorescent signal sensed by the fluorescent sensor 232 may be different according to the corresponding filter 231.

10 is a diagram showing a fluorescence sensor of the present invention.

Referring to FIG. 10, the fluorescent sensor 232 according to the present invention may include a substrate 233, an electrode 234 formed on the substrate 233, and a zinc oxide nanorods layer 235. .

The substrate 233 may be a semiconductor substrate, a conductive substrate, a non-conductive substrate, a silicon on insulator (SOI), or the like. A metal substrate, a conductive organic compound substrate, or the like may be used as the conductive substrate, and a glass substrate, a polymer compound substrate, or the like may be used as the non-conductive substrate.

The electrode 234 may be formed on the substrate 233. The electrode 234 may have various other shapes, such as a rectangle, an ellipse, a circle, a polygon, or a combination thereof, but the two electrodes 234 are formed in an IDT (Inter Digital Transducer) structure formed by staggering each other. desirable. The electrode 234 is made of two or more kinds of metals, such as gold (Au), nickel (Ni), platinum (Pt), palladium (Pd), iridium (Ir), silver (Ag), and rhodium (Rh). ), ruthenium (Ru), stainless steel, aluminum (Al), molybdenum (Mo), chromium (Cr), copper (Cu), tungsten (W), and a combination thereof. For silver, a gold (Au) electrode and a nickel (Ni) electrode may be used.

After the electrode 234 is formed, the zinc oxide nanorod layer 235 may be grown. The zinc oxide nanorod layer 235 is formed between the electrodes 234 and may be formed in a zigzag shape. In addition, the zinc oxide nanorod layer 235 is in contact with a portion of one electrode 234 and a portion of the other electrode 234 among the two electrodes 234 formed alternately, and one nanorod and the other It is preferred that the nanorods can be in electrical contact with each other.

By the structure of the fluorescence sensor 232, a high-sensitivity sensor that maximizes the surface area to area can be used, and the shape of the material or nanostructure can be changed according to the fluorescence wavelength region desired to be measured.

Subsequently, the circuit unit 240 is positioned below the second light source unit 220 and the detection unit 230 and outputs a difference in conductivity of the fluorescent signal detected by the detection unit 230 as a ratio.

11 is a circuit diagram showing a circuit part of the present invention.

Referring to FIG. 11, the circuit unit 240 includes a fluorescent sensor resistance Rs connected in parallel with each other and a load resistance RL connected in series with a fluorescent sensor resistance Rs to correspond to the fluorescent sensor resistance Rs. I can. In addition, the fluorescence sensor resistance Rs and the load resistance RL of the circuit unit 240 may have a form of a Wheatstone bridge in order to output a difference in conductivity of the fluorescence signals detected by the detection unit 230, respectively. have. Due to this Wheatstone bridge structure, even weak signal differences occurring from two or more sensors can be detected largely.

As an example, the configuration of the circuit unit 240 shown in FIG. 11 shows a circuit configuration in the case where the overlapping wavelength points 10 are two. That is, an example when two fluorescence sensors 232 are used is shown.

Here, RS1 represents the resistance of the first fluorescent sensor, RS2 represents the resistance of the second fluorescent sensor, and RL1 and RL2 represent the load resistance corresponding to the sensor resistance, respectively.

If the conductivity of RS1 and RS2 is the same, 0V is applied to the output VG because the potentials of the A and B points are the same. However, when the difference in the intensity of the fluorescent signal received by the two fluorescent sensors 232 increases and the resistance of RS1 decreases, the potential of point A is higher than the potential of point B, so that VG is applied with a positive voltage, and points A and B The larger the potential difference of, the larger the voltage applied to VG becomes. This can be applied equally to two or more sensors according to the general Wheatstone bridge principle, and by setting the operating voltage and load resistance value, etc., taking into account the initial value of the sensor resistance and the range of fluctuations, the output signal according to the difference in conductivity between the two sensors is generated. Can be maximized.

The difference in conductivity between the two fluorescence sensors 232 measured in this way is output as a ratio and compared with the fluorescence signal ratio information for each biological tissue collected by the fluorescence information collection unit 100, so that the type of biological tissue can be easily determined in real time. have.

12 is a flow chart showing an analysis method using the real-time biological tissue analysis system of the present invention.

Referring to FIG. 12, in the analysis method using the real-time biological tissue analysis system according to the present invention, measuring a unique fluorescence signal according to the type of biological tissue for each wavelength after spectroscopy using the fluorescence information collecting unit 100 (S310 ), setting the overlapping wavelength point 10 at which each peak of the fluorescence signals measured for each wavelength overlaps (S320), storing the fluorescence signal ratio information for the overlapping wavelength point 10 (S330) And filtering the wavelength of the fluorescence signal through the biological tissue analyzer 200 based on the stored fluorescence signal ratio information to determine the type of the biological tissue in real time (S340).

First, the first light source unit 110 of the fluorescence information collection unit 100 transmits monochromatic light having a wavelength shorter than that of the fluorescence wavelength region generated in the biological tissue to the reaction unit 120 in which the biological tissue is disposed. The light transmitted to the reaction unit 120 is irradiated to a living tissue, and a fluorescent signal in the form of a complex light excited by the living tissue is transmitted to the detection unit 130. The fluorescence signal transmitted to the detection unit 130 is sensed for each wavelength after spectroscopy. (S310)

In the fluorescence signal measured for each wavelength, the overlapping peak is checked to set the overlapping wavelength point 10, and the reference overlapping point 11 corresponding to the first overlapping peak among the overlapping peaks and after the reference overlapping point 11 Partial overlapping points 12 corresponding to the peaks overlapped in are set (S320).

When the overlapping wavelength points 10 are respectively set, the entire fluorescence signal is collected by blocking the light based on the reference overlapping point 11, and the partial fluorescence signal is collected by blocking the light based on the partial overlapping points 12. The fluorescence signal ratio information data obtained by extracting the difference in the ratio of the partial fluorescence signal to the total fluorescence signal is stored (S330).

When the fluorescence signal ratio information data is stored in the fluorescence information collection unit 100, the biological tissue analysis unit 200 irradiates light to the biological tissue to be measured through the second light source unit 220, and The excited fluorescence signal is received through the detection unit 230. At this time, the number of detection units 230 is arranged to be the same as the number of overlapping wavelength points 10, and the filter 231 included in the detection unit 230 is so that the wavelength corresponding to the overlapping wavelength points 10 is filtered. Each is set so that the filtered fluorescence signal is sensed through the fluorescence sensor 232.

Each fluorescence signal sensed through the fluorescence sensor 232 outputs the difference in conductivity of the fluorescence signals as a ratio through the circuit unit 240 and compares the fluorescence signal rate information for each biological tissue collected by the fluorescence information collection unit 100. The type of biological tissue is determined in real time (S340).

As described above, in the real-time biological tissue analysis system and method according to the present invention, the fluorescence signal of the biological tissue generated in the form of complex light is measured for each wavelength, and the fluorescence signal ratio is analyzed using the overlapping wavelength of the measured signals. By doing so, it is possible to determine the type of biological tissue in real time. Accordingly, it is possible to solve problems such as a time delay, a physical burden of a patient, and an additional cost according to a conventional biopsy.

In addition, it is not limited to the biopsy module in the form of an endoscope, and the identification of tissue cells in real time, such as the identification of tissue cells that are difficult to identify with the naked eye in an incision surgery for confirming and removing tumors, etc. It can be used wherever it is necessary to check whether or not there is an abnormality. Therefore, after the conventional physical collection, if it is based on general material analysis methods such as incision, dyeing, culture, observation, etc., and if the fluorescence wavelength signal generated from each material is converted into data and used for real-time analysis, not only the medical field, but also agriculture, aerospace , Environmental analysis, etc. The range that can be applied is wide.

On the other hand, the embodiments of the present invention disclosed in the specification and drawings are merely presented specific examples to aid understanding, and are not intended to limit the scope of the present invention. In addition to the embodiments disclosed herein, it is apparent to those of ordinary skill in the art that other modified examples based on the technical idea of the present invention may be implemented.

100: fluorescence information collection unit 110: first light source unit
120: reaction unit 130: detection unit
131: spectroscopic unit 132: sensor unit
140: control unit 150: optical fiber unit
151: first optical fiber 152: second optical fiber
200: biological tissue analysis unit 210: body
220: second light source unit 230: detection unit
231: filter 232: fluorescence sensor
240: circuit part

Claims (20)

Fluorescence information for storing the ratio information data of the fluorescence signal using the overlapping wavelength point where the unique fluorescence signal according to the type of biological tissue is spectralized and measured for each wavelength, and each peak of the fluorescence signals measured for each wavelength overlaps Collection unit; And
A real-time biological tissue analysis system comprising a biological tissue analysis unit configured to determine a type of biological tissue in real time by filtering a wavelength of the fluorescent signal based on information stored in the fluorescence information collecting unit. The method of claim 1, wherein the overlapping wavelength point is
A reference overlapping point corresponding to the first overlapping peak among the overlapping peaks; And
Real-time biological tissue analysis system including partial overlapping points corresponding to overlapping peaks after the reference overlapping point. The method of claim 2, wherein the fluorescence signal information,
A total fluorescence signal collected by blocking light based on the reference overlapping point; And
Includes a partial fluorescence signal collected by blocking light based on the partial overlapping points,
The real-time biological tissue analysis system that is information stored by extracting the difference in the ratio of the partial fluorescence signal to the entire fluorescence signal. The method of claim 1, wherein the fluorescence information collection unit,
A first light source unit for irradiating light to the biological tissue;
A reaction unit configured to block external interference light to detect the fluorescent signal;
A sensing unit that detects a fluorescence signal excited from the biological tissue; And
A real-time biological tissue analysis system comprising a control unit for storing unique fluorescence information according to the biological tissue. The method of claim 4, wherein the detection unit,
A spectroscopic unit that adjusts the wavelength of the fluorescent signal incident on the sensing unit; And
A real-time biological tissue analysis system comprising a sensor unit for sensing a fluorescence signal whose wavelength is adjusted by the spectroscopic unit. The method of claim 5,
The spectroscopic unit is a real-time biological tissue analysis system including a spectroscope (Monochromator) or an optical system (Optical System). The method of claim 4,
The first light source unit and the detection unit is a real-time biological tissue analysis system to be connected to the reaction unit using an optical fiber unit. The method of claim 7, wherein the optical fiber unit,
A first optical fiber connecting the first light source unit and the reaction unit; And
A real-time biological tissue analysis system comprising a plurality of second optical fibers connected to the sensing unit and the reaction unit, and arranged to surround the first optical fiber. The method of claim 1, wherein the biological tissue analysis unit,
Body;
A second light source unit disposed at the center of the body and transmitting light for generating the fluorescent signal to the biological tissue;
A sensing unit disposed around the second light source unit, sensing the fluorescence signal, and detecting a fluorescence signal blocked based on the overlapping wavelength point; And
A real-time biological tissue analysis system comprising a circuit unit positioned below the second light source unit and the sensing unit and outputting a difference in conductivity of the fluorescent signal detected by the sensing unit as a ratio. The method of claim 9, wherein the detection unit,
A filter disposed on the body and filtering a wavelength corresponding to the overlapping wavelength point; And
A real-time biological tissue analysis system comprising a fluorescence sensor disposed under the filter and configured to sense the fluorescence signal incident through the filter. The method of claim 10,
The filter is a real-time biological tissue analysis system that is formed so that the amount of light of the fluorescence signal incident on the fluorescence sensor is different. The method of claim 1,
The fluorescence information collection unit or the biological tissue analysis unit is arranged to be in contact with the biological tissue when measuring the biological tissue. The method of claim 9,
The real-time biological tissue analysis system, wherein the number of detection units has the same number as the number of overlapping wavelength points. The method of claim 13,
The sensing units are each disposed so that the same distance is spaced apart from the second light source unit. The method of claim 10, wherein the fluorescence sensor,
Board;
An electrode formed on the substrate and formed in an IDT structure; And
A real-time biological tissue analysis system comprising a zinc oxide nanorods (ZnO Nanorods) layer formed on the electrode and formed in a zigzag shape. The method of claim 9, wherein the circuit unit,
Fluorescence sensor resistors having the same number as the number of detection units and connected in parallel with each other; And
And a load resistor connected in series with each of the fluorescent sensor resistors so as to correspond to the fluorescent sensor resistance,
The fluorescence sensor resistance and the load resistance are in the form of a Wheatstone Bridge in order to output a difference in conductivity of the fluorescence signal detected by the detection unit, respectively. Measuring a unique fluorescence signal according to the type of biological tissue for each wavelength after spectroscopy using a fluorescence information collection unit;
Setting an overlapping wavelength point at which each peak of the fluorescence signals measured for each wavelength overlaps;
Storing fluorescence signal ratio information for the overlapping wavelength points; And
And determining the type of biological tissue in real time by filtering the wavelength of the fluorescent signal through a biological tissue analysis unit based on the stored fluorescence signal ratio information. The method of claim 17, wherein the setting of the overlapping wavelength point comprises:
Checking the overlapping peaks for the fluorescence signals measured for each wavelength;
Setting a reference overlapping point corresponding to the first overlapping peak among the overlapping peaks; And
And setting partial overlapping points corresponding to the overlapping peaks after the reference overlapping point. The method of claim 17, wherein the storing of the fluorescence signal information comprises:
Blocking light based on the reference overlapping point to collect the entire fluorescence signal;
Collecting a partial fluorescence signal by blocking light based on the partial overlapping points; And
The method of analyzing a biological tissue analysis system further comprising the step of extracting and storing a difference in the ratio of the partial fluorescence signal to the total fluorescence signal. The method of claim 17, wherein determining the type of biological tissue in real time,
Detecting a fluorescence signal excited in the living tissue;
Filtering and sensing the detected fluorescence signal based on the overlapping wavelength point; And
The analysis method of the biological tissue analysis system further comprising the step of determining the type of the biological tissue by comparing the sensed fluorescence signal with the stored fluorescence signal information.

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