JP4242633B2 - Discrimination method and discrimination device - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a discrimination method for discriminating the size of a particle, a discrimination method for discriminating a state of a cell, and a discrimination apparatus for performing the discrimination method, and in particular, by observing backscattered light from a scatterer. The present invention relates to a discriminating method for discriminating the size, a discriminating method for discriminating the state of cells using the discriminating method, and a discriminating apparatus.
[0002]
[Prior art]
A technique for observing cell information on a living tissue by detecting scattered light when the living tissue is irradiated with light is known. For example, Patent Document 1 describes a technique for determining the size of a cell nucleus based on the difference in wavelength spectrum of scattered light. Patent Document 2 and Non-Patent Document 1 disclose a technique for detecting a change in the structure of a cell organ using the scattering angle characteristic of scattered light.
[0003]
Since canceration of cells often occurs in the epithelium, it is required to be able to observe information on cells in the surface layer of living tissue. When cells become cancerous, the structure of the cells changes. As shown in Patent Document 1, particularly in the cell nucleus, the size increases, the size variation increases, and the refractive index slightly increases. Therefore, the scattering characteristics of single scattered light by the cell nucleus are different between normal cells and cancer cells. By utilizing this, the state of the cell can be diagnosed.
[0004]
However, since the biological tissue is a multiple scatterer, when scattered light from the biological tissue is detected, the background component due to multiple scattering occupies most of the information of single scattering due to the intracellular structure. become. Therefore, in order to remove multiple scattered light and detect only a single scattered component due to the intracellular structure, Patent Document 1 irradiates linearly polarized light and detects two of the parallel polarized light component and the orthogonal polarized light component of the scattered light. Thus, a method of calculating the difference is used.
[0005]
[Patent Document 1]
US Pat. No. 6,404,497
[0006]
[Patent Document 2]
JP-A-6-98892
[0007]
[Non-Patent Document 1]
IEEE J. Select. Topics Quantum Electron., Vol. 7,
pp.887-893 (2001)
[0008]
[Problems to be solved by the invention]
In the method of observing the wavelength spectrum of scattered light as disclosed in Patent Document 1, it is necessary to detect the spectrum shape in a relatively wide wavelength region, and thus a plurality of wavelength selection filters or spectrometers are required. As a result, the apparatus configuration becomes large and complicated. Moreover, in the thing of patent document 1, in order to remove the multiple scattering component by a biological tissue, polarized light is utilized. In this case, a polarizing element is used to change the illumination light into linearly polarized light or to detect a polarization component. In this case, since only a specific polarization component is limited, the light utilization efficiency is lowered. Furthermore, when the technique of Patent Document 1 is applied to endoscopic observation, the arrangement of polarizing elements becomes a problem. When observing a living tissue using an optical fiber or the like, it is necessary to arrange a polarizing element between the optical fiber and the living tissue, or to use a polarization-preserving optical fiber. This is disadvantageous in that the degree of freedom in device design is limited and complicated adjustment work is indispensable. In addition, the endoscope needs to be able to withstand severe environments such as high temperature and high humidity including use and disinfection work. In this respect, the resistance of the polarizing element becomes a problem.
[0009]
On the other hand, in Patent Document 2 or the like that uses the scattering angle characteristic of scattered light, in order to obtain a scattering angle profile, a large number of detectors or detectors with a large number of pixels are actually required. In addition, there is no description regarding the setting of the optimum observation angle range for efficient measurement.
[0010]
The present invention has been made in view of such problems of the prior art, and its purpose is to measure the size of particles on the surface of a scatterer with a relatively easy apparatus configuration when observing backscattered light from the scatterer. It is an object of the present invention to provide a method and an apparatus capable of determining the thickness. Another object of the present invention is to provide a method and apparatus that can more easily discriminate the state of cells on the surface of a tissue than in the prior art, particularly when observing living tissue.
[0011]
[Means for Solving the Problems]
In order to achieve the above object, the discriminating method according to the present invention discriminates the size of particles on the surface of the scatterer by illuminating the scatterer with light from a light source and observing backscattered light from the scatterer. A way to
Detecting scattered light in a first scattering angle range, including a scattering angle range that is within a scattering angle range of 175 ° or more and 180 ° or less and that gives a maximum of an angular distribution of scattered light intensity by the particles;
Unlike the first scattering angle range, the step of detecting scattered light in the second scattering angle range in which the scattered light intensity does not change depending on the diameter of the particles relative to the first scattering angle range. When,
By calculating the difference or ratio between the intensity of the scattered light in the first scattering angle range and the intensity of the scattered light in the second scattering angle range, respectively detected in the step, the particles on the surface of the scatterer Determining the size of
It is the method characterized by having.
[0012]
The discriminating apparatus according to the present invention for achieving the above object discriminates the size of particles on the surface of the scatterer by illuminating the scatterer with light from a light source and observing backscattered light from the scatterer. A discrimination device for
A first light receiving unit for detecting scattered light in a first scattering angle range including a scattering angle range that is within a scattering angle range of 175 ° or more and 180 ° or less and that gives the maximum of the angular distribution of scattered light intensity by the particles. When,
Unlike the first scattering angle range, the scattered light intensity in the second scattering angle range is detected in which the scattered light intensity does not change relative to the particle diameter relative to the first scattering angle range. Two light receiving units;
By calculating the difference or ratio between the intensity of the scattered light in the first scattering angle range and the intensity of the scattered light in the second scattering angle range, respectively detected by each light receiving unit, the surface of the scatterer An arithmetic unit for determining the size of particles of
It is characterized by having.
[0013]
Another discriminating apparatus according to the present invention discriminates the state of cells on the surface of the living tissue containing particles by illuminating the living tissue with light from a light source and observing backscattered light from the living tissue. Because
A first light receiving unit for detecting scattered light in a first scattering angle range including a scattering angle range that is within a scattering angle range of 175 ° or more and 180 ° or less and that gives the maximum of the angular distribution of scattered light intensity by the particles. When,
Unlike the first scattering angle range, the second detecting the scattered light in the second scattering angle range in which the scattered light intensity does not change relative to the particle diameter relative to the first scattering angle range. The light receiving unit,
By calculating the difference or ratio between the intensity of the scattered light in the first scattering angle range and the intensity of the scattered light in the second scattering angle range, respectively detected by each light receiving unit, the biological tissue surface A computing device for determining the state of the cells of
It is characterized by having.
[0014]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, embodiments and examples of a discrimination method for discriminating the size of particles, a discrimination method for discriminating the state of a cell, and a discrimination apparatus for carrying out these discrimination methods according to the present invention will be described.
[0015]
The discrimination method of the present invention is:
Illuminating light from a light source onto a scatterer and observing backscattered light from the scatterer to determine the size of particles on the scatterer surface,
Scattered light is detected in a first scattering angle range including at least a part of a scattering angle range that gives a maximum of an angular distribution of scattered light intensity by the particles with respect to a range of the diameter of the particles;
Detecting scattered light in a second scattering angle range different from the first scattering angle range;
The size of the particles on the scatterer surface is calculated by calculating the difference or ratio between the intensity of the scattered light detected in the first scattering angle range and the intensity of the scattered light detected in the second scattering angle range. It is a method characterized by discriminating the thickness.
[0016]
Another discrimination method of the present invention is as follows.
Illuminating light from a light source onto a living tissue, and observing backscattered light from the living tissue to determine the state of cells on the surface of the living tissue,
In a first scattering angle range including at least a part of a scattering angle range that gives a maximum of an angular distribution of scattered light intensity by the particles with respect to a range of particle diameters associated with changes in the state of cells of the particles constituting the cells. Detect scattered light,
Detecting scattered light in a second scattering angle range different from the first scattering angle range;
Determining the state of the cell by calculating a difference or ratio between the intensity of the scattered light detected in the first scattering angle range and the intensity of the scattered light detected in the second scattering angle range. It is a characteristic method.
[0017]
Below, the reason and effect | action which take the said structure in this invention are demonstrated.
[0018]
FIG. 5 shows the result of calculating the angular distribution of the scattering intensity of the backscattered light by the spherical particles based on the Mie scattering theory. Here, the average value of the particle diameter is 4 μm, 6 μm, 10 μm, and 14 μm, the refractive indexes of the particles are 1.38, 1.39, and 1.4, and the refractive index around the particles is 1.33. In the graph of FIG. 5, for example, “E-11” is “× 10”. ^-11 ”Means the same.
[0019]
FIG. 5 shows vertical and horizontal 4 × 3 = 12 graphs. Here, the numerical value on the uppermost graph is the refractive index of the particle. The numerical value on the side (left side) of the left graph is the average value of the particle diameters. At each intersection of these numerical values in the horizontal direction and the vertical direction, the state of the angular distribution of the backscattered light scattering intensity in the particles having the respective particle diameter and refractive index is shown.
[0020]
In these graphs, the particle size variation follows a normal distribution. In addition, for particles having an average diameter of 4 μm and 6 μm, the standard deviation of the particle diameter is 1 μm. On the other hand, for particles having an average diameter of 10 μm and 14 μm, the standard deviation of the particle diameter is 1.5 μm. In each graph, the average intensity per particle in such a particle size distribution is shown. As shown in FIG. 6, the scattering angle on the horizontal axis of each graph is defined as an angle represented by θ in the figure with respect to the orientation of the scattered light 102 when the incident light 101 enters the particle 103. . Further, in each graph of FIG. 5, the scattering intensity when the wavelength of light is 400 nm, 600 nm, and 800 nm is superimposed and displayed.
[0021]
As shown by the arrow a in FIG. 5, it can be seen from each graph that the scattering intensity becomes maximum at a scattering angle of 175 ° to 180 ° with respect to light having a wavelength of 400 nm. Moreover, it turns out that the intensity | strength peak appears notably, so that a particle diameter is large. FIG. 7 is a plot of the angle giving the maximum value with respect to the particle diameter when the wavelength is 400 nm and the refractive index of the particle is 1.39.
[0022]
On the other hand, as shown by b in FIG. 5, in the range where the scattering angle is 175 ° or less, the scattering intensity does not change greatly with respect to the scattering angle regardless of the particle diameter, refractive index, and wavelength. Recognize. Moreover, it turns out that the relative intensity with respect to b of the intensity peak shown by a becomes large, so that a particle diameter becomes large.
[0023]
Therefore, for example, a sample including spherical particles is irradiated with light having a wavelength of 400 nm, and backscattered light from the sample is detected. The backscattered light from the sample includes single scattered light and multiple scattered light. Here, the single scattered light is light scattered only by the surface of the sample. On the other hand, the multiple scattered light is light that is diffused many times by particles in the sample as the light diffuses inside the sample.
[0024]
In the present invention, for example, the first scattering angle range is set to 178 ° to 179 °, the second scattering angle range is set to 170 ° to 175 °, and the back scattered light from the sample is detected in each scattering angle range. . From the results shown in FIG. 5, it is considered that the angle distribution of the multiple scattered light does not change greatly in the vicinity of the first and second scattering angle ranges. On the other hand, in single scattering, a difference in relative scattering intensity between the first and second scattering angle ranges occurs according to the size of the particle. Therefore, the multiple scattered light component is canceled by calculating the difference or ratio between the intensity of the scattered light detected in the first scattering angle range and the intensity of the scattered light detected in the second scattering angle range. It is. As a result, it is possible to extract a scattered signal based on only a single scattered light component. This calculation result represents the size relationship of the scattering particles in the sample. Therefore, for example, the particle size can be estimated from the calculation result and the simulation result. Moreover, the size of the scattering particles can be determined by relatively comparing the calculation results between different samples.
[0025]
In cells of living tissue, typical values of cell nuclei are approximately 5 μm to 15 μm. On the other hand, the relative refractive index of the cell nucleus relative to the cytoplasm is 1.035 to 1.05. Each graph in FIG. 5 shows the scattering intensity for the scattering particles in the range of those values.
[0026]
Therefore, for example, light with a wavelength of 400 nm is irradiated onto the living tissue, and backscattered light from the living tissue is detected. The back scattered light from the living tissue includes single scattered light and multiple scattered light. Here, the single scattered light is light scattered by cells only on the surface of a living tissue such as an epithelial cell layer. On the other hand, the multiple scattered light is light that is diffused many times by particles in the tissue as the light diffuses inside the living tissue.
[0027]
Here, similarly to the above, the first scattering angle range is set to 178 ° to 179 °, the second scattering angle range is set to 170 ° to 175 °, and the back scattered light from the living tissue is set in the respective scattering angle ranges. To detect. Then, by calculating the difference or ratio between the intensity of the scattered light detected in the first scattering angle range and the intensity of the scattered light detected in the second scattering angle range, the multiple scattered light component is canceled out. It is. As a result, it is possible to extract a scattered signal based on only a single scattered light component. Therefore, the size of the scattering particles can be determined by relatively comparing the calculation results on different living tissues. Since the calculation result varies depending on the size of the cell nucleus assumed as the scattering particle, the state of the cell can be determined based on this.
[0028]
In these discrimination methods of the present invention, the wavelength of light from the light source is preferably 500 nm or less. In the case of backscattering by a scatterer, particularly cells, when the scattering by the cell nucleus is the main scattering, as shown in FIG. 5, the change in the scattering angle with respect to the size of the scattering particles becomes more significant as the wavelength of the light source is shorter. . Therefore, it is desirable to use a wavelength of 500 nm or less, which is a short wavelength particularly in the visible light region.
[0029]
In addition, another determination method according to the present invention is as follows.
Illuminating light from a light source onto a scatterer and observing backscattered light from the scatterer to determine the size of particles on the scatterer surface,
The light from the light source includes a plurality of wavelengths;
Scattered light is detected in a first scattering angle range including at least a part of a scattering angle range that gives a maximum of an angular distribution of scattered light intensity by the particles with respect to a range of the diameter of the particles;
Detecting scattered light in a second scattering angle range different from the first scattering angle range;
The relative intensity between the plurality of wavelengths of scattered light detected in the first scattering angle range and the relative intensity between the plurality of wavelengths of scattered light detected in the second scattering angle range In this method, the size of particles on the surface of the scatterer is determined by calculating a difference or a ratio.
[0030]
Furthermore, another discrimination method according to the present invention is as follows:
Illuminating light from a light source onto a living tissue, and observing backscattered light from the living tissue to determine the state of cells on the surface of the living tissue,
The light from the light source includes a plurality of wavelengths;
In a first scattering angle range including at least a part of a scattering angle range that gives a maximum of an angular distribution of scattered light intensity by the particles with respect to a range of particle diameters associated with changes in the state of cells of the particles constituting the cells. Detect scattered light,
Detecting scattered light in a second scattering angle range different from the first scattering angle range;
The relative intensity between the plurality of wavelengths of scattered light detected in the first scattering angle range and the relative intensity between the plurality of wavelengths of scattered light detected in the second scattering angle range It is a method characterized by discriminating the state of a cell by calculating a difference or a ratio.
[0031]
When there is a large variation in the density of the particles of the scatterer, the scattering intensity varies depending on the number of particles that contribute to the scattering. Therefore, in the present invention, a plurality of wavelengths of light illuminating the scatterer are used. In this way, a scattering signal based on the particle diameter can be obtained regardless of the density of the particles. In the example shown in FIG. 5, the scattering intensity at the wavelength of 800 nm does not change greatly with respect to the scattering angle as compared with the wavelength of 400 nm. Therefore, the relative intensity of the scattered light at a wavelength of 400 nm with respect to the scattered light at a wavelength of 800 nm is obtained for each of the first scattering angle range and the second scattering angle range. By comparing these relative intensities, a signal corresponding to the size of the particles can be obtained without being influenced by the density of the scattering particles.
[0032]
In these other discrimination methods of the present invention, it is desirable that at least one of the plurality of wavelengths is 500 nm or less and the other at least one wavelength is 500 nm or more. As shown in FIG. 5, the change in the scattering angle distribution with respect to the size of the scattering particles is more remarkable as the wavelength of the light source is shorter. On the other hand, regardless of the particle size, the longer the wavelength, the less the intensity changes with respect to the scattering angle. Therefore, it is desirable to use a short wavelength of 500 nm or less and a long wavelength of 500 nm or more, particularly in the visible light region.
[0033]
In these other determination methods of the present invention, it is desirable that the second scattering angle range is set in the vicinity of the first scattering angle range. In order to extract only a single scattered light component efficiently, it is desirable that the scattering state by the multiple scattered light is substantially the same in the first scattering angle range and the second scattering angle range. Therefore, the first and second scattering angle ranges are preferably set in the vicinity of each other.
[0034]
In these other determination methods of the present invention, the first scattering angle range includes at least part of 176 ° to 180 °, and the second scattering angle range includes at least part of 170 ° to 176 °. It is desirable to be set to. In particular, in the case of backscattering by cells, when scattering by cell nuclei is the main scattering, the scattering angle distribution of scattered light has the properties as shown in FIG. For this reason, the first and second scattering angle ranges are preferably set as described above.
[0035]
In these other determination methods of the present invention, it is desirable that at least one of the first scattering angle range and the second scattering angle range be adjustable. It is preferable that the scattering angle range to be detected can be adjusted according to the type of scattering particles and cells.
[0036]
Next, with reference to the drawings, an embodiment of a discrimination method for discriminating the size of particles, a discrimination method for discriminating the state of cells, and a discrimination apparatus for carrying out these methods will be described.
[0037]
[Example 1]
FIG. 1A shows the configuration of an example of an apparatus for discriminating the state of cells of a biological sample using the method of the present invention. In FIG. 1A, light from a light source 1 passes through an optical fiber 2 and is irradiated onto a biological sample 4 by a probe 3.
[0038]
FIG. 1B shows a front view of the tip of the probe 3. Light that has passed through the optical fiber 2 is irradiated from an illumination end face (illumination region) 11 of the optical fiber 2. Scattered light from the biological sample 4 includes a light receiving end face (light receiving area) 12 of the optical fiber 5 for receiving the first scattering angle range and a light receiving end face (light receiving) of the optical fiber 6 for receiving the second scattering angle range. (Region) 13 receives light. The light received by the light receiving end faces 12 and 13 passes through the optical fibers 5 and 6, respectively, and is detected by the detector 7 and the detector 8, respectively. The outputs of the detectors 7 and 8 are input to the calculator 9, and the difference or ratio between the output values of the detectors 7 and 8 is calculated.
[0039]
The illumination end face 11 and the light receiving end faces 12 and 13 may each be a single optical fiber end face, or may be composed of a plurality of optical fiber end faces. The first scattering angle range and the second scattering angle range are shown in FIG. These ranges are the distance d1 between the probe 3 and the biological sample 4, the diameter and NA (numerical aperture) of the illumination end face 11, the light receiving end faces 12 and 13, the distance d2 between the illumination end face 11 and the light receiving end face 12, and the illumination end face 11 and It is determined by the distance d3 to the light receiving end face 13.
[0040]
In the backscattering by the biological sample 4, when the scattering by the cell nucleus of the cells constituting the biological sample 4 is the main scattering, the scattering angle characteristics as shown in FIG. Set as follows.
[0041]
Diameter of illumination end face 11: 1.0 mm, NA: 0.2
Diameter of light receiving end face 12: 1.5 mm, NA: 0.2
Diameter of light receiving end face 13: 1.5 mm, NA: 0.2
d1: 40 mm
d2: 1.25 mm
d3: 6 mm
At this time, the scattering angle ranges of the scattered light received by the light receiving end face 12 and the light receiving end face 13 are the ranges indicated by a1 and b1 in FIG. 1C, respectively. Therefore, in the range indicated by a1, the intensity peak indicated by a in the graph having a large particle diameter in FIG. 5 is detected. On the other hand, in the range indicated by b1, scattered light in the vicinity shown by b in FIG. 5 is detected. In this way, observation is performed within the sample surface in different scattering angle ranges. When a region where the result calculated by the computing unit 9 is different from the surroundings is detected, it can be understood that the size of the cell nucleus is different from the surroundings in the region, so that knowledge such as cell canceration is obtained. be able to.
[0042]
[Example 2]
FIG. 2A shows the configuration of another example of an apparatus for discriminating the state of cells of a biological sample using the method of the present invention. In FIG. 2A, the light source 21 includes semiconductor lasers 21a and 21b having wavelengths of 400 nm and 800 nm. Lights from the respective semiconductor lasers 21 a and 21 b are synthesized by the optical coupler 22 and guided to the optical fiber 23. Light from the optical fiber 23 is applied to the biological sample 4 by the probe 3.
[0043]
FIG. 2B shows a front view of the distal end portion of the probe 3. Light that has passed through the optical fiber 23 is irradiated from the illumination end face 31. Scattered light from the biological sample 4 is detected by the light receiving end faces 32a and 32b for receiving the first scattering angle range of the present invention and the light receiving end faces 33a and 33b for receiving the second scattering angle range. The The light received by the light receiving end faces 32a and 32b passes through the optical fiber bundle 25 and is detected by the detectors 27a and 27b, respectively. The light received by the light receiving end faces 33a and 33b passes through the optical fiber bundle 26 and is detected by the detectors 28a and 28b, respectively. The detectors 27a and 28a detect scattered light having a wavelength of 400 nm. The detectors 27b and 28b detect scattered light having a wavelength of 800 nm. Note that a wavelength selection filter or the like can be inserted immediately before each detector for detection of each wavelength. The outputs of the detectors 27a, 27b, 28a, 28b are input to the calculator 29.
[0044]
The light receiving end faces 32a and 32b are arranged so that the scattering angles of scattered light received by the respective end faces are the same. The same applies to the light receiving end faces 33a and 33b. Similar to the first embodiment, the spatial arrangement of the respective light receiving end faces is set so that scattered light having scattering angles indicated by a and b in FIG. 5 can be received.
[0045]
In this embodiment, the relative scattering intensity at a wavelength of 400 nm with respect to the scattering intensity at a wavelength of 800 nm is compared in the first scattering angle range and the second scattering angle range. Therefore, the arithmetic unit 29 first calculates the output ratio of the detectors 27a and 27b and the output ratio of the detectors 28a and 28b. Next, the difference or ratio between the former output ratio and the latter output ratio is calculated. From this calculation result, the size of the cell nucleus in the observation region on the biological sample 4 can be determined.
[0046]
Example 3
FIG. 3A shows the configuration of another example of an apparatus for discriminating the state of cells of a biological sample using the method of the present invention. In FIG. 3A, the light from the light source 1 is irradiated to the biological sample 4 by the probe 3 through the optical fiber 2.
[0047]
FIG. 3B shows a front view of the tip of the probe 3. Light that has passed through the optical fiber 2 is irradiated from the illumination end face 51. Scattered light from the biological sample 4 is received by the light receiving end face 52 of the optical fiber bundle 45 and the light receiving end face 53 of the optical fiber bundle 46. The light receiving end faces 52 and 53 are arranged side by side in a direction in which a plurality of optical fiber end faces are separated from the illumination end face 51 so that scattered light in a wide scattering angle range can be received. In order to increase the light receiving area, it may be arranged in a ring shape. The light received by the light receiving end faces 52 and 53 passes through the optical fiber bundles 45 and 46, respectively, and is detected by the detector 47 and the detector 48 having a plurality of detection elements. In the detectors 47 and 48, only the output of the optical fiber in the range matching the first scattering angle range and the second scattering angle range of the present invention is selected from the outputs from the plurality of optical fibers and input to the calculator 49. To do. In each of the detectors 47 and 48, the output from one optical fiber may be selected, or the outputs of a plurality of optical fibers may be selected. The calculator 49 calculates the difference or ratio between the output values of the detectors 47 and 48.
[0048]
In the configuration of this embodiment, the first scattering angle range and the second scattering angle range are adjusted by the detectors 47 and 48 according to the type of the biological sample 4 and the type of cells constituting the biological sample 4. Can do.
[0049]
Example 4
FIG. 4A is a diagram showing an example in which a device for discriminating the state of cells of a biological sample using the method of the present invention is combined with a colposcope. In FIG. 4A, the colposcope 61 is for observing the image of the sample captured by the objective lenses 63a and 63b with the eyepiece lenses 62a and 62b. In the present embodiment, a probe 64 of an apparatus for discriminating the state of a cell using the method of the present invention is provided on the surface on which the objective lenses 63a and 63b are arranged.
[0050]
FIG. 4B shows a configuration of the probe 64, and FIG. 4C shows a configuration of an apparatus related to the method of the present invention. The light beam 65 emitted from the laser light source 72 is reflected by the scanning mirrors 66a and 66b provided on the probe 64 to illuminate the sample surface. The scattered light from the sample is received by the optical fiber 75 that receives the first scattering angle range and the optical fiber 76 that receives the second scattering angle range of the present invention. Lights from the optical fibers 75 and 76 are input to detectors 77 and 78, and outputs from the detectors 77 and 78 are input to a calculator 79. The calculator 79 calculates the difference or ratio between the output values of the detectors 77 and 78.
[0051]
The scanning mirrors 66 a and 66 b scan the sample surface two-dimensionally by the scanning control device 80. The computer 81 synchronizes driving of the scanning control device 80 and signal input from the computing unit 79. The image display device 82 displays a two-dimensional distribution of calculation results in the scanning range on the sample surface. The presence of abnormal cells can be detected if a region in which the value of the calculation result is different from the surroundings is detected within the scanning range.
[0052]
The discrimination method and discrimination device of the present invention described above can be configured as follows, for example.
[0053]
[1] A method of determining the size of particles on the surface of the scatterer by illuminating the scatterer with light from a light source and observing backscattered light from the scatterer,
Scattered light is detected in a first scattering angle range including at least a part of a scattering angle range that gives a maximum of an angular distribution of scattered light intensity by the particles with respect to a range of the diameter of the particles;
Detecting scattered light in a second scattering angle range different from the first scattering angle range;
The size of the particles on the scatterer surface is calculated by calculating the difference or ratio between the intensity of the scattered light detected in the first scattering angle range and the intensity of the scattered light detected in the second scattering angle range. A discrimination method characterized by discriminating the thickness.
[0054]
[2] The discrimination method according to [1], wherein the wavelength of light from the light source is 500 nm or less.
[0055]
[3] The light from the light source includes a plurality of wavelengths, and is detected in the relative intensity between the plurality of wavelengths of the scattered light detected in the first scattering angle range and in the second scattering angle range. 2. The discrimination method according to claim 1, wherein the size of the particles on the surface of the scatterer is discriminated by calculating a difference or ratio between the relative intensity of the scattered light and the plurality of wavelengths.
[0056]
[4] The discrimination method according to the above item 3, wherein at least one of the plurality of wavelengths is 500 nm or less and another at least one wavelength is 500 nm or more.
[0057]
[5] The discrimination method according to any one of [1] to [4], wherein the second scattering angle range is set in the vicinity of the first scattering angle range.
[0058]
[6] The first scattering angle range includes at least part of 176 ° to 180 °, and the second scattering angle range is set to include at least part of 170 ° to 176 °. 6. The discrimination method according to any one of 1 to 5 above.
[0059]
[7] The discrimination method according to any one of the above items 1 to 6, wherein at least one of the first scattering angle range and the second scattering angle range is adjustable.
[0060]
[8] A method of determining a state of a cell on the surface of the living tissue by illuminating the living tissue with light from a light source and observing backscattered light from the living tissue,
In a first scattering angle range including at least a part of a scattering angle range that gives a maximum of an angular distribution of scattered light intensity by the particles with respect to a range of particle diameters associated with changes in the state of cells of the particles constituting the cells. Detect scattered light,
Detecting scattered light in a second scattering angle range different from the first scattering angle range;
Determining the state of the cell by calculating a difference or ratio between the intensity of the scattered light detected in the first scattering angle range and the intensity of the scattered light detected in the second scattering angle range. A distinction method.
[0061]
[9] The discrimination method according to 8 above, wherein the wavelength of light from the light source is 500 nm or less.
[0062]
[10] The light from the light source includes a plurality of wavelengths, and is detected in the relative intensity between the plurality of wavelengths of the scattered light detected in the first scattering angle range and in the second scattering angle range. 9. The discrimination method according to claim 8, wherein the state of the cell is discriminated by calculating a difference or ratio of relative intensity of the scattered light between the plurality of wavelengths.
[0063]
[11] The discrimination method according to 10 above, wherein at least one of the plurality of wavelengths is 500 nm or less, and at least one other wavelength is 500 nm or more.
[0064]
[12] The discrimination method according to any one of 8 to 11, wherein the second scattering angle range is set in the vicinity of the first scattering angle range.
[0065]
[13] The first scattering angle range includes at least part of 176 ° to 180 °, and the second scattering angle range is set to include at least part of 170 ° to 176 °. 13. The discrimination method according to any one of 8 to 12 above.
[0066]
[14] The discrimination method according to any one of 8 to 13, wherein at least one of the first scattering angle range and the second scattering angle range is adjustable.
[0067]
[15] a light source;
An illumination unit for guiding light from the light source to an object;
A first light receiving unit disposed at a first distance from the illumination unit;
A second light receiving unit disposed at a second interval from the illumination unit;
A discriminator comprising an arithmetic unit for calculating a difference or ratio of signals from the first light receiving unit and the second light receiving unit;
The first interval is an interval including a scattering angle range in which an angular distribution of scattering intensity by particles constituting the object is maximized,
The discriminating apparatus, wherein the second interval is an interval different from the first interval.
[0068]
【The invention's effect】
As is clear from the above description, according to the discrimination method and discrimination device of the present invention, when observing backscattered light from a scatterer, by performing observation in an efficient scattering angle range based on theoretical calculation. The particle size on the surface of the scatterer can be determined with a relatively easy apparatus configuration. Further, according to the discrimination method and the discrimination apparatus of the present invention, the state of cells on the surface of a living tissue can be discriminated more efficiently and simply than before.
[Brief description of the drawings]
1A and 1B are diagrams illustrating an apparatus according to a first embodiment, in which FIG. 1A is a diagram illustrating an overall configuration of the apparatus, FIG. 1B is a front view of a distal end portion of a probe, and FIG. The figure which shows the scattering angle range of the scattered light to perform, (d) is a figure which shows the 1st scattering angle range and the 2nd scattering angle range.
FIGS. 2A and 2B are diagrams illustrating an apparatus according to a second embodiment, in which FIG. 2A is a diagram illustrating an overall configuration of the apparatus, and FIG.
3A and 3B are diagrams illustrating an apparatus according to a third embodiment, in which FIG. 3A is a diagram illustrating an overall configuration of the apparatus, and FIG.
4A and 4B are diagrams illustrating an apparatus according to a fourth embodiment, where FIG. 4A is a diagram illustrating a colposcope, FIG. 4B is a diagram illustrating a configuration of a probe, and FIG. 4C is a diagram illustrating an overall configuration of the apparatus.
FIG. 5 is a diagram showing a result of calculating an angular distribution of scattering intensity of backscattered light by spherical particles by Mie scattering theory.
FIG. 6 is a diagram showing a definition of a scattering angle.
FIG. 7 is a graph plotting an angle giving a maximum value with respect to a particle diameter.
[Explanation of symbols]
1 ... Light source
2 ... Optical fiber
3 ... Probe
4 ... Biological sample
5, 6 ... Optical fiber
7, 8 ... Detector
9 ... Calculator
11 ... Lighting end face
12, 13 ... Light receiving end face
21 ... Light source
21a, 21b ... Semiconductor laser
22 ... Optical coupler
23 ... Optical fiber
25, 26 ... Optical fiber bundle
27a, 27b, 28a, 28b ... detector
29 ... Calculator
31 ... Lighting end face
32a, 32b, 33a, 33b ... light receiving end face
45, 46 ... Optical fiber bundle
47, 48 ... Detector
49 ... Calculator
51 ... Lighting end face
52, 53 ... Light receiving end face
61 ... Colposcope
62a, 62b ... eyepieces
63a, 63b ... objective lens
64 ... Probe
65 ... Light
66a, 66b ... scanning mirror
72 ... Laser light source
75, 76 ... optical fiber
77, 78 ... Detector
79 ... Calculator
80. Scanning control device
81 ... Computer
82. Image display device
101: Incident light
102 ... scattered light
103 ... Particles

Claims (10)

Illuminating light from a light source onto a scatterer and observing backscattered light from the scatterer to determine the size of particles on the scatterer surface,
Detecting scattered light in a first scattering angle range, including a scattering angle range that is within a scattering angle range of 175 ° or more and 180 ° or less and that gives a maximum of an angular distribution of scattered light intensity by the particles;
Unlike the first scattering angle range, the step of detecting scattered light in the second scattering angle range in which the scattered light intensity does not change depending on the diameter of the particles relative to the first scattering angle range. When,
By calculating the difference or ratio between the intensity of the scattered light in the first scattering angle range and the intensity of the scattered light in the second scattering angle range, respectively detected in the step, the particles on the surface of the scatterer Determining the size of
A discrimination method characterized by comprising:   The method according to claim 1, wherein the wavelength of light from the light source is 500 nm or less.   The light from the light source includes a plurality of wavelengths, and the relative intensity of the scattered light detected in the first scattering angle range and the scattered light detected in the second scattering angle range. The method according to claim 1, wherein the size of the particle on the surface of the scatterer is determined by calculating a difference or a ratio with a relative intensity between the plurality of wavelengths.   The discriminating method according to claim 3, wherein at least one of the plurality of wavelengths is 500 nm or less, and at least one other wavelength is 500 nm or more.   The determination method according to claim 1, wherein the second scattering angle range is set in the vicinity of the first scattering angle range.   The determination method according to claim 1, wherein the second scattering angle range is set to include 170 ° or more and 175 ° or less.   The determination method according to claim 1, wherein at least one of the first scattering angle range and the second scattering angle range is adjustable. Illuminating light from a light source onto a scatterer, and determining the particle size of the scatterer surface by observing backscattered light from the scatterer,
A first light receiving unit for detecting scattered light in a first scattering angle range including a scattering angle range that is within a scattering angle range of 175 ° or more and 180 ° or less and that gives the maximum of the angular distribution of scattered light intensity by the particles. When,
Unlike the first scattering angle range, the scattered light intensity in the second scattering angle range is detected in which the scattered light intensity does not change relative to the particle diameter relative to the first scattering angle range. Two light receiving units;
By calculating the difference or ratio between the intensity of the scattered light in the first scattering angle range and the intensity of the scattered light in the second scattering angle range, respectively detected by each light receiving unit , the surface of the scatterer An arithmetic unit for determining the size of particles of
A discriminating apparatus characterized by comprising: A discrimination device that illuminates a living tissue with light from a light source and discriminates the state of cells on the surface of the living tissue including particles by observing backscattered light from the living tissue,
A first light receiving unit for detecting scattered light in a first scattering angle range including a scattering angle range that is within a scattering angle range of 175 ° or more and 180 ° or less and that gives the maximum of the angular distribution of scattered light intensity by the particles. When,
Unlike the first scattering angle range, the second detecting the scattered light in the second scattering angle range in which the scattered light intensity does not change relative to the particle diameter relative to the first scattering angle range. The light receiving unit,
By calculating the difference or ratio between the intensity of the scattered light in the first scattering angle range and the intensity of the scattered light in the second scattering angle range, respectively detected by each light receiving unit , the biological tissue surface A computing device for determining the state of the cells of
A discriminating apparatus characterized by comprising:   The discrimination device according to claim 9, wherein the particle is a cell nucleus of the cell.

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