JP2019090611A - Cancer inspection device - Google Patents

Abstract

To provide a cancer inspection device that can execute a cancer testing method that does not depend on proficiency of an inspection operator.SOLUTION: A cancer inspection device includes: an acquisition unit that acquires an amount of amino acids or proteins and an amount of nucleic acids in a region of interest; and an information output unit that outputs information according to a ratio of the amount of nucleic acids acquired by the acquisition unit to the amount of amino acids or proteins acquired by the acquisition unit. The cancer inspection device may further include a detection unit that irradiates excitation light to the region of interest and acquires the amount of amino acids or proteins and the amount of nucleic acid in the region of interest on the basis of a spectrum of the region of interest.SELECTED DRAWING: Figure 4

Description

  The present invention relates to a cancer inspection apparatus.

The examination of the presence or absence of a cancer cell is performed by a doctor's visual inspection or using a tumor marker (see, for example, Patent Document 1).
Patent Document 1: JP-A-2015-180212

  Visual inspection varies in accuracy depending on the experience of the doctor. Tumor markers have problems in sensitivity and organ specificity.

  In the first aspect of the present invention, the amount of the nucleic acid obtained by the obtaining unit relative to the amount of the amino acid or protein obtained by the obtaining unit for obtaining the amount of amino acid or protein and the amount of nucleic acid in the region of interest And an information output unit that outputs information according to the ratio of

  In the second aspect of the present invention, the obtaining step for obtaining the amount of amino acid or protein and the amount of nucleic acid in the region of interest, and the nucleic acid obtained by the obtaining unit relative to the amount of amino acid or protein obtained in the obtaining step. There is provided a cancer examination program which causes a processing device to execute an output step of outputting information according to the ratio of the amount of

  The above summary of the invention does not enumerate all of the features of the present invention. Subcombinations of these features may also be inventive.

It is a schematic diagram explaining the structure of the inspection apparatus 100. As shown in FIG. 5 is a block diagram of a control unit 170. FIG. 5 is a flowchart showing an inspection procedure in the inspection apparatus 100. It is a figure which illustrates the display in the display image 301. FIG. FIG. 6 is a view exemplifying display in a display image 302. It is a figure which illustrates the display in the display image 303. FIG. It is a figure which illustrates the display in the display image 304. FIG. It is a figure which illustrates the display in the display image 305. FIG. FIG. 6 is a view illustrating display in a display image 306; FIG. 16 is a diagram illustrating display in a display image 307; It is a figure which illustrates the display in the display image 308. FIG. It is an example of the display image 309 which shows a detection position. It is an example of the display image 310 which shows a detection position. 10 is a flowchart showing another inspection procedure in the inspection device 100.

  Hereinafter, the present invention will be described through embodiments of the invention. The following embodiments do not limit the claimed invention. Not all combinations of features described in the embodiments are essential to the solution of the invention.

  The inventors focused on the ratio of the amount of amino acid or protein to the amount of base in cells, and found that there is a difference between the ratio in cancer cells and the ratio in normal cells. Therefore, in this embodiment, the amount of amino acids or proteins and the amount of bases are detected in the cells to be tested, and the ratio of the amount of amino acids or proteins detected to the amount of bases is calculated and compared with the prepared threshold value. In this case, a cancer test method has been developed to determine that cancer cells are included in a region of interest having a ratio above the threshold.

  FIG. 1 is a schematic view showing an overall configuration of an inspection apparatus 100 that can be used for the above-described cancer inspection method. The inspection apparatus 100 includes a stage 110, an objective optical system 120, a light source device 130, an irradiation optical system 140, a front detection system 150, a rear detection system 160, and a control unit 170.

  The stage 110 supports the sample 101 to be inspected by the inspection apparatus 100 at the peripheral portion of the container in which the sample 101 is accommodated. Further, the stage 110 has an opening that exposes the lower surface of the sample 101 in the drawing. Thereby, the sample 101 placed on the stage 110 can be observed also from the lower side in the drawing.

  Stage 110 is coupled to stage scanner 111. The stage scanner 111 drives the stage 110 parallel and perpendicular to the plane on which the sample 101 is placed, as indicated by arrows xyz in the figure. Thereby, in the inspection apparatus 100, the three-dimensional area in the sample 101 can be made into the object area of observation or an inspection, with the optical axis of the optical system and the optical axis of the excitation light fixed. Hereinafter, the region to be observed or inspected may be referred to as a region of interest.

  The objective optical system 120 has a front objective lens 121 and a rear objective lens 122 disposed symmetrically with respect to the sample 101 on opposite sides of the stage 110. In the illustrated inspection apparatus 100, the front objective lens 121 also plays a role of condensing excitation light, illumination light and the like irradiated to the sample 101.

  The light source device 130 includes a plurality of light sources 131 and 132 that generate different types of illumination light, and a combiner 139. At least one of the light sources 131 and 132 may be excitation light used when measuring the Raman spectrum of the sample 101, for example, a laser light source generating laser light with a wavelength of 532 nm. In addition, one of the plurality of light sources 131 and 132 may be a light source of illumination light used when observing a microscopic image of the sample 101.

  In addition, it is preferable that the irradiation light irradiated to the sample 101 is light of a long wavelength which is hard to invade cells, whether it is excitation light or illumination light. Furthermore, more specifically, the wavelength range of 400 nm to 800 nm can be exemplified.

  The illumination light emitted from the light sources 131 and 132 becomes a beam passing through a single optical path by the combiner 139. Thus, the irradiation light generated by the plurality of light sources 131 and 132 can be irradiated to the same position of the sample 101.

  The irradiation optical system 140 has a galvano scanner 141 and a scan lens 142. The galvano scanner 141 comprises a pair of reflecting mirrors swinging around two swinging axes which are not parallel to each other. As a result, the optical path of the excitation light incident on the galvano scanner is two-dimensionally displaced in the direction intersecting the optical axis.

  The scan lens 142 focuses the excitation light emitted from the galvano scanner 141 on a predetermined primary image surface 143. Thereby, the excitation light emitted from the light source device 130 can be irradiated to any region of interest set in the sample 101.

  The front detection system 150 includes a dichroic mirror 151, relay lenses 152 and 153, a band pass filter 154, and a spectroscope 155. The dichroic mirror 151 efficiently transmits the excitation light emitted toward the sample 101.

  The dichroic mirror 151 reflects the Raman scattered light generated in the sample 101 and guides it to the relay lenses 152 and 153. The band pass filter 154 transmits the Raman scattered light generated from the sample 101 to be incident on the spectroscope 155 while absorbing or reflecting the excitation light and the Rayleigh scattered light. Thus, the spectroscope 155 efficiently detects the Raman scattered light reflected by the sample 101 and outputs a spectral image.

  The rear detection system 160 includes a reflecting mirror 161, relay lenses 162 and 163, a band pass filter 164, and a spectroscope 165. The reflecting mirror 161 reflects the Raman scattered light generated in the sample 101 and guides it to the relay lenses 152 and 153, the band pass filter 164, and the spectroscope 165. In place of the reflecting mirror 161, a dichroic mirror that selectively reflects the wavelength of the Raman scattered light may be provided.

  The band pass filter 164 transmits the Raman scattered light generated from the sample 101 to be incident on the spectrometer 165 while absorbing or reflecting the Rayleigh scattered light and the excitation light. Thus, the spectrometer 165 efficiently detects the Raman spectrum of the transmitted light of the sample 101.

  The control unit 170 includes a processing device 171, a mouse 172, a keyboard 173, and a display unit 174. The mouse 172 and the keyboard 173 are connected to the processing device 171, and are operated when inputting an instruction of the user to the processing device 171. The display unit 174 returns feedback to the user's operation by the mouse 172 and the keyboard 173, and displays the image or character string generated by the processing device 171 to the user. Furthermore, in the illustrated inspection apparatus, the display unit 174 also displays an observation image obtained by optically observing the sample 101, characters or an image representing an inspection result, and the like.

  The processing device 171 controls the operation of the light source device 130, the stage scanner 111, and the galvano scanner 141. Further, the processing device 171 determines the state of the sample 101 based on the spectral images acquired from the spectroscopes 155 and 165 of the front detection system 150 or the rear detection system 160, that is, whether the sample 101 contains cancer cells or not. inspect. Furthermore, the processor 171 can process the spectra obtained from the spectrometers 155, 165 to improve the detection accuracy of amino acids or proteins and nucleic acids.

  The Raman scattered light detected by the front detection system 150 disposed on the same side as the irradiation optical system 140 with respect to the sample 101 is the published Raman scattered light which is also reflected by the sample 101. On the other hand, the Raman scattered light detected by the rear detection system 160 disposed on the side opposite to the irradiation optical system 140 with respect to the sample 101 is forward Raman scattered light which transmits the sample 101 as well.

  FIG. 2 is a block diagram schematically showing an internal structure of the processing device 171 of the control unit 170 in the inspection apparatus 100. As shown in FIG. As illustrated, the processing device 171 includes a detection unit 271 and an information output unit 276. Furthermore, the information output unit 276 includes a calculation unit 272, a comparison unit 273, a storage unit 274, and an output unit 275.

  The detection unit 271 detects the amount of protein and the amount of nucleic acid present in one region of interest in the sample 101, based on the spectra acquired from the spectrometers 155 and 165. The amount of protein and the amount of nucleic acid can be detected as the height of a particular peak in the spectrum of sample 101, respectively.

  In the inspection apparatus 100, the amount of amino acids in the region of interest may be detected as an indicator of the amount of protein in the region of interest of the sample 101. Furthermore, as described later, an aromatic amino acid of the amino acids, more specifically, at least one of tryptophan and phenylalanine may be detected as an indicator of the amount of protein. In the test apparatus 100, the amount of nucleic acid bases may be detected as an index of the amount of nucleic acids in the sample 101. Furthermore, as described later, of the nucleic acids, at least one of cytosine and adenine may be detected.

  The calculating unit 272 calculates the ratio of the detected amount of protein detected by the detecting unit 271 to the detected amount of nucleic acid. The ratio calculated here has a value reflecting the state of cells in the region of interest. That is, the ratio of the amount of protein to the amount of nucleic acid in the nucleus of cells is different in normal cells and cancer cells.

  Therefore, by comparing the value of the ratio calculated by the calculation unit 272 with a predetermined threshold value in advance, it can be easily and reliably inspected whether the sample cell 101 to be examined contains a cancer cell. When the amount of protein and the amount of nucleic acid are detected by peak values in the spectrum, the calculated ratio can be calculated as the ratio of peak values in the spectrum.

  A plurality of regions of interest included in a region of interest wider than one region of interest may be set for one sample 101, and the inspection of the sample 101 may be performed in each of the regions of interest. This reduces the probability of missing locally generated cancer cells.

  The comparison unit 273 executes an inspection that compares the value of the ratio calculated by the calculation unit 272 with the threshold value stored in advance in the threshold storage unit 274. The result of the examination is communicated to the user through the output unit 275. Specifically, for example, the inspection result may be displayed on the display unit 174 as a character string or an image. Further, the inspection result may be output by voice or the like. Furthermore, test results may be accumulated to allow the user to refer at any time. Thus, the information output unit 276 outputs information according to the ratio of the amount of nucleic acid acquired by the detection unit 271 to the amount of protein acquired by the detection unit 271.

  FIG. 3 is a flowchart showing an inspection procedure using the inspection apparatus 100. First, a threshold is stored in the threshold storage unit 274 of the inspection apparatus 100 (step S101). The threshold to be stored is obtained, for example, by examining a cell sample whose state is known with the examination apparatus 100.

  Next, a sample 101 to be inspected is prepared and loaded into the inspection apparatus 100 (step S102). Subsequently, a microscopic image obtained by optically enlarging the sample 101 is observed, and a region of interest to be inspected is set (step S103). The microscopic image of the sample 101 including the region of interest observed in step S103 may be captured and stored (step S104).

  Also, multiple regions of interest in the sample 101 may be set for one sample 101. This reduces the probability that a locally generated cancer cell will be missed in a test.

  Next, in the region of interest, the region of interest is irradiated with excitation light, and the spectrum of the sample 101 is obtained by the spectrometers 155 and 165 (step S105). Furthermore, the spectrum detected by the spectroscopes 155 and 165 is processed by the processing device 171 to detect the amount of protein and the amount of nucleic acid in the region of interest, for example, as the light intensity of the corresponding peak in the spectrum (step S106).

  Next, in the processing device 171, the ratio of the detected amount of protein to the amount of nucleic acid is calculated, for example, as a ratio of peak values (step S107). Next, the value of the calculated ratio is compared with the threshold stored in the threshold storage unit 274 (step S108). As a result, depending on whether the value of the calculated ratio exceeds the threshold value, a test result as to whether or not the corresponding region of interest includes cancer cells can be obtained. The obtained inspection result is recorded in the inspection apparatus 100 or output to the user (step S109). Furthermore, the control unit 170 of the inspection apparatus 100 checks whether or not there is a region of interest that has not been inspected yet while being set for the sample 101 in step S103 (step S110).

  If there is a region of interest that has not been examined in step S110 (step S110: YES), the control unit 170 returns the process of the examination apparatus 100 to step S105 and executes the examination again. Also, in this way, the examination apparatus 100 examines whether or not cancer cells exist in the region of interest of the sample 101. If it is determined in step S110 that there is no remaining region of interest that has not been examined (step S110: NO), the examination in the examination apparatus 100 ends.

  The spectrum of the sample 101 can be obtained without staining the sample 101. However, the nucleus of the cells contained in the sample 101 may be stained to make the microscopic image clearer, thereby facilitating the work such as setting of the region of interest.

[Example]
As an example of determining the threshold value, a sample 101 known to contain cancer cells was inspected by the above-described inspection apparatus 100, and the value of the threshold value stored in the threshold storage unit 274 was examined. Sample 101 was made from unstained sections of human stomach.

  First, a microscopic image of the sample 101 was obtained using an objective lens with a magnification of 100 ×. Next, laser light with a wavelength of 532 nm was irradiated to the region of interest as excitation light, and a spectrum due to Raman scattering was detected.

  In addition, in order to improve the efficiency of Raman scattering, the beam diameter of the excitation light was reduced to, for example, 1 μm or less smaller than that of cells. Therefore, in each of the 10 μm × 10 μm regions of interest in the sample 101, excitation light was applied to the 121 detection spots in each of the regions of interest for 10 seconds to detect the spectrum of 121 points. Furthermore, the detection values detected from the spectrum at 121 points were averaged to obtain the detection value of the amount of protein or nucleic acid in the region of interest. Thereby, the detection value is set to a value having a spatial spread.

In addition, image processing was performed in the processor 171 to remove in advance the known glass peak from the detected spectrum, and the influence of the container of the sample 101 was eliminated. In addition, by approximating with a fifth order polynomial, the influence of autofluorescence was eliminated. From the spectra thus obtained, the light amounts of the peaks corresponding to the nucleic acids shown in Table 1 below and the proteins shown in Table 2 below were detected as the amounts of nucleic acid and protein.

Example 1
FIG. 4 is a view showing an example of a graph included in the display image 301 displayed on the display unit 174 as a result of the examination. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

In the graph shown, the value of the horizontal axis, the amount of adenine detected from the peak light quantity of the Raman shift 725 cm ^-1 in the Raman spectroscopic image as an example of a nucleic acid, likewise detected from the peak light quantity of the Raman shift 756cm ^-1 as an example of a protein Ratio with the amount of tryptophan. Further, the value on the vertical axis is the ratio of the amount of adenine to the amount of protein with the α-helix structure detected from the peak light amount of Raman shift 1263 cm ⁻¹ as an index.

  Focusing on the distribution of normal cells and cancer cells plotted in the graph, it can be seen that only cancer cells are distributed in the range where the ratio of adenine to tryptophan is 0.85 or more. Therefore, by setting the threshold value 0.85 for the ratio of the amount of adenine to the amount of tryptophan in step S101 shown in FIG. 3, the cancer is detected based on the ratio of the amounts of adenine and tryptophan detected from the Raman spectroscopy image. It can check for the presence of cells. Although normal cells and cancer cells are distinguished by black and white dots in FIG. 4, the visibility can be improved in the display unit 174 by distinguishing them by the color and shape of the dots.

Example 2
FIG. 5 is a view showing an example of a graph included in another display image 302 displayed on the display unit 174 as a result of the examination. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

In the graph shown, the value on the horizontal axis is the ratio of the amount of adenine, which is an example of a nucleic acid, to the amount of tryptophan, which is an example of an amino acid. The value on the vertical axis is the ratio of the amount of adenine to the amount of phenylalanine detected from the peak light amount of Raman shift 1002 cm ⁻¹ as an example of a protein.

  Focusing on the distribution of normal cells and cancer cells plotted in the graph, it can be seen that only cancer cells are distributed in the range where the ratio of adenine to tryptophan is 0.85 or more. Therefore, by setting the threshold value 0.85 for the ratio of the amount of adenine to the amount of tryptophan in step S101 shown in FIG. 3, the cancer is detected based on the ratio of the amounts of adenine and tryptophan detected from the Raman spectroscopy image. It can check for the presence of cells.

[Example 3]
FIG. 6 is a diagram showing an example of a graph included in the display image 303 displayed on the display unit 174 as a result of the examination. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

In the illustrated graph, the value on the horizontal axis is the ratio of the amount of cytosine detected from the peak light amount of Raman shift 782 cm ⁻¹ in a Raman spectroscopic image as an example of a nucleic acid to the amount of tryptophan as an example of an amino acid. The value on the vertical axis is the ratio of the amount of cytosine, which is an example of a nucleic acid, to the amount of phenylalanine, which is an example of a protein.

  Focusing on the distribution of normal cells and cancer cells plotted in the graph, only the cancer cells are distributed in the range where the ratio of cytosine to tryptophan is 1.2 or more on the horizontal axis. Can be seen. In addition, it can be seen that only cancer cells are distributed in the range where the ratio of cytosine to phenylalanine exceeds 0.3 on the vertical axis.

  Therefore, in step S101 shown in FIG. 3, a threshold of 1.2 is set for the ratio of the amount of cytosine to the amount of tryptophan, and a threshold of 0.3 for the ratio of the amount of cytosine to the amount of phenylalanine. Thus, the presence of cancer cells can be examined based on the ratio between the amount of each nucleic acid detected from the Raman spectroscopic image and the amount of protein.

Example 4
FIG. 7 is a diagram showing an example of a graph included in the display image 304 displayed on the display unit 174 as a result of the examination. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

In the graph shown, the value on the horizontal axis is the ratio of the amount of cytosine, which is an example of a nucleic acid, to the amount of tryptophan, which is an example of a protein. Also, the value on the vertical axis is the amount of amide bond derived from the protein having the secondary structure of the β sheet detected from the amount of cytosine which is an example of nucleic acid and the peak light amount of Raman shift 1250 cm ⁻¹ “The amount of amide bond”. The amount of amide bonds gives an indication of the amount of protein.

  Focusing on the distribution of normal cells and cancer cells plotted in the graph, for the horizontal axis, cancer cells can be identified by the threshold value 1.2, but for the vertical axis, the threshold value at which cancer cells can be identified. It turns out that it has risen to 0.43. Therefore, by setting the threshold value 0.43 in step S101 shown in FIG. 3, the presence of cancer cells can be examined based on the ratio of the amount of cytosine to the amount of amide bond.

[Example 5]
FIG. 8 is a diagram showing an example of a graph included in the display image 305 displayed on the display unit 174 as a result of the examination. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

  In the graph shown, the value on the horizontal axis is the ratio of the amount of cytosine, which is an example of a nucleic acid, to the amount of symmetrical tryptophan, which is an example of a protein. The value on the vertical axis is the ratio of the amount of adenine, which is an example of a nucleic acid, to the amount of symmetrical tryptophan, which is an example of a protein.

  Focusing on the distribution of normal cells and cancer cells plotted in the graph, cancer cells can be identified by the threshold value 1.3 for the horizontal axis, and cancer cells at the threshold value 0.9 for the vertical axis. I see that I can do it. Therefore, by setting the threshold value 1.3 in step S101 shown in FIG. 3, the presence of cancer cells can be examined based on the ratio of cytosine to the amount of symmetrical tryptophan. The presence of cancer cells can be examined based on the ratio between the amount of and the amount of symmetrical tryptophan.

[Example 6]
FIG. 9 is a diagram showing an example of a graph included in the display image 306 displayed on the display unit 174 as a result of the examination. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

  In the graph of the figure, the value on the horizontal axis is the ratio of the amount of cytosine in the sample 101, which is an example of a nucleic acid, to the amount of protein whose index is the α-helix structure. The value on the vertical axis is the ratio of the amount of adenine, which is an example of a nucleic acid, to the amount of protein whose index is the α-helix structure.

  Focusing on the distribution of normal cells and cancer cells plotted in the graph, cancer cells can be identified by the threshold value 0.5 for the horizontal axis, but cancer cells are distributed alone for the vertical axis. It can be seen that there is no area.

  Therefore, in examining the presence of cancer cells in step S101 shown in FIG. 3, the threshold value 0 is better than calculating the ratio between the amount of adenine which is a nucleic acid and the amount of protein based on the α-helix structure. It is preferable to set .5 and test based on the ratio of the amount of cytosine, which is a nucleic acid, to the amount of a protein whose index is the α-helix structure.

[Example 7]
FIG. 10 is a diagram showing an example of a graph included in the display image 307 displayed on the display unit 174 as a result of the inspection. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

  In the graph shown, the value on the horizontal axis is the ratio of the amount of cytosine in the sample 101 as an example of a nucleic acid to the amount of amide bond as an example of a protein. The value on the vertical axis is the ratio of the amount of adenine, which is an example of a nucleic acid, to the amount of amide bond, which is an example of a protein. Focusing on the distribution of normal cells and cancer cells plotted in the graph, the cancer cells can be identified by the threshold of 0.45 for the horizontal axis, but cancer cells are distributed alone for the vertical axis. It can be seen that there is no area.

  Therefore, by setting the threshold value of 0.45 in step S101 shown in FIG. 3, the presence of cancer cells can be examined based on the ratio between the amount of cytosine and the amount of amide bond, but the amount of adenine, Even calculating the ratio to the amount of amide bond can not examine the presence of cancer cells.

[Example 8]
FIG. 11 is a diagram showing an example of a graph included in the display image 308 displayed on the display unit 174 as a result of the examination. In the illustrated graph, the ratio of the peak value of the nucleic acid to the peak value of the protein, which is output by the calculation unit 272, is plotted.

  In the graph shown, the value on the horizontal axis is the ratio of the amount of cytosine in the sample 101, which is an example of a nucleic acid, to the amount of phenylalanine, which is an example of a protein. The value on the vertical axis is the ratio of the amount of adenine, which is an example of a nucleic acid, to the amount of phenylalanine, which is an example of a protein. Focusing on the distribution of normal cells and cancer cells plotted in the graph, cancer cells can be identified by the threshold value 0.28 for the horizontal axis, but cancer cells are distributed alone for the vertical axis It can be seen that there is no area.

  Therefore, by setting the threshold value 0.28 in step S101 shown in FIG. 3, the presence of cancer cells can be examined based on the ratio of the amount of cytosine to the amount of β sheet-type amide, but the amount of adenine Even if the ratio of phenylalanine to the amount of phenylalanine is calculated, the presence of cancer cells can not be examined.

  From the examination of the test results shown in FIG. 4 to FIG. 11, in the sample 101 used in the above example, the ratio of the amount of cytosine, which is a nucleic acid, to the amount of phenylalanine, which is a protein, It was found that the difference with the cells was remarkable. Also, as shown in each graph, when different ratios are assigned to the vertical axis and the horizontal axis, the peak ratio of adenine and phenylalanine is assigned to one axis, and the peak ratio of cytosine to phenylalanine is assigned to the other axis It was found that the difference between normal cells and cancer cells is remarkable.

[Example 9]
FIG. 12 is a view showing a display image 309 showing the result of the examination by the examination apparatus 100 on the sample 101 containing cancer cells. The microscopic image of the cells in the figure is one that was imaged and stored in step S104 of FIG.

  The numerical values shown in the figure correspond to the order in which the examination was performed, and the positions of the numerals indicate the position of the region of interest in the examination. In addition, the numbers shown in white indicate that the test results at the relevant places were normal cells. On the other hand, the numbers shown in black indicate that the test results at the relevant places were cancer cells. As shown, in the sample 101 known to contain cancer cells, it can be seen that the peak ratio specific to the cancer cells is detected in a specific region of interest.

[Example 10]
FIG. 13 is a view showing a display image 310 showing the result of the examination by the examination apparatus 100 on the sample 101 which does not contain cancer cells. The microscopic image of the cells in the figure is one that was imaged and stored in step S104 of FIG. Also, the meanings of the numerals shown in the figure are the same as in FIG.

  As shown, in this sample 101, the peak ratio specific to cancer cells is not detected, and only the peak ratio of normal cells is detected. Thus, it can be seen that the sample 101 does not contain cancer cells.

  The numbers shown in italics in FIG. 12 and FIG. 13 indicate that the test results at the corresponding part could not be judged as normal cells or cancer cells. If all the regions of interest in the entire sample 101 result in such a test result, it means that the selection of the type of nucleic acid and protein for detecting the peak value was not appropriate.

  Therefore, in this case, other types of nucleic acids and proteins may be selected to detect peak values of the spectral waveform, and a test may be performed by setting a threshold adapted to such nucleic acids and proteins. In addition, significant detection results may be obtained by changing the detection conditions such as the wavelength of the excitation light to be irradiated to the sample 101 and performing the inspection.

  FIG. 14 is a flowchart showing another inspection procedure. In FIG. 14, the same steps as those in FIG. 3 are denoted by the same reference numerals and redundant description will be omitted.

  In the illustrated procedure, in step S111, in addition to the threshold to be compared in step S108, a second threshold regarding the width of the difference between the calculation result and the threshold is set. In addition, after the step S108, although the detection result exceeds the threshold set in the step S101, when the second threshold is not exceeded, the same for the purpose of improving the certainty of the cancer cell detection The step of executing another re-examination in which the examination condition has been changed is added to the region of interest (step S112).

  As a result, the region of interest for which the determination based on the examination result is suspended can be reduced, and the examination accuracy can be further improved. In addition, as a test | inspection condition changed in step S112, the change etc. of the nucleic acid used as the object which calculates peak ratio, and a protein can be illustrated, for example. In addition, the procedure may be further changed to change the wavelength of excitation light for detecting a spectrum.

  The above inspection method can be executed if a spectrum can be acquired, and therefore, may be incorporated into an existing spectrometer as a program for executing the inspection method. Also, in the examples described so far, the amounts of nucleic acid and protein were detected from the spectrum of Raman scattering. However, a specific measurement method can be selected from any measurement method that can detect the amount of amino acid or protein and calculate it as the ratio of the amount of nucleic acid to the amount of amino acid or protein. For example, spectra may be detected from CARS light generated by the CARS process (Coherent Anti-Stokes Raman Scattering). Infrared spectroscopy may be used instead of or in addition to using Raman spectroscopy or CARS light.

  Further, as described above, when calculating the ratio of the amount of nucleic acid used for the test to the amount of protein, the ease of determination may differ depending on the type of nucleic acid to be detected and the type of protein. . Therefore, the test apparatus 100 may switch the types of the nucleic acid and the protein whose amount is to be detected by the operation of the user.

  Furthermore, in the inspection apparatus 100, the method of displaying the inspection result on the display unit 174 may be switched. The display format may be graphed as shown in FIGS. 4 to 11, for example, or may be displayed superimposed on a microscopic image as shown in FIGS. In addition, character strings, colors, sounds, etc. may be displayed in combination. Furthermore, the display format may be selected by the user's operation.

  Still further, although the inspection apparatus 100 described above is formed by combining an optical microscope and a spectroscope, the spectrum can be detected noninvasively by, for example, an endoscope. Therefore, the cancer examination can be performed noninvasively by combining the endoscope which can perform spectrometry and the processing device 171.

  Furthermore, in the above example, the examination of the regions of interest one by one was taken as an example. However, the inspection apparatus 100 may be configured to simultaneously irradiate excitation light to a plurality of regions of interest and inspect the plurality of regions of interest in parallel. This can reduce the time required for cancer screening.

  As mentioned above, although this invention was demonstrated using embodiment, the technical scope of this invention is not limited to the range as described in the said embodiment. It is apparent to those skilled in the art that various changes or modifications can be added to the above embodiment. It is also apparent from the scope of the claims that the embodiments added with such alterations or improvements can be included in the technical scope of the present invention.

  The execution order of each process such as operations, procedures, steps, and steps in the apparatuses, systems, programs, and methods shown in the claims, the specification, and the drawings is particularly “before”, “preceding” It is to be noted that “it is not explicitly stated as“ etc. ”and can be realized in any order as long as the output of the previous process is not used in the later process. With regard to the flow of operations in the claims, the specification and the drawings, even if it is described using “first,” “next,” etc. for convenience, it means that it is essential to carry out in this order. It is not a thing.

Reference Signs List 100 inspection apparatus, 101 sample, 110 stage, 111 stage scanner, 120 objective optical system, 121 front objective lens, 122 rear objective lens, 130 light source device, 131, 132 light source, 139 combiner, 140 irradiation optical system, 141 galvano scanner , 142 scan lens, 143 primary image plane, 150 front detection system, 151 dichroic mirror, 152, 153, 162, 163 relay lens, 154, 164 band pass filter, 155, 165 spectroscope, 160 rear detection system, 161 reflection Mirror, 170 control unit, 171 processing unit, 172 mouse, 173 keyboard, 174 display unit, 271 detection unit, 272 calculation unit, 273 comparison unit, 274 storage unit, 275 output unit, 276 information output unit, 301, 302, 03,304,305,306,307,308,309,310 display image

Claims (19)

An acquisition unit for acquiring the amount of amino acids or proteins in the region of interest and the amount of nucleic acid;
An information output unit configured to output information according to a ratio of the amount of nucleic acid acquired by the acquisition unit to the amount of protein acquired by the acquisition unit.   The detection unit according to claim 1, further comprising: a detection unit configured to irradiate the excitation light to the region of interest and detect the amount of protein and the amount of nucleic acid in the region of interest based on the spectrum of the region of interest. Cancer testing device. And a threshold storage unit for storing a first predetermined threshold.
The information output unit has a comparison unit that compares the ratio with the first threshold, and a cancer cell based on whether the ratio exceeds the first threshold according to the comparison by the comparison unit. The cancer inspection apparatus according to claim 1, wherein the presence or absence is output as the information. The acquisition unit acquires the amount of amino acids in the region of interest as an indicator of the amount of protein, and acquires the amount of nucleic acid bases as an indicator of the amount of nucleic acid,
The cancer inspection apparatus according to any one of claims 1 to 3, wherein the information output unit outputs information according to a ratio of a detection amount of a nucleic acid base to a detection amount of an amino acid.   The cancer inspection apparatus according to claim 4, wherein the amino acid comprises an aromatic amino acid, the nucleobase comprises a cyclic amine, and the ratio is a ratio of a detected amount of the cyclic amine to a detected amount of the aromatic amino acid.   The cancer test apparatus according to claim 5, wherein the amino acid comprises at least one of tryptophan and phenylalanine, and the cyclic amine comprises at least one of cytosine and adenine.   The cancer inspection apparatus according to any one of claims 1 to 6, wherein at least one of the amount of amino acid or protein and the amount of nucleic acid is an amount detected by Raman spectroscopy.   The cancer test apparatus according to claim 7, wherein at least one of the amount of the amino acid or protein and the amount of the nucleic acid is detected by CARS microscopy.   The amount of nucleic acids comprises the amount of nucleic acids of a first type and the amount of nucleic acids of a second type different from the first type, the ratio being the ratio of the first to the amount of the amino acids or proteins. 9. The method according to any one of claims 1 to 8, comprising a first ratio which is the ratio of the amount of nucleic acids of the type and a second ratio which is the ratio of the amount of the nucleic acids of the second type to the amount of the amino acids or proteins. The cancer inspection device according to one item.   Detection of the amount of the nucleic acid of the second type and calculation of the second ratio when the calculated ratio is larger than a predetermined second threshold which is larger than a predetermined first threshold The cancer inspection apparatus according to claim 9, which omits.   The cancer inspection apparatus according to any one of claims 1 to 10, comprising optically observing the region of interest with light in the infrared band.   At least one of the amount of the amino acid or protein and the amount of the nucleic acid is detected by an average value of measurements in a plurality of small regions located inside the region of interest, narrower than the region of interest, and separated from each other The cancer inspection device according to any one of claims 1 to 11. The amount of the amino acid or protein and the amount of the nucleic acid may be determined in each of a plurality of regions of interest included in a region of interest wider than the region of interest.
Calculating the ratio of the amount of said detected nucleic acid to the amount of said detected amino acid or protein,
The cancer inspection apparatus according to any one of claims 1 to 12, wherein the presence or absence of a cancer cell is inspected based on whether or not the calculated ratio exceeds a predetermined first threshold.   The said acquisition part irradiates excitation light to several places of a biological body, and acquires the quantity of several amino acids or proteins based on several spectrum, and the quantity of several nucleic acids in any one of Claim 1 to 13 Cancer inspection equipment.   The cancer inspection apparatus according to any one of claims 1 to 14, further comprising a display unit that displays the information output by the information output unit.   The cancer inspection apparatus according to claim 15, wherein the display unit displays a two-axis graph indicating two different ratios of at least one of the nucleic acid and the amino acid or protein.   The cancer inspection device according to claim 14 or 16, wherein the display image of the display unit identifies whether the value of the ratio is larger than a previously prepared threshold value.   The cancer inspection apparatus according to any one of claims 14 to 17, wherein the display unit displays a plurality of output values of the information output unit in a distinguishable state. Obtaining an amount of amino acid or protein and an amount of nucleic acid in a region of interest of the living body;
And an output step of outputting information according to a ratio of the amount of nucleic acid acquired in the acquisition step to the amount of amino acid or protein acquired in the acquisition step.