JP2004354346A - Measuring device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a measuring device capable of independently optimizing a statistic analysis value acquiring optical path and a microscopic image acquiring optical path, and further acquiring a statistic analysis value and a microscopic image at the same time. <P>SOLUTION: This device comprises a light detector 6 for detecting the intensity of fluorescence from a sample 5 and outputting a light intensity value, a statistic analysis device 7 for statistically analyzing the light intensity value detected by the detector 6, and a microscopic image measuring device 9 for acquiring the microscopic image of the sample 5. The statistic analysis value acquiring optical path and the microscopic image acquiring optical path for acquiring the microscopic image from the sample 5 to the statistic analysis device 7 are provided independently. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a measurement device for statistically analyzing a measurement signal relating to a specific portion of a sample.
[0002]
[Prior art]
2. Description of the Related Art Conventionally, as a measurement device that statistically analyzes a measurement signal relating to a specific portion of a sample, a measurement device using, for example, fluorescence correlation spectroscopy that performs measurement using fluorescence is known.
[0003]
Fluorescence correlation spectroscopy using this method is discussed in, for example, Non-Patent Document 1, Non-Patent Document 2, and Patent Document 1.
[0004]
Such a fluorescence correlation spectroscopy device includes, for example, a light source device, a spectroscopy device, a light collection device, a sample holding device, a light detection device, and a statistical analysis device.
[0005]
In such a configuration, when excitation light is emitted from the light source device, the excitation light is applied to the sample via the spectroscopic device and the light condensing device, and the sample generates fluorescence. Further, the fluorescence from the sample enters the light detection device via the light condensing device and the spectroscopic device, and causes the light detection device to output a light intensity value indicating the intensity of the fluorescence. Then, this light intensity value is input to a statistical analysis device, statistical analysis is performed, and output as a light intensity statistical analysis value.
[0006]
Conventionally, statistical analysis of the fluorescence intensity from a sample has been performed in this way.Furthermore, in the fluorescence correlation analysis, the fluctuation of the fluorescence intensity of the particles is measured, and the result is analyzed to obtain an autocorrelation function. The diffusion time, concentration, size, and the like of the target particle are estimated from the autocorrelation function.
[0007]
By the way, such a measuring device is used in combination with a microscope so as to enhance the accuracy of a measurement position of a statistical analysis value on a sample. That is, when actually performing a measurement for obtaining a statistical analysis value, it may be necessary to arrange a measurement point at a specific position on a sample. For example, when a cell is used as a sample, the center of the nucleus of the cell may be measured. In such a case, first set the sample roughly on the optical path for statistical analysis value measurement, fix the measurement point of the statistical analysis value at the center of the field of view, switch from this state to the optical path for microscope observation, and change the microscope image. The sample was moved while watching so as to adjust the nucleus of the cell to the center of the visual field, and from this state, the optical path was switched again to the statistical analysis value measurement to measure the statistical analysis value.
[0008]
[Patent Document 1]
Japanese Unexamined Patent Publication No. Hei 11-502608
[Non-patent document 1]
"" Fluorescence correlation spectroscopy "R. Rigler, ES Elson (eds.) Springer (Berlin)"
[0010]
[Non-patent document 2]
Masataka Kaneshiro, “Protein Nucleic Acid Enzyme” (1999) Vol. 9, P1433-137
[0011]
[Problems to be solved by the invention]
However, in the conventional apparatus, when trying to measure a statistical analysis value at a specific position of a sample, an extremely cumbersome procedure must be taken, which causes a problem that work efficiency becomes extremely poor.
[0012]
Further, in the conventional apparatus, it is necessary to switch the respective optical paths between the measurement of the statistical analysis value and the observation with the microscope, so that the microscope image cannot be observed during the measurement of the statistical analysis value. For this reason, even if the intended position in the sample deviates from the measurement point due to movement of the sample during the measurement of the statistical analysis value, this cannot be known, and the measurement of the statistical analysis value during this time is not possible. There is also a problem that the accuracy becomes accurate and a highly accurate statistical analysis value cannot be measured.
[0013]
Furthermore, since the optical path for acquiring the statistical analysis value and the optical path for acquiring the microscope image overlap each other, in this overlapping portion, it is necessary to optimize for acquiring the statistical analysis value, optimize for acquiring the microscope image, or perform the optimization. There is also a problem that the accuracy of either the statistical analysis value or the microscope image has to be given up to some extent because the choice of whether to adopt a compromise is made.
[0014]
The present invention has been made in view of the above circumstances, the optical path for obtaining a statistical analysis value and the optical path for obtaining a microscope observation image can be independently optimized, and the statistical analysis value and the microscope observation image can be obtained simultaneously. It is an object of the present invention to provide a measuring device that can perform the measurement.
[0015]
[Means for Solving the Problems]
The invention according to claim 1 is an optical detection unit that optically detects a measurement signal related to a specific portion of a sample, a statistical analysis unit that statistically analyzes a measurement value detected by the optical detection unit, A microscope observation image acquisition unit for acquiring a microscope observation image, and a microscope analysis image acquisition optical path for acquiring the microscope analysis image from the sample and the statistical analysis value acquisition optical path from the sample to the statistical analysis unit, respectively. It is characterized by being provided.
[0016]
According to a second aspect of the present invention, in the first aspect of the present invention, the microscope observation image obtaining means includes an imaging means for imaging the sample image.
[0017]
According to a third aspect of the present invention, in the first aspect of the present invention, the microscope observation image acquiring unit detects light intensity of the fluorescence from the sample and outputs a light intensity value, and light on the sample is obtained. A measuring point moving means for moving the position of the measuring point of the intensity and outputting position data of the measuring point; and a microscope based on the measuring point position data of the measuring point moving means and the light intensity value from the light intensity detecting means. A microscope observation image generating means for generating an observation image.
[0018]
As a result, according to the present invention, the measurement signal relating to the specific portion of the sample goes to the optical path to the optical detection means, and the other part goes to the optical path of the microscope observation image acquisition means. The optical detection means outputs a measurement value relating to the specific part. The statistical analysis means analyzes the measured value and outputs a light intensity statistical analyzed value which is a statistical analyzed value based on the measured value. On the other hand, a microscope observation image of the sample is output from the microscope observation image acquisition means. In this case, the light intensity statistical analysis value and the microscope observation image can be measured simultaneously. At this time, the optical path for obtaining a statistical analysis value from the sample to the statistical analysis means and the optical path for obtaining a microscope image for obtaining a microscope observation image are independent of each other, and do not overlap. Therefore, the optical path for acquiring the statistical analysis value and the optical path for acquiring the microscope image can be independently optimized.
[0019]
Further, according to the present invention, the microscope observation image generation means generates a scanning fluorescence image as a microscope observation image based on the light intensity value from the light intensity detection means and the measurement point position data from the measurement point moving means. Also in this case, the optical path for obtaining a statistical analysis value from the sample to the statistical analysis means and the optical path for obtaining a microscope image for obtaining a microscope observation image are independent of each other, and do not overlap. Therefore, the optical path for acquiring the statistical analysis value and the optical path for acquiring the microscope image can be independently optimized.
[0020]
BEST MODE FOR CARRYING OUT THE INVENTION
Hereinafter, embodiments of the present invention will be described with reference to the drawings.
[0021]
(First Embodiment)
FIG. 1 shows a schematic configuration of a measuring apparatus to which the first embodiment of the present invention is applied.
[0022]
In the drawing, reference numeral 1 denotes a light source device, and this light source device 1 uses, for example, an argon laser having an oscillation wavelength of 488 mm as an excitation light generation source.
[0023]
On the optical path of the light emitted from the light source device 1, a spectroscopic device 2 is arranged. For the spectroscopic device 2, for example, a dichroic mirror that reflects light having a wavelength of 500 mm or less and transmits light having a wavelength of 500 mm or more is used.
[0024]
A first light condensing device 3 and a sample holding device 4 are arranged on the reflection-side optical path of the spectroscopic device 2. Here, for the first light collecting device 3, for example, an objective lens having a magnification of 40 and a numerical aperture of 1.2 is used. The sample holding device 4 is a sample stage for a microscope, and a sample 5 is placed on the sample holding device 4.
[0025]
In the transmission side optical path of the spectroscopic device 2, a light detection device 6 as an optical detection means is disposed. The photodetector 6 uses an avalanche photodiode. The light detection device 6 optically detects a measurement signal relating to a specific portion of the sample 5 incident through the spectroscopic device 2. Here, the light detection device 6 detects the fluorescence from the sample 5, and the intensity of the fluorescence at this time. Is output according to the light intensity.
[0026]
The photodetector 6 is connected to a statistical analyzer 7 as a statistical analyzer. The statistical analysis device 7 is provided with a correlation function analysis board for performing a statistical analysis, and statistically analyzes the light intensity value input from the photodetection device 6 to obtain a statistical analysis value. Output.
[0027]
In this case, the optical path from the sample 5 through the photodetector 6 to the statistical analyzer 7 is defined as an optical path A for acquiring a statistical analysis value.
[0028]
A second light collecting device 8 is disposed at a position facing the first light collecting device 3 with the sample holding device 4 interposed therebetween. For the second light collecting device 8, for example, an objective lens with a magnification of 10 times is used. Further, a microscope image measuring device 9 as a microscope observation image acquiring means is arranged in an optical path passing through the second light collecting device 8. The microscope image measuring device 9 uses an imaging unit for imaging a sample image, for example, a CCD camera. The microscope image measuring device 9 measures a microscope observation image of the sample 5 obtained through the second light collecting device 8.
[0029]
In this case, the optical path from the sample 5 to the microscope image measuring device 9 via the second light condensing device 8 is the optical path B for acquiring a microscope image.
[0030]
On the other hand, a memory 21 and a monitor 24 are connected to the microscope image measuring device 9. An XYZ drive device 22 is connected to the sample holding device 4. A CPU 23 is connected to the memory 21, the monitor 24, and the XYZ drive device 22. An input device 25 such as a mouse or a touch panel is connected to the monitor 24.
[0031]
In this embodiment, when cells are observed as the sample 5, for example, the inside of the cells is labeled with a rhodamine green dye, and the cell membrane is labeled with rhodamine green modified with an alkyl chain having about 18 carbon atoms. Can be In addition, observation may be performed with a fusion protein such as GFP or YFP. Further, optical elements such as lenses, pinholes, mirrors, and various filters (not shown) are appropriately arranged in the optical path A for acquiring a statistical analysis value and the optical path B for acquiring a microscope image. Further, it is desirable that the optical system for detecting the fluorescence intensity by the light detection device 6 is a confocal optical system.
[0032]
Next, an embodiment configured as described above will be described.
[0033]
When the excitation light is emitted from the light source device 1, the excitation light is reflected by the spectroscopic device 2, passes through the first light condensing device 3, and irradiates the sample 5. The sample 5 is excited by irradiation with the excitation light and emits fluorescence. Fluorescence emitted from the sample 5 passes through the first light condensing device 3, passes through the spectroscopic device 2, and enters the light detection device 6.
[0034]
The light detection device 6 outputs a light intensity value corresponding to the intensity of the incident fluorescence. This light intensity value is sent to the statistical analysis device 7.
[0035]
The statistical analysis device 7 statistically analyzes the light intensity value using the correlation function analysis board to obtain a light intensity statistical analysis value, and outputs the statistical analysis value to the outside.
[0036]
On the other hand, a microscope observation image of the sample 5 is taken out via the second light condensing device 8 and measured by the microscope image measuring device 9. Then, the microscope observation image of the sample 5 measured by the microscope image measurement device 9 is displayed on the monitor 24.
[0037]
In this way, the optical path A for obtaining a statistical analysis value from the sample 5 via the photodetector 6 to the statistical analyzer 7 and the microscopic image measuring device 9 from the sample 5 via the second condenser 8 Can be formed independently so that the optical path B for acquiring a microscopic image does not overlap at all. Therefore, it is possible to optimize the optical path A for acquiring a statistical analysis value and the optical path B for acquiring a microscopic image independently. it can.
[0038]
This means that, for example, in the case of fluorescence correlation measurement, the first light condensing device 3 having a large aperture diameter is required. For this reason, the magnification becomes large. However, since the first light collecting device 3 is independent of the second light collecting device 8, the second light collecting device 8 is No limitation by 3 Thereby, for example, if the microscope observation image is acquired by the microscope image measurement device 9 using the objective lens with a small magnification for the second light collection device 8, the sample 5 can be observed in a wide range. .
[0039]
Next, the case where the observation of the microscope and the measurement of the light intensity statistical analysis value are performed in association with each other will be described. First, an observation image of the sample 5 is acquired through the optical path B for acquiring a microscope image and displayed on the monitor 24. Here, the position of the sample 5 on the sample holding device 4 and the position of the sample screen on the monitor 24 are stored in the scale 21. Next, one point to be measured for the sample 5 on the monitor 24 is designated by the input device 25. The designated position information on the monitor 24 is managed by the CPU 23 together with the information in the memory 21, and controls the XYZ drive device 22. As a result, the sample holding device 4 is moved in the XYZ directions, but stops when the focal position of the first light condensing device 3 matches the measurement point specified by the input device 25. Then, light detection at the measurement point is performed by the statistical analysis value acquisition optical path A. Here, when the designated measurement points are two or more, the light detection by the light detection device 6 and the light detection by the sample holding device 4 are performed so that the light detection in the statistical analysis value acquisition optical path A is sequentially performed at each measurement point. The CPU 23 controls so that the alignment of the measurement points is coordinated.
[0040]
Thereafter, the analysis value of the statistical analyzer 7 is displayed on the monitor 24 via the CPU 23 together with the position information and / or image information from the memory 21.
[0041]
In this manner, observation of the microscope and measurement of the light intensity statistical analysis value can be performed in association with each other. Further, even in such a case, all of the fluorescent light collected by the first light collecting device 3 can be guided to the light detecting device 6, and at the same time, the fluorescent light collected by the second light collecting device 8 can be obtained. Can be guided to the microscope image measuring device 9 as a microscope observation image, so that at the time of these measurements, in particular, it is possible to secure a sufficient light amount for the measurement of the light intensity statistical analysis value, and to obtain a highly accurate measurement result. Obtainable.
[0042]
Here, as a specific example, a case where the sample 5 is a cell and the fluidity of a cell membrane is measured will be described. In this case, a fluorescence image (microscope observation image) of the sample 5 is acquired by the microscope image measurement device 9 via the second light condensing device 8, and the position of the cell membrane is confirmed from the fluorescence image. Then, the intensity of the fluorescence is detected at the confirmed position by the light detection device 6 via the first light condensing device 3, and the fluorescence correlation measurement is performed using the statistical analysis device 7. In this case, the excitation light is condensed at the measurement point of the fluorescence correlation, and strong fluorescence and scattered light are generated from the measurement point. Therefore, the position of the measurement point is determined from the fluorescence image acquired by the microscope image measurement device 9. You can easily check. Then, the fluidity of the cell membrane can be obtained while confirming the position of the cell membrane and the position of the measurement point of the fluorescence correlation from the fluorescence image.
[0043]
Further, since the focus position of the excitation light for acquiring the light intensity statistical analysis value can be known from the microscope observation image acquired by the microscope image measuring device 9, the statistical analysis value acquiring optical path A and the microscope image acquiring optical path B Can be easily adjusted.
[0044]
Furthermore, since the optical path A for acquiring a statistical analysis value and the optical path B for acquiring a microscope image are independent of each other, the angles and positions of both optical paths can be set variously. Thereby, for example, the amount of light incident on the optical path B for acquiring a microscope image from the light source device 1 can be adjusted.
[0045]
Note that the configuration of the above-described embodiment can be variously modified and changed. For example, illumination for photographing a microscope observation image may be added to the optical path B for acquiring a microscope image. Alternatively, when the sample 5 emits sufficient fluorescence, the light source device 1 and the spectroscopic device 2 can be omitted. Further, a special effect can be expected by using a plurality of excitation optical systems or a plurality of detection optical systems, or by processing detected signals in various ways.
[0046]
As described above, according to the first embodiment, the light intensity statistical analysis value and the microscope observation image can be measured simultaneously, and the statistical analysis value acquisition optical path from the sample 5 to the light detection device 6 can be obtained. A and the optical path B for acquiring a microscope image from the sample 5 to the microscope image measuring device 9 can be prevented from overlapping at all, and the optical path A for acquiring a statistical analysis value and the optical path B for acquiring a microscope image can be independently optimized. it can.
[0047]
(Second embodiment)
Next, a second embodiment of the present invention will be described.
[0048]
FIG. 2 shows a schematic configuration of the second embodiment, and the same parts as those in FIG. 1 are denoted by the same reference numerals.
[0049]
In this case, the microscope observation image acquisition means is configured as follows.
[0050]
In the figure, reference numeral 10 denotes a second light source device, and this light source device 10 also uses, for example, an argon laser having an oscillation wavelength of 488 mm as an excitation light generation source.
[0051]
On the optical path from the light source device 10, a second spectroscopic device 11 is arranged. A dichroic mirror that reflects light having a wavelength of 500 mm or less and transmits light having a wavelength of 500 mm or more, for example, is also used in the spectroscopic device 11.
[0052]
In the reflected light path of the spectroscopic device 11, a second measuring point moving device 12 as a measuring point moving means, the second condensing device 8 described in the first embodiment, and the sample holding device 4 are arranged. . The second measuring point moving device 12 moves (scans) the position of the measuring point of the light intensity on the sample 5 and outputs the position data of the measuring point. For example, a galvano scanner is used.
[0053]
In the transmitted light path of the spectroscopic device 11, a second light detecting device 13 is disposed as light intensity detecting means. The photodetector 13 uses a photomultiplier tube. The light detection device 13 detects the fluorescence from the sample 5 incident through the spectroscope 11, and outputs a light intensity value c2 indicating the intensity of the fluorescence at this time.
[0054]
An image processing device 14 is connected to the light detection device 13 as a microscope observation image generation unit. Further, the second measurement point moving device 12 is connected to the image processing device 14.
[0055]
As the image processing device 14, a personal computer is used. The image processing device 14 generates a scanning fluorescence image as a microscope observation image based on the measurement point position data of the second measurement point moving device 12 and the light intensity value from the light detection device 13.
[0056]
On the other hand, a first measuring point moving device 31 is connected between the first light collecting device 3 and the spectroscopic device 2.
[0057]
A monitor 33 is connected to the image processing device 14. Further, a CPU 32 is connected to the monitor 33, the first measuring point moving device 31, and the second measuring point moving device 12. An input device 34 such as a mouse or a touch panel is connected to the monitor 33.
[0058]
Also in this case, the optical path from the sample 5 to the statistical analysis device 7 via the photodetection device 6 is defined as an optical path A for acquiring a statistical analysis value, and the second measurement point moving device 12 and the light detection device 13 The optical path to the image processing apparatus 14 via the optical path is a microscope image acquiring optical path B.
[0059]
In this embodiment, when cells are observed as the sample 5, for example, the inside of the cells is labeled with a rhodamine green dye, and the cell membrane is labeled with rhodamine green modified with an alkyl chain having about 18 carbon atoms. Used. In addition, optical elements such as lenses, pinholes, mirrors, and various filters are appropriately disposed in each optical path such as an optical path for statistical analysis value measurement.
[0060]
Others are the same as FIG.
[0061]
Next, an embodiment configured as described above will be described.
[0062]
In this case, when the excitation light a1 is emitted from the light source device 1, the excitation light a1 is reflected by the spectroscopic device 2, passes through the first light condensing device 3, and irradiates the sample 5. The sample 5 is excited by irradiation with the excitation light and emits fluorescence b1. The fluorescent light b1 emitted from the sample 5 passes through the first light condensing device 3, passes through the spectroscopic device 2, and is incident on the light detecting device 6.
[0063]
The light detection device 6 outputs a light intensity value c1 corresponding to the intensity of the incident fluorescence b1. This light intensity value is sent to the statistical analysis device 7.
[0064]
The statistical analysis device 7 statistically analyzes the light intensity value using the correlation function analysis board to obtain a light intensity statistical analysis value, and outputs the statistical analysis value to the outside.
[0065]
On the other hand, when the excitation light a2 is emitted from the light source device 10, the excitation light a2 is reflected by the spectroscopy device 11, passes through the second condensing device 8, and irradiates the sample 5. The sample 5 is excited by irradiation with the excitation light a2 and emits fluorescence b2. The fluorescent light b2 emitted from the sample 5 passes through the second light condensing device 8, passes through the spectroscopic device 11, and enters the light detecting device 13. The light detection device 13 outputs a light intensity value c2 corresponding to the intensity of the fluorescence.
[0066]
In this case, the measurement position on the sample 5 is moved (scanned) by the second measurement point moving device 12. Then, the intensity of the fluorescence at each of the moved positions is detected by the light detection device 13, and the corresponding light intensity value c2 is output.
[0067]
Thereby, the image processing device 14 generates and outputs a scanning fluorescence image as a microscope observation image based on the measurement point position data of the second measurement point moving device 12 and the light intensity value c2 from the light detection device 13. .
[0068]
Also in this case, the optical path for statistical analysis value measurement from the sample 5 to the photodetector 6 and the optical path for scanning fluorescent image observation from the sample 5 to the photodetector 13 can be configured not to overlap at all. The optical path for statistical analysis value measurement and the optical path for scanning fluorescent image observation can be independently optimized.
[0069]
This means that, for example, in the case of fluorescence correlation measurement, the first light condensing device 3 having a large aperture diameter is required. For this reason, the magnification becomes large. However, since the first light-collecting device 3 is independent of the second light-collecting device 8, scanning fluorescent image observation is not performed by the first light-collecting device 3. Not subject to restrictions. Thus, for example, if the scanning fluorescence image is acquired by the image processing device 14 using an objective lens having a small magnification as the second light condensing device 8, the sample 5 can be observed in a wide range.
[0070]
Next, the case where the observation of the scanning fluorescent image and the measurement of the light intensity statistical analysis value are performed in association with each other will be described. First, the monitor 33 displays a scan image of the sample 5 acquired based on the measurement point position data of the second measurement point moving device 12 on the microscope image acquisition optical path B. Next, one point to be measured for the sample 5 on the monitor 33 is designated by the input device 34. The designated position information on the monitor 33 is managed by the CPU 32 and controls the first measuring point moving device 31. As a result, the focal position of the first light condensing device 3 is moved to the designated measurement point, and light detection at the measurement point is performed by the statistical analysis value acquisition optical path A.
[0071]
In this way, the observation of the scanning fluorescent image and the measurement of the light intensity statistical analysis value can be performed in association with each other. Further, even in such a case, all of the fluorescent light collected by the first light collecting device 3 can be guided to the light detecting device 6, and at the same time, the fluorescent light collected by the second light collecting device 8 can be obtained. Can be guided to the photodetector 13. Thereby, at the time of these measurements, in particular, a light amount sufficient for measuring the light intensity statistical analysis value can be secured, and a highly accurate measurement result can be obtained.
[0072]
Here, as a specific example, a case where the sample 5 is a cell and the fluidity of a cell membrane is measured will be described. In this case, the excitation light is condensed at the measurement point of the fluorescence correlation, and strong fluorescence and scattered light are generated from the measurement point. Therefore, the position of the corresponding measurement point is determined from the scanning fluorescence image acquired from the image processing device 14. You can easily check. Then, the fluidity of the cell membrane can be obtained while confirming the position of the cell membrane and the position of the measurement point of the fluorescence correlation from the scanned fluorescence image.
[0073]
Further, since the focal position of the excitation light for acquiring the light intensity statistical analysis value can be known from the scanning fluorescence image acquired from the image processing device 14, the optical path A for acquiring the statistical analysis value and the optical path B for acquiring the microscope image can be determined. Alignment can be easily performed.
[0074]
Furthermore, since the optical path A for acquiring a statistical analysis value and the optical path B for acquiring a microscope image are independent of each other, the angles and positions of both optical paths can be set variously. Thereby, for example, the amount of light incident on the optical path B for acquiring a microscope image from the light source device 1 can be adjusted.
[0075]
Note that the configuration of the above-described embodiment can be variously modified and changed. For example, it is useful to optimize the components of the optical path A for acquiring a statistical analysis value and the components of the optical path B for acquiring a microscope image to different dyes. When the sample 5 emits sufficient fluorescence, the light source device 1 and the spectroscopic device 2 can be omitted. Furthermore, a special effect can be expected by using a plurality of excitation optical systems or a plurality of detection optical systems, or by processing detected signals in various ways.
[0076]
As described above, according to the second embodiment, the light intensity statistical analysis value and the scanning fluorescence image can be measured simultaneously, and the statistical analysis value acquisition optical path from the sample 5 to the photodetector 6 can be obtained. A and the optical path B for acquiring a microscope image from the sample 5 to the photodetector 13 can be prevented from overlapping at all, so that the optical path A for acquiring a statistical analysis value from the sample 5 to the statistical analyzer 7 via the photodetector 6 Then, the optical path B for acquiring a microscope image from the sample 5 to the image processing device 14 via the second measuring point moving device 12 and the light detecting device 13 can be independently optimized.
[0077]
In addition, the present invention is not limited to the above-described embodiment, and can be variously modified in an implementation stage without departing from the spirit of the invention.
[0078]
Further, the embodiments include inventions at various stages, and various inventions can be extracted by appropriately combining a plurality of disclosed constituent features. For example, even if some components are deleted from all the components shown in the embodiments, the problem described in the column of the problem to be solved by the invention can be solved, and the problem described in the column of the effect of the invention can be solved. In the case where the effect described above is obtained, a configuration from which this configuration requirement is deleted can be extracted as an invention.
[0079]
【The invention's effect】
As described above, according to the present invention, the optical path for acquiring a statistical analysis value and the optical path for acquiring a microscope observation image can be independently optimized, and the measurement can also simultaneously acquire the statistical analysis value and the microscope observation image. Equipment can be provided.
[Brief description of the drawings]
FIG. 1 is a diagram showing a schematic configuration of a first embodiment of the present invention.
FIG. 2 is a diagram showing a schematic configuration of a second embodiment of the present invention.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Light source device 2 ... Spectroscopy device 3 ... First condensing device 4 ... Sample holding device 5 ... Sample 6 ... Photodetection device 7 ... Statistical analysis device 8 ... Second condensing device 9 ... Microscope image measuring device 10 ... Light source device 11 Spectroscopic device 12 Second measuring point moving device 13 Photodetector 14 Image processing device 21 Memory 22 XYZ drive device 23 CPU
24 monitor 25 input device 31 first measurement point moving device 32 CPU
33 monitor 34 input device A optical path for obtaining statistical analysis values B optical path for obtaining a microscope image

Claims (3)

Optical detection means for optically detecting a measurement signal relating to a specific portion of the sample,
Statistical analysis means for statistically analyzing the measurement value detected by the optical detection means,
Microscope observation image acquisition means for acquiring a microscope observation image of the sample,
A measuring apparatus, wherein an optical path for obtaining a statistical analysis value from the sample to the statistical analysis means and an optical path for obtaining a microscope image for obtaining the microscope observation image are provided independently of each other. The measurement apparatus according to claim 1, wherein the microscope observation image acquisition unit includes an imaging unit that captures the sample image. The microscope observation image acquisition means includes:
Light intensity detecting means for detecting the intensity of fluorescence from the sample and outputting a light intensity value, and measuring point moving means for moving the position of the light intensity measurement point on the sample and outputting position data of the measurement point 2. A microscope observation image generating means for generating a microscope observation image based on measurement point position data of the measurement point moving means and a light intensity value from the light intensity detection means. Measuring device.

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