Optical microscope and performance assessment method

Abstract

This optical microscope includes an objective lens, a light source that outputs excitation light, and an excitation output measurement unit that measures excitation output, the optical microscope comprising: an image-formation performance measurement unit that measures image-formation performance; and an assessment unit that assesses the performance of the optical microscope on the basis of the results of measuring the image-formation performance and the results of measuring the excitation output.

Description

Optical microscope and performance evaluation method

[0001] The disclosure herein relates to an optical microscope and a performance evaluation method.

[0002] While images acquired by optical microscopes have primarily been used to observe morphology, they are also increasingly being used for quantitative analysis. In recent years, advances in computer analysis technologies such as AI (Artificial Intelligence) have led to the increasing use of image data in omics analysis and other fields. As a result, the quantitative nature of the data used in analysis has become increasingly important.

[0003] In a fluorescence microscope, the intensity of the excitation light source (excitation output) and detection sensitivity are measured, and the excitation light source intensity and image signals are corrected based on the measured values, thereby quantifying the data.

[0004] For example, a known technique adjusts the light source intensity based on the measurement value of a power meter and adjusts the exposure time based on the measured luminance value of a calibration sample (see, for example, Patent Document 1). Another known technique measures fluctuations in light source intensity using a reflective surface attached to a filter cube, measures fluctuations in the optical path using a calibration sample, and corrects the fluorescence image luminance by image processing as necessary (see, for example, Patent Document 2).

[0005] JP 2022-119771 A Patent No. 5593221 A

[0006] Although detection sensitivity can also be reduced by abnormalities (including degradation) in imaging performance (spatial resolution), conventional techniques have evaluated the reduction in detection sensitivity without taking abnormalities in imaging performance into consideration. In other words, the evaluation was not conducted separately from abnormalities in imaging performance. As a result, it was not possible to accurately identify the cause of device abnormalities affected by imaging performance.

[0007] An object according to one aspect of the present invention is to provide a technique that makes it possible to isolate and evaluate the influence of abnormalities in imaging performance, and to accurately identify the cause of the problem.

[0008] An optical microscope according to one aspect of the present invention is an optical microscope including an objective lens, a light source that outputs excitation light, and an excitation output measurement unit that measures the excitation output, and further includes an imaging performance measurement unit that measures imaging performance, and a judgment unit that judges the performance of the optical microscope based on the measurement results of the imaging performance and the measurement results of the excitation output.

[0009] An optical microscope according to another aspect of the present invention is an optical microscope including an objective lens, a detector that detects light from a specimen, and a detection sensitivity measurement unit that measures detection sensitivity, and further includes an imaging performance measurement unit that measures imaging performance, and a judgment unit that judges the performance of the optical microscope based on the measurement results of the imaging performance and the measurement results of the detection sensitivity.

[0010] A method according to one aspect of the present invention is a performance evaluation method for evaluating the performance of an optical microscope, and includes measuring excitation output, measuring imaging performance, and evaluating the performance of the optical microscope based on the measurement results of the excitation output and the measurement results of the imaging performance.

[0011] A method according to another aspect of the present invention is a performance determination method for determining the performance of an optical microscope, which includes measuring the detection sensitivity of a detector that detects light from a specimen, measuring imaging performance, and determining the performance of the optical microscope based on the measurement results of the detection sensitivity and the measurement results of the imaging performance.

[0012] A method according to another aspect of the present invention is a performance evaluation method for evaluating the performance of an optical microscope, which includes measuring excitation output, measuring the detection sensitivity of a detector that detects light from a specimen, measuring imaging performance, and evaluating the performance of the optical microscope based on the measurement results of the excitation output, the measurement results of the detection sensitivity, and the measurement results of the imaging performance.

[0013] An optical microscope according to another aspect of the present invention comprises a light source that outputs excitation light, an objective lens, an imaging performance measurement unit that measures imaging performance, and an excitation output measurement unit that measures excitation output, and the excitation output measurement unit is built into the microscope body of the optical microscope.

[0014] An optical microscope according to another aspect of the present invention comprises an objective lens, a detector that detects light from a specimen, an imaging performance measurement unit that measures imaging performance, and a detection sensitivity measurement unit that measures detection sensitivity, and the detection sensitivity measurement unit is built into the microscope body of the optical microscope.

[0015] According to the above-described aspect, it is possible to evaluate the influence of abnormalities in imaging performance separately, and to accurately identify the cause of the problem.

[0016] FIG. 1 is a diagram illustrating the configuration of an optical microscope according to a first embodiment. FIG. 2 is a diagram illustrating a functional configuration related to a performance determination function of an optical microscope according to the first embodiment. FIG. 3 is a diagram illustrating an example of an abnormality cause (NG cause) stored in a storage unit according to the first embodiment. FIG. 4 is a diagram illustrating an example of a functional configuration related to a performance determination function of an optical microscope according to a second embodiment. FIG. 5 is a diagram illustrating an example of an abnormality cause (NG cause) stored in a storage unit according to the second embodiment. FIG. 6 is a diagram illustrating an example of a functional configuration related to a performance determination function of an optical microscope according to a third embodiment. FIG. 7 is a flowchart illustrating processing related to the performance determination function performed in an optical microscope. FIG. 8 is a diagram illustrating determination results for each measurement result, abnormality causes (NG causes) selected based on the determination results, and measures to be taken for the abnormality causes, according to a third embodiment. FIG. 9 is a diagram illustrating an example of a microscope main body and an excitation output measurement unit. FIG. 10 is a diagram illustrating an example of a detection sensitivity measurement unit. FIG. 11 is a diagram illustrating an example of an imaging performance measurement unit. FIG. 12 is a diagram illustrating an example of measuring imaging performance using an edge chart specimen. FIG. 13 is a diagram illustrating an edge chart specimen. FIG. 14 is a diagram illustrating an example of a display of evaluation results. 1 is a diagram illustrating a determination result for each measurement result, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to a first modification of the third embodiment. FIG. 2 is a diagram illustrating a determination result for each measurement result, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to a first modification of the second embodiment. FIG. 3 is a diagram illustrating a determination result for each measurement result, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to a first modification of the first embodiment. FIG. 4 is a diagram illustrating a determination result for each measurement result of irradiation density and excitation output, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to a second modification of the second or third embodiment. FIG. 5 is a diagram illustrating a determination result for each measurement result of irradiation density and detection sensitivity, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to a second modification of the third embodiment. FIG. 10 is a diagram illustrating the judgment results for each measurement result of the irradiation density and the imaging performance, the cause of abnormality (NG cause) selected based on the judgment results, and the countermeasure for the cause of abnormality, according to the second modification of the second or third embodiment.10 is a diagram illustrating an example of a determination result for each measurement result of irradiation density, imaging performance, and excitation output, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to a second modification of the second or third embodiment.

[0017] Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

[0018] 1 is a diagram illustrating the configuration of an optical microscope according to a first embodiment. The optical microscope 1 illustrated in FIG. 1 includes a microscope body 2 and a computing device 3, and may further include a display device 4.

[0019] The microscope main body 2 includes an objective lens, a light source that outputs excitation light, a detector that detects light from the specimen, etc. Note that the microscope main body 2 is shown in a simplified form in Fig. 1, but its details will be described later. The calculation device 3 is, for example, a general-purpose computer, and controls the microscope main body 2 and generates specimen images based on signals obtained by the microscope main body 2. The display device 4 is, for example, a liquid crystal display, and displays the specimen images generated by the calculation device 3.

[0020] In addition, the optical microscope 1 has a function for determining the performance of the optical microscope 1, and the calculation device 3 also performs performance determination of the optical microscope 1, and the display device 4 also displays the determination results (e.g., "detection sensitivity: OK" or "imaging performance: NG").

[0021] 2 is a diagram illustrating a functional configuration related to a performance determination function of an optical microscope according to the first embodiment. The optical microscope 1 illustrated in FIG. 2 includes a detection sensitivity measurement unit 11, an imaging performance measurement unit 12, a storage unit 13, a determination unit 14, and a display unit 15.

[0022] The detection sensitivity measurement unit 11 measures the detection sensitivity of the microscope main body 2 (detector). The imaging performance measurement unit 12 measures the imaging performance of the microscope main body 2. Note that the measurement results (measured values) of the detection sensitivity and imaging performance may be absolute values ​​or relative values. Details of the detection sensitivity measurement unit 11 and the imaging performance measurement unit 12 will be described later.

[0023] The memory unit 13 stores a detection sensitivity threshold value for determining whether the measurement result of the detection sensitivity is normal or abnormal, an imaging performance threshold value for determining whether the measurement result of the imaging performance is normal or abnormal, and causes of abnormality corresponding to combinations of judgment results for the measurement result of the detection sensitivity and judgment results for the measurement result of the imaging performance.

[0024] The determination unit 14 determines the performance of the optical microscope 1 based on the measurement results of the detection sensitivity and the measurement results of the imaging performance. Specifically, the determination unit 14 compares the measurement results of the detection sensitivity with a detection sensitivity threshold stored in the storage unit 13, and determines whether the measurement results of the detection sensitivity are normal or abnormal based on the comparison results. The determination unit 14 also compares the measurement results of the imaging performance with an imaging performance threshold stored in the storage unit 13, and determines whether the measurement results of the imaging performance are normal or abnormal based on the comparison results. The determination unit 14 then outputs the determination results for the measurement results of the detection sensitivity and the measurement results of the imaging performance to the display unit 15. At this time, the measurement results of the detection sensitivity and the measurement results of the imaging performance may also be output to the display unit 15.

[0025] Furthermore, based on the judgment result for the measurement result of the detection sensitivity and the judgment result for the measurement result of the imaging performance, the judgment unit 14 selects a corresponding abnormality cause from the abnormality causes stored in the storage unit 13. Then, the judgment unit 14 outputs the selected abnormality cause to the display unit 15, and also outputs to the display unit 15 a countermeasure to be taken by the user of the optical microscope 1 based on the selected abnormality cause.

[0026] The display unit 15 displays one or more of the measurement results of the detection sensitivity and the imaging performance, the judgment results for each measurement result, the cause of the abnormality, and the countermeasures, which are output by the judgment unit 14. The display unit 15 corresponds to the display device 4.

[0027] FIG. 3 is a diagram illustrating examples of abnormality causes (abnormality causes) stored in a storage unit according to the first embodiment. FIG. 3 illustrates examples of abnormality causes (NG causes) corresponding to combinations in which one or both of the judgment results for the measurement results of the detection sensitivity and the judgment results for the measurement results of the imaging performance are abnormal (NG). For example, a decrease in brightness and a decrease in sensitivity due to aberrations in the objective lens and / or the excitation / detection optical path (the optical system on the excitation / detection optical path) are shown as abnormality causes corresponding to a combination in which the judgment result for the measurement results of the imaging performance is abnormal (NG for imaging) and the judgment result for the measurement results of the detection sensitivity is abnormal (NG for sensitivity). In the past, this combination was simply considered to be a decrease in sensitivity because the imaging performance was not taken into consideration. Furthermore, for example, a decrease in brightness due to aberrations in the objective lens and / or the excitation optical path (the optical system on the excitation optical path) is shown as an abnormality cause corresponding to a combination in which the judgment result for the measurement results of the imaging performance is abnormal (NG for imaging) and the judgment result for the measurement results of the detection sensitivity is normal (OK for sensitivity). In the case of this combination, in the past, imaging performance was not taken into consideration, so the abnormality was not judged and was overlooked.

[0028] The details of the processing related to the performance determination function performed by the optical microscope 1 according to the first embodiment will be described later.

[0029] According to the first embodiment, the detection sensitivity and imaging performance are each measured, each measurement result is judged to be normal or abnormal, and the cause of the abnormality is identified based on each judgment result. Therefore, it is possible to distinguish between abnormality causes caused by detection sensitivity and abnormality causes caused by imaging performance, and it is possible to present the user of the optical microscope 1 with the exact cause of the abnormality and also to present an appropriate countermeasure.

[0030] Second Embodiment The optical microscope according to the second embodiment differs from the optical microscope 1 according to the first embodiment in that it includes an excitation output measurement unit instead of the detection sensitivity measurement unit 11. Accordingly, the contents stored in the storage unit 13 and the judgment performed by the judgment unit 14 are also different. The second embodiment will be described below, focusing on the differences. Note that the same elements as those in the first embodiment are denoted by the same reference numerals, and detailed explanations thereof will be omitted.

[0031] Fig. 4 is a diagram illustrating a functional configuration related to the performance determination function of an optical microscope according to the second embodiment. The optical microscope 1 illustrated in Fig. 4 has a configuration in which an excitation output measurement unit 16 is provided instead of the detection sensitivity measurement unit 11 in the optical microscope 1 illustrated in Fig. 2. The excitation output measurement unit 16 measures the excitation output of the microscope main body 2. Note that the measurement result (measurement value) of the excitation output may be an absolute value or a relative value. Details of the excitation output measurement unit 16 will be described later.

[0032] In addition, in the second embodiment, the memory unit 13 stores an excitation output threshold value for determining whether the measurement result of the excitation output is normal or abnormal, an imaging performance threshold value for determining whether the measurement result of the imaging performance is normal or abnormal, and causes of abnormality corresponding to combinations of the judgment results for the measurement result of the excitation output and the judgment results for the measurement result of the imaging performance.

[0033] In the second embodiment, the determination unit 14 determines the performance of the optical microscope 1 based on the measurement results of the excitation output and the measurement results of the imaging performance. Specifically, the determination unit 14 compares the measurement results of the excitation output with an excitation output threshold stored in the storage unit 13, and determines whether the measurement results of the excitation output are normal or abnormal based on the comparison results. The determination unit 14 also compares the measurement results of the imaging performance with an imaging performance threshold stored in the storage unit 13, and determines whether the measurement results of the imaging performance are normal or abnormal based on the comparison results. The determination unit 14 then outputs the determination results for the measurement results of the excitation output and the measurement results of the imaging performance to the display unit 15. At this time, the measurement results of the excitation output and the measurement results of the imaging performance may also be output to the display unit 15.

[0034] Furthermore, based on the judgment result for the measurement result of the excitation output and the judgment result for the measurement result of the imaging performance, the judgment unit 14 selects a corresponding abnormality cause from the abnormality causes stored in the storage unit 13. Then, the judgment unit 14 outputs the selected abnormality cause to the display unit 15, and also outputs to the display unit 15 a countermeasure to be taken by the user of the optical microscope 1 based on the selected abnormality cause.

[0035] In addition, in the second embodiment, the display unit 15 displays one or more of the measurement results of the excitation output and imaging performance output by the judgment unit 14, the judgment results for each measurement result, the cause of the abnormality, and the countermeasures.

[0036] FIG. 5 is a diagram illustrating examples of abnormality causes (NG causes) stored in a storage unit according to the second embodiment. FIG. 5 illustrates examples of abnormality causes (NG causes) corresponding to combinations in which one or both of the judgment results for the measurement results of the excitation output and the judgment results for the measurement results of the imaging performance are abnormal (NG). For example, a decrease in brightness due to aberrations in the objective lens and / or the excitation / detection optical path and a decrease in output are shown as abnormal causes corresponding to a combination in which the judgment result for the measurement results of the imaging performance is abnormal (NG for imaging) and the judgment result for the measurement results of the excitation output is abnormal (NG for output). In the past, this combination was considered to be simply a decrease in output because the imaging performance was not taken into consideration. Furthermore, for example, a decrease in brightness due to aberrations in the objective lens and / or the excitation / detection optical path is shown as an abnormal cause corresponding to a combination in which the judgment result for the measurement results of the imaging performance is abnormal (NG for imaging) and the judgment result for the measurement results of the excitation output is normal (OK for output). In the case of this combination, in the past, imaging performance was not taken into consideration, so the abnormality was not judged and was overlooked.

[0037] The details of the processing related to the performance determination function performed by the optical microscope 1 according to the second embodiment will be described later.

[0038] According to the second embodiment, the excitation output and the imaging performance are each measured, each measurement result is judged to be normal or abnormal, and the cause of the abnormality is identified based on each judgment result. Therefore, it is possible to distinguish between abnormalities caused by the excitation output and abnormalities caused by the imaging performance, and it is possible to present the user of the optical microscope 1 with the exact cause of the abnormality and also to present an appropriate countermeasure.

[0039] <Third embodiment> The optical microscope according to the third embodiment differs from the optical microscope 1 according to the first embodiment in that it further includes the excitation output measuring unit 16 described in the second embodiment. Accordingly, the contents stored in the storage unit 13 and the judgment performed by the judgment unit 14 are also different. Below, the third embodiment will be described, focusing on the differences. Note that the same elements as those in the first and second embodiments are assigned the same reference numerals, and detailed explanations thereof will be omitted.

[0040] Fig. 6 is a diagram illustrating a functional configuration related to a performance determination function of an optical microscope according to a third embodiment. The optical microscope 1 illustrated in Fig. 6 has a configuration in which an excitation output measurement unit 16 is further provided in addition to the optical microscope 1 illustrated in Fig. 2.

[0041] In addition, in the third embodiment, the memory unit 13 stores an excitation output threshold value for determining whether the measurement result of the excitation output is normal or abnormal, a detection sensitivity threshold value for determining whether the measurement result of the detection sensitivity is normal or abnormal, an imaging performance threshold value for determining whether the measurement result of the imaging performance is normal or abnormal, and causes of abnormality corresponding to combinations of the judgment result for the measurement result of the excitation output, the judgment result for the measurement result of the detection sensitivity, and the judgment result for the measurement result of the imaging performance.

[0042] In the third embodiment, the determination unit 14 determines the performance of the optical microscope 1 based on the measurement results of the excitation output, the detection sensitivity, and the imaging performance. Specifically, the determination unit 14 compares the measurement results of the excitation output with an excitation output threshold stored in the storage unit 13 and determines whether the measurement results of the excitation output are normal or abnormal based on the comparison results. The determination unit 14 also compares the measurement results of the detection sensitivity with a detection sensitivity threshold stored in the storage unit 13 and determines whether the measurement results of the detection sensitivity are normal or abnormal based on the comparison results. The determination unit 14 also compares the measurement results of the imaging performance with an imaging performance threshold stored in the storage unit 13 and determines whether the measurement results of the imaging performance are normal or abnormal based on the comparison results. The determination unit 14 then outputs the determination results for the measurement results of the excitation output, the determination results for the detection sensitivity, and the determination results for the measurement results of the imaging performance to the display unit 15. At this time, the measurement results of the excitation output, the measurement results of the detection sensitivity, and the measurement results of the imaging performance may also be output to the display unit 15.

[0043] Furthermore, based on the judgment result for the measurement result of the excitation output, the judgment result for the measurement result of the detection sensitivity, and the judgment result for the measurement result of the imaging performance, the judgment unit 14 selects a corresponding abnormality cause from the abnormality causes stored in the storage unit 13. Then, the judgment unit 14 outputs the selected abnormality cause to the display unit 15, and also outputs to the display unit 15 a countermeasure to be taken by the user of the optical microscope 1 based on the selected abnormality cause.

[0044] In addition, in the third embodiment, the display unit 15 displays one or more of the measurement results of excitation output, detection sensitivity, and imaging performance output by the judgment unit 14, the judgment results for each measurement result, the cause of the abnormality, and a countermeasure.

[0045] The details of the processing related to the performance determination function performed by the optical microscope 1 according to the third embodiment will be described later.

[0046] According to the third embodiment, the three major factors related to fluorescence intensity, namely excitation output, detection sensitivity, and imaging performance, are each measured, and each measurement result is judged to be normal or abnormal. The cause of the abnormality is identified based on each judgment result. Therefore, it is possible to distinguish between abnormalities caused by excitation output, abnormalities caused by detection sensitivity, and abnormalities caused by imaging performance, and it is possible to present the user of the optical microscope 1 with the exact cause of the abnormality and also to present an appropriate countermeasure.

[0047] For example, if the judgment result for the measurement result of the imaging performance is abnormal and the judgment result for the measurement result of the excitation output is normal, then as shown in the example of Figure 5, the cause of the abnormality would be a decrease in brightness due to aberrations in the objective lens and / or the excitation / detection optical path. However, if the judgment result for the measurement result of the detection sensitivity is also normal, then the cause of the abnormality can be narrowed down to a decrease in brightness due to aberrations in the objective lens and / or the excitation optical path.

[0048] Furthermore, in the third embodiment, when excitation light is used to measure detection sensitivity, the measurement value at the detector can be normalized by the measurement value of the excitation output, which makes it possible to improve the measurement accuracy of detection sensitivity and simplify the measurement unit.

[0049] <Processing Related to Performance Determination Function Performed by Optical Microscope 1> In each of the first to third embodiments, the processing related to the performance determination function performed by the optical microscope 1 is the processing illustrated in FIG.

[0050] 7 is a flowchart illustrating a process related to the performance evaluation function performed by the optical microscope. Note that, before starting this process, it is assumed that each threshold value, each abnormality cause, and each correction possibility threshold value are stored in the storage unit 13.

[0051] More specifically, in the first embodiment, a detection sensitivity threshold and an imaging performance threshold are stored as the thresholds, an anomaly cause corresponding to each combination of an abnormality when one or both of the judgment results for the measurement results of the detection sensitivity and the judgment results for the measurement results of the imaging performance are abnormal is stored as the abnormality cause, and a detection sensitivity correction threshold is stored as the correction possibility threshold. The detection sensitivity correction possibility threshold is a threshold for determining whether the detection sensitivity can be corrected based on the measurement result of the detection sensitivity that is judged to be abnormal.

[0052] In the second embodiment, an excitation output threshold and an imaging performance threshold are stored as the thresholds, an abnormality cause corresponding to each combination of an abnormality in the case where one or both of the judgment result for the measurement result of the excitation output and the judgment result for the measurement result of the imaging performance are abnormal is stored as the abnormality cause, and an excitation output correction threshold is stored as the correction possibility threshold. The excitation output correction possibility threshold is a threshold for determining whether the excitation output can be corrected based on the measurement result of the excitation output that is judged to be abnormal.

[0053] In the third embodiment, an excitation output threshold, a detection sensitivity threshold, and an imaging performance threshold are stored as each threshold, and an abnormality cause corresponding to each combination of an abnormality when one or more of the judgment results for the measurement results of the excitation output, the judgment results for the measurement results of the detection sensitivity, and the judgment results for the measurement results of the imaging performance are abnormal is stored as each correction feasibility threshold, and an excitation output correction feasibility threshold and a detection sensitivity correction feasibility threshold are stored as each correction feasibility threshold.

[0054] 7 , first, each measurement unit performs measurement (step S1). More specifically, in the first embodiment, the detection sensitivity measurement unit 11 measures the detection sensitivity, and the imaging performance measurement unit 12 measures the imaging performance. In the second embodiment, the excitation output measurement unit 16 measures the excitation output, and the imaging performance measurement unit 12 measures the imaging performance. In the third embodiment, the excitation output measurement unit 16 measures the excitation output, the detection sensitivity measurement unit 11 measures the detection sensitivity, and the imaging performance measurement unit 12 measures the imaging performance.

[0055] Next, the determination unit 14 compares each measurement result (each measurement value) with a corresponding threshold value to determine whether each measurement result is normal or abnormal (step S2). Specifically, in the first embodiment, the determination unit 14 compares the measurement result of detection sensitivity with the detection sensitivity threshold value and the measurement result of imaging performance with the imaging performance threshold value to determine whether each measurement result is normal or abnormal. In the second embodiment, the determination unit 14 compares the measurement result of excitation output with the excitation output threshold value and the measurement result of imaging performance with the imaging performance threshold value to determine whether each measurement result is normal or abnormal. In the third embodiment, the determination unit 14 compares the measurement result of excitation output with the excitation output threshold value, the measurement result of detection sensitivity with the detection sensitivity threshold value, and the measurement result of imaging performance with the imaging performance threshold value to determine whether each measurement result is normal or abnormal.

[0056] If all of the measurement results are determined to be normal in step S2 (OK in step S2), the determination unit 14 determines that there is no abnormality and determines to continue use as is (step S3). At this time, the determination unit 14 may output the determination result (normal) for each measurement result to the display unit 15, and the display unit 15 may display it.

[0057] On the other hand, if none of the measurement results are determined to be normal in step S2 (NG in step S2), the process proceeds to step S4. In step S4, the determination unit 14 selects a corresponding abnormality cause (NG cause) from the abnormality causes stored in the storage unit 13 based on the determination results for each measurement result. In more detail, in the first embodiment, the corresponding abnormality cause is selected based on the determination result for the measurement result of detection sensitivity and the determination result for the measurement result of imaging performance. In the second embodiment, the corresponding abnormality cause is selected based on the determination result for the measurement result of excitation output and the determination result for the measurement result of imaging performance. In the third embodiment, the corresponding abnormality cause is selected based on the determination result for the measurement result of excitation output, the determination result for the measurement result of detection sensitivity, and the determination result for the measurement result of imaging performance.

[0058] In step S4, the judgment unit 14 may output the selected cause of the abnormality to the display unit 15, which may then display it, or the judgment unit 14 may further output each measurement result and / or the judgment result for each measurement result to the display unit 15, which may then display it.

[0059] Next, the determination unit 14 calculates the fluctuation range (difference) between the measurement result (measurement value) determined to be abnormal in step S2 (excluding the measurement result (measurement value) of the imaging performance) and the corresponding threshold (step S5), and determines whether the fluctuation range is less than the corresponding correction feasibility threshold (i.e., whether the fluctuation range is correctable) (step S6). More specifically, in the first embodiment, when the measurement result of the detection sensitivity is determined to be abnormal, the determination unit 14 calculates the fluctuation range between the measurement result of the detection sensitivity and the detection sensitivity threshold (detection sensitivity fluctuation range), and determines whether the detection sensitivity fluctuation range is less than the detection sensitivity correction feasibility threshold. In the second embodiment, when the measurement result of the excitation output is determined to be abnormal, the determination unit 14 calculates the fluctuation range between the measurement result of the excitation output and the excitation output threshold (excitation output fluctuation range), and determines whether the excitation output fluctuation range is less than the excitation output correction feasibility threshold. In the third embodiment, when the measurement result of the excitation output and / or the measurement result of the detection sensitivity are determined to be abnormal, the excitation output fluctuation range and / or the detection sensitivity fluctuation range are calculated, and it is determined whether the excitation output fluctuation range is less than the threshold value for whether the excitation output can be corrected and / or whether the detection sensitivity fluctuation range is less than the threshold value for whether the detection sensitivity can be corrected.

[0060] If the determination results for the measurement results other than the imaging performance are normal, step S5 is skipped. If the determination result for the measurement result of the imaging performance is abnormal or if the optical microscope 1 is not equipped with a correction mechanism (a mechanism for performing correction), the determination result in step S6 is processed as NO, and the process proceeds to step S8. As a result, if the determination result for the measurement result of the imaging performance is abnormal, the process proceeds to step S8 rather than step S7. This is because priority is given to addressing the cause of the abnormality in the imaging performance, and details will be described later.

[0061] In step S6, if the judgment result for the measurement result of the imaging performance is normal and the optical microscope 1 is equipped with a correction mechanism, and if it is judged that the fluctuation range calculated in step S5 is a fluctuation range that can be corrected (step S6 is YES), proceed to step S7; if not (step S6 is NO), proceed to step S8.

[0062] In step S7, the determination unit 14 determines a correction method as a method for dealing with the abnormality cause selected in step S4, outputs the correction method to the display unit 15, and displays it. Once the correction method is determined, the correction is then performed. The correction may be performed automatically by a correction function provided in the optical microscope 1, or manually by a user of the optical microscope 1. The excitation output is corrected, for example, by adjusting the excitation output by controlling an AOM (Acousto Optic Modulator) provided downstream of the light source or by controlling the LD (Laser Diode) drive current of the light source. The detection sensitivity is corrected by adjusting the amount of light incident on the detector to a degree that does not significantly affect imaging performance (spatial resolution), for example, by adjusting the incident optical axis to the confocal pinhole using a beam shifter provided upstream of the confocal pinhole or by adjusting the pinhole diameter of the confocal pinhole.

[0063] In step S8, the determination unit 14 determines a maintenance / repair solution as a solution to the cause of the abnormality selected in step S4, and outputs the maintenance / repair solution to the display unit 15, which then displays it. Once the maintenance / repair solution is determined, the maintenance / repair is then carried out. Maintenance includes cleaning the objective lens, cleaning the chart specimen (wiping off the immersion liquid (oil), etc.), adjusting the correction collar of the objective lens, etc. Repairs are performed for issues that cannot be addressed without contacting the manufacturer's service, such as a decrease in brightness caused by the excitation light path or detection light path.

[0064] According to the processing related to the performance evaluation function as described above, countermeasures for dealing with the cause of the abnormality can be displayed on the display unit 15, so that the user of the optical microscope 1 can check the displayed content and take appropriate measures without hesitation.

[0065] 8 is a diagram illustrating the judgment results for each measurement result, the causes of abnormality (NG causes) selected based on the judgment results, and countermeasures for the causes of abnormality according to the third embodiment. However, in FIG. 8, when the judgment results for the measurement results of the imaging performance, excitation output, and detection sensitivity are all normal (imaging OK, output OK, sensitivity OK), "No abnormality" is displayed as the cause of abnormality, and "No abnormality, use as is" is displayed as the countermeasure.

[0066] As illustrated in FIG. 8, when the judgment result for the measurement result of the imaging performance is normal (imaging OK), and the judgment result for one or both of the measurement results of the excitation output and the detection sensitivity is abnormal (NG), the remedy is to correct (execute the correction function) or repair (contact service).

[0067] On the other hand, if the judgment result for the measurement result of the imaging performance is abnormal (imaging NG), the remedy is maintenance (cleaning the objective lens, adjusting the correction collar) or repair (contacting service), regardless of the judgment results for the measurement results of the excitation power and the detection sensitivity. In this case, even if the judgment result for one or both of the measurement results of the excitation power and the detection sensitivity is abnormal (NG), the remedy does not include correction. This is because if the judgment result for one or both of the measurement results of the excitation power and the detection sensitivity is abnormal, the cause of the abnormality may be due to an abnormality in the imaging performance. Therefore, in such a case, priority is given to addressing the cause of the abnormality in the imaging performance, so the remedy is maintenance or repair, not correction. This is the same in the first and second embodiments. If the judgment result for the measurement result of the imaging performance and the judgment results for the other measurement results are both abnormal, priority is given to addressing the cause of the abnormality in the imaging performance, so the remedy is maintenance or repair, not correction.

[0068] <Microscope main body 2 and excitation output measuring unit 16> Fig. 9 is a diagram illustrating an example of the microscope main body and the excitation output measuring unit 16. In this example, it is assumed that the optical microscope 1 is a laser scanning confocal fluorescence microscope.

[0069] The microscope body 2 illustrated in Figure 9 is a laser scanning confocal fluorescence microscope body, and includes a plurality of objective lenses 202 (202a, 202b, 202c) attached to a revolver 201, a laser light source 203 that outputs excitation light, and a plurality of detectors 204 (204a, 204b, 204c, 204d) that detect light (fluorescence) from the specimen.

[0070] In the microscope main body 2, light emitted from the laser light source 203 passes through a beam splitter 205, a dichroic mirror 206, a pair of galvanometer mirrors 207, a lens 208, a dichroic mirror 209, a lens 210, a fluorescence cube 211, and an objective lens 202, and is then irradiated as excitation light onto a specimen (not shown) placed on a stage 212. The optical path along which the light emitted from the laser light source 203 reaches the specimen is the excitation optical path. An AOM may also be provided in the optical path between the laser light source 203 and the beam splitter 205.

[0071] Fluorescence generated by irradiating the specimen with excitation light passes through the objective lens 202, the fluorescence cube 211, the lens 210, the dichroic mirror 209, the lens 208, the pair of galvanometer mirrors 207, the dichroic mirror 206, the lens 213, and the confocal pinhole 214, and is then detected by detectors 204a and / or 204b via dichroic mirrors 215 and 216, and / or by detectors 204c and / or 204d via dichroic mirrors 215, 217, and 218. The optical path along which the fluorescence from the specimen reaches the detector 204 is the detection optical path. A beam shifter may be further provided in the optical path between the lens 213 and the confocal pinhole 214.

[0072] The microscope main body 2 also includes a transmission detector 219, which can detect light that has passed through a specimen placed on the stage 212. In this case, the light that has passed through the specimen passes through a condenser lens 220, mirrors 221 and 222, and a lens 223, and is then detected by the transmission detector 219. The microscope main body 2 also includes a transmission illumination light source 224, which can illuminate a specimen placed on the stage. In this case, the mirror 222 flips up, and illumination light from the transmission illumination light source 224 passes through the mirror 221 and the condenser lens 220 to illuminate the specimen.

[0073] In the microscope main body 2, the excitation output measuring unit 16 is realized using an excitation light intensity monitor 225. The excitation light intensity monitor 225 is, for example, a photodiode. The excitation light intensity monitor 225 measures the intensity of the light (excitation light) from the laser light source 203 that is split by the beam splitter 205. The measurement result at this time becomes the measurement result of the excitation output measuring unit 16.

[0074] The excitation light intensity monitor 225 may be provided in another position. For example, the excitation light intensity monitor 225 may be built into the laser light source 203. Alternatively, the excitation light intensity monitor 225 may be provided in a position that can be inserted and removed from the main optical path, such as an objective lens attachment hole of the revolver 201. Alternatively, the excitation light intensity monitor 225 may be provided near the specimen surface, such as on the back surface of the stage 212. Alternatively, the excitation light intensity monitor 225 may be provided on the transmission side of the specimen surface. In this case, the excitation light intensity monitor 225 may be provided near the condenser lens 220, for example. As described above, the excitation light intensity monitor 225 may be provided in the optical path closer to the laser light source 203 than the objective lens 202, near the focal plane of the objective lens 202, or on the optical path opposite the objective lens 202 across the focal plane of the objective lens 202. Alternatively, the transmission detector 219 may be used as the excitation light intensity monitor 225. Note that no specimen is placed on the stage 212 during excitation output measurement. In this way, if the excitation light intensity monitor 225 is provided at another position, the beam splitter 205 can be eliminated.

[0075] The excitation light intensity monitor 225 and the transmission detector 219 used as the excitation light intensity monitor 225 can also be called an excitation output measuring device that measures the excitation output.

[0076] <Detection Sensitivity Measurement Unit 11> FIG. 10 is a diagram illustrating an example of the detection sensitivity measurement unit.

[0077] The detection sensitivity measurement unit 11 is realized using a detector 204. More specifically, as illustrated in FIG. 10 , a reflecting element 227 is provided in the fluorescent cube 211, the dichroic mirror 206 is replaced with a partial reflecting mirror 226, and the pinhole diameter of the confocal pinhole 214 is adjusted to optimize the amount of light incident on the detector 204 during detection sensitivity measurement. Then, light emitted from the laser light source 203 is reflected by the reflecting element 227 via the partial reflecting mirror 226 or the like, and the reflected light is detected by the detector 204 via the partial reflecting mirror 226 or the like. This realizes the detection sensitivity measurement unit 11. The detection result of the detector 204 at this time becomes the measurement result of the detection sensitivity measurement unit 11. The reflecting element 227 may be a flat reflecting element or a corner cube prism. The laser light source 203 may further include a reference light source that emits light for detection sensitivity measurement during detection sensitivity measurement.

[0078] The reflecting element 227 may be provided anywhere in the optical path between the partial reflection mirror 226 and the objective lens 202. For example, it may be provided in an objective lens mounting hole of the revolver 201 or in a differential interference observation prism mounting portion that is removably provided in the optical path. Furthermore, the reflecting element 227 may be provided on the stage 212 when the objective lens 202 is removed.

[0079] Alternatively, the detection sensitivity measurement unit 11 may be realized as follows: a phosphor is placed at the primary image position P in the optical path, and the phosphor is irradiated with light from the laser light source 203, and the fluorescence generated by the phosphor is detected by the detector 204, thereby realizing the detection sensitivity measurement unit 11. In this case, there is no need to replace the dichroic mirror 206 with the partial reflection mirror 226.

[0080] Alternatively, the detection sensitivity measurement unit 11 may be realized as follows: A reference specimen such as a fluorescent specimen or a reflective specimen is fixed near the specimen surface, such as on the rear surface of the stage, and a reference objective lens (e.g., objective lens 202a) is inserted into the optical path. The detection sensitivity measurement unit 11 may then be realized by irradiating the reference specimen with light from the laser light source 203 and detecting light (fluorescence or reflected light) from the reference specimen with the detector 204. Note that when a fluorescent specimen is used as the reference specimen, there is no need to replace the dichroic mirror 206 with the partial reflection mirror 226.

[0081] Alternatively, the detection sensitivity measurement unit 11 may be realized as follows: A reference light source is fixed to the surface of the specimen or the rear surface of the stage, a reference objective lens (e.g., the objective lens 202 a) is inserted into the optical path, and the light from the reference light source is detected by the detector 204, thereby realizing the detection sensitivity measurement unit 11.

[0082] Alternatively, the detection sensitivity measurement unit 11 may be realized as follows: After flipping up the mirror 222, the detection sensitivity measurement unit 11 may be realized by detecting light from the transmitted illumination light source 224 with the detector 204.

[0083] In this way, the detection sensitivity measurement unit 11 may be realized by including a reflecting element 227 or a fluorescent material provided in the optical path on the laser light source 203 side of the objective lens 202, a reference specimen or a reference light source arranged in the vicinity of the focal position of the objective lens 202, or a light source provided on the opposite side of the objective lens 202 across the focal position of the objective lens 202. The detector 204 used in measuring the detection sensitivity can also be referred to as a detection sensitivity measurement device that measures the detection sensitivity.

[0084] <Imaging Performance Measurement Unit 12> FIG. 11 is a diagram illustrating an example of the imaging performance measurement unit.

[0085] The imaging performance measurement unit 12 is realized using a detector 204 and a computing device 3. More specifically, as illustrated in Fig. 11 , a fluorescent bead specimen 228 or a pinhole specimen 229 is placed on a stage 212 (if a pinhole specimen 229 is placed, the dichroic mirror 206 is further replaced with a partial reflection mirror), and the confocal pinhole 214 is set to about 2 AU (Airy Unit) instead of 1 AU to make it easier to monitor out-of-focus aberrations. Then, light from a laser light source 203 is irradiated onto the fluorescent bead specimen 228 or the pinhole specimen 229 while being scanned two-dimensionally by a pair of galvanometer mirrors 207, and light (fluorescence or reflected light) from the fluorescent bead specimen 228 or the pinhole specimen 229 is detected by a detector 204 (e.g., 204a). Then, based on the detection result, the calculation device 3 generates an image of the fluorescent bead specimen or an image of the pinhole specimen, and calculates a PSF (Point Spread Function) based on the fluorescent bead specimen image or the pinhole specimen image. This realizes the imaging performance measurement unit 12. The PSF calculated by the calculation device 3 at this time becomes the measurement result of the imaging performance measurement unit 12. In this way, the imaging performance measurement unit 12 may be realized by acquiring an image of the reference specimen (fluorescent bead specimen 228 or pinhole specimen 229) placed at the focal position of the objective lens 202 and calculating a PSF based on the image.

[0086] Alternatively, the imaging performance measuring unit 12 may be realized as follows: a wavefront sensor is provided on the stage 212, light from the laser light source 203 is irradiated onto the wavefront sensor, and the wavefront of the irradiated light is measured by the wavefront sensor. Then, the arithmetic unit 3 calculates a PSF based on the measurement results. In this way, the imaging performance measuring unit 12 may be realized. In this way, the imaging performance measuring unit 12 may be realized by measuring the wavefront of the excitation light at the focal plane of the objective lens 202 and calculating a PSF based on the measured wavefront.

[0087] Alternatively, the imaging performance measurement unit 12 may be realized as follows: An edge chart sample, which is an example of an edge sample, is placed on the stage 212, the dichroic mirror 206 is replaced with a partial reflection mirror, and the confocal pinhole 214 is set to approximately 2 AU. Light from the laser light source 203 is then irradiated onto the edge chart sample while being scanned two-dimensionally by a pair of galvanometer mirrors 207, and light (reflected light) from the edge chart sample is detected by a detector 204 (e.g., 204a). The calculation unit 3 then generates an image of the edge chart sample based on the detection result, and calculates an LSF (Line Spread Function) based on the edge chart sample image. The imaging performance measurement unit 12 may be realized in this manner. The LSF calculated by the calculation unit 3 at this time becomes the measurement result of the imaging performance measurement unit 12. Alternatively, the calculation device 3 may calculate a correlation coefficient between the calculated LSF and a theoretical LSF (LSF in the case of no aberration), which is a theoretical value, and use this as the measurement result of the imaging performance measurement unit 12. In this way, the imaging performance measurement unit 12 may be realized by acquiring an image of the edge sample placed at the focal position of the objective lens 202 and calculating the LSF (or the correlation coefficient with the theoretical LSF) based on the image.

[0088] FIG. 12 is a diagram showing an example of measuring imaging performance using an edge chart specimen. In the example shown in FIG. 12, Z-stack imaging is performed on the reflected light from the edge chart specimen 230, edge responses are obtained in four directions indicated by arrows, and LSFs in the four directions are calculated. The horizontal axis of each LSF indicates the Z direction (the optical axis direction of the objective lens 202 in the optical path), and the vertical axis indicates the X direction (or Y direction). The correlation coefficient between each of the LSFs in the four directions and the theoretical LSF is then calculated as imaging performance. Note that Z-stack imaging is performed to later eliminate the effects of image blur due to focus deviation and to determine aberrations from the blur in the Z direction. The edge responses are acquired in at least four directions, and may be acquired in eight directions around 360°. This also makes it possible to evaluate the degree of LSF degradation depending on the direction.

[0089] FIG. 13 is a diagram illustrating an edge chart specimen. The edge chart specimens illustrated in (a), (b), and (c) of FIG. 13 all have a radial pattern. By using such an edge chart specimen, it is possible to evaluate imaging performance in various directions. Note that the edge chart specimen may have a radial pattern with edges in eight or more directions around a 360° circumference.

[0090] Furthermore, when measuring the imaging performance using an edge chart sample, the LSF may be further decomposed into aberration amounts, and the aberration amounts and aberration levels may be displayed as evaluation results on the display unit 15 (display device 4). In this case, the aberration amounts may be classified into coma aberration, spherical aberration, and astigmatism and displayed. Furthermore, the presence or absence of aberration, or a value quantified in any unit, may be displayed. The aberration level may be determined based on a comparison between the aberration amount and a threshold value. In this case, the threshold value may be, for example, the Strehl ratio, FWHM (Full Width at Half Maximum), or wavefront aberration RMS (Root Mean Square), or may be set by AI.

[0091] 14 is a diagram showing an example of a display of the evaluation results. In the display example shown in Fig. 14, the evaluation results show that there is aberration, that the spherical aberration is at an NG level and the coma aberration and astigmatism are at an OK level, and the wavefront aberration RMS of the spherical aberration, coma aberration, and astigmatism are displayed on the display device 4.

[0092] <Matters common to measurements of excitation output, detection sensitivity, and imaging performance> In each measurement of excitation output, detection sensitivity, and imaging performance, measurements are performed using one excitation wavelength and one or more detection wavelengths. When measurements of excitation output, detection sensitivity, and imaging performance are performed at approximately the same time, and when the light source intensity value of the laser light source 203 is used to measure detection sensitivity, the order of measurements is as follows: measurement of excitation output and detection sensitivity comes first, and measurement of imaging performance comes after. This is because measurement of imaging performance always requires the use of a sample, which takes time, and during that time, there is a possibility that the two performances, excitation output and detection sensitivity, may fluctuate.

[0093] <Variation 1 of each of the first to third embodiments> When measuring the detection sensitivity in the optical microscope 1 according to each of the first and third embodiments, the following measurements may be further performed, and when measuring the imaging performance in the optical microscope 1 according to each of the first to third embodiments, the following measurements may be further performed.

[0094] In measuring the detection sensitivity, the incident optical axis to the confocal pinhole 214 may be further shifted using a beam shifter or the like, and the detection sensitivity for each shift amount may be measured. The calculation device 3 may then measure the amount of deviation of the incident optical axis to the confocal pinhole 214 based on the measurement result of the detection sensitivity when the incident optical axis is not shifted (the measurement result of the detection sensitivity as before) and the measurement result of the detection sensitivity when the incident optical axis is shifted. In this case, for example, by determining whether the deviation of the incident optical axis to the confocal pinhole 214 is equal to or greater than a threshold, it is possible to determine whether the incident optical axis to the confocal pinhole 214 is normal or abnormal. This makes it possible to identify whether the cause of the abnormality in the measurement result of the detection sensitivity is due to the detector 204 and / or the detection optical path, or due to the deviation of the incident optical axis to the confocal pinhole 214.

[0095] The measurement of the imaging performance may further be performed using each of the plurality of objective lenses 202. In this case, the determination unit 14 may determine whether each of the measurement results of the imaging performance using each of the plurality of objective lenses 202 is normal or abnormal. This makes it possible to more accurately identify the cause of the abnormality in the measurement result of the imaging performance based on each measurement result. For example, if the determination result for the measurement result of the imaging performance is abnormal and the determination results for the measurement results of the imaging performance using each of the plurality of objective lenses 202 are also abnormal, it is unlikely that all of the objective lenses 202 are abnormal. Therefore, it can be determined that the cause of the abnormality in the measurement result of the imaging performance is a decrease in brightness due to aberrations in the excitation / detection optical paths, rather than the objective lenses 202.

[0096] FIG. 15 is a diagram illustrating an example of a determination result for each measurement result, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to the first modification of the third embodiment.

[0097] 15, the judgment results further include a judgment result as to whether the incident optical axis to the confocal pinhole 214 is normal or abnormal, and a judgment result for the measurement results of the imaging performance using each of the multiple objective lenses 202. "PH incident optical axis OK" indicates that the incident optical axis to the confocal pinhole 214 is judged to be normal, and "PH incident optical axis NG" indicates that the incident optical axis to the confocal pinhole 214 is judged to be abnormal. "Multiple objective lenses ALL NG" indicates that all of the measurement results of the imaging performance using each of the multiple objective lenses 202 are judged to be abnormal, and "Multiple objective lenses 1 OK" indicates that the measurement results of the imaging performance using each of the multiple objective lenses 202 are judged to be normal. In addition, if the judgment result for the measurement results of the imaging performance is normal (imaging OK), no judgment is made for the measurement results of the imaging performance using each of the multiple objective lenses 202, and this is shown as ``multiple (no judgment).''

[0098] According to the example shown in FIG. 15, when the judgment result for the measurement result of detection sensitivity is abnormal (sensitivity NG) or when the judgment result for the measurement result of imaging performance is abnormal (imaging NG), it is possible to select a more accurate cause of the abnormality and present a more appropriate countermeasure.

[0099] Although not shown in Figure 15, if the judgment result for the measurement result of the detection sensitivity is normal (sensitivity OK), for example, it is possible to proceed in the same manner as illustrated in Figure 8 without making a judgment as to whether the incident optical axis to the confocal pinhole 214 is normal or abnormal and without making a judgment on the measurement result of the imaging performance using each of the multiple objective lenses 202.

[0100] FIG. 16 is a diagram illustrating an example of a determination result for each measurement result, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to the first modification of the second embodiment.

[0101] 16, the judgment result further includes a judgment result for the measurement result of the imaging performance using each of the plurality of objective lenses 202. According to this example, when the judgment result for the measurement result of the imaging performance is abnormal (imaging NG), it is possible to select a more accurate cause of the abnormality and to present a more appropriate countermeasure.

[0102] FIG. 17 is a diagram illustrating an example of a determination result for each measurement result, an abnormality cause (NG cause) selected based on the determination result, and a countermeasure for the abnormality cause, according to the first modification of the first embodiment.

[0103] 17, the judgment result further includes a judgment result for the measurement result of the imaging performance using each of the plurality of objective lenses 202. According to this example, when the judgment result for the measurement result of the imaging performance is abnormal (imaging NG), it is possible to select a more accurate cause of the abnormality and to present a more appropriate countermeasure.

[0104] In the first modification, the judgment unit 14 judges whether the incident optical axis to the confocal pinhole 214 is normal or abnormal (judgment of "PH incident optical axis OK" or "PH incident optical axis NG") and judges the measurement results of the imaging performance using each of the multiple objective lenses 202 (judgment of "multiple lenses ALL NG" or "multiple lenses, one lens OK"). Also, causes of abnormality corresponding to combinations of each judgment result are stored in advance in the storage unit 13. Then, based on each judgment result, the judgment unit 14 selects the corresponding cause of abnormality.

[0105] In the first modification, the detection sensitivity may be measured using each of multiple detection channels (e.g., multiple detectors 204), and the results of the detection sensitivity measurements using each of the multiple detection channels may be used to identify the cause of an abnormality when the determination result for the detection sensitivity measurement is abnormal. For example, if the determination result for the detection sensitivity measurement is abnormal, and all of the determination results for the detection sensitivity measurements using each of the multiple detection channels are also abnormal, and the incident optical axis to the confocal pinhole 214 is determined to be normal, the cause of the abnormality can be identified as a decrease in brightness due to aberrations in the detection optical path (the optical system on the detection optical path) and / or a decrease in sensitivity in the detection optical path other than the detector. Furthermore, if the determination result for the imaging performance measurement is normal at this time, the decrease in brightness due to aberrations in the detection optical path can be excluded from the causes of the abnormality, and the cause of the abnormality can be narrowed down to a decrease in sensitivity in the detection optical path other than the detector. In this way, the cause of the abnormality when the determination result for the detection sensitivity measurement is abnormal can be more accurately identified. The determination unit 14 determines whether each of the measurement results of the detection sensitivity using each of the multiple detection channels is normal or abnormal.

[0106] <Variation 2 of the Second and Third Embodiments> The optical microscope 1 according to the second and third embodiments may further include an irradiation density measuring unit that measures the irradiation density on the specimen surface based on the measurement result (excitation output) of the excitation output measuring unit 16 and the measurement result (imaging performance) of the imaging performance measuring unit 12.

[0107] Specifically, the irradiation density measurement unit calculates the irradiation density using formula (1). Irradiation density = excitation output / imaging area formula (1) The excitation output used in formula (1) is the objective lens output. When the measurement result of the excitation output measurement unit 16 is not the objective lens output, the objective lens output may be calculated by multiplying the measurement result of the excitation output measurement unit 16 by the transmittance of the optical system from the measurement position to the specimen surface. The imaging area used in formula (1) is calculated based on the PSF or LSF, which is the measurement result of the imaging performance measurement unit 12. Note that the calculated value of the irradiation density using formula (1) may be a relative value (unitless) or an absolute value (W / mm ２ ) is also acceptable.

[0108] The storage unit 13 may further store an irradiation density threshold value for determining whether the irradiation density measurement result is normal (OK) or abnormal (NG). The storage unit 13 may also store an abnormality cause corresponding to a combination of a determination result for the irradiation density measurement result and a determination result for another measurement result. Specifically, in a second modification of the second embodiment, the storage unit 13 may further store an abnormality cause corresponding to a combination of a determination result for the imaging performance measurement result and / or a determination result for the excitation output measurement result and a determination result for the irradiation density measurement result. In a second modification of the third embodiment, the storage unit 13 may further store an abnormality cause corresponding to a combination of a determination result for the irradiation density measurement result and one or more of the determination result for the imaging performance measurement result, the determination result for the detection sensitivity measurement result, and the determination result for the excitation output measurement result.

[0109] The determination unit 14 may further compare the measurement result of the irradiation density with an irradiation density threshold stored in the storage unit 13, and determine whether the measurement result of the irradiation density is normal or abnormal based on the comparison result. Then, the determination result may be output to the display unit 15, which may display it.

[0110] Furthermore, the determination unit 14 may select a corresponding abnormality cause from among the abnormality causes stored in the storage unit 13 based on the determination result for the measurement result of the irradiation density and the determination results for other measurement results. Specifically, in a second modification of the second embodiment, the determination unit 14 may select a cause of abnormality based on the determination result for the measurement result of the imaging performance and / or the determination result for the measurement result of the excitation output, and the determination result for the measurement result of the irradiation density. In a second modification of the third embodiment, the determination unit 14 may select a cause of abnormality based on the determination result for the measurement result of the irradiation density, and one or more of the determination result for the measurement result of the imaging performance, the determination result for the measurement result of the detection sensitivity, and the determination result for the measurement result of the excitation output. The determination unit 14 may then output the selected abnormality cause to the display unit 15, and may also output a countermeasure to be taken by a user of the optical microscope 1 to the display unit 15 based on the selected abnormality cause.

[0111] Since the irradiation density has a high correlation with the change in fluorescence intensity that accompanies a change in imaging performance, it becomes possible to determine whether the fluorescence intensity change is normal or abnormal with higher accuracy.

[0112] FIG. 18 is a diagram illustrating an example of a judgment result for each measurement result of irradiation density and excitation output, an abnormality cause (NG cause) selected based on the judgment result, and a countermeasure for the abnormality cause according to Modification 2 of the second or third embodiment. FIG. 19 is a diagram illustrating an example of a judgment result for each measurement result of irradiation density and detection sensitivity, an abnormality cause (NG cause) selected based on the judgment result, and a countermeasure for the abnormality cause according to Modification 2 of the third embodiment. FIG. 20 is a diagram illustrating an example of a judgment result for each measurement result of irradiation density and imaging performance, an abnormality cause (NG cause) selected based on the judgment result, and a countermeasure for the abnormality cause according to Modification 2 of the second or third embodiment. As illustrated in FIGS. 18 to 20 , the judgment results for two measurement results including irradiation density enable the selection of an abnormality cause and the presentation of a countermeasure for the abnormality cause.

[0113] FIG. 21 illustrates an example of the judgment results for the measurement results of the irradiation density, imaging performance, and excitation output, the cause of anomaly (NG cause) selected based on the judgment results, and a countermeasure for the abnormality cause, according to Modification 2 of the second or third embodiment. As illustrated in FIG. 21 , the judgment results for three measurement results, including the irradiation density and imaging performance, enable the selection of the cause of anomaly and the presentation of a countermeasure for the abnormality cause. In addition, in the example shown in FIG. 21 , if the judgment results for the measurement results of the imaging performance and excitation output are normal (imaging OK, output OK) but the judgment result for the measurement result of the irradiation density is abnormal (density NG), it is considered that although the imaging performance and excitation output have not yet been judged to be abnormal, the irradiation density is beginning to be affected. Therefore, in this case, as a countermeasure, a warning can be issued indicating signs of deterioration in the imaging performance and excitation output, thereby further improving the accuracy of the countermeasure.

[0114] <Modification 3 of the Second and Third Embodiments> In the second and third embodiments, the excitation output may be measured using each of a plurality of light sources, and the measurement results of the excitation output using each of the plurality of light sources may be used to identify the cause of the abnormality when the determination result of the excitation output measurement is abnormal. For example, when the determination result of the excitation output measurement is abnormal, a decrease in light source output and / or a decrease in transmittance of the excitation light path may be identified as the cause of the abnormality. However, when the determination results of the excitation output measurement using each of the plurality of light sources are also abnormal, the cause of the abnormality may be narrowed down to a decrease in transmittance of the excitation light path. In this way, when the determination result of the excitation output measurement is abnormal, the cause of the abnormality can be more accurately identified. Note that the determination of whether each of the measurement results of the excitation output using each of the plurality of light sources is normal or abnormal is performed by the determination unit 14.

[0115] <Hardware Configuration of Arithmetic Device 3> The arithmetic device 3 may be realized by a computer 300 illustrated in Fig. 22. Fig. 22 is a diagram illustrating an example of the hardware configuration of the computer.

[0116] The computer 300 illustrated in FIG. 22 comprises a processor 301, a memory 302, a storage device 303, a portable storage medium drive device 304, a communication interface 305, and an input / output interface 306, each of which is connected to a bus 307 and is capable of sending and receiving data to and from each other.

[0117] The processor 301 may be, for example, a single processor, a multi-processor, or a multi-core processor. The processor 301 executes programs such as an operating system (OS) and applications to perform various processes (for example, the processes illustrated in FIG. 7 ).

[0118] The memory 302 includes a RAM (Random Access Memory) and a ROM (Read Only Memory). The RAM temporarily stores parts of the programs executed by the processor 301. The RAM is also used as a working storage area for the processor 301. The ROM stores the programs executed by the processor 301 and various data required for executing the programs.

[0119] The storage device 303 is a device for storing data, such as a hard disk drive (HDD) or a solid state drive (SSD).

[0120] The portable storage medium drive device 304 drives a portable storage medium 304a, accesses its storage contents, and reads and writes data. The portable storage medium 304a is a memory device, a flexible disk, an optical disk, a magneto-optical disk, etc. The portable storage medium 304a also includes a CD-ROM (Compact Disc Read Only Memory), a DVD (Digital Versatile Disc), a Blu-ray Disc, a USB (Universal Serial Bus) memory, an SD card memory, etc.

[0121] The communication interface 305 is connected to a network via a wired or wireless connection, and communicates with external devices connected to the network.

[0122] The input / output interface 306 is connected to an external device and performs input / output of data between the external device and the input / output interface 306. The external device connected to the input / output interface 306 is, for example, the microscope body 2 and the display device 4, and may further be connected to an input device. The input device may be a keyboard, a mouse, a joystick, a touch panel, or the like.

[0123] In such a computer 300, the programs executed by the processor 301 and various data required for executing the programs may be stored in the storage device 303 or the portable storage medium 304a, not limited to the memory 302. Furthermore, the programs executed by the processor 301 and various data required for executing the programs may be stored in the storage device 303 or the portable storage medium 304a via the communication interface 305 from an external device connected to the network.

[0124] Furthermore, the computer 300 is not limited to the one illustrated in FIG. 22, and may be configured to include multiple components that are part of the components illustrated in FIG. 22, or may be configured to omit some of the components that are illustrated in FIG. 22.

[0125] The computer 300 may also be configured to include hardware such as a microprocessor, a DSP, an ASIC (Application Specific Integrated Circuit), a PLD (Programmable Logic Device), an FPGA (Field-Programmable Gate Array), etc. For example, the processor 301 may be implemented using at least one of these pieces of hardware.

[0126] 22, the functions of a part of the imaging performance measurement unit 12, the determination unit 14, and the irradiation density measurement unit are realized by a processor 301. The functions of the storage unit 13 are realized by one or more of a memory 302, a storage device 303, and a portable storage medium 304a.

[0127] Although the embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments, and various improvements and modifications can be made without departing from the spirit and scope of the present invention.

[0128] REFERENCE SIGNS LIST 1 Optical microscope 2 Microscope body 3 Calculation device 4 Display device 11 Detection sensitivity measurement unit 12 Imaging performance measurement unit 13 Memory unit 14 Determination unit 15 Display unit 16 Excitation output measurement unit 201 Nosepiece 202, 202a, 202b, 202c Objective lens 203 Laser light source 204, 204a, 204b, 204c, 204d Detector 205 Beam splitter 206 Dichroic mirror 207a, 207b Galvanometer mirror 208 Lens 209 Dichroic mirror 210 Mirror 211 Fluorescence cube 212 Stage 213 Lens 214 Confocal pinhole 215, 216, 217, 218 Dichroic mirror 219 Transmission detector 220 Condenser lens 221, 222 Mirror 223 Lens 224 Transmitted illumination light source 225 Excitation light intensity monitor 226 Partially reflecting mirror 227 Reflecting element 228 Fluorescent bead specimen 229 Pinhole specimen 230 Edge chart specimen 300 Computer 301 Processor 302 Memory 303 Storage device 304 Portable storage medium drive device 304a Portable storage medium 305 Communication interface 306 Input / output interface 307 Bus

Claims

1. An optical microscope including an objective lens, a light source that outputs excitation light, and an excitation output measurement unit that measures the excitation output, characterized in that the optical microscope is further equipped with an imaging performance measurement unit that measures imaging performance, and a judgment unit that judges the performance of the optical microscope based on the measurement results of the imaging performance and the measurement results of the excitation output.

2. The optical microscope according to claim 1, further comprising a memory unit that stores a threshold value for determining whether the measurement result is normal or abnormal, and the determination unit compares the measurement result with the threshold value stored in the memory unit and determines whether the measurement result is normal or abnormal.

3. The optical microscope according to claim 2, characterized in that the memory unit further stores an abnormality cause corresponding to a combination of a judgment result for the measurement result of the imaging performance and a judgment result for the measurement result of the excitation output, and the judgment unit selects the abnormality cause based on the judgment result for the measurement result of the imaging performance and the judgment result for the measurement result of the excitation output.

4. The optical microscope according to claim 3, characterized in that the judgment unit outputs a countermeasure to be taken by a user of the optical microscope based on the selected cause of the abnormality.

5. The optical microscope according to claim 4, characterized in that, when the judgment result for the measurement result of the imaging performance is abnormal, the judgment unit outputs a countermeasure that prioritizes countermeasures for the imaging performance as the countermeasure.

6. An optical microscope as described in claim 1, further comprising an irradiation density measuring unit that measures irradiation density, the irradiation density measuring unit calculating the irradiation density based on the measurement results of the imaging performance and the measurement results of the excitation output.

7. An optical microscope as described in claim 6, further comprising a memory unit that stores a threshold value for determining whether the measurement result is normal or abnormal, and the determination unit compares the measurement result with the threshold value stored in the memory unit and determines whether the measurement result is normal or abnormal.

8. The optical microscope according to claim 7, characterized in that the memory unit further stores an abnormality cause corresponding to a combination of a judgment result for the measurement result of the imaging performance and / or a judgment result for the measurement result of the excitation output and a judgment result for the measurement result of the irradiation density, and the judgment unit selects the abnormality cause based on the judgment result for the measurement result of the imaging performance and / or the judgment result for the measurement result of the excitation output and a judgment result for the measurement result of the irradiation density.

9. An optical microscope including an objective lens, a detector that detects light from a specimen, and a detection sensitivity measurement unit that measures detection sensitivity, characterized in that the optical microscope further includes an imaging performance measurement unit that measures imaging performance, and a judgment unit that judges the performance of the optical microscope based on the measurement results of the imaging performance and the measurement results of the detection sensitivity.

10. An optical microscope as described in claim 9, further comprising a memory unit that stores a threshold value for determining whether the measurement result is normal or abnormal, and the determination unit compares the measurement result with the threshold value stored in the memory unit and determines whether the measurement result is normal or abnormal.

11. The optical microscope according to claim 10, characterized in that the memory unit further stores an abnormality cause corresponding to a combination of a judgment result for the measurement result of the imaging performance and a judgment result for the measurement result of the detection sensitivity, and the judgment unit selects the abnormality cause based on the judgment result for the measurement result of the imaging performance and the judgment result for the measurement result of the detection sensitivity.

12. The optical microscope according to claim 11, wherein the judgment unit outputs a countermeasure to be taken by a user of the optical microscope based on the selected cause of the abnormality.

13. The optical microscope according to claim 12, characterized in that, when the judgment result for the measurement result of the imaging performance is abnormal, the judgment unit outputs, as the countermeasure, a countermeasure that prioritizes countermeasures for the imaging performance.

14. The optical microscope according to claim 9, further comprising a light source that outputs excitation light and an excitation output measurement unit that measures the excitation output, wherein the judgment unit judges the performance of the optical microscope based on the measurement results of the imaging performance, the measurement results of the detection sensitivity, and the measurement results of the excitation output.

15. An optical microscope as described in claim 14, further comprising a memory unit that stores a threshold value for determining whether the measurement result is normal or abnormal, and the determination unit compares the measurement result with the threshold value stored in the memory unit and determines whether the measurement result is normal or abnormal.

16. The optical microscope according to claim 15, characterized in that the memory unit further stores an abnormality cause corresponding to a combination of a judgment result for the measurement result of the imaging performance, a judgment result for the measurement result of the detection sensitivity, and a judgment result for the measurement result of the excitation output, and the judgment unit selects the abnormality cause based on the judgment result for the measurement result of the imaging performance, the judgment result for the measurement result of the detection sensitivity, and the judgment result for the measurement result of the excitation output.

17. The optical microscope according to claim 16, characterized in that the judgment unit outputs a countermeasure to be taken by a user of the optical microscope based on the selected cause of the abnormality.

18. The optical microscope according to claim 17, characterized in that, when the judgment result for the measurement result of the imaging performance is abnormal, the judgment unit outputs, as the countermeasure, a countermeasure that prioritizes countermeasures for the imaging performance.

19. An optical microscope as described in claim 14, further comprising an irradiation density measuring unit that measures irradiation density, the irradiation density measuring unit calculating the irradiation density based on the measurement results of the imaging performance and the measurement results of the excitation output.

20. An optical microscope as described in claim 19, further comprising a memory unit that stores a threshold value for determining whether the measurement result is normal or abnormal, and the determination unit compares the measurement result with the threshold value stored in the memory unit and determines whether the measurement result is normal or abnormal.

21. The optical microscope of claim 20, wherein the memory unit further stores an abnormality cause corresponding to a combination of one or more of the judgment result for the measurement result of the imaging performance, the judgment result for the measurement result of the detection sensitivity, and the judgment result for the measurement result of the excitation output, and a judgment result for the measurement result of the irradiation density, and the judgment unit selects the abnormality cause based on one or more of the judgment result for the measurement result of the imaging performance, the judgment result for the measurement result of the detection sensitivity, and the judgment result for the measurement result of the excitation output, and a judgment result for the measurement result of the irradiation density.

22. The optical microscope according to any one of claims 1 to 21, characterized in that the optical microscope is a laser scanning fluorescence microscope.

23. An optical microscope as described in any one of claims 3 to 5, 8, 11 to 13, 16 to 18, and 21, further comprising a display unit that displays at least one of the measurement results, the judgment results, and the cause of the abnormality.

24. The optical microscope according to any one of claims 9 to 21, characterized in that the optical microscope is a confocal microscope, and further, measures the amount of deviation of the optical axis incident on a confocal pinhole.

25. An optical microscope as claimed in any one of claims 1 to 21, characterized in that the imaging performance measurement unit measures the imaging performance using each of a plurality of objective lenses, and the judgment unit judges whether each of the measurement results of the imaging performance using each of the plurality of objective lenses is normal or abnormal.

26. An optical microscope as described in any one of claims 9 to 21, characterized in that the detection sensitivity measurement unit measures the detection sensitivity using each of a plurality of detectors, and the judgment unit judges whether each of the measurement results of the detection sensitivity using each of the plurality of detectors is normal or abnormal.

27. An optical microscope as described in any one of claims 1 to 8 and 14 to 21, characterized in that the excitation output measurement unit measures the excitation output using each of a plurality of light sources, and the judgment unit judges whether each of the measurement results of the excitation output using each of the plurality of light sources is normal or abnormal.

28. The optical microscope according to claim 8, characterized in that the judgment unit outputs a countermeasure to be taken by a user of the optical microscope based on the selected cause of the abnormality.

29. The optical microscope according to claim 21, characterized in that the judgment unit outputs a countermeasure to be taken by a user of the optical microscope based on the selected cause of the abnormality.

30. The optical microscope according to any one of claims 4, 12, 17, 28 and 29, characterized in that the judgment unit outputs, as the countermeasure, a countermeasure by executing a correction function, correction, or maintenance / repair.

31. An optical microscope as described in any one of claims 1 to 8 and 14 to 21, characterized in that the excitation output measuring unit is equipped with an intensity monitor that measures the intensity of the excitation light, and the intensity monitor is provided in the optical path on the light source side of the objective lens, near the focal plane of the objective lens, or on the optical path on the opposite side of the objective lens across the focal plane of the objective lens.

32. An optical microscope as claimed in any one of claims 9 to 21, characterized in that the detection sensitivity measurement unit comprises: a reflective element or phosphor provided in the optical path on the light source side of the objective lens; a reference specimen or reference light source arranged in the vicinity of the focal position of the objective lens; or a light source provided on the opposite side of the objective lens across the focal position of the objective lens.

33. An optical microscope according to any one of claims 1 to 21, characterized in that the imaging performance measurement unit acquires an image of a reference specimen placed at the focal position of the objective lens, and calculates a PSF based on the image.

34. An optical microscope according to any one of claims 1 to 21, characterized in that the imaging performance measurement unit measures the wavefront of the excitation light at the focal plane of the objective lens and calculates a PSF based on the measured wavefront.

35. An optical microscope according to any one of claims 1 to 21, characterized in that the imaging performance measurement unit acquires an image of an edge specimen placed at the focal position of the objective lens, and calculates an LSF based on the image.

36. The optical microscope according to claim 35, wherein the edge specimen is a chart specimen having a radial pattern with edges in eight or more directions around a full 360° circumference.

37. A performance evaluation method for evaluating the performance of an optical microscope, comprising: measuring excitation output; measuring imaging performance; and evaluating the performance of the optical microscope based on the measurement results of the excitation output and the measurement results of the imaging performance.

38. A performance evaluation method for evaluating the performance of an optical microscope, comprising: measuring the detection sensitivity of a detector that detects light from a specimen; measuring imaging performance; and evaluating the performance of the optical microscope based on the measurement results of the detection sensitivity and the measurement results of the imaging performance.

39. A performance evaluation method for evaluating the performance of an optical microscope, comprising: measuring excitation output; measuring the detection sensitivity of a detector that detects light from a specimen; measuring imaging performance; and evaluating the performance of the optical microscope based on the measurement results of the excitation output, the measurement results of the detection sensitivity, and the measurement results of the imaging performance.

40. An optical microscope comprising: a light source that outputs excitation light; an objective lens; an imaging performance measuring unit that measures imaging performance; and an excitation output measuring unit that measures excitation output, wherein the excitation output measuring unit is built into the microscope body of the optical microscope.

41. An optical microscope comprising: an objective lens; a detector that detects light from a specimen; an imaging performance measuring unit that measures imaging performance; and a detection sensitivity measuring unit that measures detection sensitivity, wherein the detection sensitivity measuring unit is built into a microscope body of the optical microscope.

42. An optical microscope according to claim 41, further comprising: a light source that outputs excitation light; and an excitation output measuring unit that measures the excitation output, the excitation output measuring unit being built into the microscope body of the optical microscope.

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