JP2009230021A - Optical microscope and spectrum measuring method - Google Patents

Abstract

An optical microscope and a spectrum measuring method capable of shortening a measuring time are provided.
An optical microscope includes a light source 11, a detector 27 that detects light from a sample 21, a confocal optical system 30 that collects light from the light source 11 in a line shape and guides the light to the sample 21, and a slit. And a diffraction grating 25 that disperses the light that has passed through 23 in a direction perpendicular to the slit 23. The detector 27 receives a plurality of signal charges transferred by the vertical transfer register in order to read the signal charges, a vertical transfer register that transfers the signal charges generated in the light receiving parts in a direction perpendicular to the slit 23, and the signal charges. And a horizontal transfer register that accumulates the light receiving portion and transfers the light collectively.
[Selection] Figure 1

Description

  The present invention relates to an optical microscope and a spectrum measuring method, and more particularly to an optical microscope having a confocal optical system for irradiating a sample with line-shaped light and a spectrum measuring method using the same.

  In a fluorescence microscope, fluorescence having a wavelength different from that of excitation light is generated by irradiating a sample with excitation light. By analyzing the spectrum of the fluorescence generated in the sample, the substance contained in the sample can be analyzed. An optical microscope for detecting such fluorescence or Raman scattered light by spectroscopic analysis is disclosed (Patent Document 1). In the optical microscope of Patent Document 1, light from a light source is converted into line-shaped light. And the Raman spectrum is measured via the line confocal optical system.

  In Patent Document 1, light that has passed through a slit is split in a direction perpendicular to the slit direction by a spectroscope. A spectroscope disperses light according to wavelength. The sample and the slit are arranged at conjugate positions. Thereby, the spectrum in a linear area | region is acquirable. That is, light is dispersed on the light receiving surface of the CCD camera according to the wavelength. Therefore, it is possible to measure the spectrum in the linear region with one frame of the CCD camera. By doing so, the measurement time can be shortened.

JP 2005-73972 A

  In the above optical microscope, light is dispersed on the light receiving surface of the CCD camera. Accordingly, light of a certain wavelength (bandwidth) is incident on the light receiving pixels of the CCD camera. That is, light with different wavelengths is incident on different light receiving pixels.

  Here, the bandwidth per light receiving pixel is determined according to the performance of the spectrometer and the size of the light receiving pixel. In order to perform finer spectrum measurement, it is necessary to increase the number of channels. When the number of channels for measuring the spectrum is increased, it is necessary to increase the number of light receiving pixels. When the number of light receiving pixels is increased, the time for reading signal charges in the CCD camera becomes longer. Therefore, the spectrum measurement time becomes long.

As described above, the conventional measurement has a problem that it takes a long time to measure spectrum data.
The present invention has been made in view of the above-described problems, and an object thereof is to provide an optical microscope and a spectrum measurement method that can shorten the measurement time of spectrum data.

  The optical microscope according to the first aspect of the present invention includes a light source, a two-dimensional array photodetector for detecting light from the sample, and condensing the light from the light source in a line to guide the sample, A confocal optical system for guiding light from the sample to a slit arranged at a position conjugate with the sample, and provided between the slit of the confocal optical system and the two-dimensional array photodetector, A wavelength dispersion element that disperses light that has passed through the slit in a direction perpendicular to the slit according to a wavelength, and the two-dimensional array photodetectors are arranged along the direction of the slit, A light receiving pixel that generates a signal charge corresponding to the intensity of light, a vertical transfer unit that transfers the signal charge generated in the light receiving pixel in a direction perpendicular to the slit, and a transfer unit that reads the signal charge in the vertical transfer unit. Is The signal charges plurality of the accumulated received pixels, collectively those having a horizontal transfer section for transferring. As a result, it is possible to shorten the image capture time and the measurement time.

  An optical microscope according to a second aspect of the present invention is the above-described optical microscope, wherein in the two-dimensional array photodetector, signal charges generated in the light receiving pixels are amplified and read out. It is what. Thereby, damage to a sample can be prevented.

  An optical microscope according to a third aspect of the present invention is the above-described optical microscope, wherein the number of light receiving pixels in the vertical direction read out collectively by the horizontal transfer unit is variable according to the readout line. is there. Thereby, a required bandwidth can be set freely. In addition, since signal charges in a wavelength band that does not need attention can be read out together, the measurement time can be further shortened.

  An optical microscope according to a fourth aspect of the present invention is the optical microscope described above, wherein the light source is a lamp light source, and a filter that passes light of a part of the wavelength from the light from the lamp light source is provided. It is what has been. Thereby, the light of a predetermined wavelength can be irradiated to a sample.

  The spectrum measurement method according to the fifth aspect of the present invention includes a light source, a two-dimensional array photodetector for detecting light from the sample, and condensing the light from the light source in a line and guiding it to the sample. A confocal optical system for guiding light from the sample to a slit disposed at a position conjugate with the sample; and provided between the slit of the confocal optical system and the two-dimensional array photodetector. A spectrum measuring method using an optical microscope comprising a wavelength dispersion element that disperses light that has passed through the slit in a direction perpendicular to the slit according to a wavelength, in the two-dimensional array photodetector, A step of generating a signal charge corresponding to the intensity of light from the sample at the light receiving pixels arranged along the direction of the slit, and a signal charge generated at the light receiving pixel in a direction perpendicular to the slit. The method comprising, for reading out the signal charge, said transfer signal charges in the vertical direction and a plurality of storage light receiving pixel row, and has a step of transferring in the horizontal direction together. As a result, it is possible to shorten the image capture time and the measurement time.

  The spectrum measurement method according to a sixth aspect of the present invention is the spectrum measurement method described above, wherein the signal charges generated in the light receiving pixels are amplified and read out in the two-dimensional array photodetector. It is what. Thereby, damage to a sample can be prevented.

  The spectrum measurement method according to a seventh aspect of the present invention is the spectrum measurement method described above, wherein the number of rows of light receiving pixels in the vertical direction that are collectively read in the horizontal direction is variable according to the readout line. is there. Thereby, a required bandwidth can be set freely. In addition, since signal charges in a wavelength band that does not need attention can be read out together, the measurement time can be further shortened.

  The spectrum measurement method according to an eighth aspect of the present invention is the spectrum measurement method described above, wherein the light source is a lamp light source, and a filter that passes light of a part of the wavelength from the light from the lamp light source. Is provided. Thereby, the light of a predetermined wavelength can be irradiated to a sample.

  ADVANTAGE OF THE INVENTION According to this invention, the optical microscope and spectrum measuring method which can shorten measurement time can be provided.

  Embodiments to which the present invention is applicable will be described below. The following description is to describe the embodiment of the present invention, and the present invention is not limited to the following embodiment. For clarity of explanation, the following descriptions are omitted and simplified as appropriate. Moreover, those skilled in the art will be able to easily change, add, and convert each element of the following embodiments within the scope of the present invention. In addition, what attached | subjected the same code | symbol in each figure has shown the same element, and abbreviate | omits description suitably.

  An optical microscope according to an embodiment of the present invention will be described with reference to FIG. FIG. 1 is a diagram schematically showing the configuration of the optical system of the optical microscope according to the present embodiment. The optical microscope is a confocal microscope using the confocal optical system 30. The optical microscope includes a light source 11, an interference filter 12, a lens 13, a slit 14, a lens 15, a beam splitter 16, a galvano mirror 17, a lens 18, a lens 19, an objective lens 20, a lens 22, a slit 23, a lens 24, and a diffraction grating 25. A lens 26 and a detector 27. Here, the description will be made assuming that the optical microscope is a fluorescence microscope that detects fluorescence generated in the sample 21.

  The light source 11 is, for example, a lamp light source. Therefore, the light source 11 emits excitation light. This excitation light is guided to the sample 21 by the confocal optical system 30. Thereby, the fluorescent substance in the sample 21 is excited and fluorescence is generated. Further, the fluorescence generated in the sample 21 is guided to the detector 27 by the confocal optical system 30. Here, the confocal optical system 30 is a line confocal optical system that makes the illumination area of light linear. In the following description, light incident on the sample 21 from the light source 11 is referred to as incident light, and light emitted from the sample 21 upon incidence of incident light is referred to as emitted light. The emitted light includes reflected light reflected from the sample 21 in addition to fluorescence. This reflected light has the same wavelength as the excitation light.

  Hereinafter, the configuration of the confocal optical system 30 will be described. White light having a predetermined spectrum is emitted from the light source 11. That is, light of a wavelength band having a predetermined width is emitted. The incident light from the light source 11 enters the interference filter 12. The interference filter 12 restricts the passage of light according to the wavelength. For example, the interference filter 12 is a band pass filter. Therefore, the interference filter 12 allows light of a certain wavelength to pass and blocks light of other wavelengths. In this case, the incident light that has passed through the interference filter 12 becomes monochromatic light. Furthermore, by exchanging the interference filter 12, the wavelength of the excitation light can be changed. That is, light having a desired wavelength can be selected by using a filter that transmits light having a predetermined wavelength. Therefore, the sample 21 can be analyzed more finely.

  Incident light that has passed through the interference filter 12 is refracted by the lens 13. The incident light condensed by the lens 13 enters the slit 14. The slit 14 is formed with a line-shaped opening. Incident light that has passed through the opening enters the lens 15. In addition, incident light incident on the outside of the opening is shielded. Therefore, the slit 14 converts incident light into line-shaped light. That is, the incident light that has passed through the slit 14 has a line shape. For example, when the direction of the opening of the slit 14 is the X direction, the longitudinal direction of the incident light is the X direction.

  The lens 15 refracts incident light from the slit 14 into a parallel light beam. The light from the lens 15 enters the beam splitter 16. The beam splitter 16 is, for example, a half mirror, and reflects about half of the incident light and transmits the other half. Accordingly, part of the incident light from the light source 11 passes through the beam splitter 16 and enters the galvanometer mirror 17. The galvanometer mirror 17 scans incident light and reflects it in the direction of the lens 18. That is, the galvanometer mirror 17 is a scanning unit that scans the light beam converted into a line shape. Incident light can be scanned by changing the angle of the galvanometer mirror 17 according to time. Thereby, the incident position of the incident light on the sample 21 can be changed. The galvanometer mirror 17 scans the incident light from the light source 11 in a direction perpendicular to the longitudinal direction of the incident light. That is, incident light is scanned in the Y direction on the sample 21. Note that the X direction and the Y direction are directions orthogonal to each other on a plane perpendicular to the optical axis.

  Incident light reflected by the galvanometer mirror 17 enters the lens 18. The incident light is refracted by the lens 18 and the lens 19 to become a parallel light beam. Incident light from the lens 19 enters the objective lens 20. The objective lens 20 condenses incident light from the light source 11 on the sample 21. The slit 14 and the sample 21 are arranged at conjugate positions. Therefore, on the sample 21, the illumination area of incident light is in a line shape. That is, the linear region of the sample 21 is illuminated by the incident light from the light source 11.

  When incident light (excitation light) enters the sample 21, fluorescence is generated from the sample 21. The sample 21 is a biological sample, for example, and generates fluorescence in response to excitation light. Of course, the sample 21 may be stained with a fluorescent material. The fluorescent substance in the sample 21 absorbs light energy and emits light having a wavelength longer than that of the excitation light. Here, the spectrum of the fluorescence generated from the sample 21 varies depending on the excitation light wavelength and the fluorescent material. That is, since the fluorescence wavelength is a value specific to the fluorescent material, even when excitation light having the same wavelength is irradiated, the fluorescence spectrum differs depending on the type of the fluorescent material. Therefore, the distribution of the fluorescent substance in the sample 21 can be analyzed by measuring the fluorescence spectrum. And the sample 21 radiate | emits the emitted light containing fluorescence according to incident light.

  Then, the emitted light generated from the sample 21 propagates in the original direction. That is, the emitted light from the sample 21 is refracted by the objective lens 20, the lens 19, and the lens 18 and enters the galvano mirror 17. The emitted light from the sample 21 includes reflected light in addition to fluorescence. The galvanometer mirror 17 reflects the outgoing light in the direction of the beam splitter 16 and descans it. Thereby, the outgoing light traveling from the galvano mirror 17 toward the beam splitter 16 propagates in the direction opposite to the traveling direction of the incident light traveling from the beam splitter 16 toward the galvano mirror 17. That is, the propagation directions of incident light and outgoing light are opposite to each other.

  Then, light emitted from the galvanometer mirror 17 enters the beam splitter 16. Since the beam is descanned by the galvanometer mirror 17, the position of the incident light and the position of the emitted light coincide on the beam splitter 16. The beam splitter 16 is, for example, a half mirror, which reflects about half of the light and transmits the other half. Accordingly, part of the emitted light passes through the beam splitter 16 and enters the lens 22. Note that a dichroic mirror may be used as the beam splitter 16. That is, it is possible to use a dichroic mirror that transmits excitation light and reflects fluorescence. Thus, the light utilization efficiency can be improved by separating the excitation light and the fluorescence according to the wavelength. That is, when the emitted light from the sample 21 includes fluorescence and Rayleigh scattered light, only the fluorescence can be reflected by using the dichroic mirror.

  The light emitted from the beam splitter 16 is refracted by the lens 22. The lens 22 condenses the emitted light on the slit 23. The slit 23 and the sample 21 are arranged in a conjugate relationship with each other. Therefore, on the slit 23, the emitted light is in a line shape. The slit 23 has a line-shaped opening whose longitudinal direction is the X direction. The outgoing light that has passed through the opening enters the lens 24. In addition, incident light incident on the outside of the opening is shielded.

  As described above, the confocal optical system 30 having slits on the incident side and the detection side is used. Therefore, light from other than the focal position is blocked by the slit 23. Thereby, the spatial resolution in the Z direction (optical axis direction) can be improved. Further, the in-focus position can be changed in the Z direction by moving the objective lens 20 up and down. The focal position on the sample 21 can be scanned in the Z direction by changing the relative distance between the objective lens 20 and the sample 21. Further, the linear light extending in the X direction is scanned in the Y direction by the galvanometer mirror 17. As a result, scanning in the XYZ directions is possible, and the three-dimensional spatial distribution of the fluorescence spectrum can be measured. In other words, the fluorescence spectrum can be measured at an arbitrary point on the three-dimensional sample 21. Even when such a three-dimensional spatial distribution of spectrum is measured, the measurement time can be shortened by illuminating in a line shape. Therefore, spectrum measurement can be performed in a short time.

  The outgoing light that has passed through the slit 23 enters the lens 24. The lens 24 refracts outgoing light. Then, light emitted from the lens 24 enters the diffraction grating 25. The diffraction grating 25 is a wavelength dispersion element, and disperses light according to the wavelength. Here, the diffraction grating 25 disperses light in a direction perpendicular to the direction of the slit 23. Therefore, the outgoing light from the diffraction grating 25 is dispersed in the Y direction. That is, the wavelength dispersion direction by the diffraction grating 25 is parallel to the Y direction. The outgoing light propagates at different angles depending on the diffraction grating 25. The light from the diffraction grating 25 enters the detector 27 through the lens 26. Instead of the diffraction grating 25, prisms or the like may be dispersed.

  The detector 27 is an area sensor in which light receiving elements are arranged in a matrix. That is, the detector 27 is a two-dimensional array photodetector such as a two-dimensional CCD in which pixels are arranged in an array. The detector 27 is an electron multiplying CCD (electron multiplying CCD). Therefore, the signal charge generated by the photoelectric conversion is multiplied and read out.

  Here, a principle configuration diagram of the detector 27 is shown in FIG. As shown in FIG. 2, the detector 27 includes light receiving units 71 arranged at a predetermined pitch in the horizontal direction (X direction) and the vertical direction (Y direction), and a vertical provided on one side of the light receiving units 71 of each column. A vertical transfer register 72 having a CCD structure extending in the direction and a horizontal transfer register 73 having a CCD structure provided at one end of each vertical transfer register 72 are provided. Each of these light receiving portions 71 serves as a light receiving pixel. In each light receiving portion 71, signal charges corresponding to the amount of received light are generated by photoelectric conversion. That is, the number of signal charges corresponding to the fluorescence intensity generated in the sample 21 is generated. Then, the signal charges are transferred to the corresponding vertical transfer registers 72. The signal charges of these vertical transfer registers 72 are transferred to the horizontal transfer register 73, and the signal charges are read out. And the detection signal according to the fluorescence intensity received by each pixel is output. In addition, it cannot be overemphasized that the imaging device concerning this invention is not limited to the detector 27 of the structure shown in FIG.

  Thus, on the light receiving surface of the detector 27, the light receiving portions 71 serving as light receiving pixels are arranged in a matrix. Here, it is assumed that the light receiving portions 71 are arranged along the X direction and the Y direction. On the light receiving surface, the X direction and the Y direction are perpendicular to each other. In FIG. 2, four light receiving parts 71 are arranged in the X direction, and six light receiving parts 71 are arranged in the Y direction. Therefore, the light receiving portions 71 of 6 rows × 4 columns are arranged.

  The light receivers 71 are arranged along the X direction and the Y direction. That is, the light receiving portions 71 are arranged in parallel with the direction of the slit 23 (X direction). Furthermore, the light receiving parts 71 are arranged along the wavelength dispersion direction (Y direction) by the diffraction grating 25. Therefore, the fluorescence dispersed by the diffraction grating 25 is detected by different light receiving portions 71 according to the position on the sample 21 and the wavelength. Therefore, on the light receiving surface of the detector 27, the X direction corresponds to the position on the sample 21, and the Y direction corresponds to the wavelength. Therefore, the light receiving portions 71 adjacent in the X direction receive fluorescence from different positions on the sample 21. In addition, the light receiving units 71 adjacent in the Y direction receive fluorescence of different wavelengths generated at the same position on the sample 21. One row of light receiving portions 71 extending in the X direction detects fluorescence having a certain bandwidth. When one line is read out in the horizontal transfer register 73, a detection signal corresponding to the fluorescence intensity of a predetermined bandwidth is output. This detection signal corresponds to a line-shaped region on the sample 21.

  And the detector 27 outputs the detection signal according to the light intensity of the emitted light received by each light-receiving part 71 to a processing apparatus (not shown). The processing device is an information processing device such as a personal computer (PC), and stores a detection signal from the detector 27 in a memory or the like. Then, the detection result is subjected to a predetermined process and displayed on the monitor. Further, the processing device controls scanning of the galvanometer mirror 17 and driving of the stage (not shown) of the sample 21. Here, the Y direction of the detector 27 corresponds to the wavelength of the emitted light. For example, the light receiving unit 71 in the first row at the upper end detects outgoing light with a long wavelength, and the light receiving unit 71 in the sixth row at the lower end detects outgoing light with a short wavelength. Of course, the opposite is also possible. Thus, the light intensity distribution in the Y direction of the detector 27 indicates a spectral distribution. Therefore, the spectrum at each point in the line-shaped region can be acquired with one frame of the detector 27.

  The detector 27 is an electron multiplying CCD. Therefore, in the horizontal transfer register 73, the signal charge is multiplied and read out. That is, electrons are multiplied at the time of horizontal transfer. Then, the multiplied signal charge is read out. An electron multiplying CCD has a quantum efficiency of about 90%, which is higher than that of a photomultiplier. Therefore, observation can be performed even when the fluorescence intensity is low. Furthermore, since the integration time can be shortened, the image capture time can be shortened. The excitation light intensity can be lowered, and damage or discoloration to the sample 21 can be prevented. Furthermore, since only one detector 27 is required, a compact optical system can be configured. By using an EMCCD with high quantum efficiency in this way, it is possible to perform measurement in a shorter time. Instead of the electron multiplying CCD, a CCD camera with an image intensifier (ICCD) may be used.

  The vertical transfer register 72 transfers signal charges in the dispersion direction, and the horizontal transfer register 73 transfers in the slit direction. Furthermore, in this embodiment, a vertical pixel mixed readout operation (binning operation) is performed. Thereby, the bandwidth (wavelength band) can be controlled for each readout line. For example, signal charges for two rows are accumulated in the horizontal transfer register 73 and read together. That is, signal charges for two rows are read as one line. The horizontal transfer of one line is completed by reading the signal charges for two rows for each column. Therefore, in the detection signals corresponding to one line, the signal charges for two pixels adjacent in the Y direction are totaled. In this case, a fluorescence intensity having a bandwidth that is approximately twice that of when one row of signal charges is read out is acquired. When there are 24 light receiving pixels, when the signal charges for two rows are read together, 12 light intensity data, which is half the number of light receiving pixels, are acquired in one frame.

  In the horizontal transfer register 73, the bandwidth increases as the number of rows to be read together increases. Further, in the horizontal transfer register 73, the bandwidth becomes narrower as the number of rows read together is reduced. Therefore, when roughly measuring spectral data, two or three pixel rows are accumulated, and signal charges are read together. Thereby, since the transfer time is shortened, the measurement time can be shortened. Of course, four or more pixel rows may be accumulated and read out together. In addition, when the spectral data is measured finely, the signal charge is read for each pixel row.

  As described above, the horizontal transfer register 73 accumulates the signal charges transferred by the vertical transfer register 72 for the plurality of light receiving portions 71 and transfers them together. By doing so, the bandwidth to be read can be widened. That is, when the binning operation is performed, when one line is read out by the horizontal transfer register 73, the fluorescence intensity of the bandwidth corresponding to the plurality of light receiving units 71 is detected. Therefore, high-speed reading can be performed and measurement time can be shortened.

  Furthermore, due to the relationship between the excitation light and the fluorescent material, it is also possible to read out signal charges for a plurality of light receiving portions 71 in a wavelength band where no fluorescence is generated. That is, in a wavelength band that does not require attention, signal charges are read out in a large number of rows. By doing in this way, measurement time can be shortened. On the other hand, in the wavelength band in which the fluorescence spectrum is to be measured finely, the signal charge is read for each light receiving pixel row. In this way, if the number of light receiving pixel rows read out in one line is variable, the bandwidth can be adjusted for each readout line. Signal charges can be read at high speed, and the measurement time can be shortened. In addition, fine spectrum data can be obtained in a notable wavelength band. Therefore, fine spectrum analysis can be performed in a short measurement time.

  As described above, in order to obtain the spectrum data, the detector 27 generates the signal charges corresponding to the intensity of the light from the sample 21 by the light receiving portions 71 arranged along the direction of the slit 23. The vertical transfer register 72 transfers the signal charge generated in the light receiving unit 71 in the direction perpendicular to the slit 23. Furthermore, in order to read out the signal charges, the signal charges transferred in the vertical direction are accumulated by the horizontal transfer register 73 for a plurality of light receiving portions 71 and transferred together.

  By performing the binning operation in the vertical direction as described above, it is possible to shorten the image capturing time. Therefore, the spectrum can be measured in a short time, and a high-speed phenomenon can be observed. Furthermore, by reading out signal charges in unnecessary wavelength bands collectively, the reading time does not become long even when performing fine spectroscopic observation. Since a CCD camera is used as the detector 27, the number of channels can be increased as compared with the photomultiplier. Further, even when the number of channels is increased, it is possible to prevent the transfer time from becoming long.

  If the number of light-receiving units 71 read together by the horizontal transfer register 73 is variable according to the read line, the bandwidth read by one line in one frame of the detector 27 can be made variable. In this case, the bandwidth can be easily controlled by adjusting the number of vertical transfer trigger signals (vertical transfer pulses) for performing vertical transfer. That is, the number of pulses of the vertical transfer trigger signal may be increased. Therefore, the necessary bandwidth can be set freely. In addition, spectral data in a wavelength band that does not require observation can be read together. Thereby, the acquisition time of spectrum data can be shortened.

  In the above description, the fluorescence microscope has been described, but the present invention is not limited to this. Any microscope may be used as long as it detects the outgoing light emitted from the sample at a wavelength different from the wavelength of the incident light. For example, a microscope that detects Raman scattered light or a microscope that detects infrared absorption may be used. Even with these microscopes, the spatial distribution of the spectrum can be measured in a short time. Further, a laser beam may be converted into a line shape using a cylindrical lens instead of the slit 14. Further, the incident position of the incident light on the sample 21 may be scanned not only by the galvanometer mirror 17 but also by an XY stage.

It is a figure which shows the structure of the optical microscope concerning this invention. It is a figure which shows typically the structure of the detector used for the optical microscope concerning this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 11 Light source 12 Interference filter 13 Lens 14 Slit 15 Lens 16 Beam splitter 17 Galvano mirror 18 Lens 19 Lens 20 Objective lens 21 Sample 22 Lens 23 Slit 24 Lens 25 Diffraction grating 26 Lens 27 Detector 30 Confocal optical system 71 Light receiving part 72 Vertical Transfer register 73 Horizontal transfer register

Claims (8)

A light source;
A two-dimensional array photodetector for detecting light from the sample;
A confocal optical system for condensing the light from the light source in a line and guiding it to the sample, and guiding the light from the sample to a slit disposed at a position conjugate with the sample;
A wavelength dispersion element that is provided between the slit of the confocal optical system and the two-dimensional array photodetector, and disperses light that has passed through the slit in a direction perpendicular to the slit according to the wavelength,
The two-dimensional array photodetector is arranged along the direction of the slit, and a light receiving pixel that generates a signal charge according to the intensity of light from the sample;
A vertical transfer unit that transfers signal charges generated in the light receiving pixels in a direction perpendicular to the slit;
An optical microscope comprising: a horizontal transfer unit that stores the signal charges transferred by the vertical transfer unit for a plurality of the light receiving pixel rows and transfers them collectively to read out the signal charge.   The optical microscope according to claim 1, wherein the two-dimensional array photodetector is an EMCCD that amplifies and reads out signal charges generated in the light receiving pixels.   The optical microscope according to claim 1 or 2, wherein the number of rows of light receiving pixels that are collectively read by the horizontal transfer unit is variable according to the readout line. The light source is a lamp light source;
The optical microscope according to any one of claims 1 to 3, wherein a filter that passes light of a part of wavelengths among the light from the lamp light source is provided. A light source;
A two-dimensional array photodetector for detecting light from the sample;
A confocal optical system for condensing the light from the light source in a line and guiding it to the sample, and guiding the light from the sample to a slit disposed at a position conjugate with the sample;
A wavelength dispersion element that is provided between the slit of the confocal optical system and the two-dimensional array photodetector and disperses light that has passed through the slit in a direction perpendicular to the slit in accordance with the wavelength. A spectrum measurement method using an optical microscope,
In the two-dimensional array photodetector, in the light receiving pixels arranged along the direction of the slit, generating a signal charge according to the intensity of light from the sample;
Transferring signal charges generated in the light receiving pixels in a direction perpendicular to the slits;
A spectrum measuring method including a step of accumulating the signal charges transferred in the vertical direction for the plurality of light receiving pixel rows and transferring them in the horizontal direction in order to read out the signal charges.   6. The spectrum measuring method according to claim 5, wherein in the two-dimensional array photodetector, signal charges generated in the light receiving pixels are amplified and read out.   The spectrum measurement method according to claim 5 or 6, wherein the number of rows of light receiving pixels that are read together in the horizontal direction is variable according to the readout line. The light source is a lamp light source;
The spectrum measurement method according to any one of claims 5 to 7, wherein a filter that passes light of a part of the wavelength from the light from the lamp light source is provided.

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