JPH1048528A - Optical scanning microscope and image acquiring method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an optical scanning microscope capable of recording plural image data in a recording medium without lowering a time resolution in the optical scanning microscope for simultaneously acquiring plural images. SOLUTION: In a microscope 10, a sample 61 is scanned by a laser beam, a light beam from the sample is converted into an electric signal by photoelectric converting means 22, 32 and image data are formed, the microscope comprises plural photoelectric converting, means and frame memories 25, 35 for storing the respective images formed by the respective photoelectric converting means, a scanner control means 13 capable of changing the set of a number of scanning lines at the time of scanning the sample 61 by the laser beam is provided, the respective image data stored in the frame memories 25, 35 are synthesized by matching with the setting change of the number of scanning lines and a read-out address generating circuit 59 for outputting the image data from the frame memories 25, 35 so as to form image data of a screen with a video rate.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an optical scanning microscope, and more particularly to a microscope capable of simultaneously acquiring and recording a plurality of images.

[0002]

2. Description of the Related Art At present, an optical scanning microscope, which scans a sample or a sample by irradiating a sample or a sample with a laser beam and converts the light from the sample or the like into an image signal by computer processing or the like, has been widely used. This optical scanning microscope uses laser irradiation means such as a laser output device. Then, the laser beam from the laser irradiating unit is shaken by an XY scanner using a mirror, an acousto-optic element, or the like, and the sample is irradiated with the laser beam so as to scan a required range of the sample in two dimensions. Further, a luminance signal is formed by photoelectric conversion means such as a photomultiplier tube from the light from the sample that has received the laser beam. Then, a video signal is formed by synthesizing the luminance signal and the horizontal and vertical synchronizing signals in accordance with the scanning timing of the laser beam.
Further, the video signal is stored in a storage medium, the video signal is read from the storage medium, and the video of the sample is displayed on the monitor device.

[0003] When scanning a sample and acquiring an image, the optical scanning microscope requires three times per second.
There are a high-speed scanning microscope that acquires images at a video rate of 0 frame, and a low-speed scanning microscope that acquires several or less images per second. In many cases, a video rate recording device is used for image storage of a high-speed scanning microscope that acquires images at this video rate. As this video rate recording device, a general video tape recorder or a magneto-optical disk is used. In addition, a bird disk or the like is often used for image storage of a low-speed scanning microscope.

[0004] At present, a sample may be stained with a fluorescent dye or the like to emit light from the sample. In this case, the wavelength of the light emitted from the sample differs depending on the type of the fluorescent dye and the wavelength of the laser beam. And, depending on the condition of the stained sample, when it is irradiated with a laser beam,
The wavelength of light emitted from the sample may be different. In this way, when the wavelength of the light emitted from the sample differs depending on the state of the sample, the light emitted from the sample when a single laser beam is irradiated is separated by wavelength to observe a more detailed state of the sample. In addition, it has come to be practiced to individually record the sample images at the separated wavelengths on a storage medium.

When a plurality of images are obtained at the same time, a high-speed scanning microscope sometimes obtains a plurality of images in real time by using a plurality of video rate recording devices in parallel. In addition, a control device incorporating a plurality of frame memories and a video rate recording device may be used to sequentially record sample images of different wavelengths in one video rate recording device. In this case, one of a plurality of image data obtained by performing one scan by laser beam irradiation is directly recorded in the video rate storage device. Then, the other image data obtained at the same time is temporarily stored in the frame memory. The scanning of the sample by the laser beam is interrupted until all other images can be stored, and the image data stored in each frame memory is transferred to the video rate recording device. Therefore, it is possible to record image data for each frame of video based on different wavelengths in one video rate storage device.

Further, a video rate recording apparatus which performs image processing at a higher speed than a normal video rate of 30 frames per second may be used. In this case, one of the image data acquired while performing continuous scanning is directly recorded in the video rate storage device, and the other image data acquired simultaneously is temporarily stored in the frame memory. And
The image data stored in each frame memory is transferred to the high-speed video rate recording device.

[0007]

As described above, when a plurality of images are obtained simultaneously, in a high-speed scanning microscope, a plurality of images are obtained in real time by using a plurality of video rate recording devices in parallel. . However, when a plurality of video rate recording devices are used, when observing the image of the sample with one monitor device, each storage medium is sequentially switched and connected to the monitor device.
Therefore, originally, the images obtained at the same time cannot be simultaneously viewed, and it has been difficult to perform detailed observation.

[0008] In addition, when a plurality of monitor devices are used to observe the recorded image of each storage medium with each monitor device, there is often a disadvantage that it is difficult to synchronize the images. Further, there is a disadvantage that the operation of each monitor device is complicated. As described above, when a plurality of images are simultaneously acquired by a high-speed scanning microscope, image data is also recorded in one video rate storage device using a frame memory and a video rate storage device. I have. In this case, the scanning of the sample by the laser beam is interrupted for each scan, the image data once stored in the frame memory is transferred to the video rate recording device, and the image data for each frame is recorded in one video rate storage device. Things.

As described above, when image data of different wavelengths is recorded for each frame in one video rate recording apparatus, it is possible to easily observe images obtained simultaneously by frame advance or the like. However, when acquiring two images simultaneously, only 15 frames can be scanned per second,
When acquiring two images simultaneously, only 10 frames can be scanned per second. That is, the time resolution is reduced, and it may not be suitable for a sample having a fast reaction or activity.

Then, using a video rate recording device higher in speed than the normal video rate, the other image data obtained simultaneously is temporarily stored in a frame memory, and the image data stored in each frame memory is transferred to the video rate recording device. Things have also been done. In this case, image data can be recorded without reducing the time resolution. However, there is a disadvantage that the recording device and the monitor device are special and expensive.

When the number of images to be simultaneously acquired also increases in a low-speed scanning microscope, a hard disk for storing image data is required to be capable of high-speed processing, and the number of images to be acquired simultaneously is limited. Become. SUMMARY OF THE INVENTION The present invention has been made to solve the above problems, and provides an optical scanning microscope capable of recording a plurality of image data in one storage medium without lowering the time resolution.

[0012]

SUMMARY OF THE INVENTION The present invention relates to a microscope which scans a sample with a laser beam and converts light from the sample into electrical signals by photoelectric conversion means to form image data. Scanner control capable of changing the setting of the number of scanning lines when scanning a sample with a laser beam in an optical scanning microscope having a plurality of frame memories each storing image data formed by each photoelectric conversion unit. Means for outputting image data from each frame memory such that the image data stored in the frame memory is combined with the setting change of the number of scanning lines to form one screen of image data at a video rate. A read address generation circuit is provided.

As described above, since the optical scanning microscope according to the present invention has a plurality of photoelectric conversion means, a plurality of image data of the same material can be formed at the same time.
In addition, since there is a frame memory for storing the image data of each photoelectric conversion unit, image data necessary for forming one screen formed by each photoelectric conversion unit can be temporarily stored.

Since there is a scanner control means capable of changing the setting of the number of scanning lines, when forming image data by each photoelectric conversion means, the number of scanning lines forming one screen is reduced, and The data amount of the image data can be reduced. Further, since there is a read address generation circuit that can combine each image data stored in the frame memory and output it as one screen of image data in accordance with the setting change of the number of scanning lines, each monitor screen has An image obtained by synthesizing the image obtained by the photoelectric conversion means can be displayed.

Further, according to the present invention, as the scanner control means, when the setting of the scanning line is changed, a setting for reducing the number of the scanning lines to 1 / N which is a positive integer with respect to the standard number of the scanning lines is provided. In some cases, scanner control means may be set to be possible and the maximum of N is set to the number of photoelectric conversion means. In this manner, by setting the number of scanning lines to 1 / N and setting the maximum of N to the number of photoelectric conversion units, the total number of scanning lines formed by each photoelectric conversion unit to be used can be reduced to the standard number of scanning lines. It can be. Therefore, it is possible to easily combine and display the images of the image data formed by the photoelectric conversion means on one screen.

Further, as the scanner control means, when setting the number of scanning lines to 1 / N, the scanner control means may set the interval between scanning lines to N times the standard. As described above, when the number of scanning lines is set to 1 / N, the amount of image data is reduced to 1 / N by using the scanner control unit that can make the interval between the scanning lines N times the standard. Even at this time, the amount of image data can be reduced without changing the scanning range.

According to the present invention, superimposing means may be provided between the photoelectric conversion means and the frame memory, and the superimposing means may cause the frame memory to store image data based on pixel data obtained by adding corresponding pixel data in a plurality of images. is there. Thus, if the microscope is provided with the superimposing means,
By adding the same image data of the same place by the superimposing means, it is possible to easily form bright image data of the sample image.

The superimposing means comprises a frame buffer and an integrator. As described above, the microscope in which the frame buffer and the integrator constitute the superimposing means can easily perform the integration of the corresponding pixel data. Furthermore,
According to the present invention, when simultaneously acquiring a plurality of images of the same object by an optical scanning microscope having a plurality of photoelectric conversion units and a frame memory, the number of scanning lines is set to a fraction of the number of standard scanning lines. This is an image acquisition method for a scanning microscope in which image data is acquired and temporarily stored in a frame memory.

As described above, the method of temporarily storing each image data in which the number of scanning lines is a fraction of a number in the frame memory is as follows.
It is possible to sequentially read out the image data stored in each frame memory and to synthesize the image data of one frame. Then, when acquiring each image data with the number of scanning lines being a fraction, and storing it in the frame memory,
Each image data may be stored in a frame memory as image data obtained by integrating the same image data a plurality of times.

As described above, by integrating a plurality of identical image data, it is possible to form clear image data. Also, image data with the number of scanning lines reduced to a fraction is
An image can be acquired in a fraction of the time of a normal one-frame image acquisition, and integrated image data can be formed within a normal one-frame time.

[0021]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A first embodiment of an optical scanning microscope according to the present invention has a fluorescence detector as a first photoelectric conversion means 22, as shown in FIG. It also has a fluorescence detector as means 32. Further, the microscope 10 has analog-to-digital converters 23, 3 for converting output signals of the respective photoelectric conversion units 22, 32 into digital signals.
3 and frame memories 25 and 35 for storing digital data
Also have two each in accordance with the number of photoelectric conversion means.

That is, the first image data storage means 21 combining the first photoelectric conversion means 22, the first analog-to-digital converter 23, and the first frame memory 35, and the second photoelectric conversion means 32 and the second This is provided as a second image data storage unit 31 in which an analog-to-digital converter 33 and a second frame memory 35 are combined. The optical scanning microscope 10 has a laser light source as a laser irradiating unit 11, similarly to a conventional optical scanning microscope. or,
The optical scanning microscope 10 has a first dichroic mirror 17 that reflects a laser beam from a laser irradiation unit 11 and sends it to an XY scanner 15 and transmits light (light from a sample) from the XY scanner 15. . And
The XY scanner 15 has a combination of a resonant scanner and a galvanometer scanner.
It enables two-dimensional scanning by a laser beam at a video rate for forming a frame, and controls the scanner control means 13 to oscillate the traveling direction of the laser beam.

Accordingly, the laser beam emitted from the laser irradiation means 11 is sent to the XY scanner 15 by the first dichroic mirror 17, and illuminates the sample 61 by the XY scanner 15 so as to scan the sample 61 at a video rate. I do. The video rate is usually 30 frames per second, and the number of horizontal scanning lines in one frame is about 420 to 525. In addition, one scan line is divided into about 512 to about 640 pixels when converted into digital pixel data.

The sample 61 has been subjected to a treatment such as staining so that it emits fluorescence or the like when irradiated with a laser beam. The light emitted from the sample 61 is scanned by the XY scanner 15, returned to the first dichroic mirror 17, passed through the first dichroic mirror 17, and sent to the second dichroic mirror 19. The second dichroic mirror 19 separates the light emitted from the sample 61 to the first photoelectric conversion means 22 and the second photoelectric conversion means 32 and sends the separated light. The first photoelectric conversion means 22 and the second photoelectric conversion means 32 output an output signal corresponding to the intensity of light of each wavelength separated by the second dichroic mirror 19.

The optical scanning microscope 10 is provided with a synchronizing signal generating circuit 51. This synchronization signal generation circuit 51
It outputs a vertical synchronization signal (VD) of about 60 Hz, a horizontal synchronization signal (HD) of about 15.6 kHz, and a pixel clock (PCLK) of about 8 MHz. And the pixel clock is the first analog-to-digital converter
23, the second analog-to-digital converter 33, as well as the write address generation circuit 57 and the read address generation circuit 59. Further, the vertical synchronizing signal and the horizontal synchronizing signal are input to the scanner control means 13, the write address generation circuit 57 and the read address generation circuit 59.

The first photoelectric conversion means 22 and the second photoelectric conversion means 32 use a photomultiplier tube or the like to convert weak light into an electric signal. Then, the first analog-to-digital converter 23 in the first image data storage unit 21 samples the output signal of the first photoelectric conversion unit 22 based on the pixel clock from the synchronizing signal generation circuit 51 and converts it into digital image data. Things. Further, the first frame memory 25 sequentially records digital image data output from the first analog-to-digital converter 23 in accordance with an address signal output from the write address generation circuit 57.

In the second image data storage means 31, the second analog-to-digital converter 33 is also used as the second photoelectric conversion means.
The 32 output signals are converted into digital image data. The second frame memory 35 stores the digital image data. The scanner control means 13 in the optical scanning microscope 10 not only controls the XY scanner 15 so that the laser beam is scanned at a video rate such as the NTSC standard, but also controls the interval, the number, and the position of the horizontal scanning lines. And the like can be changed by the input setting means 55.

That is, the number of scanning lines can be reduced by half by the input setting means 55, and a range necessary for observation can be designated as shown in FIG. Therefore, when the number of scanning lines is set to one half and the scanning range is set to a specific range of the sample 61, the scanner control means 13 sets the oscillation width of the Y-axis mirror to drive control of the XY scanner 15. Under the control, the scanning by the laser beam is performed only on the observation region B which is set to a half of the full image region A, and the laser beam waits outside the sample 61 for the remaining horizontal scanning time.

In other words, the number of scanning lines of the obtained image can be reduced to half the number of scanning lines of the full image area A by the scanner control means 13. The image data of the observation area B is converted into an electric signal by the first photoelectric conversion means 22 and the second photoelectric conversion means 32 at the same time and output. The output signal of the first photoelectric conversion means 22 is converted into digital image data by the first analog-to-digital converter 23, and the second photoelectric conversion means 32
Are converted into digital image data by the second analog-to-digital converter 33.

At this time, an address designation start signal is input from the scanner control means 13 to the write address generation circuit 57, and the write address generation circuit 57 is synchronized with the timing at which the observation area B is scanned by the laser beam.
57 to the first frame memory 25 and the second frame memory 35
Output an address signal. Therefore,
First frame memory in first image data storage means 21
25 and the second frame memory 35 of the second image data storage means 31 can sequentially store the image data of the observation region B acquired at the same time. The data amount of the image data stored in the first frame memory 25 and the second frame memory 35 is one half of the data amount of the image data in the full image area A.

The read address generation circuit 59 changes the observation area B to the full image area A by the scanner control means 13.
When the value is 1/2, an address signal for reading out the image data of the first frame memory 25 and the image data of the second frame memory 35 together is output. That is, as a read address, first, an address signal is output to the first frame memory 25, and a half of the normal data amount is read, thereby reading image data of the observation area B recorded in the first frame memory 25. And the image data is output from the first frame memory 25 to the outside. Then, after that, an address signal is output to the second frame memory 35 to read out a half of the normal data amount, whereby the observation area B recorded in the second frame memory 35 is read.
Image data can be read out.

Therefore, as shown in FIG.
One frame of image data for displaying the image obtained by the photoelectric conversion unit 22 and the image obtained by the second photoelectric conversion unit 32 in one screen can be formed. As described above, when the number of scanning lines is reduced by limiting the observation area B with respect to the full image area A and reducing the scanning range of the laser beam, the read address generation circuit
59 includes a first frame memory 25 and a second frame memory
The first frame memory 25 and the second frame memory 25 are combined so as to form one frame of image data by combining the two image data stored in 35.
The address signal is output to the frame memory 35. Therefore, the image data having a data amount of one half of the image data for forming one frame can be stored in the first frame memory 25 and the second frame memory in one frame time of 1/30 second.
The data can be output from the frame memory 35 and combined with one frame of image data.

Accordingly, in the optical scanning microscope 10, the scanning range of the laser beam can be limited, and the range in which the sample 61 is damaged by the laser beam can be the minimum range necessary for observation. Further, it is possible to observe the reaction by different reagents together on one screen, and observe different states such as the reagent concentration and the stained state by the reagent on one screen.

The input setting means 55 can also make settings for changing the scanning interval by the laser beam. In this case, based on the operation of the input setting means 55,
The scanner control means 13 controls the moving speed of the Y-axis mirror when driving and controlling the XY scanner 15, and as shown in FIG. 3, when scanning the full image area A with a laser beam, the scanning line interval is set to two. Can be doubled. Further, the scanner control means 13 doubles the interval between the scanning lines, reduces the number and time of the scanning lines by half, and makes the laser beam stand by outside the sample 61 for the remaining horizontal scanning time. It controls the scanner 15.

Also in this case, the write address generating circuit 57 outputs the address signal of the write address to the first frame memory 25 and the second frame memory 35 in synchronization with the scanning timing by the laser beam. Therefore, predetermined digital image data can be sequentially stored in the first frame memory 25 and the second frame memory 35. The read address generation circuit 59 first outputs an address signal to the first frame memory 25 as a read address, reads out a half of the normal data amount, and then outputs the address signal to the second frame memory 35. The data is output to read out a half of the normal data amount. That is, reading out the image data of the observation area B recorded in the first frame memory 25 and the image data of the observation area B recorded in the second frame memory 35 is the same as described above.

Accordingly, as shown in FIG. 3 (2), it is possible to form image data for displaying different images obtained at the same time in one screen as images compressed in half in the vertical direction. it can. In the above description, two screens are acquired at the same time. However, as shown in FIG. 1, a third photoelectric conversion unit 42 for converting the transmitted light from the sample 61 into an electric signal may be further provided.

In this case, a third analog-to-digital converter for digitally converting the output signal of the third photoelectric conversion means 42 and a third frame memory are added together with the third photoelectric conversion means 42 so that the third image data storage means It is assumed that. Then, from the input setting means 55, it is also possible to set a range designation of one third of the full image area A as the observation area B.
It is also possible to specify that the scanning line interval is tripled and set the scanning of the full image area A.

Further, in this case, the write address generating circuit 57 outputs an address signal to the third frame memory together with the first frame memory 25 and the second frame memory 35 in accordance with the scanning timing. And
When the observation area B is set to one third of the full image area A or when the scanning line interval is specified to be three times, the read address generation circuit 59 sets the second frame memory next to the first frame memory 25. After outputting the address signal to 35, the address signal is further output to the third frame memory.

As described above, the optical scanning microscope 10 is provided with a plurality of sets of image data accumulating units each including a photoelectric conversion unit, an analog / digital converter, and a frame memory. Then, control for setting the scanner control means 13 is performed by the input setting means 55, thereby enabling adjustment to limit the scanning range of the laser beam or to increase the interval between scanning lines by the laser beam. Therefore, the amount of image data formed at one frame timing by the photoelectric conversion means can be reduced in accordance with the number of image data storage means. Further, the image data with the reduced data amount is temporarily stored in the frame memory of each image data storage means, and the data stored in the frame memory of each image data storage means can be sequentially read out in one frame time. It is.

Therefore, when forming image data of one screen, image data of one frame can be formed by different image data of the same sample 61 obtained at the same time. The photoelectric conversion units 22 and 32 are not limited to those that output analog signals, such as a photomultiplier tube, and may use a solid-state imaging device and an amplifier. Further, as the photoelectric conversion means, a means for outputting a digitalized output signal may be used. In this case, as the image data storage means, it is sufficient to combine a frame memory for storing digital signals output from the photoelectric conversion means and the photoelectric conversion means.

Further, it is also possible to increase the number of dichroic mirrors and add photoelectric conversion means, and to use four or five sets of image data storage means. In this case, as the setting by the input setting means 55, the number of scanning lines can be appropriately set in multiple stages according to the number of image data storage means, such as 1/4, 1/5, etc. To control. In addition to making the number of image data storage means plural, the read address generation circuit 59 sequentially outputs the read address to all the frame memories, and in accordance with the reduction rate of the scanning line among the plurality of frame memories. Thus, it is possible to appropriately set the address signal to be output to two or three frame memories.

As described above, in the optical scanning microscope 10 provided with a plurality of image data storage means including the photoelectric conversion means,
It is possible to set a numerical value that is set to an arbitrary fraction of an integral number that sets the maximum number to the number of image data storage units, and to operate the frame memory of the image data storage unit by this number, so that images with different required number of image data can be set. Can be obtained at the same time, and synthesized and observed on one screen.

Accordingly, the optical scanning microscope 10 can form image data of one screen by making an image acquired and formed by a plurality of photoelectric conversion means simultaneously equal to the image data amount of one frame at the video rate. . That is, it is possible to simultaneously display and observe different images obtained simultaneously using a general monitor device. Furthermore, the irradiation amount of the laser beam to the sample 61 can be set to the minimum irradiation amount necessary for image formation, and there is an advantage that damage to the sample 61 can be reduced.

As another embodiment, as shown in FIG. 4, image data storage means 21 and 31 are provided with superimposing means 27 and 37 each including a frame buffer and an integrator in front of each frame memory. Sometimes. The optical scanning microscope 10 according to this embodiment, when the observation area B is set to half of the full image area A by the input setting means 55, or when the interval between the scanning lines is set to twice, Control means 13
Thus, the XY scanner 15 is controlled so as to scan the same portion twice during one frame time of 1/30 second.

Then, the first frame buffer 28 and the second
The frame buffer 38 stores the image data when the first scan is performed by the XY scanner 15 based on the control signal from the input setting means 55 and based on the address signal from the write address generation circuit 57. And When the second scanning is performed by the XY scanner 15, the image data stored in the first scanning is sequentially transferred from the first first frame buffer 28 to the first integrator based on the address signal from the write address generating circuit 57. Similarly, the image data is sequentially output from the second frame buffer 38 to the second integrator 39. And
The first integrator 29 integrates the digital image data from the first frame buffer 28 and the digital image data from the first analog-to-digital converter 23. That is, image data of a value obtained by averaging the value of the digital image data from the first frame buffer 28 and the value of the digital image data from the first analog-to-digital converter 23 is output. Also, the write address generation circuit 57
Is to control the XY scanner 15 to output a write address signal to the first frame memory 25 and the second frame memory 35 when the second scan is performed.

Therefore, in the superimposing means 27 and 37, the digital image data obtained by the first scan is temporarily stored in the frame buffer, and the digital image data obtained by the second scan and the first digital image data sequentially read from the frame buffer are stored. The addition can be performed by the integrator, and the image data of the added value can be stored in the frame memory.

As described above, when the scanning time for forming one frame is reduced by changing the scanning range or the scanning interval, the same portion is scanned again in the remaining time and the image data is integrated. Image data that makes the image of the sample 61 clear by reducing the disturbance can be easily formed.

[0048]

According to the first aspect of the present invention, there are provided a plurality of photoelectric conversion means, a frame memory for storing each image data formed by each photoelectric conversion means, and
Optical scanning having scanner control means capable of changing the setting of the number of scanning lines by a laser beam, and having a read address generation circuit for synthesizing each image data stored in the frame memory to form image data of one screen at a video rate. It is a type microscope.

Therefore, when performing image acquisition, it is possible to store image data of a plurality of images with reduced scanning lines of one screen in the frame memory. Then, an image obtained by synthesizing the plurality of images can be displayed on one screen of the monitor device, and observation of the sample can be performed easily and accurately. Further, the invention described in claim 2 enables setting to reduce the number of scanning lines to 1 / N (N is a positive integer) with respect to the standard number of scanning lines. Is an optical scanning microscope provided with scanner control means for matching the number of photoelectric conversion means provided in the microscope.

Therefore, it is possible to divide the screen into a maximum number of divisions corresponding to the number of photoelectric conversion means, and it is possible to display different images based on the image data obtained simultaneously on one screen. Further, a microscope can be provided in which a sample is irradiated with a laser beam only in a range necessary for display and the sample is not damaged unnecessarily. According to a third aspect of the present invention, there is provided an optical scanning microscope having scanner control means capable of setting the interval between scanning lines to N times the standard when setting the number of scanning lines to 1 / N. Things.

Therefore, it is possible to display the image divided on the screen corresponding to the number of the photoelectric conversion means on one screen while maintaining the scanning range wide. Further, the invention described in claim 4 is:
An optical scanning microscope having superimposing means between the photoelectric conversion means and the frame memory is provided. Therefore, a clear image of the sample can be formed.

According to a fifth aspect of the present invention, the superimposing means is an optical scanning microscope constituted by a frame buffer and an integrator. Therefore, the superimposing means can be easily formed, and clear image data can be easily obtained. Further, according to the invention described in claim 6, when simultaneously acquiring a plurality of images of the same object by an optical scanning microscope having a plurality of photoelectric conversion means and a frame memory, the number of scanning lines is reduced to the number of standard scanning lines. This is an image acquisition method for a scanning microscope in which each fraction of image data is acquired and temporarily stored in a frame memory.

Therefore, a plurality of images can be obtained simultaneously and a plurality of images can be displayed on a single screen at a video rate.
This is an image acquisition method that irradiates a beam only to a necessary minimum range and does not damage the sample unnecessarily. And Claim 7
According to the invention described in (1), when acquiring each image data with the number of scanning lines being a fraction and storing it in the frame memory, each image data is stored in the frame memory as image data obtained by integrating a plurality of identical image data. This is an image acquisition method using a scanning microscope.

Therefore, when a plurality of images are simultaneously obtained and a plurality of images are displayed on a single screen at a video rate, clear images can be displayed.

[Brief description of the drawings]

FIG. 1 is a block diagram showing a main part of an optical scanning microscope according to the present invention.

FIG. 2 is a diagram showing a scanning range and a state of an output image in the optical scanning microscope according to the present invention.

FIG. 3 is a diagram showing a scanning range and a state of an output image in the optical scanning microscope according to the present invention.

FIG. 4 is a main part block diagram showing another embodiment of the optical scanning microscope according to the present invention.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 10 Optical scanning microscope 11 Laser irradiation means 13 Scanner control means 15 XY scanner 17 1st dichroic mirror 19 2nd dichroic mirror 22 1st photoelectric conversion means 23 1st analog / digital converter 25 1st frame memory 27 1st superposition means 28 Frame memory 29 Integrator 32 Second photoelectric conversion means 33 Second analog-to-digital converter 35 Second frame memory 37 Second superimposing means 38 Frame memory 39 Integrator 42 Third photoelectric conversion means 51 Synchronous signal generation circuit 55 Input setting means 57 Write address generator 59 Read address generator 61 Sample

Claims (7)

[Claims] 1. A microscope that scans a sample with a laser beam and converts light from the sample into an electric signal by a photoelectric conversion unit to form image data, the microscope having a plurality of the photoelectric conversion units, In an optical scanning microscope having a frame memory for storing each image data formed by each photoelectric conversion means, a scanner control means capable of changing the setting of the number of scanning lines when scanning the sample or the like with a laser beam, A read address generation circuit for outputting image data from the frame memory so as to form image data of one screen at a video rate by synthesizing each image data stored in the frame memory in accordance with the setting change of the number of scanning lines An optical scanning microscope, comprising: 2. The method according to claim 1, wherein the scanner control unit changes the setting of the scanning line by 1 / N of a standard number of scanning lines.
2. The optical scanning microscope according to claim 1, wherein the number of scanning lines can be reduced to the number of (N is a positive integer), and the maximum of N is set to the number of the photoelectric conversion units. 3. The scanner control means sets a scan line interval to a standard N when setting the number of scan lines to 1 / N.
3. The optical scanning microscope according to claim 2, wherein the magnification is doubled. 4. A superimposing means between the photoelectric conversion means and the frame memory, wherein the superimposing means causes the frame memory to store image data based on pixel data obtained by adding corresponding pixel data in a plurality of images. An optical scanning microscope according to any one of claims 1 to 3, characterized in that: 5. An optical scanning microscope according to claim 4, wherein said superimposing means comprises a frame buffer and an integrator. 6. When simultaneously acquiring a plurality of images of the same object by an optical scanning microscope having a plurality of photoelectric conversion units and a frame memory, the number of scanning lines is reduced to a fraction of the number of standard scanning lines. An image acquisition method for a scanning microscope, wherein each image data is acquired and temporarily stored in the frame memory. 7. When acquiring each image data with the number of scanning lines reduced to a fraction and storing it in the frame memory, each image data is stored in the frame memory as image data obtained by integrating a plurality of identical image data. 7. The method for acquiring an image of a scanning microscope according to claim 6, wherein: