JP2005315574A - Electronic camera - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an electronic camera excelling in image reproducibility and also being low-cost, and a scanning laser microscope. <P>SOLUTION: As to this electronic camera, a specimen is illuminated by using light with at least one selected wavelength to photograph reflected light or transmitted light by the electronic camera. The specimen is illuminated by using light with a wavelength other than that of the illumination light to photograph reflected light of the light or transmitted light thereof by this electronic camera. Images thus obtained are used to observe the specimen. This scanning laser microscope is equipped with a modulator for modulating a scanning laser beam to control power per hour of the laser beam so to keep it constant on the specimen, a pulse generator for driving an actuator for causing the laser beam to scan, and an estimator for estimating the scanning speed of the laser beam caused to scan by the actuator. The scanning speed, which is an output of the estimator, is inputted into the modulator. The laser beam outputted from the modulator is pulse-width modulated, pulse-frequency modulated, or pulse-amplitude modulated. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention uses a light-emitting element such as a laser or LED (Light Emitting Diode) as a light source, irradiates the sample by changing the wavelength of the light source, and separates light reflected from or transmitted through the sample according to the type of wavelength. The present invention relates to an electronic camera that inputs to a memory device or an electronic camera that scans a sample by amplitude modulation of a light source or a light beam from the light source, and has a CCD (Charge Coupled Device), CMOS (Complementary Metal-Oxide Semiconductor) sensor as a light receiving element .

A conventional imaging device using a microscope color CCD camera (electronic camera) separates light incident on the CCD into a color signal and a luminance signal using an optical color filter, for example, an RGB signal (red, green, Signal).
For this reason, since a plurality of CCDs are used or a color filter is provided on the incident surface of the CCD, it is necessary to use a CCD having a large number of pixels in order to obtain a predetermined resolution.
Further, when a color filter is provided on the CCD entrance surface so as to match the pixels of the CCD, the pixel positions for obtaining the luminance information and the color information are different, and there is a problem that the image of the subject cannot be reproduced faithfully. For this reason, three CCDs are used in the field of medical images that require high fidelity, R, G, and B color filters are provided on the entrance surface of each CCD, and color information is detected separately. A color image was obtained by synthesis. However, at this time, positional accuracy between the three CCDs is required, and high-accuracy alignment is required, and when the alignment is shifted, there is a problem in image reproducibility. Further, when a light beam emitted from a laser beam is used as the light for irradiating the sample, the optical system for scanning the laser beam is complicated and expensive, thereby preventing the spread.

FIG. 11 illustrates an imaging apparatus using a conventional electronic camera.
A three-color separation prism is provided after the imaging lens, and is divided into three primary colors of R, G, and B. R, G, B signals can be obtained by providing CCDs of the same size at the image forming positions of the respective rays.
In FIG. 11, 1101 is a sample to be observed, 1102 is a light source, and usually an incandescent light bulb or the like is used. Then, the light is converted into parallel light by the condenser lens 1103, and the sample is irradiated by the objective lens 1105 via the beam splitter 1 1104.
The reflected light of the irradiated sample is supplied to the electronic camera unit 1114 through the beam splitter 1 1104, the imaging lens 1106, and the beam splitter 2 1107. In the electronic camera 1114, 1109 is a beam splitter 3 that gives red (R) from incident light to the CCD 1 of 1110, 1111 also gives blue (B) to the CCD 2 of 1112, and green (G ) Is shown. The signals obtained from each CCD are processed separately, and finally used as standard signals (RGB, NTSC (National Television Standards Committee), etc.) Also, laser scanning that scans and irradiates a sample with a laser beam For example, a microscope has a mechanism for rotating a disk having a microarray lens and scanning a sample with a laser beam spot, and has required very high accuracy.

In the conventional system, the beam splitter used for color separation is expensive, and it is necessary to increase the accuracy of the mechanism for positioning between the CCDs and to accurately place the CCD at the imaging position of the optical image. If the alignment is deviated, there is a problem because the image of the target sample is displaced in the output of the CCDs 1, 2, and 3 in FIG. Also, it was necessary to use three CCDs, which was also expensive. Further, when a scanning laser is used as a light source, the overall structure is complicated and very expensive.
An object of the present invention is to provide an electronic camera and a scanning laser microscope that are excellent in image reproducibility and can be reduced in price.

The present invention relates to the following inventions.
1. An image observed by an optical system for observing a sample is converted into an electrical signal, and the electrical signal is converted into a digital signal, and then the electronic camera that takes in the memory device is used to irradiate the sample to be observed A plurality of wavelengths of light can be selected, and the sample is irradiated with light having at least one selected wavelength, and reflected light or transmitted light of the light irradiated on the sample is photographed by the electronic camera. Using an image and light having a wavelength different from the irradiation light, irradiating the sample, and observing the sample using an image obtained by photographing reflected light or transmitted light of the light irradiated on the sample with the electronic camera An electronic camera characterized by
2. In a scanning laser microscope that scans a sample with a beam spot irradiated from a laser and observes reflected light or transmitted light of the scanned laser beam spot, in order to control the power per time of the scanned laser beam on the sample constant, A modulator for modulating a scanning laser beam; a pulse generator for driving an actuator for scanning the laser beam; an estimator for estimating a scanning speed of the laser beam scanned by the actuator; and the estimator for the modulator. The scanning laser microscope is characterized in that a scanning speed which is an output of the above is input, and the laser beam output from the modulator is subjected to pulse width modulation, pulse frequency modulation or pulse amplitude modulation.

Light sources with different wavelengths are sequentially turned on and irradiated, the reflected light or transmitted light is taken into the same CCD camera or CMOS sensor, and input to different memories for each wavelength so that they can be observed. Thus, there is no positional deviation between the obtained images. Further, since it is not necessary to use different optical filters for each wavelength, the price can be reduced.
When a laser is used as a light source, since the wavelength spread of the laser light source is small, the purity of the color is significantly improved as compared with the LED, so that the quality of the obtained image is improved.
Further, a laser microscope focused and scanned with a laser beam can be simplified in configuration as compared with a conventional laser microscope. At this time, since the laser beam output is subjected to pulse interval (pulse frequency), pulse width, or pulse amplitude modulation, the irradiation energy per unit time of the scanned laser beam on the sample can be controlled to be constant. As a result, the brightness of the image in which the reflected light or transmitted light is detected is constant, and the sample is not damaged.

The invention according to claim 1 of the present invention discloses an electronic camera that changes the wavelength of a light source that irradiates a subject and captures a photographed image of the subject for each wavelength.
The electronic camera proposed here has a configuration in which an image observed by an optical system for observing a subject is converted into an electric signal, and the electric signal is converted into a digital signal and then taken into a memory device.

  An image obtained by changing the wavelength of the light source, for example, sequentially irradiating the subject with light having each wavelength of R (red), G (green), and B (blue), and irradiating the subject with the wavelength of R The image R is taken from the image obtained when the subject is irradiated with the wavelength of G, the image G is taken from the image obtained when the subject is irradiated with the wavelength of B, and the image B is taken from the image obtained when the subject is irradiated with the wavelength of B. Each captured image is stored in an image memory, and the stored image is converted into a standard image format that can be displayed on a display and then observed. In the present invention, there is only one imaging device such as a CCD for converting an optical image into an electronic image, and an optical filter for converting an optical image into color signal information is unnecessary, which is very simple. The structure has an effect that a color image can be obtained.

  According to an eighth aspect of the present invention, in a scanning laser microscope for irradiating a sample with a scanning laser beam, a laser beam irradiation block in which a laser and an aperture lens are integrated is provided, and a beam spot irradiated to the sample is set. Since scanning is performed by an actuator, a scanning laser microscope can be realized with a simple configuration.

Hereinafter, examples specifically showing the best mode for carrying out the present invention will be described with reference to the drawings.
Hereinafter, the first embodiment of the present invention will be described with reference to FIGS. 1, 2, 3 and 4. FIG. 1 is a block diagram for photographing the light reflected by the sample with a CCD camera.

In FIG. 1, reference numeral 101 denotes a synchronization signal generator which generates a frame synchronization signal at the timing shown in FIG. The frame synchronization signal is input to the LEDs 102a, 102b and 102c or the laser drive amplifier, and the red LED (R), the green LED (G) and the blue LED (B) shown in 103a, 103b and 103c are turned on. Each LED irradiates the sample 105. As shown in FIG. 3, the timing of lighting is such that 301 R pulse signal is ON from time t1 to t2, similarly 302 G pulse signal is ON from time t2 to t3, and similarly 303 B pulse signal. Becomes ON from time t3 to t4, and then the signal repeats the operations of 301 to 303.
Although the synchronization signal is continuously generated, the synchronization operation actually required is only three frames of RGB in an arbitrary period.

  A synchronizing signal generator 104 generates a 1/30 second synchronizing pulse signal, that is, the frame synchronizing signal described above, in the vertical synchronizing signal 104b to 101 of the 104 B / W (monochrome) CCD camera. The image of the sample 105 irradiated by the LED is picked up by the CCD of the B / W camera 104, and a video output signal of 104a is obtained. The output signal is sent to the switcher 106 to be combined with the R, G, B frame images, and is set to the specified signal level by the video amplifier 107. The signal output 108 is transferred to the next image processing unit.

FIG. 2 shows a laser light source block for illuminating the sample. Reference numeral 202 denotes a laser 1, 203 denotes a collimating lens 1, 204 denotes a laser 2, 205 denotes a collimating lens 2, 206 denotes a laser 3, and 207 denotes a collimating lens 3. The light emitted from each laser is converted into parallel light by each collimating lens, enters the X-cube prism 201, and mixed light is obtained as an illumination light output 208.
For example, when a device that emits R (red), G (green), and B (blue) light is used for the three types of lasers 202, 204, and 206, the illumination light 208 becomes white light. Light 208 output from the block irradiates the sample, and reflected light or transmitted light is guided to the optical sensor via the optical system. In FIG. 1, the illumination light source has been described using an LED. However, the use of a laser as the light source is preferable for observation with a microscope because the wavelength of the light source is small and high-purity light is obtained. The wavelength of each laser 1, 2, 3 can be selected according to the type of sample.

FIG. 4 is a block diagram showing the principle of I / F (interface) and image processing of the present invention.
In FIG. 4, an RGB signal indicated by an output 108 from the circuit unit in FIG. 1 is input to an input terminal 401, 401 passes through an input I / F 402, and is stored in a memory 403 in a format such as a bitmap. The R, G, and B images called from the memory by frame switching 404 are further extracted with color channels 405 of R (red) channel, G (green) channel, and B (blue) channel, and images of 405a, 405b, and 405c, respectively. It is input to the memory, and 406 RGB images are obtained. This image can be confirmed on the display device 407.

Next, the operation will be described.
A sample to be observed with a microscope requires a dedicated light source because the amount of light is insufficient because natural light in a laboratory or the like is subject to magnification observation. In the present invention, the sample is irradiated with red (R), green (G), and blue (B) light sources separately at different timings. The irradiated image is captured by a B / W (black and white) camera, and the luminance when red, green, and blue are irradiated is converted into an electrical signal through a CCD element and sent to an image processing unit to be sent to each color density. In color composition expression.
In addition, since the present invention is not a system that performs visual inspection with an eyepiece, it does not directly observe continuous RGB light, but leads the result to the display device. It does not give visual harm.

The present invention is a system capable of reproducing a red image on a display device as a result even if a B / W camera is used, for example, when a red LED (or laser) irradiates a sample.
For this purpose, it is important that the synchronizing signal generator 101 shown in FIG. 1 emits the LED at an accurate timing and simultaneously obtains an image of the light emitting section.
As the B / W camera 104, for example, a CCD video camera can be used. In the embodiment of FIG. 1, this B / W camera uses a TV video signal output, and one frame of 1/30 second can be obtained by the vertical synchronization signal 104b. At the same time, the R, G, and B LEDs 103a, 103b, and 103c are sequentially turned on by being supplied with current by the drive amplifiers 102a, 102b, and 102c at the same timing. In the switcher 106, the synchronization signal controls the video output signal 104a, and the LED lighting and the frame are synchronized. The processed video signal is adjusted to a prescribed level by a video amplifier, and sent to the next image processing unit as a parallel output 108 of R video, G video, and B video.

  4, an RGB parallel input 401 from the circuit unit of FIG. 1 passes through an input I / F (interface) 402 that is a physical interface, and a still image image such as a bitmap that can be read in the system as one frame as one frame. And stored in the memory 403. The R, G, and B images called out by the frame switching 404 from the memory further extract the color channels 405 of the R (red) channel, the G (green) channel, and the B (blue) channel, and the memories of 405a, 405b, and 405c, respectively. Accumulated in. The output is a 406 RGB image which is a full color display. This image can be monitored by a display device 407 such as an image monitor.

The second embodiment of the present invention will be described below with reference to FIGS.
FIG. 5 shows a block diagram illustrating an embodiment of the present invention.
In FIG. 5, reference numeral 501 designates a counter for generating an output with different intervals depending on the clock signal, 502 is a clock signal generator for LED lighting test, and its cycle can be varied by a flicker adjustment trimmer 502a. . The clock timing is the same as in FIG. 3 described in the first embodiment.
RGB LEDs are arranged in two places, an upper reflected light LED block 508 and a lower transmitted light LED block 509, and switching is performed by an upper / lower LED changeover switch 503.
Each LED block is supplied with a drive current by an upper reflected light LED driver 504 and a lower transmitted light LED driver 505, and each LED has a luminous intensity by an upper reflected light intensity adjustment trimmer 506 and a lower transmitted light intensity adjustment trimmer 507. Adjustment is possible.
The sample 511 irradiated by the LED block 508 or 509 is photographed by the 510 B / W camera, the video signal is output at 510a, and the vertical synchronization signal is output at 510b. The sync signal 510b is halved by the ½ divider of 510c. The frequency-divided signal is switched by 502 test clock generator and 512 LED lighting test switch and sent to the counter 501. In addition, when the white light generation switch for focus adjustment 513 is used for direct current lighting unrelated to the clock, the RGB LEDs are simultaneously lighted, so that white light is obtained, which is convenient for focus adjustment.

The video output signal 510a switches the video switch 514 by the signal of the counter 501, and obtains an output synchronized with the signal for driving the LED. The output is adjusted to a specified level by a 515 video output level trimmer and sent to an interface 517 through a video amplifier 516. The output 518 is connected to the image processing unit as an RGB image output signal.
Also, a composite sync signal 520 is obtained from the video output through the video sync signal separation circuit 519 and used for the synchronization of the next stage I / F.

  The second embodiment shown in FIG. 5 is a configuration obtained by developing the first embodiment, and includes a reflection / transmission light selection function, an LED light intensity individual adjustment function, an LED arbitrary period setting function, an LED irradiation test function, and a focus adjustment function. It has a user interface that conforms to actual use, such as a white light generation function and a video signal individual adjustment function.

  FIG. 6 is a block diagram showing the principle of I / F and image processing of the present invention. The signal output 518 from FIG. 5 is converted from a 641 input to a digital signal by an A / D converter 642 and temporarily stored in a 643 frame memory. The image signal is sent to the system by the 644 PCI bus master I / F, and the frames are sequentially selected by the frame switching 645 and the images are taken in. The images are stored in the memory 648 by the image controller 1 646. The image file is read from the memory 648 by the image controller 2 of 647, and the R, G, B images are further 649 colors of the R (red) channel of 649a, the G (green) channel of 649b, and the B (blue) channel of 649c. Sent to the channel. The color image can be analyzed with a desired parameter such as a color distribution state by an image analysis apparatus 650. The analysis result, the color image, and the color channel can be observed with the display device 651. Of course, RGB color images can be used as they are.

The present invention is a system that can reproduce a red image on a display device as a result even if a B / W camera is used, for example, when a red LED irradiates a sample.
For this purpose, it is important to perform a synchronous operation in which the counter 501 in FIG. 5 causes the LED to emit light at an accurate timing and at the same time obtain an image of the light emission section with the 510 B / W camera. The B / W camera is not a digital camera for obtaining a still image, but a so-called CCD video camera, and has an EIA standard TV video signal output and video video output signal 510a. One frame of 1/30 second can be obtained by dividing the vertical synchronizing signal 510b obtained by the camera with a ½ divider of 510c and can be used for blinking of the LED blocks 508 and 509. And can be synchronized.
In the counter 501, ON / OFF is repeated at a clock of 1/30 seconds, and the signals are output on channels for R, G, and B at the timing shown in FIG. Next to the output of RGB, a reset works, and the operation returns to RGB again. The time required for each timing (each interval of t1-t2-t3-t4) is about 0.1 microsecond or less and does not adversely affect the operation.
The lighting test clock generator 502 generates a period other than a 1/30 second clock by a timer operation and can be used for individual LED light emission tests, system checks, and light intensity measurements. The period can be varied by the flicker frequency adjustment trimmer 502a. The signal from the camera and the signal from the test clock generator can be switched and used with 512 LED lighting switches.

The light source LED is convenient for sample observation by being equipped with two light sources for upper reflected light and lower transmitted light. The two LED light sources can be switched with a 503 up / down LED selector switch. The output of the counter for three RGB channels is given to the LED driver for reflected light 504 and the LED driver for transmitted light 505, R, G, B (504a, 504b, 504c and 505a, 505b, 505c), respectively. Each LED current is applied at the timing shown in FIG.
Since the current of each LED affects the luminous intensity, a reflected light luminous intensity adjustment trimmer 506 and a transmitted luminous intensity adjustment trimmer 507 are provided to adjust the current balance for each individual LED to adjust the RGB balance. The LED is composed of an LED block for upper reflected light 508 and an LED block for lower transmitted light 509, which are R-LED chip 508a, G-LED chip 508b, B-LED chip 508c, R-LED chip 509a, and G-LED, respectively. A chip 509b and a B-LED chip 509c are included.

  The white light generation switch for focus adjustment 513 changes the clock generated by the 510c counter or the 502 lighting test clock generator to a continuous high-level direct current, and causes the LEDs to emit light at the same time to mix the colors of RGB. As a result, a white light source is obtained. This facilitates the adjustment of the focus except for the visual detriment caused by the blinking of R, G, and B appearing on the display device described later. The white light also has a function of adjusting the mixing gain when the switch is turned on in order to make the brightness visually different from that when RGB blinks. Also, color balance can be easily adjusted by adjusting the light quantity of each LED with a trimmer of 506 and 507 so that the light source can surely obtain a white color with this function.

The image of the sample 511 irradiated by the LED block is obtained by the 510 B / W camera, the 510a video image output signal is obtained, sent to the video switch 514, and the frequency of 501 obtained by frequency division from the synchronizing signal of the 510b. It is switched by the clock of the counter and assigned every frame. As a result, each frame corresponding to R, G, B irradiation is obtained. The signal after the switch is sent to a video amplifier 516, converted to a specified level, and output 518 by an image signal output interface 517. By mixing the R, G, and B signals at a ratio of light corresponding to a relative visibility curve that associates a human-recognizable color with a spectral characteristic as a physical quantity, a visually correct color can be felt.
This relationship is expressed as E _k = 0.3Er + 0.59Eg + 0.11Eb
However,
E _k : Voltage value of luminance signal Er, Eg, Eb: Since it is a voltage value of RGB three primary colors, R is a 515a trimmer, G is a 515b trimmer, B is a 515b trimmer, and B is a 515c trimmer. The voltage value can be adjusted.

In the detailed block diagram of the I / F and image processing unit in FIG. 6, the video signal from the circuit unit in FIG. FIG. 6 is processed using an electronic computer system except for the display device 651.
Since the input is an analog RGB signal, it is first digitally converted so that it can be processed in the computer.
This signal is stored in a frame buffer or frame memory 643 which is a storage device corresponding to the designated pixel and used for image display. The data is taken into the system through the internal bus interface of the electronic computer, the bus master 644, and a necessary frame is selected and switched by switching the frame 645, and one frame is filed and managed as one image. The image is captured by the image controller 1 646, stored in the memory 648 by the image format file save function, and a corresponding file is read from the memory by the image file load function of the image controller 2 647 for image processing.
Three types of image files, R, G, and B, are read out, and each file stores 649 color channels as an R color channel 649a, a G color channel 649b, and a B color channel 649c.
The observation result is displayed on a 651 display device such as a display, and an RGB image, each color channel image, and an analysis result can be displayed. In addition, real-time image observation, focus confirmation adjustment, color adjustment, and the like can be performed using the image on the display device. Although the above description has been given taking a microscope as an example, any imaging device having the same configuration can be applied to a camera used in a studio or the like and is not limited to a microscope.

FIG. 7 is a block diagram showing one embodiment of a scanning laser microscope according to the third embodiment.
In FIG. 7, reference numeral 702 denotes a laser scanning unit that irradiates a sample 701. The laser beam scanning unit integrally configures a laser light source 702c and an objective lens 702d, and scans a laser beam emitted from the objective lens 702d in the two-dimensional directions of X and Y on the sample 701 by a laser scanning actuator 702b. . Reference numeral 701 denotes a sample to be observed, and reference numeral 703 denotes a white light source. The light emitted from the white light source 703 is converted into parallel light by the condenser lens 704, and irradiates the sample with the objective lens 706 via the beam splitter 1 705. The reflected light of the irradiated sample passes through the beam splitter 1 705, passes through the optical filter 710, and the sample 701 is photographed by the CCD camera 711.

Next, the operation will be described with reference to FIGS.
FIG. 8 is a block diagram of a laser scanning microscope. Reference numeral 801 denotes a pulse generator, reference numeral 802 denotes a modulator, and reference numeral 803 denotes a laser drive amplifier. Reference numeral 805 denotes a synchronization signal generator which is the same type as that used in the CCD camera of FIG. A frequency divider 806 divides the output of the periodic signal generator 805 and inputs it to an actuator X-direction drive amplifier 807 to drive the laser scanning actuator 702b and displace it in the X direction. Similarly, the output of the synchronization signal generator 805 is divided and input to the actuator Y-direction drive amplifier 809, and the laser scanning actuator 702b is driven and displaced in the Y-direction.
In this embodiment, the scanning frequency in the X direction is, for example, several hundred Hz, and the scanning frequency in the Y direction is several Hz. This scanning frequency is preferably adapted to the characteristics of the actuator. Reference numeral 808 denotes a speed estimator, which differentiates the drive input in the actuator X direction of 807 and estimates the scanning speed of the actuator by the CPU 804. A CPU 804 controls the above-described operation.

FIG. 9 is a timing chart showing the output of the modulator 802 of FIG. 8 as the modulator output waveform of 903. The output waveform of the actuator drive amplifier of 807 is the actuator drive waveform of 901, and the output waveform of the speed estimator of 808 is , Shown in waveform 902.
In FIG. 9, the horizontal axis indicates time 904 and the vertical axis indicates signal amplitude 905. The output of the modulator 802 is proportional to the output of the velocity estimator 808 and changes the pulse interval 906 or the pulse width and pulse amplitude 905. Here, when the pulse interval is changed, it is called pulse frequency modulation. The speed estimator used here is obtained, for example, by using a differentiator that differentiates 901 actuator drive waveforms. It is also possible to measure the speed by providing a speed detector with a combination of a magnetic coil and a magnet in the actuator movable part.

The pulse interval 906 of the modulator output 903 is changed so as to be in inverse proportion to the output of the speed estimator 808. That is, when the scanning speed is high, the pulse interval is set short and the pulse frequency is set high.
By controlling the modulation waveform as described above, the laser beam scanned by the actuator and the laser beam energy per unit time when the sample is scanned can be made constant. By doing so, the sample can be prevented from being damaged by the energy of the laser beam applied to the sample.
That is, when the actuator is displaced in the X-axis direction, the relative speed of the laser beam with respect to the sample becomes zero at the start and end points of the displacement. Further, the relative speed becomes maximum at the intermediate point between t1 and t3 shown in FIG. 9, and the unit time energy of the scanning laser power becomes minimum (indicated by t2 in FIG. 9). Therefore, when the laser beam pulse is not modulated, the sample is damaged by heat due to the energy of the laser beam. In order to avoid this, in the present invention, the laser beam is subjected to pulse width modulation and amplitude modulation, and the laser irradiation energy per unit time when the unit time sample is scanned is controlled to a constant value.

  Next, when the laser beam scans the sample, it is necessary to detect the scanning position. For this reason, a glass plate for calibration having grooves in the XY directions shown in FIG. 10 is placed at the position where the sample is arranged, the laser beam is scanned in the XY direction before observing the sample, and the output of the CCD is AD converted, Store on the semiconductor memory.

FIG. 10 shows a calibration glass plate. In FIG. 10, reference numeral 1001 denotes a glass plate, and 1002 denotes a pre-groove in the X-axis direction. On this glass plate, grooves called pregrooves are provided at intervals of 5 μm in the X-axis direction. 1003 is also a pre-groove in the Y-axis direction, and grooves are provided, for example, at intervals of 10 μm.
When this calibration glass plate is placed at the sample position of the microscope and scanned with a laser beam prior to sample observation, it is displayed as a streak in the X and Y axis directions on the CCD screen, and the laser beam scanning position is known. I can do it. Of course, the sample may be placed on a glass plate for calibration and observed. At that time, the size of the sample to be observed can be known.

The operation when using the scanning laser microscope of FIG. 7 will be described.
First, the white light source 703 is turned on, and the light reflected from the sample 701 is photographed by the CCD camera 711 via the optical filter 710. Next, the laser of 702c is turned on, the target portion of the sample of 701 is irradiated, the transmitted light passes through the objective lens 706 and the imaging lens 707, and the optical filter of 710 is scanned by the CCD camera of 711. Set so that it can be detected by, and observe. It is also possible to observe an image via the beam splitter 2 708 with the eyepiece 709.
At this time, since the laser beam spot can be set to several μm or less, the details of the sample can be observed. Of course, when the sample is colored with a phosphor, the light generated by the phosphor can be selectively observed with the optical filter 710. Although not shown in FIG. 7, it is possible to perform focus control by displacing the objective lens 702d in a direction perpendicular to the sample. At this time, the distance between the objective lens 702d and the sample 701 can be measured, and the distance can be automatically controlled to be constant.

FIG. 2 is a block diagram illustrating the principle of photographing the light reflected by a sample with a CCD camera, which is the principle of the present invention. It is the block diagram which showed the illumination part in the electronic camera of this invention, and the laser light source which illuminates a sample. It is a timing chart of the LED lighting pulse of R (red), G (green), and B (blue). The principle of the I / F and the image processing unit of the present invention is shown in a block diagram. It is a detailed block diagram of one Example of this invention. It is the block diagram which showed the principle of I / F of this invention, and an image process part. FIG. 3 illustrates a configuration of a laser scanning microscope according to a third embodiment of the present invention. FIG. It is a block diagram of a laser scanning microscope. It is a timing chart which shows the actuator drive waveform 901, speed estimator output 902, and modulator output 903 of a laser scanning microscope. 1 is a diagram illustrating a glass plate for position calibration for a laser scanning microscope. 1 is a block diagram of a conventional electronic camera optical system.

Explanation of symbols

DESCRIPTION OF SYMBOLS 101 Synchronous signal generator 102 LED driver 102a LED (R) drive amplifier 102b LED (G) drive amplifier 102c LED (B) drive amplifier 103 LED block 103a LED (R)
103b LED (G)
103c LED (B)
104 B / W camera 104a Video output signal 104b Vertical synchronization signal 105 Sample 106 Switcher 107 Video amplifier 108 Output 201 X-Cube prism 202 Laser 1
203 Collimating lens 1
204 Laser 2
205 Collimating lens 2
206 Laser 3
207 Collimating lens 3
208 Illumination light output 301 R pulse signal 302 G pulse signal 303 B pulse signal 401 Input 402 Input I / F
403 Memory 404 Frame switcher 405 Color channel 405a Image memory (R)
405b Image memory (G)
405c Image memory (B)
406 RGB image 407 Display device 501 Counter 502 Lighting test clock generator 502a Flicker frequency adjustment trimmer 503 Upper / lower LED changeover switch 504 Upper reflected light LED driver 504a R-LED driver 504b G-LED driver 504c B-LED driver 505 Lower transmission LED driver for light 505a R-LED driver 505b G-LED driver 505c B-LED driver 506 Upper reflected light intensity adjustment trimmer 506a R-LED trimmer 506b G-LED trimmer 506c B-LED trimmer 507 Lower transmitted light intensity Adjustment trimmer 507a R-LED trimmer 507b G-LED trimmer 507c B-LED trimmer 508 Upper reflected light LED block 508a R-LED chip 508b G-LED chip 5 08c B-LED chip 509 Lower transmitted light LED block 509a R-LED chip 509b G-LED chip 509c B-LED chip 510 B / W camera 510a Video image output signal 510b Vertical synchronization signal 510c 1/2 divider 511 Sample 512 LED lighting test switch 513 Focus adjustment white light generation switch 514 Video switch 514a R image selection video switch 514b G image selection video switch 514c B image selection video switch 515 Image output level adjustment trimmer 515a R image trimmer 515b G image trimmer 515c B image trimmer 516 Video amplifier 516a R image signal video amplifier 516b G image signal video amplifier 516c B image signal video amplifier 517 Image signal output interface 518 Image output signal 519 Video sync signal separation circuit 520 Composite sync signal 641 Input 642 A / D converter 643 Frame FIFO memory 644 PCI bus Master I / F
645 Frame switcher 646 Image controller 1
647 Image controller 2
648 Memory 649 Color channel 649a R color channel 649b G color channel 649c B color channel 650 Image analysis device 651 Display device 701 Sample 702 Laser scanning unit 702a Focus actuator 702b Laser scanning actuator 702c Laser 702d Objective lens 703 White light source 704 Condensing lens 705 Beam splitter 1
706 Objective lens 707 Imaging lens 708 Beam splitter 2
709 Eyepiece lens 710 Optical filter 711 CCD camera 801 Pulse generator 802 Modulator 803 Laser drive amplifier 804 CPU
805 Synchronization signal generator 806 Divider 807 Actuator X direction drive amplifier 808 Speed estimator 809 Actuator Y direction drive amplifier 901 Actuator X direction drive waveform 902 Speed estimator output 903 Modulator output 904 Time 905 Signal amplitude 906 Pulse interval 1001 Glass Plate 1002 X-axis pre-groove 1003 Y-axis pre-groove 1101 Sample 1102 White light source 1103 Condensing lens 1104 Beam splitter 1
1105 Objective lens 1106 Imaging lens 1107 Beam splitter 2
1108 Eyepiece 1109 Beam splitter 3
1110 CCD1
1111 Beam splitter 4
1112 CCD2
1113 CCD3
1114 Electronic camera part

Claims (9)

An image observed by an optical system for observing a sample is converted into an electrical signal, and the electrical signal is converted into a digital signal, and then the electronic camera that takes in the memory device is used to irradiate the sample to be observed A plurality of wavelengths of light can be selected, and the sample is irradiated with light having at least one selected wavelength, and reflected light or transmitted light of the light irradiated on the sample is photographed by the electronic camera. Using an image and light having a wavelength different from the irradiation light, irradiating the sample, and observing the sample using an image obtained by photographing reflected light or transmitted light of the light irradiated on the sample with the electronic camera An electronic camera characterized by In the electronic camera that captures an image observed by an optical system for observing a sample, the wavelength of light for irradiating the sample to be observed can be selected, and the selected first wavelength is Scanning with the electronic camera, then irradiating with light having a second wavelength, scanning, then irradiating with light having a third wavelength, and scanning the same An electronic camera that captures an image in a memory device by a camera, reads a digitized image captured in the memory device from the memory device, transfers the image to a display device, and displays the image. A white LED using an optical semiconductor is used as a light source for irradiating light to the observation sample, and the white LED is composed of three colors RGB, and each RGB color is selectively switched for irradiation. The electronic camera according to claim 1. As a light source for irradiating the observation sample with light, at least three types of lasers having different wavelengths are used. The three types of laser output light are converted into parallel light by a collimating lens and then incident on a beam splitter. The electronic camera according to claim 1, wherein the sample is irradiated after mixing the output light. The electronic camera according to claim 4, wherein the laser light is selectively switched for irradiation. As a light source for irradiating light to the observation sample, a white LED using an optical semiconductor or a white light source using a laser light source is used, and the white light source is composed of three colors RGB, and the RGB light sources are simultaneously turned on. 2. The electron according to claim 1, wherein focus adjustment of an image observed by an optical system for observing the sample is performed, and after the focus adjustment is completed, each of the RGB colors is selectively switched and irradiated. camera. 2. The electronic camera according to claim 1, wherein a current value supplied to an LED or a laser emitting light of different wavelengths can be individually adjusted, and a wavelength sensitivity difference between a CCD or CMOS sensor receiving the light is corrected. In a scanning laser microscope that scans a sample with a beam spot irradiated from a laser and observes reflected light or transmitted light of the scanned laser beam spot, the power per time on the sample of the scanned laser beam is controlled to be constant. A modulator for modulating a scanning laser beam; a pulse generator for driving an actuator for scanning the laser beam; an estimator for estimating a scanning speed of the laser beam scanned by the actuator; and the estimator for the modulator. The scanning laser microscope is characterized in that a scanning speed which is an output of the above is input, and the laser beam output from the modulator is subjected to pulse width modulation, pulse frequency modulation or pulse amplitude modulation. The scanning laser microscope characterized in that the laser beam pulse width or pulse height output from the modulator is proportional to the scanning speed of the actuator during laser beam scanning.

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