JP2005173487A - Fluorescence microscope and computer program - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fluorescent microscope and a computer program which enables a light source to be successfully controlled. <P>SOLUTION: Light from a plurality of LEDs 4 and 14 constituting the light source is condensed on a fluorescent sample S through an objective lens 3, and an eyepiece 10 or a CCD camera 11 is provided which observes fluorescence emitted from the fluorescent sample, and the plurality of LEDs 4 and 14 are controlled by a control part 20 for controlling lighting/extinction of the plurality of LEDs 4 and 14 so that their lighting start times are mutually shifted. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a fluorescence microscope and a computer program using a small light emitting element such as a plurality of LEDs as a light source.

  Fluorescence observation is widely known as being capable of visualizing specific molecules of biological cells and observing the distribution of the molecules.

  Conventionally, as a microscope for performing fluorescence observation, there is a fluorescence microscope for irradiating a specimen with illumination light (excitation light) having only a specific wavelength component and observing fluorescence generated in the specimen. In such a fluorescence microscope, a mercury lamp, a halogen lamp, or the like is used as a light source of illumination light, and a specific wavelength component of the illumination light is used by using a wavelength selection filter called an excitation filter for the illumination light emitted from these light sources. Is extracted, and this is irradiated to the specimen as excitation light.

  Recently, it has been proposed to use a high-intensity light-emitting diode (LED) as illumination light as a small light-emitting element as disclosed in, for example, Patent Document 1. Since these LEDs are monochromatic and have various emission wavelengths, good fluorescence observation is possible by generating excitation light using an LED having an emission wavelength most effective for fluorescence. In addition, compared to mercury lamps, halogen lamps, and the like, LEDs can be expected to have effects such as system miniaturization, low power consumption, low heat generation, and long life.

  On the other hand, when an LED is used as illumination light, dimming is required to adjust the light intensity. However, with respect to LEDs, if the applied voltage is simply increased or decreased, the intensity distribution peak of the emission wavelength shifts (wavelength shift). For this reason, when an LED is used as illumination light, if a wavelength shift occurs due to the dimming of the LED, excitation light having an optimum wavelength for fluorescence observation cannot be obtained, and fluorescence observation may not be possible.

Therefore, as disclosed in Patent Document 2 and Patent Document 3, conventionally, PWM (Pulse Width Modulation) for efficiently causing the LED to emit light has been considered. This PWM repeatedly generates a pulse train (several tens to several MHz) for turning on (turning on) / off (extinguishing) the LED, and dimming is possible by the duty ratio of these ON and OFF. Is used, even if the brightness of the LED is changed by changing the duty ratio, the applied voltage is always the same when the LED is turned on, and thus stable light control without causing a wavelength shift is possible.
JP 2002-321093 A Japanese translation of PCT publication No. 2003-522393 JP 2001-153808 A

  By the way, in a general microscope using LEDs as illumination light, a light source unit incorporating an LED called a lamp house, a PWM control circuit for controlling the LED, and a power supply unit incorporating a power source have different configurations. These are connected via a cable of several tens of centimeters to several meters. In addition, the LED consumes less power than a mercury lamp or a halogen lamp, but a current of about several hundred mA is passed at a rated current for efficient lighting.

  However, in the case of such a configuration, electrical noise may be mixed with the current flowing through the LED due to the periodic ON / OFF of the PWM via the cable. This is because, in fluorescence observation, since a plurality of excitation lights having different wavelengths are used, when a single color LED is used as a light source, a plurality of LEDs having different wavelengths may be turned on simultaneously. By turning ON / OFF at the same timing, a larger electrical noise is generated, which may affect the surrounding environment.

  In addition, when observing living cells, light of a specific wavelength is irradiated for a specific time, and subsequent cell changes are observed. In such a case as well, the LED is electrically controlled. The irradiation time of the excitation light can be set with high accuracy because it can be turned on and off at high speed only.

  However, in this case as well, there is a problem that it is difficult to know how much light is actually irradiated to the cells when the excitation light is easily combined with PWM. .

  The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a fluorescence microscope and a computer program capable of controlling a good light source.

  The invention according to claim 1 is a light source having a plurality of small light emitting elements, an objective lens for condensing the light from the light source on the specimen, an observation means for observing the fluorescence generated from the specimen, and the light source Blinking control means for controlling blinking of the plurality of small light emitting elements, wherein the blinking control means controls the lighting start times of the plurality of small light emitting elements to be shifted from each other.

  According to a second aspect of the present invention, in the first aspect of the invention, the blinking control unit calculates a ratio of lighting and extinguishing according to a brightness setting value set for the plurality of small light emitting elements. Calculating a phase delay time of a pulse train generated based on the ratio of turning on and off when the plurality of light emitting elements are turned on simultaneously, and outputting a pulse train for turning on the plurality of light emitting elements from these calculation results It is characterized by.

  According to a third aspect of the present invention, there is provided a light source having a plurality of small light emitting elements, an objective lens for condensing light from the light source on the specimen, an observation means for observing fluorescence generated from the specimen, Blinking control means for controlling blinking of a plurality of small light emitting elements, and the blinking control means instructs the irradiation time of the specimen by the light from the light source to the plurality of small light emitting elements. The ratio of lighting and extinguishing is calculated according to the set brightness value, and the instructed lighting time is compared with the calculated lighting time, and a pulse train corresponding to the lighting time is determined based on the comparison result. It is characterized by determining whether to generate or to generate a single pulse.

  The invention according to claim 4 is a computer program for controlling a light source having a plurality of small light emitting elements of a fluorescence microscope for observing fluorescence of a specimen, and is a brightness setting value set for the plurality of small light emitting elements And calculating the ratio of lighting and extinction according to the above, and calculating the phase delay time of the pulse train generated by the ratio of lighting and extinction at the time of simultaneous lighting of the plurality of light emitting elements, from the calculation results A pulse train for lighting the light emitting element is output.

  The invention according to claim 5 is a computer program for controlling a light source having a plurality of small light emitting elements of a fluorescence microscope for observing fluorescence of a specimen, and suspends an indication of an irradiation time of the specimen by light from the light source. The ratio of lighting and extinguishing is calculated according to the brightness setting value set for the plurality of small light emitting elements, and the indicated lighting time is compared with the calculated lighting time. Thus, it is characterized in that a pulse train corresponding to the lighting time or a single pulse is determined and output.

  As a result, according to the present invention, lighting of a plurality of small light emitting elements does not start at the same timing, and reduction of electric noise generation and power saving can be realized.

  Further, according to the present invention, the observer can irradiate for a specified time without being aware of the control of the irradiation time of the small light emitting element, and the operability can be improved.

  Furthermore, according to the present invention, it is possible to reduce the generation of electrical noise and save power by controlling the small light emitting elements not to start lighting at the same timing. Can be simplified

  According to the present invention, in a fluorescent microscope using a plurality of small light emitting elements such as LEDs as a light source, even when a plurality of these small light emitting elements are turned on, lighting does not start at the same timing. Generation can be reduced, the influence on the surrounding environment can be minimized, and further power saving can be realized.

  In addition, since the lighting time of the small light emitting element can be controlled by specifying the lighting time of the specimen, the observer can irradiate the specimen with illumination light at a preset brightness without being aware of switching of the lighting control of the LED. Thus, it is possible to improve the operability, and in particular, it is possible to realize a fluorescence microscope excellent in observation with living cells.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings.

(First embodiment)
FIG. 1 shows a schematic configuration of a fluorescence microscope to which the present invention is applied. In the first embodiment, an example is shown in which two LEDs, which are small light-emitting elements, are used as the illumination light source.

  In FIG. 1, reference numeral 1 denotes a stage, and a fluorescent sample S is placed on the stage 1. Further, the stage 1 can be moved up and down in the optical axis direction (arrow direction in the figure), and the fluorescent sample S can be focused.

  Above the stage 1, the objective lens 3 is disposed in the vicinity of the fluorescent sample S. A plurality of objective lenses 3 having different magnifications (only two are shown in the drawing) are held by the revolver 2 and are selectively positioned on the observation optical path a by the operation of the revolver 2. ing.

  Reference numeral 4 denotes an LED (A) as a small light emitting element which is a light source for illumination, and a collector lens 5 and an excitation filter 6 are arranged in an optical path of light emitted from the LED (A) 4. The collector lens 5 converts the light from the LED (A) 4 into parallel light. The excitation filter 6 transmits wavelength light in a band necessary for exciting the fluorescent sample S among the light transmitted through the collector lens 5.

  A dichroic mirror 7 is disposed in the optical path of the light transmitted through the excitation filter 6 at a position intersecting the observation optical path a. The dichroic mirror 7 has a characteristic that reflects the wavelength light in the band necessary for exciting the fluorescent sample S that has passed through the excitation filter 6 and transmits the fluorescence emitted from the fluorescent sample S.

  The objective lens 3 described above is disposed in the reflection optical path of the dichroic mirror 7, and the excitation light reflected by the dichroic mirror 7 is condensed and irradiated on the fluorescent sample S, and is longer than the wavelength of the excitation light by the fluorescent sample S. Light shifted to the wavelength side is generated.

  An optical path switching unit 9 is disposed in the reflected optical path of the dichroic mirror 7 via a barrier filter 8. The barrier filter 8 cuts light outside the fluorescent band and acquires an observation image with a good S / N. An optical path switching motor 13 is connected to the optical path switching unit 9. The optical path switching unit 9 can switch the optical path of the observation image to an eyepiece lens 10 or a CCD camera 11 as observation means by an optical path switching motor 13.

  On the other hand, reference numeral 14 denotes an LED (B) as a small light emitting element which is another illumination light source, and a collector lens 15 and a half mirror 16 are arranged in an optical path of light emitted from the LED (B) 14. The collector lens 15 converts the light from the LED (B) 14 into parallel light. The half mirror 16 is disposed at a position intersecting the optical path of the light from the LED (A) 4, transmits the light from the LED (A) 4, reflects the light from the LED (B) 14, and each light. Is made incident on the excitation filter 6.

  In this case, similarly to the light from the LED (A) 4, the light from the LED (B) 14 transmits wavelength light in a band necessary for exciting the fluorescent sample S by the excitation filter 6, and the dichroic mirror 7 transmits the light. After the reflection, the light is condensed and irradiated on the fluorescent specimen S through the objective lens 3, and light shifted from the fluorescent specimen S to the longer wavelength side than the wavelength of the excitation light is generated.

  An LED drive unit (A) 17 is connected to the LED (A) 4, and an LED drive unit (B) 18 is connected to the LED (B) 14. These LED drive unit (A) 17 and LED drive unit (B) 18 perform drive control by PWM control on LED (A) 4 and LED (B) 14, respectively.

  A camera control unit 19 is connected to the CCD camera 11. The camera control unit 19 executes various controls for the CCD camera 11.

  A control unit 20 is connected to the LED drive unit (A) 17, the LED drive unit (B) 18, and the camera control unit 19. The control unit 20 includes a known CPU circuit. As shown in FIG. 2, the control unit 20 includes a CPU main body 201, a ROM 202 that stores a program for controlling the system, a volatile memory that stores data necessary for control, and the like. RAM 203, I / O port 204 for inputting / outputting control signals, I / F 205 for communicating with external host computer 21, power source 206, oscillator (not shown) necessary for controlling the entire control unit 20, address decoder, etc. It has a well-known peripheral circuit. The CPU main body 201, ROM 202, RAM 203, I / O port 204, and I / F 205 are connected by a data bus 207.

  An LED driving unit (A) 17 and an LED driving unit (B) 18 are connected to the I / O port 204. Further, the LED drive unit (A) 17 and the LED drive unit (B) 18 are connected to the power source 206. The power source 206 is for supplying electric power for lighting the LED (A) 4 and the LED (B) 14 via the LED driving unit (A) 17 and the LED driving unit (B) 18.

  A dimming volume (A) 22 and a dimming volume (B) 23 are connected to the I / O port 204. These dimming volume (A) 22 and dimming volume (B) 23 are the brightness of LED (A) 4 and LED (B) 14 via LED driving unit (A) 17 and LED driving unit (B) 18. Is to set

  Here, two dimming volumes (A) 22 and dimming volume (B) 23 are prepared corresponding to LED (A) 4 and LED (B) 14, but one using a switching method is adopted. The dimming volume may be used for operation. In addition, the dimming volume (A) 22 and the dimming volume (B) 23 are of analog type. For example, the dimming value is obtained by counting the rotation angle of the knob using a rotary encoder or the like. It is also possible to use a digital type in which the light control value is counted up or down by pressing a push switch or the like.

  The I / F 205 can communicate with the host computer 21 using a known communication method such as RS-232C, and the dimming of the LED (A) 4 and the LED (B) 14 is performed by a command transmitted from the host computer 21. It is also possible to do.

  Such a control unit 20 may be a dedicated unit or a general personal computer such as the host computer 21.

  Next, the operation of the embodiment configured as described above will be described.

  In this case, when light is emitted from the LEDs (A) 4 and LEDs (B) 14 of the plurality of illumination light sources, the light from the LED (A) 4 is converted into parallel light by the collector lens 5, and the half mirror 16 is The light is transmitted and incident on the excitation filter 6, the wavelength light in the band necessary for excitation of the fluorescent sample S is transmitted, reflected by the dichroic mirror 7, and irradiated onto the surface of the fluorescent sample S through the objective lens 3. Similarly, the light from the LED (A) 14 is also converted into parallel light by the collector lens 15, reflected by the half mirror 16 and incident on the excitation filter 6, and wavelength light in a band necessary for exciting the fluorescent sample S. The light is transmitted, reflected by the dichroic mirror 7, and irradiated on the fluorescent sample S surface through the objective lens 3.

  Then, the light generated from the fluorescent sample S by the excitation light passes through the dichroic mirror 7 and enters the barrier filter 8, and light other than the fluorescent band is cut off by the eyepiece 10 via the optical path switching unit 9. Visual observation of the observation image or imaging of the observation image by the CCD camera 11 is performed.

  From this state, dimming of the LED (A) 4 and the LED (B) 14 by the control unit 20 is executed by the program shown in FIG. FIG. 3 is a flowchart showing the LED dimming control program stored in the ROM 202 of the control unit 20.

  First, in step S1, setting values from the dimming volume (A) 22 and dimming volume (B) 23 are read. That is, in step S1, the set values of the dimming volume (A) 22 and dimming volume (B) 23 corresponding to the brightness of the LED (A) 4 and LED (B) 14 set by the observer are set to I. Read separately via / O port 204.

  Next, in step S2, the PWM duty for the set values of the dimming volume (A) 22 and dimming volume (B) 23 is calculated. In this case, the CPU main body 201 calculates the PWM duty from the set values read from the dimming volume (A) 22 and the dimming volume (B) 23.

  Here, PWM will be briefly described with reference to FIG. The waveform W1 is a pulse train in which a rectangular wave with a period TPWM is repeated. If the LED is turned on when the pulse train becomes a high level, and the LED is turned off when the pulse train becomes a low level, the LED is turned on by the waveform W1. The apparent brightness can be changed by repeatedly turning off and turning on, and changing the ratio of turning on and off, that is, the duty.

  The PWM control is to change the ratio of the high level time TON and the low level time TOFF in one cycle of the pulse train. Then, when the maximum value VMAX of the volume is the maximum brightness of the LED, the high level, that is, the LED lighting time TON can be calculated by the equation 1 based on the volume setting value Vc. However, since one cycle is divided by the number of divisions (for example, 4 to 16 bits), in the calculation in the CPU main body 201, it is calculated as a ratio of the number of divisions.

TON = TPWM × (Vc / VMAX) ...... Equation 1
Next, in step S3, when both LED (A) 4 and LED (B) 14 are turned on, the phase delay time of the pulse train generated by PWM is calculated. In this case, the CPU main body 201 calculates the phase delay time of the pulse train of each LED when the LED (A) 4 and the LED (B) 14 are turned on simultaneously.

  Here, a method of calculating the phase delay time will be described with reference to FIG. The waveform W2 drives the LED (A) 4 and is a pulse train with a period TPWM, and the waveform W3 drives the LED (B) 14 and is a pulse train with a period TPWM similar to the waveform W2. Further, when the high level of each waveform, that is, the LED lighting time is TAON and TBON, respectively, and the LOW level, that is, the LED extinguishing time is TAOFF and TBOFF, the difference between the rising levels PA and PB of the high level of each waveform. That is, the phase delay time TDelay can be obtained by Equation 2.

TDelay = (TAOFF / 2) + TAON− (TBON / 2) …… Equation 2
Next, in step S4, a pulse train generated by PWM is output from the I / O port 204. In this case, the waveform of the pulse train generated based on the results calculated in step S2 and step S3 is output from the I / O port 204, and the LED (A) is output by the LED drive unit (A) 17 and the LED drive unit (B) 18. 4. Turn on the LED (B) 14.

  Therefore, if the waveform W3 is delayed from the waveform W2 by the phase delay time TDelay in this way, the turn-on time TBON of the LED (B) 14 enters during the turn-off time TAOFF of the LED (A) 4. Since each LED (A) 4 and LED (B) 14 are not lit simultaneously, the current consumption of the entire circuit can be reduced, and power saving can be realized. In addition, by preventing the LEDs (A) 4 and LED (B) 14 from starting lighting at the same timing, it is possible to prevent inrush currents for the two LEDs (A) 4 and LED (B) 14. The electrical noise that accompanies it can be greatly reduced, and the influence on the surrounding environment can be minimized.

  Further, as shown in FIG. 6, when the turn-off time TAOFF of the LED (A) 4 of the waveform W4 is smaller than the turn-on time TBON of the LED (B) 14 of the waveform W5, the LED (A) 4, There may be times when the LEDs (B) 14 are turned on at the same time. In this case as well, the LED (A) 4 and the LED (B) 14 do not start lighting at the same timing. 4. Inrush current for two LEDs (B) 14 can be prevented.

(Modification)
Next, a modification of the first embodiment will be described.

  In the first embodiment, two LEDs of LED (A) 4 and LED (B) 14 have been described as illumination light sources. However, three or more LEDs may be provided depending on the configuration of the lamp house. In this case also, the phase delay time TDelay can be calculated.

  FIG. 7 shows waveforms (waveforms W6, W7, W8, and W8) when four LEDs (LED (A), LED (B), LED (C), and LED (D)) as illumination light sources are turned on. W9).

  Also in this case, it can be obtained by equally dividing the TAOFF time and assigning the TBON, TCON, and TDON times in the same manner as the calculation of the phase delay time described above. This is expressed as follows.

TBDelay = (TAOFF / 4) + TAON− (TBON / 2) …… Equation 3
TCDelay = (TAOFF / 2) + TAON− (TCON / 2) ...... Formula 4
TDDelay = (TAOFF × (3/4)) + TAON− (TDON / 2) Equation 5
In this case, when there is an LED that is not lit, the calculation may be performed only by the number of LEDs that are lit.

  Therefore, even when a plurality of LEDs (A), LED (B), LED (C), and LED (D) are turned on in this way, the plurality of LEDs are prevented from starting to light at the same timing. Inrush current when the LED is turned on can be prevented, and electrical noise can be reduced.

(Second Embodiment)
Next, a second embodiment of the present invention will be described.

  In this case, since the hardware configuration of the second embodiment is the same as that of FIGS. 1 and 2 described in the first embodiment, these drawings are incorporated.

  FIG. 8 is a flowchart showing the LED irradiation time control (one-shot) program stored in the ROM 202 of the control unit 20 described in FIG.

  Here, the program shown in FIG. 8 is executed when the control unit 20 is activated and the LED can be dimmed.

  First, in step S11, input of irradiation time is awaited. That is, in step S11, it waits until an observer inputs the value of irradiation time. The observer inputs the irradiation time of light to be irradiated on the specimen. The irradiation time may be input by inputting a command from the external host computer 21 or by connecting an input device (not shown) to the I / O port 204 or I / F 205.

  When the observer inputs the irradiation time from this state, the set value from the light control volume (A) 22 is read out in step 12. Here, only the LED (A) 4 is dimmed, but when a plurality of LEDs are used, the set value is read from the dimming volume of each LED.

  Next, in step S13, the PWM duty for the set value of the dimming volume (A) 22 is calculated. That is, in step S13, the PWM duty is calculated based on the set value of the dimming volume (A) 22, as described in step S2 shown in FIG.

  Next, in step S14, it is determined whether LED lighting time <irradiation time. That is, in step S14, the LED lighting time TON and the irradiation time Ti are compared.

  Here, a comparison between the LED lighting time TON and the irradiation time Ti will be described with reference to FIG. First, when the irradiation time Ti is longer than the LED lighting time TON, that is, when Ti = Ti1, the process proceeds to step S15, the pulse train generated by PWM is output from the I / O port 204 for the irradiation time Ti1, and the pulse is stopped. . That is, as shown by the waveform W10 in FIG. 9, the pulse train having the duty calculated in step S13 is output during the irradiation time Ti1.

  On the other hand, if the irradiation time Ti is shorter than the LED lighting time TON, that is, if Ti = Ti2, the process proceeds to step S16, and the I / O port 204 is set to Low after being set to High for the irradiation time Ti2. That is, as shown by the waveform W11 in FIG. 9, the I / O port 204 is set to High for the irradiation time Ti2, and then irradiation is performed by returning to Low.

  Next, in step S17, the total irradiation amount is calculated and displayed. That is, in step S17, the total irradiation amount of the light irradiated on the specimen is calculated, and the value is transmitted to the external host computer 21 for display, or a display (not shown) connected to the I / O port 204 or I / F 205 is displayed. Display on the device. The calculation of the total irradiation amount here is different between the case where the processing in step S15 is performed and the case where the processing is performed in step S16. If the total irradiation amounts are E1 and E2, respectively, Is as follows.

First, in the case of the processing of step 15 (PWM control pulse train),
E1 = (dw / dt) × (TON / TPWM) × Ti1 Equation 6
In the case of the process of step 16,
E2 = (dw / dt) × Ti2 Equation 7
However, (dw / dt): Irradiation amount per unit time when PWM is not performed. Therefore, in this way, illumination light from the LED can be irradiated for the irradiation time instructed by the observer to the specimen. A person can improve the operability such as being able to irradiate the specimen with illumination light at a preset brightness without being aware of switching of LED lighting control.

  Thus, for example, in live cell observation, there is a case where illumination light is irradiated to a cell for a specific time, and then the reaction of the cell is observed. In such observation, LED illumination is performed using mercury. Compared with lamps, halogen lamps, etc., the response time is fast, the lighting and extinguishing can be controlled with high accuracy in terms of time, and irradiation is possible for a very short time, which is extremely effective.

(Modification)
Next, a modification of the second embodiment will be described.

  When performing a series of observations with a fluorescence microscope, the order of observation can be stored and executed as a macro in the control unit 20 or the external host computer 21. For example, when the macro is started as shown in FIG. 10, the LED (A) 4 having the waveform W12 is set to (TON / TPWM) × 100 = 10% LED lighting time, that is, 10% of the maximum brightness. Turn on for irradiation time TAi1 hour. On the other hand, as shown in the waveform W14, the macro is started and the CCD camera 11 is controlled at the same time, and the fluorescent image from the fluorescent specimen S is photographed and recorded for the time TS1.

Next, as shown by the waveform W13, the LED (B) 14 is irradiated for TBi2 irradiation time (TBi2 <TPWM) after TBP time from the start of the macro. Further, on the other hand, after taking a picture for only TS1 as shown in the waveform W14, after taking a picture for TS2 time, taking a picture for TS3 time, and taking a fluorescent image from the fluorescent specimen S obtained by the LED (B) 14 Record.
Accordingly, if the imaging order such as LED brightness, irradiation time setting, camera control, etc. can be created as a macro by the observer, the observer can perform the steps shown in FIG. 8 during actual observation. It is not necessary to set the irradiation time and the dimming volume as described in S11 and S12, and the fluorescent image can be automatically observed.

  In addition, this invention is not limited to the said embodiment, In the implementation stage, it can change variously in the range which does not change the summary.

  Furthermore, the above embodiments include inventions at various stages, and various inventions can be extracted by appropriately combining a plurality of disclosed constituent elements. For example, even if some constituent requirements are deleted from all the constituent requirements shown in the embodiment, the problem described in the column of the problem to be solved by the invention can be solved, and is described in the column of the effect of the invention. If the above effect is obtained, a configuration from which this configuration requirement is deleted can be extracted as an invention.

The figure which shows schematic structure of the 1st Embodiment of this invention. The figure which shows schematic structure of the control part used for 1st Embodiment. The flowchart for demonstrating operation | movement of 1st Embodiment. The figure for demonstrating PWM applied to 1st Embodiment. The figure for demonstrating operation | movement of 1st Embodiment. The figure for demonstrating operation | movement of 1st Embodiment. The figure for demonstrating the operation | movement of the modification of the 1st Embodiment of this invention. The flowchart for demonstrating the operation | movement of the 2nd Embodiment of this invention. The figure for demonstrating operation | movement of 2nd Embodiment. The figure for demonstrating operation | movement of the modification of the 2nd Embodiment of this invention.

Explanation of symbols

S ... fluorescent specimen, 1 ... stage, 2 ... revolver 3 ... objective lens, 4.14 ... LED
DESCRIPTION OF SYMBOLS 5 ... Collector lens, 6 ... Excitation filter 7 ... Dichroic mirror, 8 ... Barrier filter 9 ... Optical path switching part, 10 ... Eyepiece lens, 11 ... CCD camera 13 ... Motor for optical path switching, 15 ... Collector lens 16 ... Half mirror, 17 ... LED drive (A)
18 ... LED drive part (B), 19 ... Camera control part 20 ... Control part,
201 ... CPU body, 202 ... ROM
203 ... RAM, 204 ... I / O port 205 ... I / F, 206 ... power supply, 207 ... data bus 21 ... external host computer 22 ... dimming volume (A), 23 ... dimming volume (B)

Claims (5)

A light source having a plurality of small light emitting elements;
An objective lens for condensing the light from the light source on the specimen;
Observation means for observing fluorescence generated from the specimen;
Blinking control means for controlling blinking of a plurality of small light emitting elements of the light source,
The fluorescent light microscope, wherein the blinking control unit controls the lighting start times of the plurality of small light emitting elements to be shifted from each other. The blinking control unit calculates a ratio of lighting and extinguishing according to a set value of brightness set for the plurality of small light emitting elements, and the lighting and extinguishing at the time of simultaneous lighting of the plurality of light emitting elements. 2. The fluorescence microscope according to claim 1, wherein a phase delay time of the pulse train generated by the ratio is calculated, and a pulse train for lighting the plurality of light emitting elements is output from the calculation results. A light source having a plurality of small light emitting elements;
An objective lens for condensing the light from the light source on the specimen;
Observation means for observing fluorescence generated from the specimen;
Blinking control means for controlling blinking of a plurality of small light emitting elements of the light source,
The blinking control unit calculates an on / off ratio according to a set value of brightness set for the plurality of small light emitting elements by instructing an irradiation time of the specimen by light from the light source. And comparing the instructed illumination time with the calculated lighting time, and determining whether to generate a pulse train corresponding to the lighting time or to generate a single pulse according to the comparison result, and outputting A characteristic fluorescence microscope. A computer program for controlling a light source having a plurality of small light emitting elements of a fluorescence microscope for observing fluorescence of a specimen,
A pulse train generated by calculating the ratio of lighting and extinguishing according to the set brightness value set for the plurality of small light emitting elements, and by the ratio of lighting and extinguishing when the plurality of light emitting elements are simultaneously lit A computer program characterized by calculating a phase delay time of and outputting a pulse train for lighting the plurality of light emitting elements from these calculation results. A computer program for controlling a light source having a plurality of small light emitting elements of a fluorescence microscope for observing fluorescence of a specimen,
The indication of the irradiation time of the specimen by the light from the light source is calculated, and the ratio of lighting and extinction is calculated according to the brightness setting value set for the plurality of small light emitting elements, and the instructed A computer program that compares an illumination time with the calculated lighting time, determines whether to generate a pulse train corresponding to the lighting time or a single pulse based on the comparison result, and outputs the pulse train.

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