JP4633100B2 - Laser scanning microscope - Google Patents

Description

  The present invention relates to a laser scanning microscope that realizes a long life of a laser light source.

  For example, some laser scanning microscopes used in the biological field use a plurality of staining dyes for a sample and enable observation of an observation object colored with these staining dyes simultaneously. In such a laser scanning microscope, a laser light source that emits excitation light having different wavelengths by combining a single laser beam or a plurality of laser beams is employed as a light source device.

  Incidentally, it is known that a laser light source generally has a short lifetime and is expensive in price. For this reason, conventionally, it has been considered to extend the life of the laser light source by turning off the laser light source or reducing the amount of output light when the laser light is not used. Such a laser light source has an external control terminal in the power source, and by inputting an appropriate signal to this external control terminal, the laser tube discharges from the normal state to the minimum current (laser that causes laser oscillation). (Standby state). Therefore, in a laser light source having such a laser standby function, by inputting a control signal to the external control terminal and minimizing the discharge current, the laser oscillation can be put into a standby state so that the laser light source can be used efficiently. ing.

  However, normally, in a laser scanning microscope, when the laser light source is not used continuously for a predetermined time, the laser light source is manually turned off, but two-dimensional scanning for obtaining a scanned image is performed. The laser light from the laser light source remains emitted for a short period when it is not. In addition, in the case of obtaining excitation light having different wavelengths by combining a plurality of laser beams as described above, it is possible to obtain excitation light having a target wavelength by appropriately inserting a shutter or a filter into the plurality of laser light sources. However, even in such a case, the laser light from the laser light source whose optical path is cut off by the shutter is still emitted, causing the life of the laser light source to be shortened.

  The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a laser scanning microscope capable of realizing a long life of a laser light source.

The invention according to claim 1 irradiates and scans an observation target using a staining dye with a laser beam having a wavelength corresponding to the staining dye used for the observation target among laser beams from a plurality of laser light sources, In a laser scanning microscope that obtains a scanning image of an observation image based on light from the observation object,
A state detection unit that detects a state for obtaining the scanned image; and a control unit that performs light amount control for reducing the output light amount of the laser light source based on the state information detected by the state detection unit. Detects whether it is an observation period for acquiring the scanned image,
Based on the state information from the state detection unit, the control unit performs control to reduce the output light amount for all laser light sources when it is not the observation period, and when it is the observation period, the observation target The laser light source that outputs laser light having a wavelength other than the wavelength selected by the used dye information used in the above is controlled to reduce the output light amount.
According to a second aspect of the present invention, in the first aspect of the invention, the state detecting means detects an open / close state of a shutter that causes the laser light to enter the means for scanning the laser light. .
According to a third aspect of the present invention, in the first or second aspect of the invention, the control means forcibly switches the laser light source to standby control when the scanning of the laser light is instructed and the scanning is not started for a predetermined time. It is characterized by having a timer means for shifting.
The invention according to claim 4 scans the object to be observed using a dye by irradiating laser light having a wavelength corresponding to the dye used for the object to be observed among laser beams from a plurality of laser light sources, A method of controlling a laser light source in a laser scanning microscope that obtains a scanned image of an observation image based on light from the observation object,
A step of detecting a state for obtaining the scanned image; and a step of performing a light amount control for reducing an output light amount of the laser light source based on the detected state information. The detecting step includes: In the controlling step, based on the information on the state obtained in the detecting step, the output light quantity for all laser light sources when it is not the observation period. Control to a state in which the output light amount is reduced with respect to a laser light source that outputs laser light having a wavelength other than the wavelength selected by the dye information used for the observation target during the observation period. It is characterized by.

  As a result, according to the present invention, the laser light source is shifted to the standby state except when there is an instruction to obtain a scanned image, thereby preventing unnecessary use of the laser light source and extending the life of the laser light source. it can.

  Further, according to the present invention, even if there is an instruction to obtain a scanned image, if a predetermined operation is not performed during a predetermined time, it is forcibly shifted to standby control so that the laser light source is wasted. Can be prevented.

  According to the present invention, the laser light source can be prevented from being wasted and the life of the laser light source can be extended by shifting the laser light source to the standby state except when there is an instruction for obtaining a scanned image.

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

(First embodiment)
FIG. 1 shows a schematic configuration of a laser scanning microscope to which the first embodiment is applied. In the figure, reference numerals 1a, 1b, and 1c denote a plurality of laser light sources, and laser power sources 2a, 2b, and 2c are connected to the laser light sources 1a, 1b, and 1c.

  These laser light sources 1a, 1b, and 1c are composed of gas lasers such as argon and krypton, and emit laser beams having different wavelengths in response to voltage supply from laser power sources 2a, 2b, and 2c. The laser power sources 2a, 2b, and 2c enable control of the output light amounts of the laser light sources 1a, 1b, and 1c, and normal outputs from the laser light sources 1a, 1b, and 1c according to an instruction signal from a laser power source control unit 63 described later. The normal control for generating the amount of light and the standby control for controlling the discharge current supplied to the laser light sources 1a, 1b, and 1c to the minimum value (standby state) can be selected.

  Laser beams from the laser power sources 2a, 2b, and 2c pass through the shutters 3a, 3b, and 3c separately and enter the laser scanning microscope main body 4 through a common optical path such as an optical fiber (not shown).

  The laser scanning microscope main body 4 includes a scan unit 41 and a microscope unit 42, and laser light incident on the scan unit 41 through a common optical path from the shutters 3a, 3b, and 3c is galvano-coupled via the incident shutter 41a. The light enters the mirror 41b, is deflected in the X direction and the Y direction by the operation of the galvano mirror 41b, is irradiated onto the sample 5 through an objective lens (not shown) of the microscope unit 42, and two-dimensionally scans the sample.

  A control unit 6 is connected to the laser power sources 2a, 2b, 2c and the laser scanning microscope main body 4. The control unit 6 includes a shutter driver 61, a galvano driver 62, a laser power source control unit 63 and a main control unit 64. The shutter driver 61 controls the opening and closing of the shutters 3a, 3b, and 3c and the entrance shutter 41a according to instructions from the main control unit 64. The shutters 3a, 3b, and 3c are not used according to the dye information used according to the observation conditions. The laser light sources corresponding to the laser light sources 1a, 1b, and 1c are controlled to be closed, and the incident shutter 41a is controlled to be opened when the scanned image is observed. The galvano driver 62 drives and controls the galvano mirror 41b in the X direction and the Y direction according to an instruction from the main control unit 64. The laser power source control unit 63 determines normal control or standby control for the laser power sources 2a, 2b, and 2c based on an instruction from the main control unit 64 to the shutter driver 61, and outputs each instruction. The state detector 63a for detecting the state of the incident shutter 41a is detected. When the state detector 63a detects that the incident shutter 41a is open, normal control is determined as the observation period of the scanned image, and the incident shutter 41a is detected. When the closed state is detected, standby control is determined, and respective instructions are output to the laser light sources 2a, 2b, and 2c.

  Reference numeral 7 denotes a monitor that displays a scanning image for the sample 5 and a menu image for instructing observation.

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

  First, the laser light sources 1a, 1b, and 1c to be used are determined based on the used dye information corresponding to the observation conditions. Here, only the laser light sources 1a and 1b are used. Then, an instruction is issued from the main controller 64, and the shutter driver 61 opens the shutters 3a and 3b corresponding to the instructed laser light sources 1a and 1b, and closes the shutter 3c corresponding to the unused laser light source 1c. .

  In this state, the main control unit 64 determines whether or not the shutter driver 61 controls to open the entrance shutter 41a, that is, whether or not it is a scanning image observation period. Here, if it is determined that the incident shutter 41a is closed and it is not the scanning image observation period, an instruction is given to the laser power source control unit 63, and the laser power sources 2a and 2b of the laser light sources 1a, 1b, and 1c are put on standby. Direct control. As a result, the current supplied from the laser power sources 2a, 2b and 2c to the laser light sources 1a, 1b and 1c is controlled to the minimum value (standby state), and the respective laser light sources 1a, 1b and 1c are shifted to the standby state. The

  Next, when the observation of the scanned image is instructed, the shutter driver 61 controls the entrance shutter 41a to be opened.

  The main control unit 64 determines from the state of the incident shutter 41a that it is the scanning image observation period. Then, an instruction is given to the laser power source control unit 63, and normal control is instructed only to the laser power sources 2a and 2b of the laser light sources 1a and 1b instructed to use. Then, the current supplied to the laser light sources 1a and 1b from the laser power sources 2a and 2b is controlled to a value necessary for generating a normal output light amount. The respective laser light sources 1a and 1b Transition to state. In this case, the laser light source 1c remains in a standby state.

  In this state, the laser light emitted from the laser light sources 1a and 1b is introduced into the scan unit 41 of the laser scanning microscope main body 4 through an optical path composed of an optical fiber, passes through the incident shutter 41a, and is then galvanomirror 41b. Is incident on. In this case, the galvanometer mirror 41b is driven and controlled in the X and Y directions by the galvanometer driver 62, and the laser light incident on the galvanometer mirror 41b is deflected in the XY direction and passes through the microscope unit 42 on the sample 5. The sample 5 is scanned two-dimensionally. Then, the fluorescence emitted from the sample 5 is received by a photodetector (not shown) via the scan unit 41, then subjected to image processing, generated as an observation image, and displayed on the monitor 7.

  Accordingly, the laser light from the laser light sources 1a to 1c can be incident on the galvano mirror 41b via the incident shutter 41a, and the sample 5 can be scanned two-dimensionally by the galvano mirror 41b. At the same time, when the state of the incident shutter 41a is detected by the state detector 63a and the open state of the incident shutter 41a is detected by a laser scanning microscope that is adapted to obtain a scanned image according to the intensity of light from the sample 5, The laser power control unit 63 determines that it is the observation period of the scanned image, performs normal control, and when the closed state of the incident shutter 41a is detected, the laser power control unit 63 determines that the scanned image is not being observed, and performs standby control. Do. From these results, the laser power sources 2a to 2c are controlled so that the output light amounts of the laser light sources 1a to 1c can be adjusted. Therefore, the laser light sources 1a to 1c can be shifted to the standby state outside the scanning image observation period. Thus, useless use of the laser light sources 1a to 1c can be prevented, and the life of the laser light sources 1a to 1c can be extended.

  Further, since the standby state is maintained for the laser light source 1c whose optical path is cut off by the shutter 3c, useless use of the laser light source 1c that is not used can be prevented.

  In the first embodiment described above, for example, another signal can be used as long as the standby control instruction is linked to the scanning state. For example, a sensor may be provided inside the laser scanning microscope main body 4 to detect the opening / closing of the shutter or the mechanical position and operating state of the galvano mirror, and this detection signal may be used. The detection method of the scanning image observation state for switching the control of the laser light sources 1a to 1c between the normal control and the standby control is, for example, the control in the optical path switching unit such as the motorized scanning image observation optical path and the visual observation optical path. A state may be determined by providing a sensor in a signal detection method or an optical path switching unit. Further, as a method of controlling the output light amount of the laser light sources 1a to 1c, a method of controlling the light amount by external control and reducing the output light amount can be used without using the standby function of the laser power sources 2a to 2c. Furthermore, in the above description, gas lasers have been mainly described as the laser light sources 1a to 1c, but solid lasers such as semiconductor lasers can also be applied.

(Second Embodiment)
In this case, since the schematic configuration of the laser scanning microscope to which the second embodiment is applied is the same as that of FIG. 1, the same figure is used, but here, the timer 63 b is provided in the laser power source control unit 63. The standby control output output from the laser power supply control unit 63 is sent only when a certain condition (time) is satisfied.

  In this way, for example, when the observation of the scanned image is instructed, and the entrance shutter 41a is controlled to be opened by the shutter driver 61 and scanning is started, the laser power supply control unit 63 performs normal operations on the laser power supplies 2a and 2b. Although control is instructed, the timer 63b is started at the same time as the scanning is stopped.

  In this case, if the drive control of the galvano mirror 41b by the galvano driver 62 for the next scanning image observation is performed before the predetermined time T set in the timer 63b elapses, the time elapse detection of the timer 63b is invalidated. The normal control for the laser power sources 2a and 2b is made effective, and the laser light sources 1a and 1b are shifted to the normal state. Further, if the drive control of the galvano mirror 41b by the galvano driver 62 is not performed before the predetermined time T elapses, the detection of the time elapse of the timer 63b is enabled and the normal control for the laser power sources 2a and 2b is disabled. At the same time, the instruction of the laser power source control unit 63 is switched to standby control and is given to the laser power sources 2a and 2b. Thereby, the current supplied to the laser light sources 1a and 1b by the laser power sources 2a and 2b is controlled to the minimum value (standby state), and the laser light sources 1a and 1b are shifted to the standby state.

  Therefore, in this way, even when a normal control instruction is given for observing the scanned image, if the predetermined operation is not performed during the predetermined time T, the standby control is forcibly switched. Therefore, useless lighting of the laser light sources 1a to 1b can be eliminated, and the life of the laser light sources 1a to 1b can be further extended.

  In addition, 2nd Embodiment is good also as a system which asks an observer whether the transfer is possible at the time of transfer to standby control after predetermined time T progress.

The figure which shows schematic structure of embodiment of this invention.

Explanation of symbols

1a. 1b. 1c Laser light source 2a. 2b. 2c ... Laser power supply 3a. 3b. 3c ... Shutter 4 ... Laser scanning microscope main body 41 ... Scan unit 41a ... Incident shutter 41b ... Galvano mirror 42 ... Microscope part 5 ... Sample 6 ... Control unit 61 ... Shutter driver 62 ... Galvano driver 63 ... Laser power supply control part 63a ... State Detection unit 63b ... Timer 64 ... Main control unit 7 ... Monitor

Claims (4)

Based on the light from the observation object, the object to be observed using the staining dye is scanned by irradiating laser light having a wavelength corresponding to the dye used for the observation object among laser lights from a plurality of laser light sources. In a laser scanning microscope that obtains a scanned image of an observation image,
State detecting means for detecting a state for obtaining the scanned image;
And control means for performing light quantity control of dropping the output light amount of by Ri said laser light source on the state information detected by the state detecting means,
The state detection means detects whether it is an observation period for acquiring the scanned image,
Based on the state information from the state detection unit, the control unit performs control to reduce the output light amount for all laser light sources when it is not the observation period, and when it is the observation period, the observation target Performing control to reduce the output light amount with respect to a laser light source that outputs laser light other than the wavelength selected by the used dye information used in
A laser scanning microscope characterized by that. It said state detecting means, the laser scanning microscope according to claim 1, characterized in that for detecting the open or closed state of the shutter to be incident laser beam to the means for scanning the laser beam.   3. The control means according to claim 1, further comprising timer means for forcibly shifting the laser light source to standby control when scanning of the laser light is instructed and the scanning is not started for a predetermined time. Laser scanning microscope. Based on the light from the observation object, the object to be observed using the staining dye is scanned with laser light having a wavelength corresponding to the dye used for the observation object among laser lights from a plurality of laser light sources. A method of controlling a laser light source in a laser scanning microscope that obtains a scanned image of an observation image,
Detecting a state for obtaining the scanned image;
Performing light amount control to reduce the output light amount of the laser light source based on the detected state information;
Comprising
In the detecting step, it is detected whether it is an observation period for acquiring the scanned image,
In the controlling step, based on the information on the state obtained in the detecting step, when it is not the observation period, control is performed to reduce the output light amount with respect to all laser light sources, and the observation period. When controlling the state in which the output light amount is reduced with respect to a laser light source that outputs laser light having a wavelength other than the wavelength selected by the used dye information used for the observation target ,
A laser light source control method in a laser scanning microscope.

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