JP3861000B2 - Scanning laser microscope - Google Patents

Scanning laser microscope Download PDF

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Publication number
JP3861000B2
JP3861000B2 JP2001392524A JP2001392524A JP3861000B2 JP 3861000 B2 JP3861000 B2 JP 3861000B2 JP 2001392524 A JP2001392524 A JP 2001392524A JP 2001392524 A JP2001392524 A JP 2001392524A JP 3861000 B2 JP3861000 B2 JP 3861000B2
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Japan
Prior art keywords
sample
scanning
laser
means
laser light
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Expired - Fee Related
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JP2001392524A
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Japanese (ja)
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JP2003195172A (en
Inventor
伸之 永沢
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オリンパス株式会社
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Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a scanning laser microscope that irradiates a sample while scanning with laser light and detects reflection or fluorescence from the sample.
[0002]
[Prior art]
As is well known, a scanning laser microscope irradiates a sample with a point light source such as a laser beam while scanning the sample in the X- and Y-axis directions via an objective lens, and again emits observation light of fluorescence or reflected light from the sample. Two-dimensional information is obtained by detection with a photodetector via an objective lens and an optical system, and the result is displayed on a monitor screen such as a CRT so that it can be observed as image information.
[0003]
In this case, the condition settings for acquiring the sample image usually include selection of the objective lens, setting of the display image size (number of vertical and horizontal pixels to be displayed on the screen), setting of the scanning speed for display on the monitor, laser Setting of the light intensity (relative value to the intensity of the laser light emitted from the laser light source) is performed. In other words, when acquiring an image of the sample, by setting these conditions, sampling is performed to detect the scanning speed and deflection angle of the galvano scanner that scans the laser light in the XY axis direction, the fluorescence emitted from the sample, and the reflected light. The frequency and the like are determined, and the resulting image signal is displayed on a monitor such as a CRT.
[0004]
[Problems to be solved by the invention]
However, with conventional scanning laser microscopes, the main purpose is to observe the fluorescence (or reflection) image of a sample with a confocal optical system by measuring the observation light emitted from the sample. When detecting the observation light emitted from the sample, the time to irradiate one point on the sample with laser light, the distance between the points sampled on the sample or the size of the stimulus (excitation) given to the sample, etc. It is not set up actively to get various data. For this reason, there has been a problem that data acquisition at a desired resolution, quantitative data acquisition, reproducible data acquisition, and the like cannot be performed.
[0005]
The present invention has been made in view of the above circumstances, and provides a scanning laser microscope capable of positively acquiring data at a desired resolution, quantitative data acquisition, reproducible data acquisition, and the like. With the goal.
[0006]
[Means for Solving the Problems]
According to the first aspect of the present invention, a plurality of objective lenses having different magnifications, objective lens switching means for selectively inserting one of the plurality of objective lenses into the observation optical axis, a laser light source, and the laser light source emit light. Intensity adjusting means for adjusting the intensity of the laser light to be scanned, scanning means for two-dimensionally scanning the laser light on the sample, and a pin disposed at a position conjugate with the focal plane of the objective lens inserted in the observation optical axis A hole, a photodetector that photoelectrically converts observation light from the sample that has passed through the pinhole, an A / D converter that converts an electric signal output from the photodetector into a digital signal, and a condition for acquiring an image of the sample. Input operation means, and the objective lens switching means, the intensity adjustment means, the scanning means, and the front based on the image acquisition conditions input to the operation means In a scanning laser microscope comprising: a microscope control device that controls the A / D converter and constructs an observation image of the sample from the digital signal; and a monitor that displays an image constructed by the microscope control device. The image acquisition conditions input to the operation means are an irradiation spot diameter and an irradiation intensity of the laser light applied to the sample.
[0007]
According to a second aspect of the present invention, in the first aspect of the present invention, the image acquisition condition input to the operation means further includes a scanning range of the laser light and a time during which the laser light is irradiated to one point of the sample. And at least one of the sampling intervals of the observation light.
[0008]
According to a third aspect of the present invention, in the first or second aspect of the present invention, the zoom control system further includes a zoom optical system that adjusts a beam diameter of the laser light incident on the objective lens. The zoom optical system is controlled so as to satisfy the condition of the irradiation spot diameter of the inputted laser beam.
[0009]
The invention according to claim 4 is the invention according to claim 1 or 2, further comprising laser power monitor means for detecting the intensity of the laser beam, wherein the microscope control device is based on a detection result of the laser power monitor means. The intensity adjusting means is controlled so as to satisfy the condition of the irradiation intensity of the laser beam input to the operating means.
[0010]
The invention according to claim 5 is the invention according to claim 1 or 2, wherein the microscope control device sets the objective lens switching means so as to satisfy a condition of an irradiation spot diameter of the laser beam input to the operation means. Controlling the intensity adjusting means so as to satisfy the condition of the irradiation intensity of the laser beam input to the operating means, and satisfying the condition of the scanning range of the irradiation spot of the laser beam input to the operating means Controlling the scanning width of the scanning means, and controlling the scanning speed of the scanning means so as to satisfy the condition of the irradiation time of one point on the sample of the laser light input to the operating means, The sampling interval of the A / D converter is controlled so as to satisfy the resolution condition of the input image.
[0011]
As a result, according to the present invention, the spot diameter of the laser beam on the sample, the irradiation intensity of the laser beam, the irradiation time of the laser beam per point, the optical data acquisition pitch, the optical data acquisition range, etc. are arbitrarily set. This makes it possible to acquire various data with resolutions according to these condition settings, so that it is possible to quantitatively control the stimulus (excitation) applied to the sample and detect the observation light of fluorescence or reflected light emitted from the sample. This enables quantitative measurement and reproducible measurement.
[0012]
According to the present invention, the spot diameter of the laser beam on the sample can be finely controlled by providing a zoom optical system and controlling the numerical aperture (NA) of the objective lens in a minute step or steplessly.
[0013]
Furthermore, according to the present invention, by providing a laser power monitor, the intensity of the laser beam irradiated on the sample can be controlled more accurately, and highly accurate quantitative measurement and reproducible measurement can be performed. it can.
[0014]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, embodiments of the present invention will be described with reference to the drawings.
[0015]
(First embodiment)
FIG. 1 shows a schematic configuration of a scanning laser microscope to which the present invention is applied.
[0016]
Reference numeral 1 denotes a laser light source, and laser light emitted from the laser light source 1 is irradiated onto the sample 5 in a spot shape via a laser power control means 2 as an intensity adjusting means, a dichroic mirror 3 and an objective lens 4. In this case, the laser power control means 2 adjusts the irradiation intensity of the laser light emitted from the laser light source 1. ) To adjust the irradiation intensity of the laser beam. A plurality of objective lenses 4 having different magnifications are provided in a revolver 4a as an objective lens switching means, and one of the plurality of objective lenses 4 is selectively inserted into the observation optical axis by the revolver 4a. It has become so.
[0017]
The spot-like laser light is scanned on the sample 5 by the scanning means 6. In this case, the scanning means 6 is configured to two-dimensionally scan the laser beam on the sample 5 in the XY axis direction in accordance with the operation control signal from the microscope control device 7. At this time, the laser beam is scanned from the sample 5. It can be stopped at a certain point above or moved at a desired speed.
[0018]
The observation light of fluorescence or reflected light emitted from the sample 5 irradiated with the laser light is confocal disposed at a position conjugate with the focal plane of the objective lens 4, the scanning means 6, the dichroic mirror 3, and the objective lens 4. The light is projected onto the photodetector 9 through the pinhole 8.
[0019]
The photodetector 9 photoelectrically converts observation light from the sample 5 that has passed through the confocal pinhole 8, and the electrical signal photoelectrically converted by the photodetector 9 is synchronized with a sampling pulse emitted from the microscope control device 7. It is converted into a digital signal by the A / D converter 10 and given to the microscope control device 7.
[0020]
The microscope control device 7 constructs an observation image of the sample 5 from the received signal and displays it on the monitor 11, and generates light amount data and displays it on the monitor 11 as a graph or a table.
[0021]
On the other hand, the operation means 12 is for the microscope observer to set conditions for acquiring an image (light intensity data).
(A) Laser wavelength irradiated to the sample 5,
(B) Spot diameter of laser beam formed on the sample 5;
(C) Illuminance intensity of laser light applied to sample 5 (d) Time to irradiate one point on sample 5 or spot movement speed of laser light scanned on sample 5 (e) emitted from sample 5 Interval for sampling fluorescence or reflected light signal (f) Scanning range for acquiring an image (light intensity data) on the sample 5;
The conditions such as can be set.
[0022]
Next, processing for such a condition setting will be described.
[0023]
First, when the laser wavelength irradiated to the sample 5 and the spot diameter of the laser beam formed on the sample 5 are set, the spot diameter of the laser beam on the sample 5 here is used as the numerical aperture (NA) of the objective lens 4. The objective lens 4 to be used is determined from the laser wavelength and the spot diameter. That is, by setting the laser wavelength irradiated to the sample 5 and the spot diameter of the laser beam formed on the sample 5 in the operation means 12, the objective lens 4 suitable for the laser wavelength is determined by the microscope control device 7. In this case, the determined objective lens 4 is automatically inserted on the observation optical axis by the revolver 4a, or the type of the objective lens 4 to be switched on the optical path is displayed on the monitor 11 to the observer. Notification is made and the revolver 4a is manually inserted on the observation optical axis.
[0024]
Next, when the irradiation intensity of the laser light applied to the sample 5 is set, based on this setting, the microscope control device 7 inserts / removes the ND filter (not shown) of the laser power control means 2 into the optical path or AOTF (acoustic optical). The irradiation intensity of the laser beam can be adjusted by the variable wavelength filter.
[0025]
Up to this point, the spot diameter of the laser beam formed on the sample 5 and the irradiation intensity of the laser beam are determined, and the irradiation intensity of the laser beam irradiated to one point on the sample 5 can be set.
[0026]
Next, when the time of the laser beam irradiated to one point on the sample is set, the moving speed of the laser beam spot on the sample 5 is determined. In this case, the time during which the laser beam is applied to one point on the sample 5 corresponds to the time during which the laser beam spot moves by the diameter of the spot, and therefore, from the value of the spot diameter and the time to irradiate the laser beam. Can be sought. Instead of setting the time for irradiating one point on the sample 5, the moving speed of the spot of the laser beam may be set directly to obtain the time for irradiating one point on the sample 5. When the moving speed at which the laser light spot moves on the sample 5 is determined, a control signal is output from the microscope control device 7 to the scanning means 6 such as a galvanometer, and the laser light spot is converted to the sample 5. Control that stops at a certain point above or moves at a certain speed is possible.
[0027]
Next, when the interval at which the fluorescence or reflected light emitted from the sample 5 is sampled is set, the degree of density (resolution) at which the fluorescence or reflected light emitted from the sample 5 is detected by this sampling interval. It is determined. For example, when the distance between the points to be sampled on the sample 5 is set, the sampling time interval is determined from the set distance and the moving speed at which the laser light spot moves. By controlling the frequency of the sampling pulse emitted from the control device 7 to the A / D converter 10, the resolution of the acquired image is determined. In this case, the frequency of the sampling pulse emitted from the microscope control device 7 to the A / D converter 10 may be controlled by directly setting a time interval for detecting fluorescence or reflected light.
[0028]
And when the range which acquires the image (light intensity data) on the sample 5 is set, it is set while controlling the phase relationship with the scanning control signal issued to the scanning means 6 by the microscope control device 7 according to this setting. Sampling pulses are output to the A / D converter 10 by the number of samplings, and only light intensity data in the set scanning range is acquired.
[0029]
Thereafter, under the image acquisition conditions set as described above, the laser beam from the laser light source 1 is scanned in the X and Y axis directions by the scanning unit 6 with respect to the sample 5 through the objective lens 4 as described above. Irradiation is performed, and the observation light from the sample 5 is detected by the photodetector 9 via the objective lens 4. In this case, the information acquired from the photodetector 9 includes the spot diameter of the laser beam on the sample 5, the irradiation intensity of the laser beam, the irradiation time of the laser beam per point, the optical data acquisition pitch, the optical data acquisition range, and the like. By arbitrarily setting, various kinds of data with a resolution corresponding to each condition setting can be acquired. Thereby, control of the stimulus (excitation) applied to the sample 5 and detection of observation light such as fluorescence or reflected light emitted from the sample 5 can be quantitatively performed, and quantitative measurement and reproducible measurement can be performed. It becomes possible.
[0030]
In the above description, all the image acquisition conditions such as the spot diameter of the laser beam on the sample 5, the irradiation intensity of the laser beam, the irradiation time of the laser beam per point, the optical data acquisition pitch, and the optical data acquisition range are all set. Although an example has been described, quantitative measurement and reproducible measurement are possible by setting at least the spot diameter of the laser beam on the sample 5 and the irradiation intensity of the laser beam.
[0031]
(Second embodiment)
FIG. 2 shows a schematic configuration of the second embodiment of the present invention, and the same parts as those in FIG.
[0032]
In this case, a zoom optical system 13 is provided in the optical path between the laser power control means 2 and the dichroic mirror 3. The zoom optical system 13 controls the beam diameter of laser light incident on the objective lens 4 in a minute step or steplessly by the zoom operation, and the effective numerical aperture (NA) of the irradiation light with respect to the numerical aperture (NA) of the objective lens 4. ) To control.
[0033]
In this way, by providing the zoom optical system 13, the numerical aperture (NA) of the objective lens 4 can be narrowed down in a minute step or steplessly.
[0034]
Usually, the numerical aperture (NA) of the objective lens 4 is determined by the type (including the magnification) of the objective lens 4. For example, the spot diameter of the laser beam is set on the sample 5 by the configuration of the first embodiment. Even if it is set, the desired spot diameter of the laser beam cannot always be realized. Therefore, in the second embodiment, the zoom optical system 13 is provided, and the beam diameter of the laser light incident on the objective lens 4 is controlled in a minute step or steplessly, and the numerical aperture (NA) of the objective lens 4 is controlled. The spot diameter of the laser beam on the sample 5 can be finely controlled by controlling the effective numerical aperture (NA) of the irradiation light. Thereby, a freedom degree can be increased more in the irradiation setting of the laser beam on the sample 5. FIG.
[0035]
As a modification of the second embodiment, a similar effect can be obtained by the same control by providing a stop with a variable aperture instead of the zoom optical system 13.
[0036]
(Third embodiment)
FIG. 3 shows a schematic configuration of the third embodiment of the present invention, and the same parts as those in FIG.
[0037]
In this case, laser power monitoring means 14 is provided in the optical path between the scanning means 6 and the objective lens 4. This laser power monitor means 14 is composed of a beam splitter 141, a diaphragm 142 and a photodetector 143. Of these, the beam splitter 141 is inserted into the optical path of the laser light incident on the objective lens 4 from the laser light source 1, and this A part of the laser light introduced to the objective lens 4 is folded by the beam splitter 141 and received by the photodetector 143 through the diaphragm 142. The light intensity detected by the photodetector 143 is converted into an electric signal and transmitted to the microscope control device 7. The aperture of the diaphragm 142 can be controlled by the microscope control device 7. In this case, the aperture diameter of the diaphragm 142 is controlled in accordance with the pupil diameter of the objective lens 4 in order to monitor the irradiation intensity of the laser light emitted from the objective lens 4 more accurately. That is, if the beam diameter of the laser light is larger than the pupil diameter of the objective lens 4, a portion of the laser light that is larger than the pupil diameter of the objective lens 4 is cut, but this corresponds to the size of the pupil diameter of the objective lens 4. By providing the diaphragm 142 to be used, it is possible to accurately monitor the amount of light applied to the sample 5 even when the objective lens 4 is switched.
[0038]
In this way, the laser power monitoring means 14 is provided to monitor the irradiation intensity of the laser light actually emitted from the objective lens 4 and to feed back the monitoring result. If the laser power control means 2 is controlled in accordance with the comparison result of the intensity of the laser beam, and even if the irradiation intensity of the laser beam emitted from the laser light source 1 fluctuates, the sample 5 is irradiated. It is possible to always match the intensity of the laser beam to the set value. Thereby, since the irradiation intensity of the laser beam irradiated onto the sample 5 can be controlled more accurately, more accurate quantitative measurement and reproducible measurement can be performed.
[0039]
The laser power monitoring means 14 may be always inserted into or removed from the optical path between the scanning means 6 and the objective lens 4. For example, the laser power monitoring means 14 may be a laser emitted from the laser light source 1. When the intensity of light irradiation is low or when the intensity of fluorescent light or reflected light emitted from the sample 5 is weak, it is necessary to monitor from the optical path except when it is necessary to monitor the intensity of the laser light to prevent the attenuation. Just remove it.
[0040]
In addition, it is possible to add the zoom optical system described in the second embodiment to the configuration of the third embodiment, and in this way, the second embodiment described above. The effect can also be expected.
[0041]
(Fourth embodiment)
By the way, the scanning laser microscope uses a method of observing the form of the sample mainly as an image, as well as visually observing the fluorescence image of the sample, and gives a stimulus to the sample. There are methods such as measuring changes such as temporal changes in fluorescence intensity.
[0042]
Therefore, in the fourth embodiment, in consideration of various utilization methods, two setting modes are prepared for the condition setting method, and these setting modes can be selectively switched. Here, an image observation mode based on a conventional condition setting method is used as the first setting mode, and a measurement mode based on the condition setting methods described in the above-described embodiments is used as the second setting mode.
[0043]
In addition, this 4th Embodiment employ | adopted the structure in any one of the 1st thru | or 3rd embodiment mentioned above, and uses drawing of these 1st thru | or 3rd embodiment here. It shall be.
[0044]
First, in the case of the image observation mode in which the form of the sample 5 in the first setting mode is observed as an image, the selection of the objective lens 4 and the display image size are performed as the conventional condition setting, that is, the condition setting for acquiring the image. Setting of (number of pixels), setting of scanning speed for monitor display (several steps), setting of irradiation intensity of laser light (relative value), etc. are set. Then, based on these condition settings, the scanning speed and deflection angle of the galvano scanner that scans the laser beam in the X and Y axis directions, the fluorescence emitted from the sample 5, the sampling frequency for detecting the reflected light, etc. are determined. The observation image of the sample 5 obtained as is displayed on the monitor 11.
[0045]
On the other hand, in the measurement mode in which a stimulus is applied to the sample 5 and a change caused by the stimulus is measured, the condition setting described in the first to third embodiments, that is, the irradiation of the laser beam applied to the sample 5 is performed. Setting the intensity, setting the spot diameter of the laser beam formed on the sample 5, setting the moving intensity of the spot of the laser beam scanned on the sample 5, sampling the fluorescence or reflected light signal emitted from the sample 5 The setting of the interval, the setting of the range for acquiring the image (light intensity data) on the sample 5 are set. Then, various data with resolution based on these condition settings are acquired, and quantitative measurement and reproducible measurement can be performed.
[0046]
Accordingly, by preparing two setting modes in this way, it is possible to set conditions suitable for various applications, and to realize desired image observation and light intensity measurement.
[0047]
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.
[0048]
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.
[0049]
Embodiments of the present invention also include the following inventions.
[0050]
(1) A plurality of objective lenses having different magnifications, an objective lens switching means for selectively inserting one of the plurality of objective lenses into the observation optical axis, a laser light source, and an intensity of laser light emitted from the laser light source Intensity adjusting means for adjusting the laser beam, scanning means for two-dimensionally scanning the laser beam on the sample, a pinhole disposed at a position conjugate with the focal plane of the objective lens inserted in the observation optical axis, and the pin A photodetector that photoelectrically converts the observation light from the sample that has passed through the hole, an A / D converter that converts an electrical signal output from the photodetector into a digital signal, and an operation unit that receives image acquisition conditions of the sample And the objective lens switching unit, the intensity adjusting unit, the scanning unit, and the A / D converter based on the image acquisition condition input to the operation unit A scanning laser microscope comprising: a microscope control device that constructs an observation image of the sample from the digital signal; and a monitor that displays an image constructed by the microscope control device,
The image acquisition conditions input to the operation means include an image observation mode for setting a display image size and a scanning speed for monitor display, an irradiation spot diameter, an irradiation intensity, and a scanning of the laser light irradiated on the sample. A scanning laser microscope characterized in that it can be set by switching a range, a time during which the laser beam is irradiated to one point of the sample, and a measurement mode for setting a sampling interval of the observation beam.
[0051]
In this way, by preparing two setting modes and selectively applying them, it is possible to set conditions suitable for various applications, and as a result, desired image observation and light intensity measurement can be performed. .
[0052]
【The invention's effect】
As described above, according to the present invention, it is possible to provide a scanning laser microscope that can actively acquire data at a desired resolution, quantitative data acquisition, reproducible data acquisition, and the like.
[Brief description of the drawings]
FIG. 1 is a diagram showing a schematic configuration of a first embodiment of the present invention.
FIG. 2 is a diagram showing a schematic configuration of a second embodiment of the present invention.
FIG. 3 is a diagram showing a schematic configuration of a third embodiment of the present invention.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Laser light source 2 ... Laser power control means 3 ... Dichroic mirror 4 ... Objective lens 4a ... Revolver 5 ... Sample 6 ... Scanning means 7 ... Microscope control device 8 ... Confocal pinhole 9 ... Photo detector 10 ... A / D converter 11 ... Monitor 12 ... Operation means 13 ... Zoom optical system 14 ... Laser power monitoring means 141 ... Beam splitter 142 ... Aperture 143 ... Photo detector

Claims (5)

  1. A plurality of objective lenses with different magnifications;
    Objective lens switching means for selectively inserting one of the plurality of objective lenses into the observation optical axis;
    A laser light source;
    Intensity adjusting means for adjusting the intensity of the laser light emitted from the laser light source;
    Scanning means for two-dimensionally scanning the laser beam on the sample;
    A pinhole disposed at a position conjugate with the focal plane of the objective lens inserted in the observation optical axis;
    A photodetector that photoelectrically converts observation light from the sample that has passed through the pinhole;
    An A / D converter that converts an electrical signal output from the photodetector into a digital signal;
    An operation means for inputting conditions for obtaining an image of the sample;
    The objective lens switching unit, the intensity adjusting unit, the scanning unit, and the A / D converter are controlled based on the image acquisition condition input to the operation unit, and an observation image of the sample is constructed from the digital signal. A microscope control device;
    A monitor for displaying an image constructed by the microscope control device;
    In a scanning laser microscope equipped with
    The scanning laser microscope characterized in that the image acquisition conditions input to the operation means are an irradiation spot diameter and an irradiation intensity of the laser light applied to the sample.
  2. The image acquisition condition input to the operation means further includes at least one of a scanning range of the laser light, a time during which the laser light is applied to one point of the sample, and a sampling interval of the observation light. The scanning laser microscope according to claim 1.
  3. A zoom optical system for adjusting a beam diameter of the laser light incident on the objective lens;
    The scanning laser microscope according to claim 1, wherein the microscope control device controls the zoom optical system so as to satisfy a condition of an irradiation spot diameter of the laser light input to the operation unit.
  4. Further comprising laser power monitoring means for detecting the intensity of the laser beam,
    The microscope control apparatus controls the intensity adjusting unit so as to satisfy a condition of irradiation intensity of the laser light input to the operation unit based on a detection result of the laser power monitoring unit. Item 3. A scanning laser microscope according to Item 1 or 2.
  5. The microscope control device includes:
    Controlling the objective lens switching means so as to satisfy the condition of the irradiation spot diameter of the laser beam input to the operation means,
    Controlling the intensity adjusting means so as to satisfy the condition of the irradiation intensity of the laser beam input to the operating means,
    Controlling the scanning width of the scanning means so as to satisfy the conditions of the scanning range of the irradiation spot of the laser beam input to the operating means,
    Controlling the scanning speed of the scanning means so as to satisfy the condition of the irradiation time of one point on the sample of the laser light input to the operating means,
    3. The scanning laser microscope according to claim 2, wherein the sampling interval of the A / D converter is controlled so as to satisfy the condition of the resolution of the image input to the operation means.
JP2001392524A 2001-12-25 2001-12-25 Scanning laser microscope Expired - Fee Related JP3861000B2 (en)

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Publication number Priority date Publication date Assignee Title
JP2005148497A (en) * 2003-11-17 2005-06-09 Olympus Corp Scanning type laser microscope system
JP4689975B2 (en) * 2004-06-10 2011-06-01 オリンパス株式会社 Microscope illumination intensity measuring device
EP1610088B1 (en) * 2004-06-22 2007-01-03 Polytec GmbH Device for optically measuring an object
JP4954615B2 (en) * 2005-06-13 2012-06-20 オリンパス株式会社 Scanning laser microscope equipment
JP4873917B2 (en) 2005-09-28 2012-02-08 オリンパス株式会社 Scanning laser microscope equipment
JP2008032524A (en) * 2006-07-28 2008-02-14 National Institute Of Advanced Industrial & Technology Laser beam machining device, and focal point detection method of laser light for measurement
EP1918752A1 (en) * 2006-11-06 2008-05-07 Institut Curie Method and apparatus for measuring optical power of a light beam produced in a microscope
JP5075602B2 (en) * 2007-12-04 2012-11-21 オリンパス株式会社 Microscope equipment
JP5725294B2 (en) * 2011-07-15 2015-05-27 横河電機株式会社 Laser microscope
JP5842652B2 (en) * 2012-02-08 2016-01-13 株式会社島津製作所 Tunable monochromatic light source
DE102012015214A1 (en) * 2012-08-03 2014-02-06 DüRR DENTAL AG Focus adjustment of the scanning laser in the device
JP6210754B2 (en) * 2013-06-24 2017-10-11 オリンパス株式会社 Scanning optical microscope
WO2015087629A1 (en) * 2013-12-10 2015-06-18 ソニー株式会社 Image acquisition device and image acquisition method
JP6363890B2 (en) 2014-07-04 2018-07-25 オリンパス株式会社 Scanning microscope apparatus and super-resolution image generation method

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