JP4507596B2 - Laser modulation confocal microscope system - Google Patents

Description

  The present invention relates to a confocal microscope system using laser light, and more particularly to a laser-modulated confocal microscope system capable of modulating the brightness of laser light.

Conventionally, as laser modulation confocal microscope systems capable of modulating the brightness of laser light, for example, those disclosed in Japanese Patent Application Laid-Open Nos. 9-329750 and 2000-35400 are known.
In the laser modulation confocal microscope system disclosed in these publications, the luminance of laser light is modulated by luminance modulation means such as an acousto-optic modulator (AOM).
Japanese Patent Laid-Open No. 9-329750 JP 2000-35400 A

However, no system has been developed that can reliably set the amount of laser light to be irradiated on the sample on the monitor screen. Laser modulation that can reliably set the amount of light on the monitor screen There was a need for a confocal microscope system.
The present invention has been made in view of such conventional circumstances, and provides a laser modulation confocal microscope system capable of reliably setting the amount of laser light to be irradiated on a sample on a monitor screen. With the goal.

The laser modulation confocal microscope system according to claim 1 is a laser beam generation unit that generates laser beam, a luminance modulation unit that modulates the luminance of the laser beam, and a laser beam that is luminance-modulated by the luminance modulation unit. Scanning means for scanning; computing means for generating an image of a sample scanned by the scanning means; display means for displaying an image of the sample generated by the computing means on a screen; and irradiation of laser light to be irradiated Input means for externally inputting the region and the amount of irradiation light to the calculation means, and the calculation means is determined based on the input of the input means when displaying the image of the sample on the display means A display showing the irradiation light amount and a display showing the irradiation region of the laser light determined based on the input of the input means in the image of the sample. Wherein the display by a display mode of the irradiation area of the laser beam based on the irradiation light amount at.

The laser modulation confocal microscope system according to claim 2 is the laser modulation confocal microscope system according to claim 1, wherein the calculation means displays a display showing the irradiation light quantity after the irradiation light quantity is determined by the input means. It is characterized by terminating .
The laser modulation confocal microscope system according to claim 3 is the laser modulation confocal microscope system according to claim 1 or 2, wherein the calculation means is a type of laser light irradiated on the sample from the laser light generation means. And the irradiation area is displayed on the display means with a color corresponding to the wavelength of the laser beam.

The laser modulation confocal microscope system according to claim 4 is the laser modulation confocal microscope system according to claim 1 or claim 2, wherein the calculation means sets the irradiation light amount of the laser light to be irradiated to the sample to the irradiation light amount. It is characterized in that the display means displays with the brightness of the wavelength color of the corresponding laser light.
The laser modulation confocal microscope system according to claim 5 is the laser modulation confocal microscope system according to claim 1 or 2, wherein the calculation means sets the irradiation light amount of the laser light to be irradiated to the sample to the irradiation light amount. The display unit displays the corresponding point density.

The laser modulation confocal microscope system according to claim 6 is the laser modulation confocal microscope system according to claim 1 or 2, wherein the calculation means sets the irradiation light amount of the laser light to be irradiated to the sample to the irradiation light amount. Correspondingly, the display means displays the image with a predetermined setting color.
The laser modulation confocal microscope system according to claim 7 is the laser modulation confocal microscope system according to claim 6, wherein the calculation means causes the display means to display a color of a frame line representing the irradiation region in the set color. It is characterized by that.

The laser modulation confocal microscope system according to claim 8 is the laser modulation confocal microscope system according to claim 1 or 2, wherein the calculation means includes a plurality of types of laser light to be irradiated on the sample, and the same irradiation region. In this case, the overlapping portion is displayed on the display means with a mixed color of the wavelength colors of the laser light.
The laser-modulated confocal microscope system according to claim 9 is the laser-modulated confocal microscope system according to claim 1 or 2, wherein the calculation unit is configured to numerically indicate an irradiation light amount of the laser light to be irradiated on the display unit It is characterized by being displayed .

  In the present invention, the laser light irradiation area and the amount of light to be irradiated on a part of the sample input from the input means are displayed on the sample image displayed on the screen corresponding to the sample image. Since the display is performed, it is possible to reliably set the irradiation light amount of the laser light to be irradiated onto the sample on the monitor screen.

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.
(First embodiment)
FIG. 1 shows a first embodiment of the laser modulation confocal microscope system of the present invention.
In FIG. 1, a laser beam generator 11 emits linearly polarized laser beam. The beam expander 13 expands the beam diameter of the laser light. The laser beam having an enlarged beam diameter passes straight through the polarization beam splitter 15 and reaches the quarter-wave plate 17. The quarter wavelength plate 17 converts this laser light into circularly polarized light and emits it to the scanning unit 19.

The scanning unit 19 scans the laser light in a two-dimensional direction using a biaxial mirror driving mechanism or the like. The objective lens 21 collects this scanning light at an observation point on the sample 25 via the relay lens 23.
The scanning light reflected by the sample 25 becomes reverse circularly polarized light. The reversely circularly polarized light traces back through the objective lens 21 and the scanning unit 19 to reach the quarter wavelength plate 17.

The quarter-wave plate 17 converts this circularly polarized light into linearly polarized light whose polarization direction is orthogonal to that at the time of illumination and emits it to the polarization beam splitter 15. The polarization beam splitter 15 reflects this linearly polarized light. The condensing lens 27 condenses the reflected linearly polarized light in the pinhole 29 a of the light shielding plate 29. The photoelectric detection element 31 receives the light passing through the pinhole 29a.
The A / D converter 33 converts the luminance output of the photoelectric detection element 31 (for example, photomultiplier (PMT)) into a digital signal.

The computing means 35 comprising a microcomputer takes in the digital signal from the A / D converter 33 with a sampling clock synchronized with the scanning operation of the scanning unit 9 and generates an image of the sample 25. This image is displayed on the display means 37 comprising a display such as a liquid crystal. Input means 39 for inputting various information to the outside is connected to the calculation means 35.
In this embodiment, the laser light generation means 11 includes a first laser light generator 41, a second laser light generator 43, and a third laser light generator 45.

  The first laser light generator 41 has a laser light source 41A and luminance modulation means 41B. The laser light source 41A generates red visible light having a wavelength of 633 nm. The luminance modulation means 41B is composed of an acousto-optic modulator (AOM or AOTF) and modulates the luminance and / or spectrum of the laser light from the laser light source 41A. The laser light whose luminance is modulated by the luminance modulation means 41B is incident on the beam expander 13 via the shutter 41C and the dichroic mirror 41D.

  The second laser light generator 43 has a laser light source 43A and luminance modulation means 43B. The laser light source 43A generates green visible light having a wavelength of 543 nm. The luminance modulation means 43B comprises an acousto-optic modulator (AOM or AOTF), and modulates the luminance and / or spectrum of the laser light from the laser light source 43A. The laser light whose luminance is modulated by the luminance modulation means 43B is incident on the beam expander 13 through the shutter 43C and the dichroic mirrors 43D and 41D.

The third laser light generator 45 has a laser light source 45A. The laser light source 45A generates blue visible light having a wavelength of 405 nm. Laser light generated by the laser light source 45A is incident on the beam expander 13 via the shutter 45C and dichroic mirrors 45D, 43D, and 41D.
FIG. 2 shows details of the calculation means 35, display means 37 and input means 39.

The calculation means 35 is composed of a microcomputer and performs various calculations. The display means 37 is composed of a display such as a liquid crystal and displays various information on the screen. The input means 39 is composed of a keyboard, and various information can be externally input.
On the left side of the display unit 37 on the screen 47, a reference image display unit 51 that displays an image 49 of the sample 25 (hereinafter referred to as a reference image) generated by the calculation unit 35 can be displayed. On the reference image 49 displayed on the reference image display unit 51, as will be described later, an irradiation area of the laser beam to be irradiated on a part of the sample 25 input from the input means 39 and the amount of irradiation light. The display can be made in correspondence with the reference image 49 via the calculation means 35.

Further, on the right side of the screen 47, the amount of light for adjusting the amount of light irradiated to the sample 25 by controlling the luminance modulation means 41B, 43B of the first laser light generator 41 or the second laser light generator 43. The adjustment unit 53 is displayed.
The light amount adjustment unit 53 includes a first slider bar 55 indicated by a symbol A and a second slider bar 57 indicated by a symbol B.

  The first slider bar 55 adjusts the amount of irradiation light from the first laser light generator 41, and the amount of irradiation light is changed by moving the slider 55A displayed on the first slider bar 55. . In this embodiment, by clicking the slider 55A with the mouse 59 and moving the slider 55A, the irradiation light amount from the first laser light generator 41 can be changed within the range of 0 to 100% of the maximum irradiation light amount. ing.

  The second slider bar 57 adjusts the amount of light emitted from the second laser light generator 43, and the amount of irradiated light is changed by moving the slider 57A displayed on the second slider bar 57. Is done. In this embodiment, by clicking the slider 57A with the mouse 59 and moving the slider 57A, the irradiation light amount from the second laser light generator 43 can be changed within the range of 0 to 100% of the maximum irradiation light amount. ing.

  On the right side of the first slider bar 55 and the second slider bar 57, check boxes 55B and 57B are displayed. When the check boxes 55B and 57B are clicked with the mouse 59, check marks are displayed in the clicked check boxes 55B and 57B, and the laser beam generators 41 and 43 on which the check marks are displayed are selected. For example, the second laser light generator 43 is selected by clicking the check box 57B on the right side of the second slider bar 57 indicated by the symbol B.

FIG. 3 is a flowchart showing the setting operation of the irradiation region and the irradiation light amount in the laser modulation confocal microscope system described above.
First, in step S1, mounting information on the laser light generators 41, 43, and 45 is detected from the laser light generating means 11. That is, when at least one laser light generator 41, 43, 45 is attached to the laser light generating means 11, the attachment information is detected.

Next, in step S2, it is determined whether or not the mounting information is detected. If the mounting information is not detected, the operation is terminated because the laser beam cannot be irradiated.
On the other hand, when the mounting information is detected, in step S3, the wavelength of the laser light of each of the mounted laser light generators 41, 43, 45 and the presence / absence of the luminance modulation means 41B, 43B are detected.

  Next, in step S4, it is determined whether or not at least one laser light generator 41, 43, 45 is provided with luminance modulation means 41B, 43B. If at least one of the laser light generators 41, 43, 45 includes the luminance modulation means 41B, 43B, the reference image 49 is displayed on the image display unit 51 as shown in FIG. Is done. Further, in this embodiment, since the first laser light generator 41 and the second laser light generator 43 are provided with the luminance modulation means 41B and 43B, at this time, the light amount adjustment unit 53 shown in FIG. The first slider bar 55 and the second slider bar 57 are displayed.

Next, in step S6, as shown in FIG. 4, the irradiation area 61 is set on the image of the reference image 49. This setting is performed by moving the cursor onto the reference image 49 with the mouse 59 and clicking the mouse 59 at the set position. The irradiation area 61 is displayed by a frame line 63, and the inside of the frame line 63 becomes the irradiation area 61.
Next, in step S7, the laser light generators 41 and 43 to be used are selected. This selection is performed by clicking the check boxes 55B and 57B displayed on the right side of the first slider bar 55 and the second slider bar 57 shown in FIG. By clicking the check box 55B on the right side of the first slider bar 55 indicated by the symbol A, the first laser light generator 41 is selected. Further, by clicking the check box 57B on the right side of the second slider bar 57 indicated by the symbol B, the second laser light generator 43 is selected.

  Next, in step S8, the irradiation light quantity irradiated to the irradiation area 61 is set. This setting is performed by clicking and moving the slider 55A displayed on the first slider bar 55 with the mouse 59 when the first laser light generator 41 is selected in step S7. When the second laser light generator 43 is selected in step S7, the slider 57A displayed on the second slider bar 57 is clicked with the mouse 59 and moved.

  Next, in step S9, the irradiation light quantity irradiated to the irradiation area 61 is displayed. As shown in FIG. 5, this display is performed by covering the irradiation region 61 with a color corresponding to the wavelength of the laser beam and a brightness corresponding to the amount of irradiation light. That is, the brightness of the color is set in advance corresponding to the amount of irradiation light. In this embodiment, when the amount of irradiation light is small, it is dark as shown in FIG. 5, and as the amount of irradiation light increases, the brightness is set to increase as shown in FIG.

For example, when the first laser light generator 41 is selected in step S7, the irradiation area 61 is displayed in red, which is the wavelength color of the laser light of the first laser light generator 41, and is set. When the amount of irradiated light is small, it is displayed in dark red, and as the amount of irradiated light increases, it is displayed in bright red.
On the other hand, when the second laser light generator 43 is selected in step S7, the irradiation area 61 is displayed in green, which is the wavelength color of the laser light of the second laser light generator 43, and is set. When the amount of irradiated light is small, it is displayed in dark green, and as the amount of irradiated light increases, it is displayed in bright green.

Next, in step S10, it is determined whether or not the adjustment of the irradiation light amount has been completed. The determination of completion is made by clicking the completion button 65 displayed on the screen 47 of the display means 37 shown in FIG. 2 with the mouse 59 or operating a predetermined key of the input means 39.
When the adjustment of the irradiation light amount is completed in step S10, the display of the irradiation light amount covering the irradiation region 61 is erased from the screen 47 in step S11, and the setting is completed. In this state, only the frame line 63 indicating the irradiation region 61 is displayed on the image of the reference image 49, and the reference image 49 in the frame line 63 is visible.

  In the laser modulation confocal microscope system described above, the reference image 49 displayed on the screen 47 shows the irradiation region 61 and the irradiation light amount of the laser light to be irradiated on a part of the sample 25 input from the input means 39. In addition, since the display is made in correspondence with the reference image 49, the irradiation light amount of the laser light to be irradiated on the sample 25 can be reliably set on the screen 47 of the display means 37 which is a monitor.

That is, since the laser light irradiation area 61 and the irradiation light amount are displayed on the reference image 49 displayed on the screen 47 in correspondence with the reference image 49, the irradiation area 61 and the irradiation light amount are intuitively visually observed. Thus, it is possible to effectively prevent erroneous setting of the irradiation region 61 and the amount of irradiation light.
Further, in the laser modulation confocal microscope system described above, the display of the irradiation light amount displayed on the reference image 49 corresponding to the reference image 49 is erased from the screen 47 after setting the irradiation light amount. It is possible to effectively prevent the recognition of the reference image 49 from being disturbed by the light amount display.

Further, in the laser modulation confocal microscope system described above, the type of laser light irradiated on the sample 25 is input from the laser light generating means 11, and the irradiation area 61 is covered and displayed in a color corresponding to the wavelength of the laser light. Since it did in this way, the relationship with the laser beam used can be recognized easily and reliably.
Further, in the laser modulation confocal microscope system described above, the amount of laser light to be irradiated on the sample 25 is displayed by the intensity of the wavelength of the laser light corresponding to the amount of irradiation. The relationship with light and the amount of irradiation light can be easily and reliably recognized.
(Second Embodiment)
FIG. 7 shows a main part of a second embodiment of the laser modulation confocal microscope system of the present invention.

In this embodiment, the configuration other than the configuration relating to the display of the amount of irradiation light is the same as that of the first embodiment. Therefore, the same elements as those of the first embodiment are denoted by the same reference numerals and detailed. Description is omitted.
In this embodiment, the display of the irradiation light amount irradiated to the irradiation region 61 in step S9 shown in FIG. 3 is displayed with the color corresponding to the wavelength of the laser light and the density of points corresponding to the irradiation light amount. That is, the density of points is set in advance corresponding to the amount of irradiation light. In this embodiment, when the irradiation light quantity is small, the density of the point P is small as shown in FIG. 7, and as the irradiation light quantity increases, the density of the point P increases as shown in FIG.

  For example, when the first laser light generator 41 is selected in step S7 shown in FIG. 3, the point of the irradiation area 61 in red, which is the wavelength color of the laser light of the first laser light generator 41. P is displayed. When the set irradiation light quantity is small, the density of the points P displayed in red in the frame line 63 is displayed small, and as the irradiation light quantity increases, the density is displayed in red in the frame line 63. The density of the point P is displayed large.

  On the other hand, when the second laser light generator 43 is selected in step S7, the point P of the irradiation area 61 is displayed in green, which is the wavelength color of the laser light of the second laser light generator 43. . When the set irradiation light quantity is small, the density of the points P displayed in green in the frame line 63 is reduced, and displayed in green in the frame line 63 as the irradiation light quantity increases. The density of the point P is displayed large.

In the laser modulation confocal microscope system of this embodiment, substantially the same effect as that of the first embodiment can be obtained, but in this embodiment, the irradiation light amount of the laser light to be irradiated on the sample 25 is changed to the irradiation light amount. Since the corresponding point density is displayed, it is possible to easily and reliably recognize the irradiation light quantity.
(Third embodiment)
FIG. 9 shows an essential part of a third embodiment of the laser modulation confocal microscope system of the present invention.

In this embodiment, the configuration other than the configuration relating to the display of the amount of irradiation light is the same as that of the first embodiment. Therefore, the same elements as those of the first embodiment are denoted by the same reference numerals and detailed. Description is omitted.
In this embodiment, the display of the irradiation light amount irradiated to the irradiation region 61 in step S9 shown in FIG. 3 is displayed in a preset color determined in advance corresponding to the irradiation light amount. That is, a display color is set in advance corresponding to the amount of irradiation light. In this embodiment, the color of the frame 63 of the irradiation area 61 is set to red when the amount of irradiation light is small, green when it is intermediate, and blue when it is large.

In the laser modulation confocal microscope system of this embodiment, substantially the same effect as that of the first embodiment can be obtained, but in this embodiment, the irradiation light amount of the laser light to be irradiated on the sample 25 is changed to the irradiation light amount. Correspondingly, the display is made with a preset color, so that the irradiation light quantity can be easily and reliably recognized.
(Fourth embodiment)
FIG. 10 shows a main part of a fourth embodiment of the laser modulation confocal microscope system of the present invention.

In this embodiment, the configuration other than the configuration relating to the display of the amount of irradiation light is the same as that of the first embodiment. Therefore, the same elements as those of the first embodiment are denoted by the same reference numerals and detailed. Description is omitted.
In this embodiment, when there are a plurality of types of laser light to be irradiated on the sample 25 and the same irradiation region is displayed, the overlapping portion is displayed in a mixed color of the wavelength colors of the laser light. .

  That is, in this embodiment, the first laser light generator 41 and the second laser light generator 43 can be selected simultaneously. When the first laser light generator 41 and the second laser light generator 43 are simultaneously selected, the irradiation area 61A of the first laser light generator 41 is displayed in red, and the irradiation of the second laser light generator 43 is performed. The region 61B is displayed in green. A portion 61C where the irradiation region 61A of the first laser light generator 41 and the irradiation region 61B of the second laser light generator 43 overlap is displayed in red-green, which is a mixed color of red and green. .

In this embodiment, the irradiation light quantity of the laser beam to be irradiated is displayed numerically corresponding to the irradiation areas 61A and 61B.
That is, in this embodiment, the lead line 65A is drawn out from the irradiation region 61A of the first laser light generator 41, and the irradiation light amount of the laser light to be irradiated is, for example, 40% near the tip of the lead line 65A. And a numerical value. Further, a leader line 65B is drawn out from the irradiation region 61B of the second laser light generator 43, and the irradiation light amount of the laser light to be irradiated is displayed as a numerical value, for example, 30% in the vicinity of the tip of the leader line 65B. The

  In the laser modulation confocal microscope system of this embodiment, substantially the same effect as that of the first embodiment can be obtained, but in this embodiment, there are a plurality of types of laser light to be irradiated on the sample 25, and the same irradiation region In the case of 61, since the overlapping portion is displayed with a mixed color of the wavelength colors of the laser light, the overlapping portion can be easily and reliably recognized.

In the laser modulation confocal microscope system of this embodiment, the irradiation light quantity of the laser light to be irradiated is displayed numerically, so that the irradiation light quantity can be recognized more accurately.
In the above-described embodiment, an example in which an acousto-optic modulator (AOM) is used as the luminance modulation unit has been described. However, the present invention is not limited to such an embodiment. For example, the electro-optic modulator (EOD) ), Luminance modulation means such as a magneto-optic modulator (MOD) may be used.

It is explanatory drawing which shows 1st Embodiment of the laser modulation confocal microscope system of this invention. It is explanatory drawing which shows the detail of the display means 37 of FIG. It is explanatory drawing which shows operation | movement of the laser modulation confocal microscope system of FIG. It is explanatory drawing which shows the state which set the irradiation area | region 61 on the reference | standard image 49 in the 1st Embodiment of this invention. It is explanatory drawing which shows the state which displayed the irradiation light quantity in the irradiation area | region 61 of FIG. It is explanatory drawing which shows the state which increased the irradiation light quantity in FIG. It is explanatory drawing which shows the state which displayed the irradiation light quantity in the irradiation area | region 61 in the 2nd Embodiment of this invention. It is explanatory drawing which shows the state which increased the irradiation light quantity in FIG. It is explanatory drawing which shows the state which displayed the irradiation light quantity in the irradiation area | region 61 in the 3rd Embodiment of this invention. It is explanatory drawing which shows the state with which the irradiation area | region 61 overlapped in the 4th Embodiment of this invention.

Explanation of symbols

11 Laser light generation means 35 Calculation means 37 Display means 39 Input means 41 First laser light generator 43 Second laser light generator 45 Third laser light generators 41B and 43B Brightness modulation means 47 Screens 61, 61A, 61B Irradiation area

Claims (9)

Laser light generating means for generating laser light;
Luminance modulation means for performing luminance modulation of the laser beam;
Scanning means for scanning a sample with laser light whose luminance is modulated by the luminance modulation means;
A calculation means for generating an image of the sample scanned by the scanning means;
Display means for displaying an image of the sample generated by the computing means on a screen;
An input means for externally inputting the irradiation area of the laser beam to be irradiated and the irradiation light amount to the calculation means;
With
When the display unit displays the sample image on the display unit , the calculation unit displays the irradiation light amount determined based on the input of the input unit, and the input of the input unit in the sample image And displaying the laser light irradiation area determined based on the display, and displaying the laser light irradiation area in the sample image in a display mode based on the irradiation light quantity. Laser modulation confocal microscope system. The laser-modulated confocal microscope system according to claim 1,
The calculation means terminates the display showing the irradiation light quantity after the irradiation light quantity is determined by the input means . The laser-modulated confocal microscope system according to claim 1 or 2,
The arithmetic means inputs a type of laser light irradiated on the sample from the laser light generating means, and displays the irradiation area on the display means in a color corresponding to the wavelength of the laser light. Laser modulation confocal microscope system. The laser-modulated confocal microscope system according to claim 1 or 2,
The laser modulation confocal microscope system characterized in that the calculation means displays the irradiation light amount of the laser light to be irradiated on the sample on the display means with the brightness of the wavelength color of the laser light corresponding to the irradiation light amount. The laser-modulated confocal microscope system according to claim 1 or 2,
The said calculating means displays the irradiation light quantity of the laser beam which should be irradiated to the said sample on the said display means with the point density corresponding to the irradiation light quantity, The laser modulation confocal microscope system characterized by the above-mentioned. The laser-modulated confocal microscope system according to claim 1 or 2,
The laser modulation confocal microscope system, wherein the calculation means displays the irradiation light amount of the laser light to be irradiated on the sample on the display means with a preset color corresponding to the irradiation light amount. The laser-modulated confocal microscope system according to claim 6,
The laser modulation confocal microscope system characterized in that the calculation means causes the display means to display the color of a frame line representing the irradiation area in the set color. The laser-modulated confocal microscope system according to claim 1 or 2,
The arithmetic unit displays a plurality of types of laser light to be irradiated on the sample and displays the overlapping portion on the display unit with a mixed color of the wavelength colors of the laser light when the same irradiation region is applied. Laser modulation confocal microscope system. The laser-modulated confocal microscope system according to claim 1 or 2,
The said calculating means displays the irradiation light quantity of the said laser beam which should be irradiated on the said display means with a numerical value, The laser modulation confocal microscope system characterized by the above-mentioned.

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