JPH10282426A - Laser microscope - Google Patents

Abstract

PROBLEM TO BE SOLVED: To optimize the respective light quantities of plural laser beams without requiring complex operation, and to optimize the output balance of the plural laser beams, and also to provide an inexpensive laser microscope. SOLUTION: As to this laser microscope provided with at least one light source 10 emitting the plural laser beams having mutually different wavelength, a ROM 41 storing data related to the optimum light quantities of the laser beams, ND filters 12 and 21 capable of individually adjusting the respective light quantities of the plural laser beams, shutters 23 and 32 detecting the light quantity of at least one laser beam among the plural laser beams, a dichroic mirror 31, a plane parallel glass 33, and an SPD 34; the CPU 40 controls the filters 12 and 21 so that the respective light quantities of the plural laser beams become the optimized light quantities stored in the ROM 41 based on a detection signal obtained from the SPD 34.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a laser microscope, and more particularly, to a laser microscope capable of always performing observation with an appropriate amount of light.

[0002]

2. Description of the Related Art FIG. 3 is a block diagram illustrating a method for adjusting a laser beam of a conventional laser microscope.

The laser microscope includes a laser light source 110 for emitting a plurality of laser beams having different wavelengths, and an excitation filter 111 disposed in front of the laser light source 110.
A total reflection mirror 112 for reflecting laser light emitted from the laser light source 110, and a silicon photodiode (hereinafter referred to as SPD) 1 for detecting the amount of laser light.
13, a parallel plane glass 114 for guiding a laser beam to the SPD 113, and an ND filter 1 arranged on an optical path L between the parallel plane glass 114 and the total reflection mirror 112.
15, the ND filter 115 and the plane-parallel glass 11
Shutter 116 that can be opened and closed on the optical path L with the shutter 4
And an amplifier 117 for amplifying a light receiving output of the SPD 113
A / D which digitizes the output by A / D conversion
D converter 118 and CPU 140 for performing processing such as arithmetic
And a light amount monitor 143 for displaying a result processed by the CPU 140 and the like.

FIG. 4 is a block diagram illustrating a method for adjusting the laser light of another conventional laser microscope. The same reference numerals are given to the same parts as those of the laser microscope of FIG.

The laser microscope includes a laser light source 110 for emitting a plurality of laser beams having different wavelengths, and an excitation filter 111 disposed in front of the laser light source 110.
A laser light source 120 that emits laser light having a wavelength different from that of the laser light source 110, a total reflection mirror 121 that reflects the laser light emitted from the laser light source 120, and a laser light that reflects the laser light emitted from the laser light source 110. A dichroic mirror 122 that transmits the laser light emitted from the laser light source 120, an ND filter 123 that can be inserted into the optical path between the excitation filter 111 and the dichroic mirror 122 as shown by an arrow a, and a laser. SPD 113 for detecting the amount of light, parallel plane glass 114 for guiding laser light to SPD 113, ND filter 115 arranged on the optical path between parallel plane glass 114 and dichroic mirror 122, and ND filter 115 A glass plate that is openably and closably disposed on the optical path between , An amplifier 117 for amplifying the received light output of the SPD 113, an A / D converter 118 for digitizing the output by A / D conversion, a CPU 140 for performing processing such as computation, and a result processed by the CPU 140. And a light quantity monitor 143 for displaying

[0006] In any of the above laser microscopes, S
The amount of light detected by the PD 113 is displayed on the monitor 143,
Dimming is performed by adjusting the light amount with a fixed or continuously variable ND filter 115 arranged in the optical path based on this light amount.

[0007]

However, in the case of the laser microscope shown in FIG. 3, the ND filter 115 must be operated every time the laser light is changed based on the value displayed on the light amount monitor 143. Dimming was troublesome and troublesome.

In the case of performing multiple excitation using a plurality of light sources as in the laser microscope of FIG. 3, since the dimming is performed using one ND filter 115 and one light amount monitor 143, each laser light source is used. Excitation lights having different wavelengths emitted from 110 and 120 are modulated by the same ND filter 115.

Therefore, in the case of a combination of laser beams whose emission outputs are considerably different, for example, the discrete ND filter 123 is placed on the optical path of a laser light source having a large emission output according to the difference between the emission outputs of the respective laser beams. It is necessary to insert and balance the output of each laser beam.

[0010] Further, the laser light source 1
When the outputs of the filters 10 and 120 decrease, another ND filter 123 having a different light transmission intensity is used so that an appropriate amount of light can be obtained.

Therefore, when multiple excitation is performed using a plurality of laser light sources, dimming of the laser light is also troublesome and troublesome.

On the other hand, for each laser light source, the SPD 113
And a light amount monitor 143 can be provided to cope with this.
The optical path becomes complicated, and the microscope becomes large and expensive.

SUMMARY OF THE INVENTION The present invention has been made in view of such circumstances, and a problem thereof is that it is possible to set each of a plurality of laser beams to an appropriate light amount without requiring a complicated operation, and to reduce the number of laser beams. An object of the present invention is to provide an inexpensive laser microscope that can make the output balance appropriate.

[0014]

According to a first aspect of the present invention, there is provided a laser microscope, comprising: at least one light source for emitting a plurality of laser beams having different wavelengths; Storage means for storing data; adjusting means for individually adjusting the light amounts of the plurality of laser lights; detecting means for detecting the light amount of at least one of the plurality of laser lights; Control means for controlling the adjusting means based on the detection signal obtained from the means so that the light quantity of each of the plurality of laser beams becomes an appropriate light quantity stored in the storage means.

The adjusting means is controlled based on the detection signal obtained from the detecting means so that the respective light amounts of the plurality of laser beams become the appropriate light amounts stored in the storage means.
When multiple excitation of a plurality of laser beams is performed, each laser beam can be set to an appropriate light amount without a complicated operation, and the output balance of the plurality of laser beams can be adjusted appropriately.

According to a second aspect of the present invention, there is provided the laser microscope according to the first aspect, wherein a plurality of divided optical paths through which the plurality of laser beams pass independently, and all of the laser beams in the plurality of divided optical paths are combined. A light path excluding all or one of the plurality of divided light paths, and a light detecting means for detecting the amount of laser light passing through the main light path in the main light path. The container is arranged.

Since the light amount of the laser beam passing through the main optical path is detected by one detector, the optical path can be simplified. Therefore, the size and cost of the microscope can be reduced.

[0018]

Embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 is a block diagram illustrating a method for adjusting a laser beam of a laser microscope according to a first embodiment of the present invention.

The laser microscope 1 has a laser light source 10
, Excitation filter 11, and ND filter (adjustment means) 1
2, laser light source 20, ND filter (adjustment means) 2
1, a total reflection mirror 22, a shutter 23, a dichroic mirror 31, a shutter 32, a plane parallel glass 33, and a silicon photodiode (light receiving means) 34
(Hereinafter referred to as “SPD”), an amplifier 35, an A / D converter 36, a CPU (control means) 40, a ROM (storage means) 41, a RAM 42, and a light amount monitor 43. The shutter 23, the dichroic mirror 31, the shutter 32, the plane parallel glass 33, and the SPD 34 constitute a detecting unit.

As the laser light source 10, an Ar laser having the largest output among visible light lasers and capable of oscillating laser light of a plurality of wavelengths centering on blue and green is used. Here, an air-cooled Ar laser that emits laser light having two wavelengths of 488 nm and 514 nm is used.

The excitation filter 11 is arranged on a disk-shaped excitation filter wheel 11A. By rotating the excitation filter wheel 11A by a motor 13, the excitation filter 11 can be positioned on the divided optical path L1.

The excitation filter wheel 11A is made of a light shielding material, and transmits only light passing through the excitation filter 11. The driving of the motor 13 is controlled by the CPU 40. The excitation filter 11 has a property of transmitting only light having a wavelength λ of 488 nm and 514 nm, for example.

The ND filter 12 is for attenuating transmitted light. As the ND filter 12, a variable transmittance filter that can change the transmittance by changing the light transmission position using a motor 14 is used.

A He—Ne laser is used as the laser light source 20 to emit light having a wavelength λ of 543.5 nm.

The ND filter 21 has the same configuration as the ND filter 12, and the light transmission position of the ND filter 21 is made variable using a motor 24.

The motors 14 and 24 are driven by the CPU 4
It is controlled by a command from 0.

The shutter 23 is arranged in the divided optical path L2 of the He-Ne laser, and is opened and closed by a motor 25.

The shutter 32 has an Ar laser and He
It is arranged in the main optical path L3 of the -Ne laser and is opened and closed by the motor 37.

The driving of each of the motor 25 and the motor 37 is controlled by a command from the CPU 40.

The total reflection mirror 22 reflects the He-Ne laser emitted from the laser light source 20, and the dichroic mirror 31 reflects the He-Ne laser emitted from the laser light source 20.
The laser transmits the laser and reflects the Ar laser emitted from the laser light source 10.

The parallel flat glass 33 is disposed in the main optical path L3, and reflects the laser light.

The SPD 34 receives the laser beam reflected by the parallel plane glass 33, converts the laser beam into an electric signal corresponding to the amount of received light, and outputs the electric signal.

The amplifier 35 amplifies this electric signal, and A / A
The D converter 36 converts an output signal of the amplifier 35 into a digital signal.

The ROM 41 stores data on the light amount balance of a plurality of laser lights, data on the appropriate light amounts a and b of the individual laser lights for the sample, and the like.

The CPU 40 comprises an A / D converter 36, a ROM 4
1 and the RAM 42 to perform arithmetic processing and the like described later.

Next, the operation of the laser microscope when adjusting the light quantity will be described.

First, before acquiring an image, the shutter 32
Is closed to block the main optical path L3, and the zero point adjustment of the SPD 34 is performed.

Next, for example, the excitation filter wheel 11A
Is rotated to close the excitation filter 11, the shutter 23 and the shutter 32 are opened, respectively, and the SPD 34 receives only the He-Ne laser light emitted from the laser light source 20 and passing through the split optical path L <b> 2 and the main optical path L <b> 3.

At this time, the motor 24 is driven to move the ND filter 21 so that the light amount becomes the appropriate light amount a, and the light of the laser light source 20 is adjusted. The appropriate light amount a may be input by an operator from an input device (not shown),
It may be called from past experimental data registered in M41 or a database (not shown).

The light quantity a after the light control is stored in the RAM 42 as the output of the He-Ne laser light.

Next, the excitation filter wheel 11A is rotated to insert the excitation filter 11 on the optical path, and the Ar laser light emitted from the light source 10 also splits the divided optical path L1 and the main optical path L1.
3 so that the light can be received by the SPD 34.

Therefore, the SPD 34 detects a light quantity c obtained by combining the He-Ne laser light and the Ar laser light.

In the CPU 40, the light amount a of the He-Ne laser is subtracted from the light amount c received by the SPD 34 to obtain the light amount of the Ar laser, and the ND filter 12 is moved by driving the motor 14 so that the appropriate light amount b is obtained. RA
It is stored in M42 and displayed on the light amount monitor 43.

Thereafter, when it is desired to change the light amount, the ND filters are driven one by one, and the light amount corresponding to the driven ND filter is calculated from the increase and decrease of the light amount c each time, and the calculation result is stored in the RAM 42. Light intensity monitor 43
To be displayed.

According to this embodiment, one SPD 34
And a single light amount monitor 43 can automatically adjust the dimming of a plurality of laser lights. Therefore, even when the laser outputs are greatly different, the laser light can be adjusted to the appropriate light amount without requiring complicated operations such as replacement of an ND filter. And,
The output balance of the laser beam can be made appropriate, the optical path can be simplified, and the microscope can be reduced in size and inexpensive.

The light control is not limited to the above method. First, the shutter 23 is closed to perform light control of the laser light source 10, and then the excitation filter 11 is closed and the shutter 23 is opened to open the laser light source 20. May be performed.

Although the first embodiment has been described in connection with the case where there are two laser light sources, the present invention can be applied to a case where there are three or more light sources. Also, the control means is not the CPU 40,
A microcomputer including a ROM 41 and a RAM 42 may be used.

FIG. 2 is a block diagram illustrating a method for adjusting the laser light of a laser microscope according to a second embodiment of the present invention. The same parts as those of the laser microscope of FIG. Is omitted.

The laser microscope 50 includes a laser light source 60
, An excitation filter 61, a dichroic mirror 62,
63, ND filters (adjustment means) 64, 65, shutters 66, 67, total reflection mirrors 68, 69, shutter 32, parallel flat glass 33, SPD (light receiving means)
34, an amplifier 35, an A / D converter 36, and a CPU
(Control means) 40, ROM (storage means) 41, RA
M42 and a light amount monitor 43 are provided. Shutter 66,
67, dichroic mirrors 62 and 63, shutter 32, plane-parallel glass 33, and SPD 34 constitute detection means.

The laser light source 60 can emit laser light having a plurality of wavelengths. As the laser light source 60, for example, a krypton / argon ion laser that emits laser light having wavelengths 488 nm, 568 nm, and 647 nm and peaks in light amount is used.

The excitation filter 61 is controlled by the CPU 40 and has a characteristic of transmitting only light having a wavelength λ of 488 nm and 568 nm, for example.

The dichroic mirrors 62 and 63 have a wavelength λ.
= 488 nm laser light, wavelength λ = 568 nm
Is transmitted.

The ND filters 64 and 65 are intended to attenuate transmitted light. These ND filters 64 and 6
As 5, a variable transmittance that can change the transmittance by changing the light transmission position using the motors 64 </ b> A and 65 </ b> A is used.

The motors 64A and 65A are driven by CP
It is controlled by a command from U40.

The shutter 66, together with the ND filter 64, is disposed in the divided optical path L5 through which the laser light having the wavelength λ = 568 nm passes, and is opened and closed by a motor 66A.

Further, the shutter 67, together with the ND filter 65, has a split optical path L through which a laser beam having a wavelength λ = 488 nm passes.
4 and is opened and closed by a motor 67A.

The motors 66A and 67A are driven by CP
It is controlled by a command from U40.

The total reflection mirrors 68 and 69 are disposed in the split optical path L4, and guide the laser light having a wavelength λ = 488 nm reflected by the dichroic mirror 62 to the dichroic mirror 63 via the ND filter 65 and the shutter 67.

Next, the operation of the laser microscope when adjusting the light quantity will be described.

First, before acquiring an image, the shutter 32
To close the main optical path L6, and the SPD (light receiving means) 34
Adjust the zero point of.

Next, for example, the shutter 66 is closed, the shutter 67 and the shutter 32 are opened, and only the laser beam having a wavelength λ = 488 nm passing through the divided optical path L4 and the main optical path L6 is received by the SPD 34.

At this time, the motor 65A is driven to move the ND filter 65 so that the light amount becomes the appropriate light amount a, and light control is performed. The appropriate amount of light a may be input by an operator from an input device (not shown), or may be called from past experimental data registered in the ROM 41 or a database (not shown).

The light amount a after the light adjustment is stored in the RAM 42 as the light amount of the laser light having the wavelength λ = 488 nm.

Next, the shutter 66 is opened, and the wavelength λ = 56.
The SPD 34 can also receive 8 nm laser light.

Therefore, in the SPD 34, the ND filter 6
The light amount c obtained by combining the laser light having the wavelength λ = 488 nm adjusted in 5 and the laser light having the wavelength λ = 568 nm adjusted by the ND filter 64 is detected.

The CPU 40 obtains the light quantity of the laser light having the wavelength λ = 568 nm by subtracting the light quantity a of the laser light having the wavelength λ = 488 nm from the light quantity c received by the SPD 34, and drives the motor 64 A so that the appropriate light quantity b is obtained. Then, the ND filter 64 is moved, and the appropriate light amount b is stored in the RAM 42 and displayed on the light amount monitor 43.

Thereafter, when it is desired to change the light amount, the ND filters are driven one by one, and each time, the light amount corresponding to the driven ND filter is calculated from the increase and decrease of the light amount c, and the calculation result is stored in the RAM 42. Light intensity monitor 43
To be displayed.

According to the second embodiment, the same effects as in the first embodiment can be exhibited.

In the second embodiment, the dimming method is not limited to the above method. First, the shutter 67 is closed and the shutter 66 is opened to set the wavelength λ.
= 568 nm laser light, then shutter 6
6 and the shutter 67 is opened, and the wavelength λ = 4
The dimming of the 88 nm laser light may be performed.

In the second embodiment, the description has been given of the case of laser light of two wavelengths, but the present invention can be similarly applied to the case of three or more wavelengths. The control means is a CPU 4
A microcomputer including a ROM 41 and a RAM 42 instead of 0 may be the same as in the first embodiment.

[0072]

As described above, according to the laser microscope of the first aspect of the present invention, when performing multiple excitation of a plurality of laser beams, each laser beam can be adjusted to an appropriate amount without complicated operation. And the output balance of the plurality of laser beams can be made appropriate. Therefore, observation can always be performed with an appropriate amount of light.

According to the laser microscope of the second aspect of the present invention, since the amount of laser light passing through the main optical path is detected by one detector, the optical path can be simplified. Therefore, the size and cost of the microscope can be reduced.

[Brief description of the drawings]

FIG. 1 is a block diagram illustrating a method for adjusting a laser beam of a laser microscope according to a first embodiment of the present invention.

FIG. 2 is a block diagram illustrating a method for dimming laser light of a laser microscope according to a second embodiment of the present invention.

FIG. 3 is a block diagram illustrating a method for adjusting a laser beam of a conventional laser microscope.

FIG. 4 is a block diagram illustrating a method of adjusting laser light of another conventional laser microscope.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Laser microscope 10, 20 Laser light source 12, 21 ND filter (light quantity adjusting means) 34 SPD (light receiving means) 40 CPU (control means) 41 ROM (storage means) 43 Light quantity monitor (display means)

Claims (2)

[Claims] At least one light source that emits a plurality of laser beams having different wavelengths from each other, a storage unit that stores data on an appropriate amount of the laser beam, and individually adjusts the amount of each of the plurality of laser beams. Possible adjusting means, detecting means for detecting the light amount of at least one of the plurality of laser lights, and each of the plurality of laser lights based on a detection signal obtained from the detecting means. A control means for controlling the adjusting means so that the light quantity becomes the proper light quantity stored in the storage means. 2. A method according to claim 1, further comprising: a plurality of divided light paths through which the plurality of laser light beams independently pass; and a main light path through which all of the laser light beams of the plurality of divided light paths are combined and passed. A light shielding means is arranged on all or one of the optical paths, and one detector for detecting the amount of laser light passing through the main optical path is arranged on the main optical path. The laser microscope according to 1.