JP2004226682A - Scanning type confocal microscope and its optical axis checking method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an optical axis checking method and an optical adjusting method using guide light going against illumination light. <P>SOLUTION: The guide light G going against the illumination light beams L1 to L3 is made incident on the exit end 7b of an optical fiber 7, and the degree in alignment of the optical path of the guide light G emitted from the incident end 7a of the optical fiber 7 with the optical paths of the illumination light beams L1 to L3 made incident on the incident end 7a of the optical fiber 7 after being emitted from laser light sources 1a to 1c is checked. And the installation states of the laser light sources 1a to 1c are adjusted so that the optical path of the guide light G may align with the optical paths of the illumination light beams L1 to L3. <P>COPYRIGHT: (C)2004,JPO&NCIPI

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a scanning confocal microscope, a method for checking an optical axis of a light source of a scanning confocal microscope, and an optical adjusting method.
[0002]
[Prior art]
In a conventional scanning confocal microscope, a specimen is irradiated with laser light (illumination light) from a laser light source, and observation is performed by two-dimensionally scanning the spot-like light irradiated on the specimen. Laser light from the laser light source is transmitted to the microscope side by one optical fiber connecting the laser light source and the microscope. When a sample is irradiated with a laser beam, reflection, absorption, fluorescence, scattering, and the like occur in the irradiation area due to the optical characteristics of the sample. The reflected light and fluorescent light generated in the irradiation area are detected by the detector after being collected by the objective lens. An electric signal from the detector is taken into a processing device such as a microcomputer, and an observation image of a specimen based on the electric signal is formed. This observation image is displayed on a CRT monitor, a liquid crystal display, or the like.
[0003]
When performing fluorescence observation with a scanning confocal microscope, usually, a specimen stained with a plurality of fluorescent reagents is simultaneously irradiated with laser beams of a plurality of wavelengths. Therefore, a plurality of laser light sources are used. Laser beams of a plurality of wavelengths emitted from a plurality of laser light sources are collected at one end of one optical fiber, are incident, and are simultaneously emitted from the other end. At this time, in order to make the laser light incident from one end of one optical fiber, it is necessary to precisely match the emission angle and the emission position of the laser light emitted from the laser light source with respect to the core of the optical fiber.
[0004]
Each tissue of the specimen is stained with a plurality of fluorescent reagents specific to the tissue. The laser light acts as excitation light for generating fluorescence. When the specimen is irradiated with laser light, each tissue stained with the fluorescent reagent emits fluorescence according to the reagent. The generated fluorescence is wavelength-separated using a filter or the like, and fluorescence having a desired wavelength is detected by a detector (for example, see Patent Document 1).
[0005]
[Patent Document 1]
JP-A-2002-221663
[Problems to be solved by the invention]
In a conventional scanning confocal microscope, a single mode fiber is almost entirely used as one optical fiber for guiding a laser beam from a laser light source to a microscope main body. The single mode fiber generally has a small core diameter of 5 μm or less and a small NA of about 0.12. Therefore, it is difficult to accurately match the emission angle and emission position of the laser light with respect to the core of the optical fiber.
[0007]
The present invention provides an optical axis checking method and an optical adjustment method using guide light that goes backward to illumination light, and a scanning confocal microscope that performs optical axis check and optical adjustment using guide light that goes backward to illumination light. Is what you do.
[0008]
[Means for Solving the Problems]
According to the first aspect of the present invention, the illumination light of a predetermined wavelength emitted from the light source is made incident on the entrance end of the optical fiber by the illumination optical system, and the sample is irradiated with the illumination light incident on the microscope main body from the emission end of the optical fiber. Applied to the optical axis checking method of a scanning confocal microscope that illuminates and observes, guide light traveling in the opposite direction to the illumination light enters the optical fiber from the output end, and the guide light exits from the input end of the optical fiber The optical axis state of the light source is checked based on the degree of coincidence between the optical path of the light source and the optical path of the illumination light incident on the optical fiber from the light source.
According to the invention of claim 2, illumination light of a predetermined wavelength emitted from the light source is made incident on the entrance end of the optical fiber by the illumination optical system, and the sample is irradiated with the illumination light incident on the microscope main body from the emission end of the optical fiber. Applied to the optical adjustment method of the scanning confocal microscope for illuminating and observing, the guide light traveling in the opposite direction to the illumination light is made incident on the optical fiber from the output end, and the guide light emitted from the input end of the optical fiber is The optical axis state of at least one of the light source and the illumination optical system is adjusted such that the optical path matches the optical path of the illumination light incident on the optical fiber from the light source.
According to the third aspect of the present invention, the illumination light of a predetermined wavelength emitted from the light source is made incident on the incident end of the optical fiber by the illumination optical system, and the sample is irradiated with the illumination light incident on the microscope main body from the emission end of the optical fiber. It is applied to a scanning confocal microscope for illuminating and observing, is provided at an emission end of an optical fiber, and is capable of connecting an auxiliary light source that causes a guide light, which is counter to the illumination light, to enter the optical fiber, and a connection portion that can be connected to the guide light. And an adjustment mechanism for adjusting at least one of the optical axis state of the light source and the illumination optical system so that the optical paths of the illumination light coincide with each other.
Further, a plurality of light sources each emitting illumination light having a different wavelength may be provided, and each illumination light emitted from the plurality of light sources may be made incident on the incident end of the optical fiber.
[0009]
BEST MODE FOR CARRYING OUT THE INVENTION
Hereinafter, embodiments of the present invention will be described with reference to the drawings. First, the overall configuration of the scanning confocal microscope of the present invention will be described with reference to FIG. FIG. 1 is a schematic diagram showing a schematic configuration of a scanning confocal microscope according to the present invention, and has a configuration basically similar to that of an incident-light fluorescence microscope. The scanning confocal microscope includes a microscope main body 10, a light source unit 1, a scanning unit 12, and a detection unit 26, and a scanning unit 12 is mounted on a lens barrel 11 of the microscope main body 10. The light source unit 1 and the scanning unit 12 are connected by a single mode optical fiber 7. The scanning unit 12 and the detection unit 26 are connected by a multi-mode optical fiber 21.
[0010]
Laser light generated by the light source unit 1 is input to the scanning unit 12 via the optical fiber 7. The laser beam scanned by the scanning unit 12 irradiates the sample 24 via the second objective lens 22 and the objective lens 23. Light from the specimen 24 is transmitted to the detection unit 26 via the objective lens 23, the second objective lens 22, the scanning unit 12, and the optical fiber 21, and is detected. The analog signal detected by the detection unit 26 is converted into a digital signal by an A / D converter provided in the processing device 30 and sent to the CPU for image processing. The observation image obtained by the image processing is displayed on a monitor (not shown).
[0011]
FIG. 2 is an enlarged view of the scanning unit 12 and shows details of the optical scanning unit 19. The laser light transmitted to the scanning unit 12 by the optical fiber 7 (see FIG. 1) is emitted from the end face of the fiber with a predetermined NA, and is converted into parallel light by the collimator lens 15 in the scanning unit 12. The parallel light from the collimator lens 15 is reflected by the dichroic mirror 18 and enters the optical scanning unit 19.
[0012]
The optical scanning unit 19 is provided with a pair of movable total reflection mirrors 191 and 192, and the inclination angle can be changed in conjunction with the total reflection mirrors 191 and 192. By changing the angles of the total reflection mirrors 191 and 192, it becomes possible to two-dimensionally scan the laser light incident on the light scanning unit 19 from the dichroic mirror 18 in two directions orthogonal to the laser light.
[0013]
The laser light emitted from the optical scanning unit 19 is imaged on a primary image plane 21 via a scanning lens 20, and then is imaged on a specimen 24 by a second objective lens 22 and an objective lens 23. The image of the laser beam formed on the specimen 24 is a point image, which is condensed to a size determined by the NA of the objective lens 23. Therefore, phenomena such as reflection, absorption, fluorescence generation, and scattering occur in the irradiation area due to the optical characteristics of the specimen 24 at this point.
[0014]
The fluorescent light and the reflected light from the specimen 24 are imaged on the primary image plane 21 by the objective lens 23 and the second objective lens 22, are converted into parallel light by the scanning lens 20, and enter the optical scanning unit 19. The fluorescence from the sample 24 is always returned to the same optical path as the laser light incident on the reflection mirror 191 of the optical scanning unit 19 from the dichroic mirror 18 by being scanned by the optical scanning unit 19, that is, by being descanned. become.
[0015]
The fluorescence emitted from the light scanning unit 19 to the dichroic mirror 18 passes through the dichroic mirror 18 and enters the total reflection mirror 17. The fluorescence reflected by the total reflection mirror 17 is imaged on the light shielding plate 14 by the condenser lens 16. Since the light-shielding plate 14 and the sample 24 are in a conjugate positional relationship, an image of the fluorescence emission point on the sample 24 is formed at the position of the opening (pinhole) 13 formed in the light-shielding plate 14.
[0016]
Then, only the light that has passed through the opening 13 is transmitted to the detection unit 26 via the optical fiber 21. That is, even if there is light generated from another point on the sample 24, the light is blocked by the light shielding plate 14 and does not reach the detection unit 26. As a result, in the scanning confocal microscope, it is possible to observe a specimen having not only high horizontal resolution but also high vertical resolution.
[0017]
Hereinafter, a method of checking an optical axis to check whether illumination light from a laser light source is accurately incident on an incident end of an optical fiber and an optical adjustment method based on the method will be described in detail. FIG. 3 is an optical path diagram showing an arrangement of a plurality of laser light sources 1a to 1c, an illumination optical system 2, an optical fiber 7, and a guide light emitting unit 8. The laser light source 1a is an air-cooled Ar laser (wavelength 488 nm), the laser light source 1b is a G / He-Ne laser (wavelength 543 nm), and the laser light source 1c is an R / He-Ne laser (wavelength 633 nm). In the optical path of each illumination light (laser light), electric shutters 3a, 3b, 3c, ND filters 4a, 4b, 4c, and dichroic mirrors 5a, 5b, 5c are arranged in order. The electric shutters 3a, 3b, 3c and the ND filters 4a, 4b, 4c are used for dimming laser light, and the dichroic mirrors 5a, 5b, 5c are used for wavelength selection and optical path synthesis.
[0018]
FIG. 4 is a graph showing the spectral characteristics of the dichroic mirror. 4A shows the spectral characteristics of the dichroic mirror 5a, FIG. 4B shows the spectral characteristics of the dichroic mirror 5b, and FIG. 4C shows the spectral characteristics of the dichroic mirror 5c. The reflectance of the dichroic mirror 5a shown in FIG. 4A becomes maximum at the wavelength of 488 nm of the laser light source 1a. It also has a reflectance of about 10% even at a wavelength of 633 nm. Similarly, the dichroic mirror 5b in FIG. 4B has a maximum reflectance at the wavelength of 543 nm of the laser light source 1b, and has a reflectance of about 10% even at the wavelength of 633 nm. The reflectance of the dichroic mirror 5c in FIG. 4C is maximized at the wavelength of 633 nm of the laser light source 1c.
[0019]
Referring again to FIG. 3, the laser light that has been subjected to wavelength selection and optical path synthesis by each of the dichroic mirrors 5 a, 5 b, and 5 c is combined into one optical path, condensed by the condenser lens 6, and incident on the optical fiber 7. The light enters the end 7a. The laser light propagates through the core of the optical fiber 7, radiates to the outside from the emission end 7b, and is guided to the scanning unit 12.
[0020]
When performing the optical axis check according to the present invention, the connection between the optical fiber 7 and the scanning unit 12 is disconnected, and the light emitting unit 8 that emits the guide light is connected to the emission end 7b of the optical fiber via the connection member 9. . Note that, without disconnecting the connection between the optical fiber 7 and the scanning unit 12, the optical path from the emission end 7b of the optical fiber is branched into two, one of which is on the light emitting unit 8 side and the other is on the scanning unit 12 side. It can also be configured to guide.
[0021]
The connection member 9 includes an FC connector 9a attached to the emission end 7b of the optical fiber and a fiber coupler 9b attached to the light emitting unit 8. The FC connector 9a and the fiber coupler 9b may be integrally connected.
[0022]
The guide light G emitted from the light emitting section 8 enters the core of the optical fiber 7 from the emission end 7b via the fiber coupler 9b and the FC connector 9a. Then, the light propagates through the core, radiates from the incident end 7 a to the outside, and is guided to the condenser lens 6. The guide light G is converted into a parallel light flux by the condenser lens 6, enters the dichroic mirrors 5a, 5b, and 5c, is partially reflected in accordance with the wavelength selection characteristics of each dichroic mirror, and travels toward each laser light source. That is, the guide light G travels in the opposite direction to the illumination light L.
[0023]
The optical axis of the guide light G and the illumination light L is checked in the following procedure. In the present embodiment, an R / He-Ne laser (wavelength: 633 nm) is used as the light emitting unit 8. The light emitting section 8 is not limited to the present embodiment as long as it has a wavelength range that reflects light from the dichroic mirror. As described above with reference to the spectral characteristics of FIG. 4, the guide light G having a center wavelength of 633 nm is reflected by any dichroic mirror, and travels toward each laser light source.
[0024]
For example, if the illumination light L1 emitted from the laser light source 1a (wavelength 488 nm) is accurately incident on the incident end 7a of the optical fiber, the illumination light L1 coincides with the optical path of the guide light G. That is, the optical path from the laser light source 1a to the incident end 7a of the optical fiber coincides with both. However, when the illumination light L1 does not accurately enter the incident end 7a of the optical fiber, the optical path of the illumination light L1 does not match the optical path of the guide light G, and the optical path of the illumination light L1 is shifted. It can be said. The same applies to the illumination lights L2 and L3 emitted from the laser light sources 1b and 1c.
[0025]
The degree of coincidence between the optical axis of the illumination light L1 and the optical axis of the guide light G can be checked anywhere in the optical path within the illumination optical system 2. When the beam of the illumination light L1 is inclined with respect to the beam of the guide light G, it can be understood that an error occurs in the angle of the laser light source 1a. If the two beams are separated or overlap and become thick, it can be determined that the laser light source 1a is displaced in the direction perpendicular to the optical axis of the guide light G.
[0026]
The optical adjustment method in the present embodiment is performed according to the following procedure. In FIG. 3, when the illumination light L1 is correctly incident on the incident end 7a of the optical fiber when the guide light G is irradiated, the optical path of the illumination light L1 and the optical path of the guide light G coincide with each other. The beam diameter is the same at any of the locations as in the case of only the illumination light L1. If it is checked that there is an error in the angle of the laser light source 1a, the optical axis of the illumination light L1 is inclined with respect to the optical axis of the guide light G, so that the optical axis directions of the two coincide. Then, both or one of the mounting angle of the laser light source 1a and the angle of the dichroic mirror 5a is adjusted.
[0027]
On the other hand, when it is checked that the laser light source 1a is displaced in the vertical direction with respect to the optical axis of the guide light G, the diameter of the beam in which the beam of the guide light G and the beam of the illumination light L1 overlap each other is equal to the illumination light. It becomes thicker than when only L1 is used. Therefore, the position of the laser light source 1a is adjusted by moving the position of the laser light source 1a in the direction perpendicular to the optical axis of the guide light G so that the diameter of the overlapped beam is reduced.
[0028]
The laser light source 1a is held by a pair of adjusting members 39a also serving as a holder. The adjustment member 39a has a mechanism for adjusting the mounting position of the laser light source 1a and the emission direction of the illumination light L1. Similar adjustment members 39b and 39c are provided for the other laser light sources 1b and 1c.
[0029]
An adjusting member 29a is provided to move the dichroic mirror 5a to adjust the angle of the optical axis of the illumination light L1. The adjustment member 29a is a rotation knob that tilts the dichroic mirror 5a. Similar adjustment members 29b and 29c are provided for the other laser light sources 1b and 1c.
[0030]
FIG. 5 is a diagram illustrating an example of a holding / adjusting mechanism of the laser light sources 1a to 1c. The laser light source 1a is fixed to the stage 41 by a jig 40. The stage 41 is mounted on the second stage 42. The stage 41 can rotate around an axis 43 with respect to the stage 42, and by rotating the stage 41, the angle of the stage 41 with respect to the stage 42 can be changed. Normally, the stage 41 is fixed to the stage 42 by tightening the bolts 44, but when adjusting the angle of the stage 41, the bolts 44 are loosened to change the angle. The second stage 42 is attached to the base (not shown) of the light source unit 1 so as to be movable in the left-right direction.
[0031]
Feed screws 45 to 48 for adjusting the positions of the stages 41 and 42 are provided at the base of the light source unit 1. The tips of the feed screws 45 and 46 are in contact with the upper side surface of the stage 42, and the ends of the feed screws 47 and 48 are in contact with the lower side surface of the stage 41. When the feed screws 45 to 48 are fed to the right by the same feed amount, the stages 41 and 42 move integrally to the right, and when they are sent to the left by the same feed amount, the stages 41 and 42 move integrally to the left.
[0032]
When the bolts 44 are loosened and the feed screws 47 and 48 are fed to the right by the same feed amount, only the stage 41 tilts to the left, and the optical path of the illumination light L1 also tilts to the left. When the feed screws 47 and 48 are fed to the left by the same feed amount, only the stage 41 tilts to the right, and the optical path of the illumination light L1 also tilts to the right. The springs 49 and 50 are springs that urge the upper part of the stage 41 and the lower part of the stage 42 to the right in the drawing.
[0033]
《Optical axis adjustment method》
Next, a method of adjusting the optical axis of the laser light source 1a using the holding / adjusting mechanism of FIG. 5 will be described with reference to FIGS. FIG. 6A is a diagram showing a state of the laser light source 1a before the adjustment. The guide light G is reflected by the dichroic mirror 5a and is applied to the vicinity of the exit of the laser light source 1a, so that the operator can observe the bright spot. As shown in FIG. 6A, the exit of the laser light source 1a is shifted to the left from the observed bright spot, and the optical path of the illumination light L is inclined to the right with respect to the guide light G. I understand.
[0034]
First, the feed screws 45, 47 are sent out to the right by the same amount so that the emission port of the laser light source 1a and the luminescent spot of the guide light G coincide with each other, and the stages 41, 42 are integrally moved by the urging forces of the springs 49, 50. Move to the right. When the emission port and the luminescent spot coincide with each other, the feed screws 46 and 48 are sent out to the right and positioned. As a result, the optical path of the illumination light L is as shown in FIG.
[0035]
Next, the bolt 44 is loosened and the feed screw 47 is fed to the right, and the inclination of the stage 41 is adjusted to correct the inclination of the optical path of the illumination light L in the direction of arrow Q. If the optical path of the illumination light L and the optical path of the guide light G match as shown in FIG. 6C due to the inclination correction, the feed screw 48 is sent to the right to position the stage 41, and then the bolt 44 is tightened. The stage 41 is fixed to the stage 42. In FIG. 6 (c), the optical path of the illumination light L and the optical path of the guide light G are shifted from each other for easy viewing, but they actually match and look like a single line.
[0036]
As described above, the adjustment mechanism shown in FIG. 5 can individually adjust the position shift and the angle shift of the laser light source 1a. Although FIG. 5 illustrates a case in which two-dimensional adjustment is performed in a plane parallel to the paper surface, three-dimensional adjustment can be performed by further adding a similar mechanism. The optical adjustment is performed in the same manner for the illumination lights L2 and L3 emitted from the laser light sources 1b and 1c.
[0037]
Also, as shown in FIGS. 7A and 7B, the positional deviation between the luminescent spot and the emission port is adjusted by adjusting the angle of the dichroic mirror 5a, and only the angle adjustment is performed by the adjustment mechanism on the laser light source 1a side. It may be performed. FIG. 7A is a diagram showing a state before adjustment, and is a diagram similar to FIG. 6A. As shown in FIG. 7B, the angle of the dichroic mirror 5a is changed in the R direction, the light path direction of the guide light G from the dichroic mirror 5a to the laser light source 1a is changed from G1 to G2, and the bright spot of the guide light G is changed. And the emission port of the laser light source 1a. Thereafter, the inclination of the laser light source 1a may be adjusted so that the optical path of the illumination light L and the optical path G2 of the guide light G match.
[0038]
Here, other optical axis checking methods and optical adjustment methods will be briefly described for comparison. FIG. 8 is an optical path diagram for explaining another optical axis checking method and optical adjustment method. The arrangement of the plurality of laser light sources 1a to 1c, the illumination optical system 2, and the optical fiber 7 is the same as that of the present embodiment shown in FIG.
[0039]
In FIG. 8, when checking whether or not the illumination light L1 emitted from the laser light source 1a (wavelength 488 nm) is accurately incident on the incident end 7a of the optical fiber, for example, a pinhole is provided near the reflection point of the dichroic mirror 5a. The target plate 31 is placed, and a target plate 32 with a pinhole is also placed near the center of the condenser lens 6. The diameter of the pinhole is about 0.1 mm.
[0040]
If the optical path of the illumination light L1 is shifted, a part or all of the spot of the illumination light L1 is generated on the target plate, so that the optical axis can be checked. That is, the optical axis check is performed using a pinhole. Similarly, when performing optical adjustment, the angle shift and the position shift of the laser light source 1a are adjusted so that the illumination light L1 passes through the two pinholes by using a point called a pinhole.
[0041]
When the adjustment is performed using the two pinholes 31 and 32 as shown in FIG. 8, for example, the position and angle of the laser light source 1a are adjusted so that the beam passes through the pinhole 31 closer to the laser light source 1a. However, the beam passing through the pinhole 31 does not always pass through the other pinhole 32. Therefore, it is necessary to adjust the position and angle of the laser light source 1a so that the beam passes through both the pinholes 31 and 32 at the same time, which is difficult and complicated. In addition, since the adjustment is performed manually, the diameters of the pinholes 31 and 32 must be considerably large to about 0.1 mm with respect to the core diameter of the optical fiber 7, and the beam is accurately aligned with the core. It was difficult.
[0042]
On the other hand, in the present embodiment, since the bright spot due to the irradiation of the guide light G is formed near the exit of the laser light source 1a, the position of the laser light source 1a is shifted so that the exit of the laser light source 1a coincides with this bright spot. Can be adjusted. After the adjustment, the angle of the laser light source 1a may be adjusted so that the optical axes of both beams coincide. Therefore, the optical adjustment of the laser light source 1a can be easily and accurately performed. Whether or not the optical paths of the illumination light L and the guide light G coincide with each other can be easily confirmed by observing the thickness and direction of the beam. In particular, the beam color of the guide light G is determined by the illumination light L1. If the beam color is different from the above, it is possible to easily distinguish the coincidence / non-coincidence of the beams.
[0043]
In the above-described embodiment, the case where three types of laser light sources are provided in the light source unit 1 has been described as an example. However, the illumination light is transmitted through the illumination optical system (dichroic mirror or lens) into the core of the optical fiber. As long as the light is incident on the light source, any number of light sources may be provided, or only one light source may be provided.
[0044]
In the embodiment described above, for example, the auxiliary light source is provided by the light emitting unit 8, the connection unit is provided by the connection member 9, the adjustment members 39a, 39b, 39c, 29a, 29b, 29c, the stages 41, 42, the bolt 44, the feed screw. An adjusting mechanism is implemented by 45 to 48 and springs 49 and 50, respectively. However, the present invention is not limited to the above embodiments at all, as long as the features of the present invention are not impaired.
[0045]
【The invention's effect】
As described above, according to the present invention, it is possible to easily and accurately check the optical axis state of the light source by comparing the optical path of the illumination light entering the optical fiber from the light source with the optical path of the guide light. . In addition, by adjusting the optical axis state of the light source such that the optical path of the illumination light matches the optical path of the guide light, accurate optical adjustment can be easily performed.
[Brief description of the drawings]
FIG. 1 is a schematic diagram showing a schematic configuration of a scanning confocal microscope according to the present invention.
FIG. 2 is an optical path diagram showing a configuration of an optical system of a scanning confocal microscope according to the present invention.
FIG. 3 is a diagram illustrating an optical axis check method and an optical adjustment method according to the present invention.
FIG. 4 is a graph showing spectral characteristics of dichroic mirrors 5a, 5b, 5c.
FIG. 5 is a diagram illustrating an example of a holding / adjusting mechanism of the laser light sources 1a to 1c.
FIG. 6 is a diagram for explaining an optical axis adjustment method, and (a) to (c) show respective procedures.
FIGS. 7A and 7B are diagrams illustrating an adjustment procedure in the case where adjustment is performed by changing the angle of the dichroic mirror 5a, wherein FIG. 7A illustrates a state before the angle adjustment and FIG. 7B illustrates a state after the angle adjustment.
FIG. 8 is a diagram illustrating a comparative example of an optical adjustment method.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 Light source part 1a-1c Laser light source 2 Illumination optical system 5a, 5b, 5c Dichroic mirror 7 Optical fiber 7a Incident end 7b Emitting end 8 Light emitting part 9 Connecting member 10 Microscope main body 12 Scanning unit 24 Sample 26 Detection unit 29a, 29b, 29c , 39a, 39b, 39c Adjusting members 41, 42 Stage 44 Bolts 45 to 48 Feed screws 49, 50 Spring L, L1 to L3 Illumination light G, G1, G2 Guide light

Claims (4)

A scan in which illumination light of a predetermined wavelength emitted from a light source is made incident on an incident end of an optical fiber by an illumination optical system, and a sample is illuminated and observed by illumination light made incident on a microscope main body from the emission end of the optical fiber. In the method of checking the optical axis of a confocal microscope,
A guide light traveling in the opposite direction to the illumination light is made incident on the optical fiber from the emission end, and an optical path of the guide light emitted from the incidence end of the optical fiber and an optical path of the illumination light incident on the optical fiber from the light source Checking the optical axis state of the light source according to the degree of coincidence with the optical axis of the scanning confocal microscope. A scan in which illumination light of a predetermined wavelength emitted from a light source is made incident on an incident end of an optical fiber by an illumination optical system, and a sample is illuminated and observed by illumination light made incident on a microscope main body from the emission end of the optical fiber. In the optical adjustment method of the confocal microscope,
A guide light traveling in the opposite direction to the illumination light is made incident on the optical fiber from the emission end, and an optical path of the guide light emitted from the incidence end of the optical fiber and an optical path of the illumination light incident on the optical fiber from the light source An optical axis state of at least one of the light source and the illumination optical system is adjusted so that? A scan in which illumination light of a predetermined wavelength emitted from a light source is made incident on an incident end of an optical fiber by an illumination optical system, and a sample is illuminated and observed by illumination light made incident on a microscope main body from the emission end of the optical fiber. In a confocal microscope,
A connection portion that is provided at an emission end of the optical fiber and that can be connected to an auxiliary light source that causes guide light that is opposite to the illumination light to enter the optical fiber.
A scanning confocal microscope, comprising: an adjusting mechanism that adjusts at least one of an optical axis state of the light source and the illumination optical system so that an optical path of the guide light coincides with an optical path of the illumination light. The scanning confocal microscope according to claim 3,
Equipped with a plurality of light sources that emit illumination light of different wavelengths,
A scanning confocal microscope, wherein the illumination optical system causes each illumination light emitted from the plurality of light sources to enter an incident end of the optical fiber.

Images