JP2625330B2 - Pinhole position control method for confocal optical system and its control device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a confocal optical system such as a laser scanning microscope in which the positions of a light source side pinhole and a detector side pinhole are in a conjugate positional relationship. And technology for automating the position adjustment of the pinhole position on the detector side.

[0002]

2. Description of the Related Art Generally, for example, a laser scanning microscope is well known as a confocal optical system.
FIG. 2 is a block diagram particularly showing a configuration of a conventional laser scanning microscope.

In this laser scanning microscope (FIG. 3), light emitted from a first light source 1 is narrowed down by a light source side pinhole 2, corrected into a parallel light by a lens 3a, and a beam splitter 4 forms an optical path at a right angle. Changed to the direction.
Then, by changing the optical path by the XY scanner 5, the light condensed by the objective lens 3 b is scanned and irradiated on the sample 6.

On the other hand, the reflected light or fluorescent light of the sample 6 is corrected to a parallel light beam by an objective lens 3b, and is condensed by a lens 3c via an XY scanner 5 and a beam splitter 4, and further, is pinhole-detected. Only the central part of the spot light narrowed down at 7 is detected by the detector 8.

Conventionally, the position of each of the pinholes 2 and 7 of the confocal optical system is detected by first placing a plane mirror in place of the sample 6 and actually turning on the first light source 1 to collect the condensing lens 3c. Alternatively, the detector-side pinhole 7 is moved on a plane orthogonal to the optical axis (an XY stage having a precise feed mechanism is often used as a moving method), and detection is performed to obtain the maximum amount of light. The position of the container side pinhole 7 was set as the optimum position.

[0006]

As described above, in the conventional confocal optical system, when an optical element such as a filter is inserted into the optical path, the pinholes on the light source side and the detector side move from the conjugate position. Since the displacement occurs, there is a problem that the pinholes must be manually aligned by placing a plane mirror instead of the sample.

SUMMARY OF THE INVENTION The present invention has been made to solve the above-described problems. In a confocal optical system in which a light source side and a detector side pinholes are located at conjugate positions, the positions of these pinholes are automatically determined. It is an object to provide a method and apparatus for adjusting.

[0008]

According to a first aspect of the present invention, there is provided a pinhole position control method for a confocal optical system, the method comprising: providing a pinhole position between a detector side pinhole and a detector in the confocal optical system; A second light source for irradiating light passing through the pinhole is provided, and light passing through the pinholes on the light source side and the detector side respectively corresponds to the positions of the pinholes on the light source side and the detector side. As a spot light, the light is alternately condensed and taken out from the optical path of the confocal optical system, and by detecting the position shift of the spot light, the position shift of the pinhole on the light source side and the detector side can be detected. Features.

According to a second aspect of the present invention, an apparatus for automatically realizing the pinhole position control method for the confocal optical system is provided between the detector side pinhole and a detector. A second light source for irradiating light passing through the container-side pinhole, and a spot light obtained by condensing and irradiating light emitted from the first and second light sources from the confocal optical system, A position detecting device for detecting the positions of the spot lights respectively corresponding to the positions of the light source side and the detector side pinholes, and reflected light from the sample to the detector via the detector side pinholes or the detector side pinholes Or, based on driving means for moving a lens for forming an image of fluorescent light in a direction perpendicular to the optical axis, and position information of spot light corresponding to the pinholes on the light source side and the detector side output from the position detection device. , It is characterized by comprising a control means for moving instructions to the serial driving means.

[0010]

According to the first aspect of the present invention, a second light source indicating the position of the detector-side pinhole is provided between the detector-side pinhole and the detector in the confocal optical system, and the light source side and the second light source are provided. The light that has passed through the respective pinholes on the detector side is alternately condensed and extracted from the optical path of the confocal optical system as spot light corresponding to the positions of the light source side and the detector side pinholes. Since the position deviation of the spot light is detected by the position detection device, the positions of the pinholes on the light source side and the detector side can be confirmed at the same position (one position detector).

According to the second aspect of the present invention, the pinhole position can be automatically controlled by providing detection means (second light source and position detector), control means and driving means for performing these operations. become.

[0012]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS One embodiment of the present invention will be described below with reference to FIGS. In the drawings, the same portions are denoted by the same reference numerals, and description thereof is omitted.

FIG. 1 is a block diagram of a confocal optical system for explaining a pinhole position control method of the confocal optical system according to the first aspect of the present invention, and shows a position of a pinhole 7 on a detector side. In addition, a second light source 9 is provided between the detector 8 and the detector-side pinhole 7, and the light emitted from the second light source 9 is changed in the optical path in the direction of the detector-side pinhole 7. , A lens 3d and a beam splitter 4a.

Light corresponding to the positions of the pinholes 2 and 7 on the light source side and the detector side from the optical path of the confocal optical system is spot light condensed by the lens 3e via the beam splitter 4c. As the position detector 10
It is configured so that it can be detected by D (Position Sensitive Detector).

Light emitted from the first light source 1 passes through the light source side pinhole 2 and is reflected at a certain ratio by the beam splitters 4b and 4c to form a spot light on the PSD 10. On the other hand, the light emitted from the second light source 9 passes through the detector-side pinhole 7 via the beam splitter 4a. The light that has passed through the detector side pinhole 7 passes through the beam splitters 4b and 4c, and similarly passes through the PSD1.
A spot light is formed on 0.

At this time, if the spot lights formed on the PSD 10 coincide with each other, it can be said that the light source side pinhole 2, the detector side pinhole 7, and the spot on the sample 6 are all at conjugate positions.

In this embodiment, the positioning is
The first and second light sources 1 and 9 are alternately turned on, but only one of them is blinked and the light source side pinhole 2 or the detector side pinhole 7 is moved so that the position output from the PSD 10 becomes equal. The same effect can be obtained even if it is performed.

Next, the second aspect of the present invention will be described with reference to FIG.

FIG. 2 is a block diagram showing a configuration of an embodiment of a pinhole position control device for a confocal optical system according to the second aspect of the present invention, which is an example applied to a confocal scanning microscope.

This pinhole position control device, as described above, uses the second light source 9 indicating the position of the detector-side pinhole 7 and the light emitted from the first light source 1 through the confocal optical system. The spot light, which is alternately condensed and extracted by the beam splitter 4c and the lens 3c, is used as the position detector PS.
D detects.

The control means 11 controls the PSD 10
The position shift amount is detected from each of the spot positions detected by the driving means 12 and the driving means 12
And issues a movement instruction to the two-axis stage.

Thus, the two-axis stage 12 is connected to the detector 8
The movable lens 3c for condensing the reflected light from the sample 6 is moved to automatically correct the pinholes 2 and 7 on the light source side and the detector side to conjugate positions.

[0023]

As described above, according to the present invention, between the detector side pinhole and the detector in the confocal optical system,
A second light source indicating the position of the detector-side pinhole is installed, and light passing through the respective light-source-side and detector-side pinholes corresponds to the positions of the light-source-side and detector-side pinholes. As a spot light, the light is alternately condensed and extracted from the optical path of the confocal optical system, and by detecting the position shift of the spot light, the position shift of the pinhole on the light source side and the detector side is detected. Therefore, there is an effect that the conjugate position of the light source side pinhole and the detector side pinhole can be easily adjusted.

Further, as a device for automatically realizing the pinhole position control method of the confocal optical system, a detection means (second light source and position detector), a control means, and a drive means are provided so that each pin is provided. Automation of hole alignment can be realized, and there is an effect that individual differences in position correction do not occur.

[Brief description of the drawings]

FIG. 1 is a configuration diagram of a confocal optical system for explaining a pinhole position control method of the confocal optical system according to the first embodiment of the present invention.

FIG. 2 is a block diagram showing a configuration of an embodiment of a pinhole position control device for a confocal optical system according to the invention of claim 2;

FIG. 3 is a configuration diagram of a laser scan microscope as a conventional confocal optical system.

[Explanation of symbols]

1 ... first light source, 2 ... light source side pinhole, 3a, 3b,
3c, 3d, 3e: lens, 4a, 4b, 4c: beam splitter, 5: XY scanner, 6: sample, 7: detector side pinhole, 8: detector, 9: second light source, 10: P
SD, 11: control means, 12: two-axis stage.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Yuji Nakajima 1126, Nomachi, Ichinomachi, Hamamatsu City, Shizuoka Prefecture Inside (72) Inventor Hirotoshi Terada 1126, Nomachi, Ichinomachi, Hamamatsu City, Shizuoka Prefecture Hamamatsu Photonics Co., Ltd. (72) Inventor Hiroyuki Matsuura 1126 Nomachi, Hamamatsu City, Shizuoka Prefecture 1 Hamamatsu Photonics Co., Ltd. (56) References JP-A-61-140914 (JP, A) JP-A-62-208017 (JP, A) JP-A-4-42117 (JP, A) JP-A-4-130919 (JP, U)

Claims (2)

(57) [Claims] 1. A conjugate light source-side pinhole for narrowing light emitted from a first light source onto a sample and a detector-side pinhole for narrowing reflected light or fluorescence from the sample detected by a detector. In a pinhole position control method of a confocal optical system for setting at a position, a second light source for irradiating light passing through the detector-side pinhole is provided between the detector-side pinhole and a detector. Installed, and light passing through the pinholes on the light source side and the detector side is alternated from the optical path of the confocal optical system as spot light corresponding to the positions of the pinholes on the light source side and the detector side. A pinhole position control method for a confocal optical system, wherein the position shift of the light source side and the detector side pinhole is detected by detecting the position shift of the spot light. 2. A conjugate light source-side pinhole for narrowing down light emitted from a first light source to a sample and a detector-side pinhole for narrowing down reflected light or fluorescence from the sample detected by a detector. In a pinhole position control device of a confocal optical system for installing at a position, a second device is provided between the detector-side pinhole and the detector, and irradiates light passing through the detector-side pinhole. A light source, and a spot light obtained by condensing light emitted from the first and second light sources from the confocal optical system and corresponding to the positions of the light source side and the detector side pinholes, respectively. A position detection device that detects the position of the spot light, and a lens that forms an image of reflected light or fluorescence from a sample on the detector via the detector-side pinhole or the detector-side pinhole, perpendicular to the optical axis. Moved in the direction A drive unit for performing a movement instruction to the drive unit based on each position information of the spot light corresponding to the light source side and the detector side pinhole output from the position detection device. Pinhole position control device for confocal optical system.