JP2001242082A - Biological sample optical scanning device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a biological sample optical scanning device greatly shortening an optical scanning time on a sample chip for efficient analysis and having excellent fluorescence detection sensitiveness of fluorescent material used for labeling the biological sample. SOLUTION: A sample chip 15 carrying multiple biological samples is optically scanned by means of the biological sample optical scanning device 1 for identifying a biological sample labeled with the fluorescent material. A sample base 13 is reciprocated. Above the sample base, light transmitted from a light source and passed through a hollow part in an electric motor is led out in the radial direction and in the vertical direction by means of a pair of reflecting mirrors opposed mutually at an angle of 45 deg. so as to be radiated onto the sample chip on the sample base. Fluorescence from the fluorescent material excited by the light radiated from an objective lens is received by means of a light receiving member via the hollow part of a rotor to output an electric signal.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a biological sample optical scanning apparatus for detecting a biological sample fluorescently labeled on a microchip or a DNA chip.

[0002]

For example, when analyzing the gene expression state (mutation such as gene expression amount and gene deletion) in cells and tissues, for example, thousands to tens of thousands of cells are laid on a 1 cm ² glass substrate. A microchip or a DNA chip (hereinafter, referred to as a DNA chip) in which DNA probes and RNA probes (hereinafter, referred to as DNA probes) are dot-arranged at a predetermined density in advance.
These are microchips. ).

As a method of analyzing gene expression using a microchip, a DNA probe prepared in advance from predetermined cells or tissues is prepared from cells or tissues to be analyzed on a microchip in which dots are arranged, Multiply analysis DNA labeled with a fluorescent substance (Hybr
idize) and DNA itself has a paired relationship, so DN
When the A probe and the DNA for analysis are the same, they are bound to each other, and when they are different, they are not bound.

[0004] In this state, the analysis DNA which is not bound on the microchip is washed away with a buffer solution or the like, and then the microchip is irradiated with light such as a laser beam to form the analysis DNA bound to the DNA probe. DNA analysis is performed by detecting the type and sequence pattern of the DNA for analysis by detecting the fluorescence from the labeled fluorescent substance.

Conventionally, to scan the microchip with light such as a laser beam, the microchip itself is moved in the two-dimensional directions of the X-axis and the Y-axis to scan the laser beam or the like. However, there is a problem that scanning of light takes time. In particular, detection of the type and sequence pattern of the DNA for analysis requires the use of a large number of microchips on which dot probes of DNA probes expected to match the DNA for analysis need to be used. And the analysis work efficiency was extremely poor.

As another scanning device, an optical lens having a size substantially matching a microchip set on the sample stage is arranged above a sample stage controlled to move in a one-dimensional direction. The output laser beam is scanned by the rotating mirror so as to be focused by each DNA probe, and the fluorescence from the fluorescent substance labeled on the DNA for analysis bound to each DNA probe of the microchip is received by the light receiving member. Although a scanning mechanism having a structure for detecting by scanning is also known, since it is necessary to scan light in the entire direction orthogonal to the moving direction of the microchip, it is necessary to increase the distance between the microchip and the optical lens. However, the fluorescence from the fluorescent substance could not be efficiently incident on the sample, and the detection sensitivity was poor.

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned conventional drawbacks, and an object of the present invention is to significantly reduce the scanning time of light on a sample chip and efficiently perform an analysis operation. It is an object of the present invention to provide a biological sample optical scanning device capable of performing the above-mentioned operations.

Another object of the present invention is to provide a biological sample optical scanning device which is excellent in the sensitivity of detecting fluorescence from a fluorescent substance labeled on the biological sample.

[0009]

SUMMARY OF THE INVENTION The present invention provides a biological sample optical scanning apparatus for scanning a sample chip on which a large number of biological samples are arranged to identify a biological sample labeled with a fluorescent substance. A sample stage on which a sample chip is mounted so as to be reciprocally movable on the main body frame, a moving member for moving the sample stage so as to be able to be numerically controlled, and a vertical axis with respect to the plane of the sample stage, and an axis inside An electric motor in which a rotor having a hollow portion extending in the direction is rotatably supported so as to be rotatable, and a hollow portion extending in a radial direction perpendicular to the rotor axis, which is attached to the rotor positioned on the sample stage side and extends in the radial direction. An arm member provided with a pair of reflecting mirrors facing each other at an angle of degree, and provided on the optical axis of one of the reflecting mirrors located on the distal end side of the arm member, and the object plane is on the plane of the sample stage. Closely opposed objectives And a light source that irradiates the sample stage with light having a predetermined beam diameter along the axis of the rotor and through the pair of reflecting mirrors and the objective lens, and a fluorescent substance excited by the light emitted from the objective lens. An object lens, a pair of reflecting mirrors and a light receiving member that receives an electric signal through the hollow portion of the rotor and outputs an electric signal. A biological sample to which a fluorescent substance is attached can be detected.

[0010]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is an overall perspective view of a biological sample optical scanning device, and FIG. 2 is an explanatory diagram of the biological sample optical scanning device.

A guide shaft 5 having an axis in the left-right direction as shown in FIG. 1 is laid on the main body frame 3 of the biological sample optical scanning device 1, and a movable table 7 is reciprocally movable in the axial direction on the main body frame 3. Supported. The movable table 7 is engaged with a feed screw 9 rotatably supported by the main body frame 3 to drive a first electric motor 11 that can be numerically controlled, such as a servomotor or a pulse motor, connected to the feed screw 9. Accordingly, the movable table 7 is moved at a predetermined speed.

The first electric motor 11 is provided with a rotation angle detector 11a such as a rotary encoder.
Rotation angle detector 11a with rotation of electric motor 11
The moving position of the movable table 7 is detected based on the electric signal output from the controller.

A sample table 13 is mounted on the upper surface of the movable table 7, and a sample chip 15 such as a microchip or a DNA chip to be optically scanned is detachably fixed on the upper surface of the sample table 13. As the sample chip 15, a DNA probe (RN) prepared from cells or tissues as described above is used.
A probe) is fixed in the form of a dot matrix at the above-described predetermined intervals, and the DNA for analysis (RNA for analysis) labeled with a fluorescent substance is adjusted for each DNA probe from cells to be analyzed. Are multiplied. In FIG. 4, the DNA probe is denoted by the symbol 、, the analysis DN
The DNA probe multiplied by A is indicated by the symbol ◎.

The body frame 3 corresponding to the upper part of the sample table 13
Has numerical control of servo motor, pulse motor, etc.
Hollow portion 17 extending in the axial direction at the center of rotor 17a
The second electric motor 17 having the letter “b” is attached.
The rotor 17a of the second electric motor 17 has a stator 17c.
A mirror housing 19 extending in the radial direction is fixed to a shaft end of a rotor 17a protruding from the center of the motor housing 17d.

A rotation angle detector 17e such as a rotary encoder is attached to the second electric motor 17 similarly to the first electric motor 11, and outputs a rotation angle detection signal as the rotor 17a rotates.

The mirror housing 19 has a rotation diameter that covers at least the entire width in the width direction orthogonal to the moving direction of the sample stage 13, and has a hollow portion 17b of the rotor 17a inside.
And a hollow portion 19a extending in the radial direction is formed,
First and second mirrors 21a and 21b are attached to the center of the hollow portion 19a and the radial end thereof at an angle of 45 degrees. The mirror housing 19 corresponding to the reflection optical axis of the second mirror 21b is mounted such that the objective surface of the objective lens 23 is close to the sample table 13.

A semi-transparent mirror 27 is inclinedly arranged at an angle of 45 degrees on the main body frame 3 in a position above the center axis of the rotor 17a. A light irradiation device 29 is provided on the main body frame 3 on the optical axis of the semi-transparent mirror 27 extending in a direction orthogonal to the central axis of the rotor 17a, and a main body on the optical axis of the semi-transparent mirror 27 which coincides with the axis of the rotor 17a. Light receiving devices 31 are attached to the frames 3 respectively.

The light irradiation device 29 is a laser light output device for irradiating a laser light having a wavelength corresponding to the fluorescence wavelength emitted by the fluorescent substance labeled on the DNA for analysis bound to the DNA probe, and a semi-transparent mirror 27 for the laser light. An optical lens (neither is shown) for condensing light at a required beam diameter at the center of the optical axis.

The light receiving device 31 includes a light receiving member (not shown) such as a photodiode or a CCD for outputting an electric signal by the fluorescent light transmitted through the semi-transparent mirror 27 and the light receiving member 3.
The optical filter 1a includes an optical filter 31a that allows only light having a fluorescence wavelength to pass therethrough.

Next, the operation of the biological sample optical scanning device 1 will be described. FIG. 3 is an explanatory diagram showing a scanning trajectory of a laser beam on a sample chip, and FIG. 4 is an explanatory diagram showing an example of a detection pattern.

With the sample chip 15 set on the sample table 13, the first electric motor 11 is rotationally driven to move the movable table 7 in the direction of the solid line arrow shown in FIG. 3, and the second electric motor 17 is rotationally driven. Mirror housing 19
Is rotated in a predetermined direction at a predetermined rotation speed.

In this state, when a laser beam of a predetermined wavelength is output from the light irradiation device 29, the laser beam is reflected downward by the semi-transparent mirror 27 so as to coincide with the axis of the rotor 17a, and passes through the hollow portion 17b of the rotor 17a. After passing, the first mirror 2
1a, the light is reflected in a radial direction orthogonal to the axis of the rotor 17a, and then reflected downward by the second mirror 21b so as to coincide with the axis of the rotor 17a. It is focused by diameter.

The upper surface of the sample chip 15 is irradiated with a laser beam along an arc locus as shown in FIG. When the laser light is irradiated on the DNA probe to which the DNA for analysis is bound among the DNA probes arranged in a dot array on the sample chip 15, the fluorescent substance labeled on the DNA for analysis is excited by the laser light. The fluorescent light is emitted, and the fluorescent light passes through the semi-transparent mirror 27 through the optical path opposite to the above, and is received by the light receiving device 31. As a result, the light receiving member 31a of the light receiving device 31 outputs an electric signal by the received fluorescence and is stored in the buffer memory of the computer.

The electrical signal of the fluorescence stored in the buffer memory is supplied to the sample stage 13 output from the rotation angle detector 11a.
Position data and rotation angle detector 1 of second electric motor 17
It is stored as arc data together with the rotation angle data output from 7e.

Then, the computer exchanges the detection data as the arc data stored in the buffer memory into the orthogonal coordinate data based on the pre-programmed circular coordinate data, and as shown in FIG. Create array pattern data.

The scanning time of the sample chip 15 is determined by the A / D conversion processing speed of the computer, the number of detected fluorescent colors, the radius of rotation of the sample chip 15, the one-dimensional movement amount of the sample chip 15,
It is determined by the number of rotations of the electric motor 17 and the like.

In the present embodiment, the scanning time can be greatly reduced as compared with the conventional two-axis moving scanning method, and the work of detecting the sample chip 15 can be performed efficiently. Further, the sample chip 15 is provided through the objective lens 23 close to the sample chip 15.
Since the light is scanned upward, the fluorescence from the sample chip 15 can be efficiently incident on the objective lens 23 to detect the fluorescence with high sensitivity.

[0028]

According to the present invention, an analysis operation can be efficiently performed by remarkably shortening a scanning time of light on a sample chip. Further, the detection can be performed with the detection sensitivity of the fluorescence from the fluorescent substance labeled on the biological sample, and the detection accuracy of the biological sample can be increased.

[Brief description of the drawings]

FIG. 1 is an overall perspective view of a biological sample optical scanning device.

FIG. 2 is an explanatory diagram of a biological sample optical scanning device.

FIG. 3 is an explanatory diagram showing a scanning locus of a laser beam on a sample chip.

FIG. 4 is an explanatory diagram showing an example of a detection pattern.

[Explanation of symbols]

1—biological sample optical scanning device, 3—body frame, 11
1st electric motor, 13-sample stage, 15-sample chip,
17-second electric motor, 17a-rotor, 17b-hollow portion, 23-objective lens, 29-light irradiation device, 29-semi-transparent mirror, 31-light receiving device

 ──────────────────────────────────────────────────続 き Continued on the front page F term (reference) 2G043 AA03 BA16 CA03 DA02 EA01 FA01 GA02 GA04 GB01 GB03 GB19 HA01 HA02 HA09 KA09 LA01 LA03 NA06 NA13 2H045 AG06 BA14 4B024 AA19 CA01 CA09 CA11 HA13 HA14 4B029 AA07 BB15 FA12 CC03 CC03 CC 4B063 QA01 QA13 QA17 QA18 QA19 QQ42 QQ52 QR32 QR35 QR38 QR56 QR84 QS33 QS34 QS35 QS36 QS39 QX02

Claims (3)

[Claims] 1. A biological sample optical scanning device for scanning a sample chip on which a large number of biological samples are arranged to specify a biological sample labeled with a fluorescent substance, so as to be reciprocally movable on a main body frame. A sample stage that is supported and to which a sample chip is attached, a moving member that moves the sample stage in a numerically controllable manner, and a hollow portion that has a vertical axis with respect to the plane of the sample stage and extends inside in the axial direction. The rotor has an electric motor which is rotatably supported on the rotor and a rotor which is attached to the rotor positioned on the sample stage side and faces the rotor at an angle of 45 degrees in a gap extending in a radial direction perpendicular to the rotor axis. An arm member provided with a pair of reflecting mirrors, and an objective lens provided on the optical axis of one of the reflecting mirrors located on the distal end side of the arm member and having an object surface close to and opposed to the plane of the sample stage; Along the rotor axis And a light source for irradiating light of a predetermined beam diameter on the sample stage via a pair of reflecting mirrors and an objective lens,
A sample stage that linearly moves, comprising a light receiving member that receives the fluorescence of the fluorescent substance excited by the light emitted from the objective lens through the objective lens, the pair of reflecting mirrors, and the hollow portion of the rotor, and outputs an electric signal; A biological sample optical scanning device capable of scanning a circular sample chip with light in an arc shape to detect a biological sample to which a fluorescent substance is attached. 2. A biological sample according to claim 1, wherein the light source and the light receiving member are arranged in a relationship orthogonal to the axis of the rotor, and a semi-transparent mirror is provided at the intersection of the optical axis from the light source and the optical axis with respect to the light receiving member. Optical scanning device. 3. The biological sample optical scanning device according to claim 1, wherein the light source comprises a laser light output device having a wavelength for exciting a fluorescent substance.