JPH1123529A - Scanner device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To read the luminous pattern of a sample by both analytical methods of a fluorescence method and chemical luminescence method by reading the luminous pattern with a line sensor while irradiating a laser beam to excite a fluorescent material when read by the fluorescence method, and when reading the luminous pattern by the chemical method, converging the luminescence position with the line sensor. SOLUTION: Fluorescence is generated from a fluorescent material, which is irradiated and excited with a laser beam. A luminous position of a light of a fluorescent pattern is converged by a one-dimensional scanning line, the luminous position is scanned, and a luminous pattern is read by a line sensor 6 and a light reception section scanning device 7. An electrophoretic gel 5a is pinched by glass plates 5b, 5c and is mounted on a stage. A moving mechanism relatively moves the reading position of a sample 5 on the line sensor 6. The light of the luminous pattern is converged on the one-dimensional scanning line. The reading action is the same when the fluorescent pattern of the sample emitted from the fluorescent material excited by the laser beam is read and when the luminous pattern emitted from the sample itself is read by a chemical luminescence method.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a scanner apparatus for reading a weakly luminescent pattern from a flat sample by scanning on a light receiving side, and more particularly, to a method for analyzing both a fluorescence method and a chemiluminescence method. The present invention relates to a scanner device capable of reading a light emission pattern of a sample from the scanner together.

[0002]

2. Description of the Related Art Conventionally, analysis methods by gel electrophoresis are often used for fractionation and structural analysis of biopolymer proteins and nucleic acids. In gel electrophoresis, analysis is performed by electrophoresis using the principle that the migration distance varies depending on the molecular weight of a sample. According to this analysis method, a very small amount of sample can be appropriately analyzed. For this reason, when this gel electrophoresis method is used, in general, the amount of a sample that can be obtained is often limited. Therefore, in that case,
In the analysis processing, particularly, high detection sensitivity is required.

[0003] Therefore, conventionally, when the amount of available samples is small, the sample to be analyzed is labeled with a radioisotope, and the sample is injected into a gel to perform electrophoresis, with an emphasis on certainty. After that, the gel was attached to an X-ray film or the like and exposed, and the pattern of exposure by the radioactive isotope transferred to the X-ray film was read to read the electrophoresis pattern of the sample.

[0004] However, radioisotopes are dangerous and their handling must be strictly controlled. Therefore, in recent years, high-sensitivity detection methods (fluorescence method, chemiluminescence method) using fluorescence or chemiluminescence have been developed.
These methods are used for various experiments such as DNA base sequence determination, Southern blotting, Western blotting, and Northern blotting.

In the fluorescence method, after electrophoresis is completed, a sample labeled with a fluorescent substance is irradiated with a laser beam to excite the fluorescent substance, and the intensity distribution of the fluorescence emitted from the fluorescent substance is measured. This is a method for determining an electrophoresis pattern. As an apparatus for reading a fluorescent pattern of a sample subjected to electrophoresis, for example, an apparatus disclosed in Japanese Patent Publication No. 8-3481 (US Pat. No. 5,069,769) can be referred to.

[0006] Further, with respect to a method for reading a light emission pattern by a chemiluminescence method, examples of the method developed so far include:
`` C. Martin, L. Bresnick, R.-R. Juo. JCVoyta, and I.
Bronstein 1991: Improved Chemiluminescent DNA Seque
ncing: Bio Techniques 8: pp.110-113 ".

[0007] In the reading of an electrophoresis pattern by a chemiluminescence method, usually, after the electrophoresis is completed, a probe or the like labeled with an enzyme involved in luminescence is hybridized so that the fluorescence pattern of the target sample is specifically read. To emit light. Exposure of the film with the emitted fluorescent pattern is performed by placing the membrane in which the sample in the light emitting state has been transferred in close contact with the high-sensitivity film, placing it in a light-shielding case, and adjusting the exposure time according to the light emission. ~
Perform for about 30 minutes.

As the high-sensitivity film used here, for example, an X-ray film for radioisotopes can be used. After developing the exposed film, the pattern is analyzed by visual observation, and the film is analyzed by various image software using an image capturing device such as a camera or an image scanner.

[0009]

Incidentally, the conventional fluorescent pattern reading apparatus is a dedicated apparatus for sample analysis by the fluorescence method, and the reading of the fluorescent pattern by the chemiluminescence method requires a manual operation. , A dedicated reading device has not been developed. Further, a device capable of reading samples with high sensitivity by both the fluorescence method and the chemiluminescence analysis method has not been developed so far.

The present invention has been made to solve such a problem, and an object of the present invention is to read both the emission patterns of a sample from both the fluorescence method and the chemiluminescence method. It is to provide a scanner device which can be used.

[0011]

In order to achieve the above object, a scanner device according to the present invention is a scanner device using a line sensor that scans and reads a light emission pattern of a sample that emits light in a flat plate shape. A stage on which a sample to be read is mounted, a line sensor for one-dimensionally condensing a position for receiving the light of the light emission pattern of the sample, a moving mechanism for relatively moving the stage and the line sensor, A light-receiving section scanning device that scans and receives a light emission pattern of a sample condensed by a line sensor and converts the received light signal into an electric signal, and converts an electric signal from the light-receiving section scanning device into a digital signal. A data processing device for performing data processing. Further, when reading a light emission pattern that excites and emits a fluorescent substance of a sample developed in a flat plate shape, a light source that emits laser light and a laser beam are scanned and irradiated on the sample. A laser beam scanning device.

In the scanner device of the present invention having such features, when reading a light emission pattern of a sample which emits light in a flat plate shape as in a chemiluminescence method, a sample to be read is placed on a stage, and a line sensor is provided. Thereby, the position (light emitting position) of receiving the light of the light emitting pattern of the sample is one-dimensionally condensed. In this case, in order to read the light emission pattern two-dimensionally, the stage and the line sensor are relatively moved by the moving mechanism. Then, the light receiving section scanning device scans and receives the light emission pattern of the sample condensed by the line sensor, and converts the received light signal into an electric signal. A data processing device performs data processing by converting an electric signal from the light receiving unit scanning device into a digital signal.

Here, since the position where the light of the light emission pattern of the sample is received by the line sensor is one-dimensionally condensed, the light receiving section scanning device scans the light emission pattern of the sample condensed by the line sensor. The light is received and the received light signal is converted into an electric signal. Thus, even if the fluorescence pattern of the sample is a fluorescence pattern or a chemiluminescence method, the light emission position of the light emission pattern is one-dimensionally scanned and condensed. The emission pattern of the sample emitting light above can be read as it is.

When reading a light emission pattern emitted from a fluorescent substance of a sample, such as a fluorescence method, a laser light scanning mechanism scans laser light from a light source to excite the fluorescent substance of the sample with laser light. Then, the fluorescent substance of the sample is irradiated with laser light to excite the fluorescent substance, and the emission pattern emitted from the excited fluorescent substance is read by the line sensor. Also in this case, the light emission position of the light of the light emission pattern of the sample is one-dimensionally condensed by the line sensor, and the light receiving portion scanning device scans and receives the light emission pattern of the sample condensed by the line sensor. And converts the received optical signal into an electrical signal. Then, to read the light emission pattern two-dimensionally, the moving mechanism relatively moves the stage and the line sensor. A data processing device performs data processing by converting an electric signal from the light receiving portion scanning device into a digital signal.

[0015]

Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is a diagram illustrating a configuration of a main part of a scanner device according to the present invention. FIG.
1, 1 is a light source, 2 is a vibrating mirror, 3 is a mirror driver, 4 is an irradiation unit scanning control device, 5 is a flat sample placed on a stage, and 6 is a one-dimensional light emission position of the light emission pattern of the sample. Line sensor that focuses light on a typical scanning line,
Reference numeral 7 denotes a light receiving unit scanning device, and reference numeral 8 denotes a data processing device.

The light source 1 is a laser light source that emits laser light for exciting a fluorescent substance. The laser light from the light source 1 is
The light is reflected by the oscillating mirror 2 and is irradiated on the surface of the sample 5. The oscillating mirror 2 is driven by a mirror driver 3. The mirror driver 3 is controlled by the irradiation unit scanning control device 4.

A laser light irradiation system composed of an irradiation section scanning control device 4, a mirror driver 3, a vibration mirror 2, and a light source 1 reads a light emission pattern of a sample which emits light in a flat plate as in a chemiluminescence method. In this case, it is turned off and not used. However, in the case of reading a fluorescent pattern emitted from the fluorescent substance of the sample as in the fluorescence method, the fluorescent substance in the sample 1 is excited by the laser beam. The driver 3 is driven to control the reflection direction of the oscillating mirror 2, scan the laser light from the light source 1, and irradiate the surface of the sample 5 with the laser light. As a result, the fluorescent substance in the electrophoresed sample 5 is excited.

Fluorescence is emitted from the excited fluorescent substance by the irradiation of the laser light, and the emission pattern is converted into light of the fluorescence pattern in the line sensor 6 by the line sensor 6 and the light receiving section scanning device 7 as described later. The light emission position is focused by a one-dimensional scanning line, and the light emission position is scanned and read. In other words, the reading operation here uses a laser light irradiation system to excite the excited fluorescent substance even when reading the emission pattern of the sample itself, such as the emission pattern of the sample by the chemiluminescence method. The same applies to the case of reading the fluorescence pattern of a sample that emits light from.

Next, a scanning system for reading a light emission pattern will be described. Electrophoresis gel 5a is glass plate 5
When a sample 5 to be read sandwiched between b and 5c is placed on a stage (not shown) and a start instruction of a reading operation is given from a console panel, a moving mechanism (not shown) moves on the stage. The reading position of the sample 5 is relatively moved on the line sensor 6. From the line sensor 6,
The light of the light emission pattern of the sample 5 is condensed on a one-dimensional scanning line, and the light receiving unit scanning device 7 scans and receives the light of the light emission pattern from the sample 5 condensed on the scanning line by the line sensor 6. And, by the photoelectric converter provided inside,
The received optical signal is converted into an electric signal. Then, the data processing device 8 converts the converted electric signal into a digital signal and performs data processing. In this case, in order to read the light emission pattern two-dimensionally, the moving mechanism relatively moves the stage on which the sample 5 is mounted and the line sensor 6.

Here, the light receiving section scanning device 7 scans the light emission pattern focused on the scanning line by the line sensor 6 and receives the light. That is, the light-emitting position of the light of the light-emitting pattern is one-dimensionally scanned and condensed by the control on the light-receiving side of the light-emitting pattern (by the line sensor 6 and the light-receiving unit scanning device 7). Whether the light emission pattern of the sample 5 emits light on a plane or the light emission pattern obtained by exciting a fluorescent substance of the sample to emit light, such as a fluorescence method, can be read as it is.

FIG. 2 is a view for explaining an example of the arrangement of a line sensor and a light receiving section scanning device when a two-dimensional CCD image sensor is used. FIG. 3 is a diagram showing the structure of a line sensor when a two-dimensional CCD image sensor is used. It is a perspective view explaining. FIG. 4 is a diagram for explaining a correspondence table in the case where the position of a one-dimensional scanning line of a line sensor is converted into an array of reading positions of a two-dimensional CCD image sensor. next,
Description will be made with reference to these figures.

As shown in FIG. 2, a line sensor 21 composed of an optical fiber bundle is provided below a stage (sample table) 20 on which a sample is mounted.
The light is condensed on a one-dimensional scanning line by 1, and reading scanning is performed by the light-receiving unit scanning device 22. In that case, a two-dimensional CCD imaging device 23 is used as the light receiving unit scanning device 22. The structure of the line sensor 21 is shown in FIG.
As shown in FIG. 3, one end of the optical fiber bundle 30 is arranged in a line so that the fluorescence emitted from the sample on the sample stage side 31 can be collected by a one-dimensional scanning line.
The other end of 0 is a light receiving portion side 32 connected to the light receiving portion scanning device 22 and has a shape (rectangular shape) that matches the shape of the light receiving portion of the light receiving device (two-dimensional CCD image sensor 23) of the photoelectric converter.
The optical fiber bundle is bundled.

Accordingly, the correspondence between the position of the pixel at the light incident position and the position of the pixel at the light emitting position on the scanning line in the line sensor 21 (light entrance and light exit for each optical fiber) is shown in FIG. The relationship is as shown in FIG. These correspondences are set as correspondence data in the correspondence table 40 as shown in FIG. 4B, and based on the correspondence data, the two-dimensional CCD is processed in the signal processing by the data processing device. The positional relationship between the pixels received by the image sensor 23 is returned to the positional relationship between the pixels on the one-dimensional scan line.

That is, as shown in FIG.
As shown in FIG. 4A, the arrangement structure of the optical fiber bundle connecting the light receiving part side 32 facing the light receiving part of the image sensor 23 and the sensor part of the sample stage side 31 is such that the one-dimensional array A ( A) Optical fiber bundles are arranged side by side in a row so that 1) to A (a), and the emission side is a two-dimensional array B (1,1) to
In order to obtain B (x, y), they are bundled in a rectangular shape of x vertically and y horizontally according to the shape of the light receiving surface of the two-dimensional CCD image sensor. Thereby, the line sensor 2
The light emitted from one optical fiber bundle is input to the light receiving surface of the two-dimensional CCD image sensor, and the light amount of the scanning line of the light emission pattern is read from the two-dimensional CCD image sensor 23. The read analog signal value of the light amount is converted into a digital signal and processed. In this data processing,
The two-dimensional pattern of the light amount read from the two-dimensional CCD image sensor is returned to one-dimensional scan line data based on the correspondence data in the correspondence table 40 as described above.

The arrangement of the optical fibers of the optical fiber bundle in the line sensor 21 is based on a one-dimensional array A (1) to A (1).
The subsequence of A (a) is strictly a two-dimensional array B (1,1) to
It does not need to correspond to the horizontal or vertical partial row of B (x, y). In other words, in this case, at the start of reading, a test scan of each pixel position is performed by scanning laser light along a scanning line using a light irradiation system for reading a sample by a fluorescence method, and a one-dimensional array A
(1) The positions of the scanning lines of A (a) and the two-dimensional array B
The correspondence between the positions of the scan lines (1, 1) to B (x, y) is read, and the correspondence is registered in the correspondence table 40 as correspondence data. Then, the reading operation is started using the registered correspondence data of the correspondence table 40.

The method of determining the position of light in the light emission pattern obtained from the sample differs as follows depending on the type of light emission pattern from the sample by the fluorescence method or the chemiluminescence method. That is, in the case of the fluorescence method, the scanning position of the laser beam for exciting the fluorescent substance is obtained from the rotation angle of the vibrating mirror and the time when the laser beam is started to be irradiated, and the light emission of one scanning line of the sample is correspondingly obtained. Find a pattern.

In the case of the chemiluminescence method, the incidence side is
The correspondence relationship of the pixels on the light receiving side of the two-dimensional CCD imaging device is obtained as follows. As an initial setting operation, light is sequentially applied to the optical fiber on the incident side of the line sensor 21 from one side, and at that time, the correspondence relationship with the pixel on the light receiving side of the two-dimensional CCD image pickup device from which the signal was obtained is represented by the correspondence data. Is registered in the correspondence table 40. In a real sample,
Based on the correspondence data in the correspondence table 40, the light emission pattern obtained from the two-dimensional CCD image pickup device is converted, and the light emission pattern of one of the scanning lines of the sample is determined.

Thus, through the line sensor 21,
The amount of light at each pixel position is read by a two-dimensional CCD image sensor, and a line sensor 2 corresponding to each pixel of the sample is read.
The intensity of a pixel on one scan line is determined, and the emission pattern of fluorescence from the sample is determined. In this way, the light emission pattern is read from the sample, and the operation of reading the light emission pattern is the same in both the fluorescence method and the chemiluminescence method, with or without using light irradiation by a laser light source. Thus, that is, by turning on / off the light irradiation system of the laser beam, the reading can be performed by either the fluorescence method or the chemiluminescence method.

Next, a case where a high-sensitivity photomultiplier (PMT) is used as the light receiving element will be described. FIG. 5 is a diagram illustrating the configuration of a line sensor and a light-receiving unit scanning device when a photomultiplier (PMT) is used, and FIG. 6 is a diagram illustrating a photomultiplier (PMT).
It is a figure explaining the structure of the line sensor in case of using T). FIG. 7 is a diagram illustrating a configuration of a driving mechanism of the light receiving unit scanning device, and FIG. 8 is a diagram illustrating a configuration of the light collecting unit. Description will be made with reference to these figures.

In FIG. 5, reference numeral 50 denotes a stage (sample stage); 51, a line sensor composed of an optical fiber bundle;
Is a light-receiving unit scanning device composed of a rotating plate provided with a slit;
3 is a condensing part for condensing the light passing through the slit of the rotating plate, 5
4 is a photomultiplier of a photoelectric conversion element, 55 is a rotating plate, 56 is a slit, and 57 is a position sensor.

The line sensor 51 is, as shown in FIG.
The optical fiber bundles are arranged in a line on the stage 50 side so that the fluorescence emitted from the sample can be collected by a one-dimensional scanning line, and the optical fiber bundles are circular on the opposite side of the light receiving unit scanning device 52. They are arranged in an arc. That is, they are arranged along the circumference of the rotating plate 55 on the side of the light receiving unit scanning device 52 so as to correspond to the positions of the slits 56 of the rotating plate 55 of the light receiving unit scanning device 52 (FIG. 6). The light-receiving unit scanning device 52 includes a rotating plate 5 provided with a slit 56.
5, a stepping motor (64: FIG. 7) for driving the rotary plate 55 to rotate, and a position sensor 57 for detecting the rotational position of the slit. The rotating plate 55 is rotated by a stepping motor in accordance with the reading of the scanning line, as will be described later, and the light passing through the slit 56 is received to scan the scanning line.

When reading the fluorescent pattern of the sample placed on the stage 50, the light of the scan line of the fluorescent pattern of the sample is guided to the light receiving unit scanning device 52 by the optical fiber bundle of the line sensor 51, and the light receiving unit scanning device is read. By rotating the rotating plate 55 of 52, scanning is performed and light is received. That is, light that has passed through the slit 56 of the rotating plate 55 is selectively received, and further guided to the photomultiplier 54 of the photoelectric conversion element through the light condensing unit 53.

The photomultiplier 54 converts the received light into an electric signal according to the amount of light received and outputs the electric signal. The output electric signal is converted into a digital signal to perform data processing. In this case, the digital signal corresponding to the amount of received light is
Based on the rotation angle of the rotating plate in the light-receiving unit scanning device 52, the emission pattern of one scanning line is determined in correspondence with one-dimensional information of the scanning line.

Thus, as in the case of using the two-dimensional CCD image pickup device described above, regardless of whether the light emission pattern of the sample is obtained by the chemiluminescence method or the fluorescence method, whether the light irradiation by the laser light source is used or not is used. That is,
The light emission pattern of each sample can be read by turning on / off the laser irradiation system.

The structure of the line sensor 51 will be described in more detail. High sensitivity photo multiplier (PM
Even in the case of using T), since the light from the light emission pattern of the sample is efficiently condensed, and the light emission position is scanned and received, as shown in FIG. On the side, the optical fiber bundle is arranged in a line so that the fluorescence emitted from the sample can be collected by a one-dimensional scanning line. On the light receiving unit scanning device 52 side, the optical fiber bundle is arranged along the arc of the rotating plate. It is arranged on the circumference. In this case, the arrangement of the optical fiber bundles is such that, as shown in the partial enlarged view on the lower side in FIG. 6, when the optical fibers are arranged on the circumference on the rotating plate side, each optical fiber is positioned as densely as possible on an arc. In other words, the odd-numbered optical fibers are alternately arranged in the lower stage and the even-numbered optical fibers are arranged in the upper stage so that the circumference of the arrangement of the optical fiber bundles becomes smaller.

In the structure of the driving mechanism of the rotating plate of the light-receiving unit scanning device 52, as shown in FIG.
4 are arranged. FIG. 7 shows the structure of the mechanical part of the rotating plate as viewed from the side. The rotating plate 55 includes a rotating shaft 63.
Is directly connected to the stepping motor 64 via the stepping motor 64, and is rotationally driven by the stepping motor 64. Also, as described above, the rotating plate 55 provided with the slit 56
In order to know the rotational position of the slit 56, a position sensor 57 is provided on the side of the rotating plate 55.

Based on the timing at which the slit 56 passes through the position of the position sensor 57, the stepping motor 64
To control the rotation angle of the rotating plate 55, control the position on the same circumference of the slit 56, scan the scanning line of the line sensor 51, and read the light emission pattern.
Then, an emission pattern of one of the scanning lines of the sample is obtained.

FIG. 8 is a diagram for explaining the configuration of the light collecting section. The light-collecting unit 53 is, as described above, a light-receiving unit scanning device 5.
The optical component is formed by an optical component that condenses the light scanned and received by the second and guides the light to the photomultiplier 54 of the photoelectric conversion element. For example, as shown in FIG. 8, light that has been configured by a lens 65 and has passed through a slit of the rotating plate 55 is condensed by the lens 65 and guided to the photomultiplier 54. Instead of the lens 65, an optical fiber bundle can be similarly used as described later (FIG. 12B).

FIG. 9 is a block diagram showing a configuration of an electric system of the scanner device. As shown in FIG. 9, the configuration of the electric system is a microprocessor (CPU) 150 for executing control processing of the scanner device, and a read-only memory (ROM) 15 for storing control software.
2, a random access memory (RAM) 151 for temporary data storage and other data processing, a light receiving unit scanning device 154, an irradiation unit scanning control device 155, a mirror driver 156 for driving a vibrating mirror, and a received optical signal A / D converter 157 for analog-to-digital conversion of a signal obtained by converting a signal into an electric signal, and a shading correction circuit 158 for correcting a fixed deviation of an optical measurement system including unevenness in the intensity of a light receiving unit and a light source. ,
A light source control device 1 that controls on / off of a laser light source in accordance with whether an object to be read is an emission pattern of a sample by a fluorescence method or an emission pattern of a sample by a chemiluminescence method.
59, a SCSI controller 153 for controlling the interface with the external data processing device 160, and the like.

An outline of the operation of the electric system will be described. When the power is turned on, first, initialize the various parts of the device,
After the initialization process is completed, the emission pattern of the sample is read. The contents of the initialization process include, for example, a read-only memory (ROM).
152 and random access memory (RAM) 151
, The operation of the light source and the light receiving unit scanning device with the laser light source turned on and off, the initialization of the interface unit of the SCSI controller 53 for controlling the interface, the operation check of the drive system, and the like.

When the initialization process is completed, the system enters a command waiting state from the host (data processing device 160). When a read instruction command or the like is entered by a user's operation instruction, the command processing is executed, and the process returns to the command waiting state again.

In the case of automatically determining the sample to be read, for example, when a reading instruction command is input, the laser light source is turned off, and the light-receiving unit scanning device detects light emission from the sample for a predetermined time. Thereafter, when it is detected that the sample to be read does not emit light, the light source control device 159 is controlled to turn on the laser light source to read the light emission pattern of the sample by the fluorescence method. Then, in order to scan and irradiate a laser beam for exciting the fluorescent substance of the sample, the irradiation unit scanning control device 155 is activated, and the mirror driver 156 is controlled to drive the vibrating mirror.

The scanning timing for scanning and irradiating the laser beam here is based on the scanning timing of the light irradiating position of the laser beam irradiating by the irradiating unit scanning controller and the light emitting pattern of the sample by the line sensor. The scanning timing is controlled by providing a predetermined phase difference with the scanning timing of the light receiving position where the light is received. By adjusting the phase difference (including the case where the phase difference is zero), in the case of receiving the light emission pattern of the sample by the fluorescence method, the background noise caused by the direct excitation light of the laser light is avoided, and the light receiving unit scanning device is used. A light emitting pattern can be received.

In the case of reading the light emission pattern of the sample by the chemiluminescence method, light from the light emission of the sample is detected from the light receiving section scanning device from the beginning.
59 is controlled to start reading with the laser light source turned off. When reading is started, main scanning is performed under the control of the light-receiving unit scanning device 154, and sub-scanning is performed by controlling a moving structure (not shown). The data processing unit 160 performs, for example, shading correction processing by the shading correction circuit 58 while acquiring data for each of the scanning lines of the line sensor by main scanning.
Through the interface of the SCSI controller 153. After transmitting one line of data, the sub-scan is performed by controlling the moving structure, and reading of the next line is started.

Next, a modified example of the scanner device of the present invention will be described. As described above, in the scanner device of the present invention, the scanning of the emission pattern is performed under the control of the light-receiving unit scanning device, and any one of the emission pattern of the sample by the fluorescence method and the emission pattern of the sample by the chemiluminescence method is performed. However, in the reading of the light emission pattern of the sample by the fluorescence method, scanning control of the irradiation light of the laser light source is indispensable under the control of the irradiation unit scanning control device.

Therefore, only in this case, that is, in reading the emission pattern of the sample by the fluorescence method, the reading is performed only by the control of the irradiation unit scanning control device without controlling the light receiving unit scanning device. Can be modified so that it can be executed.

Next, such a modified example will be described. In a modified example in this case, the light receiving section scanning device is modified so as to guide all of the received light amount of the light emitting pattern to the photoelectric conversion element without controlling the scanning of the scanning line. When a two-dimensional CCD image pickup device is used as the photoelectric conversion device, detection may be performed by adding all the light amounts obtained from the line sensor.

When a high-sensitivity photomultiplier (PMT) is used as the light receiving element, the shape of the rotating plate of the light receiving section scanning device is modified, and the rotation angle of the rotating plate is fixed at a predetermined position. All light amounts obtained from the line sensor are guided to a photomultiplier (PMT).

FIG. 10 is a view showing a rotating plate of a light receiving section scanning device according to a modification of the present invention. In FIG. 10, reference numeral 72 denotes a rotating plate, 74 denotes a slit, 75 denotes a position sensor, and 76 denotes a notch. 1/3 of the circumference of the rotating plate 72 is a cutout portion 76, and has a smaller radius than the remaining portion. One-third of the circumference is the notch 7
In correspondence with No. 6, the light outlet from the optical fiber bundle of the line sensor 51 described above is configured to be provided at one third of the circumference. In other words, when the rotating plate 72 is at the position of the rotation angle as shown in FIG. 10A, the light from the optical fiber bundle of the line sensor 51 is cut into one-third of its circumference by the cutout portion 76.
Through the light-collecting unit, and to a photomultiplier (PMT).

The notch 76 provided at one third of the circumference as described above is provided with the slit 74 formed by the rotation of the rotary plate 72.
Notch 76 so as not to affect the scanning at
The positional relationship between the slit 74 and the slit 74 is set to be 120 ° apart from each other. Thus, when an optical signal at the scanning position of the scanning line is obtained from the slit 74 by the rotation of the rotating plate 72, the cutout 76 of one-third of the circumference has no effect.

Assuming that the rotating plate 72 scans the scanning line clockwise, as shown in FIG. 10B, in the case where there is a slit 74 at the scanning start position of the scanning line, as shown in FIG. The notch 76 has no effect, and even when the slit 74 is located at the scan end position of the scan line rotated clockwise by 120 ° from the scan start position of the scan line, A cutout 76 of one third of the circumference has no effect.

That is, when reading the emission pattern of the sample by the fluorescence method, the rotating plate 72 is stopped in a state as shown in FIG. In this state, the optical fiber bundle of the line sensor is all visible from the front, and all light from one of the scanning lines of the sample can be received. At this time, the laser light source is turned on, and under the control of the irradiation unit scanning control device, it is possible to read the excited fluorescence by irradiating the laser light. The position of the light in the light emission pattern of the sample is obtained from the rotation angle of the vibrating mirror (control signal of the irradiation unit scanning control device).

When reading the light emission pattern of the sample by the chemiluminescence method, the rotating plate 72 is rotated in order to selectively receive a part of the light from the scanning line of the line sensor. The rotating plate 72 has a slit 74 at 120 ° from the end point of the notch 76, and the slit 72 selects and receives a part of the light on the scanning line. The position on the line where the selected light falls
It is obtained from the rotation angle of the rotating plate 72 (the rotation angle of the stepping motor). The reference position of the rotating plate 72 is obtained by the position sensor 85.

FIG. 11 is a diagram for explaining the configuration of a line sensor and a light-receiving unit scanning device when a photomultiplier (PMT) of a modified example is used. In FIG. 11, 7
3 is a rotating plate, 74 is a slit, 75 is a position sensor, 76
Is a notch of 1/3 of the circumference, 80 is a stage (sample stage), 81 is a line sensor composed of an optical fiber bundle, 82
Is a light receiving unit scanning device including a rotary plate provided with slits and cutouts, 83 is a light collecting unit that collects light passing through the slits and cutouts of the rotary plate, and 84 is a photomultiplier of a photoelectric conversion element. is there.

The line sensor 81 is, as shown in FIG.
The optical fiber bundle is arranged in a row on the stage 80 side so that the fluorescence emitted from the sample can be collected by a one-dimensional scanning line, and the optical fiber bundle rotates on the opposite side of the light receiving unit scanning device 82. The plate 73 is arranged in an arc shape of ３ of the circumference of the plate 73. That is, on the side of the light receiving unit scanning device 82, the slit 7 of the rotating plate 73 of the light receiving unit scanning device 82
The rotary plates 73 are arranged in an arc of ３ of the circumference of the rotary plate 73 so as to correspond to the positions of the notches 4 and the notches 76. The light-receiving unit scanning device 82 further includes a stepping motor that drives the rotary plate 73 to rotate, and a position sensor 75 that detects the rotational position of the slit. This rotating plate 73
By the stepping motor, it is rotated corresponding to the scanning line reading, and receives the light passing through the slit 74,
Scanning of the scanning line is performed.

When reading the light emission pattern of the sample placed on the stage 80, the light of the scan line of the light emission pattern of the sample is guided to the light receiving unit scanning device 82 by the optical fiber bundle of the line sensor 81, and the light receiving unit scanning device is read. By rotating the rotating plate 73 of 82, scanning is performed and light is received. Due to the rotation of the rotating plate 73, the light that has passed through the slit 74 of the rotating plate 73 is selectively received, and further guided to a photomultiplier 84 of a photoelectric conversion element through a condensing unit 83.

The photomultiplier 84 converts the received light into an electric signal in accordance with the amount of received light and outputs the electric signal. The output electric signal is converted into a digital signal to perform data processing. In this case, the digital signal corresponding to the amount of received light is
Based on the rotation angle of the rotating plate in the light-receiving unit scanning device 82, the emission pattern of one scanning line is determined in correspondence with one-dimensional information of the scanning line.

FIG. 12 is a diagram showing the structure of the light collecting section.
The condensing unit here is, as described above, a light-receiving unit scanning device 8.
The optical component is configured to guide the light scanned and received by the photodetector 2 to the photomultiplier 84 of the photoelectric conversion element. In this case, for example, as shown in FIG. 12A, a configuration in which light is condensed by a lens 85 and guided to a photomultiplier 84 is used, or as shown in FIG. A configuration in which light is condensed by 86 and guided to the photomultiplier 84 can be used.

As shown in FIG. 12B, in the configuration in which light is condensed using the optical fiber bundle 86, the light incident side of the optical fiber bundle 86 is The partial scanning devices 82 are arranged so as to face each other with the rotating plate therebetween, and the other end of the light emission side is bundled according to the size of the light entrance of the photomultiplier 84. In this case, the light that has passed through the rotating plate is received by the optical fiber 86, propagates through the optical fiber bundle 86, and is guided to the photomultiplier 84.

As described above, in the scanner device of the modified example, when reading the emission pattern of the sample by the fluorescence method, the control of the scanning system of the reading position is performed without using the light receiving unit scanning device. The rotating plate may be fixed so that the light irradiation system of the laser light source can be controlled.

That is, in the scanner device of the modified example, the rotating plate 73 of the light-receiving unit scanning device 82 is provided with 1 / one of the circumference.
Since the three notches 76 are provided, the notch 76
By rotating the rotary plate 73 at an appropriate position so that the position of
Light emitted from the entire scanning line can be received at once. Therefore, in this state, the emission pattern of the sample by the fluorescence method can be read. In this case, the light emitting pattern is determined by associating the rotation angle of the vibrating mirror with the position of the light.

[0062]

As described above, according to the scanner apparatus of the present invention, it is possible to efficiently receive weak light such as chemiluminescence by a simple scanner method and visualize the pattern. In addition, the present invention has an advantage that light loss can be reduced because light is condensed by an optical fiber in close contact with light from a light emitting source. Also, to avoid using a large amount of expensive light receiving elements,
The cost has also been improved.

[Brief description of the drawings]

FIG. 1 is a diagram illustrating a configuration of a main part of a scanner device of the present invention.

FIG. 2 is a diagram illustrating a configuration example of a line sensor and a light receiving unit scanning device when a two-dimensional CCD imaging device is used;

FIG. 3 is a perspective view illustrating the structure of a line sensor when a two-dimensional CCD imaging device is used,

FIG. 4 is a view for explaining a correspondence table when a position of a one-dimensional scanning line of a line sensor is converted into an array of reading positions of a two-dimensional CCD image sensor;

FIG. 5 is a diagram illustrating a configuration of a line sensor and a light-receiving unit scanning device when a photomultiplier (PMT) is used;

FIG. 6 is a diagram illustrating a structure of a line sensor when a photomultiplier (PMT) is used;

FIG. 7 is a diagram illustrating a configuration of a drive mechanism of the light-receiving unit scanning device;

FIG. 8 is a diagram illustrating a configuration of a light collecting unit.

FIG. 9 is a block diagram illustrating a configuration of an electric system of the scanner device.

FIG. 10 is a diagram showing a rotating plate of a light receiving unit scanning device according to a modification of the present invention;

FIG. 11 is a photomultiplier (PMT) of a modified example.
Diagram illustrating the configuration of a line sensor and a light receiving unit scanning device when using the

FIG. 12 is a diagram illustrating a configuration of a light collecting unit according to a modification.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... Light source, 2 ... Vibrating mirror, 3 ... Mirror driver, 4 ... Irradiation part scanning control device, 5 ... Flat sample, 6 ... Line sensor, 7 ... Light receiving part scanning device, 8 ... Data processing device, 20 ... Stage (Sample stage), 21: line sensor, 22: light receiving unit scanning device, 23: two-dimensional CCD image pickup device, 30: optical fiber bundle, 31: sample stage side, 32: light receiving side, 40: correspondence table, 50 ... Stage (sample stage), 51 ... Line sensor, 52 ... Light receiving unit scanning device, 53 ... Condenser, 54 ... Photomultiplier, 55 ... Rotating plate, 56 ... Slit, 57 ... Position sensor, 63 ... Rotating axis, Reference numeral 64: Stepping motor, 65: Lens, 72: Rotating plate, 74: Slit, 75: Position sensor, 76: Notch, 80: Stage (sample stage), 81: Line sensor, 8 ... receiving unit scanning device, 83 ... condensing unit, 84 ... photomultiplier 85 ... lens, 86 ... optical fiber bundle,

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[Procedure amendment]

[Submission date] July 16, 1997

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0007

[Correction method] Change

[Correction contents]

[0007] In the reading of an electrophoresis pattern by a chemiluminescence method, usually, after the electrophoresis is completed, a probe or the like labeled with an enzyme involved in luminescence is hybridized so that the fluorescence pattern of the target sample is specifically read. To emit light. Then, exposure of the film of the emitted light - emitting patterns, the membrane was transferred samples in the light emission state, in close contact with high speed film, placed in a light-shielding case, while adjusting the exposure time corresponding to the emission 10
Perform for about 30 minutes.

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name] 0009

[Correction method] Change

[Correction contents]

[0009]

[SUMMARY OF THE INVENTION Incidentally, the fluorescence pattern reading apparatus to which a dedicated device for sample analysis by fluorescence method, the light emission pattern reading by the chemiluminescence method, with manual operation, In particular, no dedicated reading device has been developed. Further, a device capable of reading samples with high sensitivity by both the fluorescence method and the chemiluminescence analysis method has not been developed so far.

[Procedure amendment 3]

[Document name to be amended] Statement

[Correction target item name] 0017

[Correction method] Change

[Correction contents]

A laser light irradiation system composed of an irradiation section scanning control device 4, a mirror driver 3, a vibration mirror 2, and a light source 1 reads a light emission pattern of a sample which emits light in a flat plate as in a chemiluminescence method. In this case, it is turned off and not used. However, for the case of reading the light emission pattern of the light emitting from the fluorescent substance of the sample, such as a fluorescence method, for the fluorescent material in the specimen is excited by the laser beam, the control of the irradiation unit scan control unit 4, a mirror By driving the driver 3 to control the reflection direction of the vibrating mirror 2,
The surface of the sample 5 is irradiated with the laser beam by scanning the laser beam from the light source 1. As a result, the fluorescent substance in the electrophoresed sample 5 is excited.

[Procedure amendment 4]

[Document name to be amended] Statement

[Correction target item name] 0018

[Correction method] Change

[Correction contents]

Fluorescence is emitted from the excited fluorescent substance by the irradiation of the laser light, and the emission pattern is converted into light of the fluorescence pattern in the line sensor 6 by the line sensor 6 and the light receiving section scanning device 7 as described later. The light emission position is focused by a one-dimensional scanning line, and the light emission position is scanned and read. In other words, the reading operation here uses a laser light irradiation system to excite the excited fluorescent substance even when reading the emission pattern of the sample itself, such as the emission pattern of the sample by the chemiluminescence method. the same applies to the case of reading the light emission pattern of the sample to be emitted from.

[Procedure amendment 5]

[Document name to be amended] Statement

[Correction target item name] 0032

[Correction method] Change

[Correction contents]

[0032] When reading the fluorescence pattern of the sample placed on the stage 50, the optical fiber bundle of the line sensor 51, guides the light scanning line of the light emission pattern of the sample receiving portion scanning device 52, the light receiving unit scan By rotating the rotating plate 55 of the device 52, scanning is performed and light is received. That is, light that has passed through the slit 56 of the rotating plate 55 is selectively received, and further guided to the photomultiplier 54 of the photoelectric conversion element through the light condensing unit 53.

[Procedure amendment 6]

[Document name to be amended] Statement

[Correction target item name] 0044

[Correction method] Change

[Correction contents]

In the case of reading the light emission pattern of the sample by the chemiluminescence method, light from the light emission of the sample is detected from the light receiving section scanning device from the beginning.
59 is controlled to start reading with the laser light source kept in the off state . When reading is started, main scanning is performed under the control of the light-receiving unit scanning device 154, and sub-scanning is performed by controlling a moving structure (not shown). Then, while acquiring the data of each line of the scan line of the line sensor by the main scanning, for example, by performing correction processing of shading by the shading correction circuit 1 58, the data processing apparatus 160, delivered through the interface of SCSI controller 153 I do. After sending one line of data,
The sub-scan is performed by controlling the moving structure, and reading of the next line is started.

[Procedure amendment 7]

[Document name to be amended] Statement

[Correction target item name] 0057

[Correction method] Change

[Correction contents]

The photomultiplier 84 converts the received light into an electric signal in accordance with the amount of received light and outputs the electric signal. The output electric signal is converted into a digital signal to perform data processing. In this case, the digital signal corresponding to the amount of received light is
Based on the rotation angle of the rotating plate in the light-receiving unit scanning device 82, the emission pattern of one scanning line is determined in correspondence with one-dimensional information of the scanning line.

[Procedure amendment 8]

[Document name to be amended] Drawing

[Correction target item name] FIG.

[Correction method] Change

[Correction contents]

FIG. 10

Continued on the front page (72) Inventor Naganori Nasu 6-81 Onoecho, Naka-ku, Yokohama-shi, Kanagawa Prefecture Hitachi Software Engineering Co., Ltd.

Claims (8)

[Claims] 1. A scanner device using a line sensor that scans and reads a light emission pattern of a sample that emits light in a flat plate shape, comprising: a stage on which a sample to be read is mounted; A line sensor for one-dimensionally condensing the position, a moving mechanism for relatively moving the stage and the line sensor, and scanning and receiving the light emission pattern of the sample condensed by the line sensor, and the received light A scanner device comprising: a light-receiving unit scanning device that converts a signal into an electric signal; and a data processing device that performs data processing by converting an electric signal from the light-receiving unit scanning device into a digital signal. 2. A scanner device using a line sensor that scans and reads a light emission pattern that emits light by exciting a fluorescent substance of a sample developed in a flat plate shape, comprising: a light source that emits laser light; A laser beam scanning device that scans and irradiates laser light and a stage on which a sample to be read is placed; a line sensor that one-dimensionally condenses a position for receiving light of a light emission pattern of the sample; A moving mechanism for relatively moving a line sensor, a light-receiving unit scanning device that scans a light-emitting pattern of a sample condensed by the line sensor and receives the light, and converts the received light signal into an electric signal; A scanner device comprising: a data processing device that converts an electric signal from a scanning device into a digital signal and performs data processing. 3. The scanner device according to claim 2, wherein the light-receiving unit scanning device scans a light irradiation position of a laser irradiation position by a laser beam scanning device and a light emission pattern of the sample by a line sensor. A scanner device wherein a scanning timing of a light receiving position for receiving light is controlled by providing a predetermined phase difference. 4. The scanner device according to claim 1, wherein the line sensor arranges the light entrances on the scanning line and matches the shape of the light receiving portion of the photoelectric converter using the light exit. An optical fiber bundle bundled in a shape, at the light exit port,
A scanner device, which is scanned and received by the light receiving unit scanning device. 5. The scanner device according to claim 4, wherein the light-receiving unit scanning device is a two-dimensional CCD image pickup device, and a light emission port of the line sensor has a rectangular shape. 6. The scanner device according to claim 4, wherein the photoelectric converter in the light-receiving unit scanning device is a photomultiplier tube, and the light emission port of the line sensor has an arc shape, and A scanner device wherein scanning is performed by a shutter mechanism. 7. The scanner device according to claim 1, wherein an electric signal from the light-receiving unit scanning device is converted into a digital signal, and a distribution of a light-emitting intensity of a light-emitting pattern of a sample received by data processing on the digital signal is first-ordered. A scanner device obtained as original information. 8. The scanner device according to claim 5, wherein the light-receiving unit scanning device is a CCD imaging device, and a correspondence relationship between a position of an optical fiber arranged on a scanning line and a positional relationship of each pixel of the CCD imaging device. A scanner device which converts a table to determine an image of a light emission pattern.