JPH10274637A - Fluorescent image visualization unit - Google Patents

Abstract

PROBLEM TO BE SOLVED: To visualize an electrophoretic image in safety by an arrangement wherein a get containing a sample labeled with a fluorescent material and subjected to electrophoresis can be mounted on a diffuser formed on a blue LED array substrate provided with a plurality of blue LEDs. SOLUTION: A plurality of blue LEDs 2 are provided on a blue LED array substrate 3 mounting a BPF 4. An opal glass, a diffuser 5 made of a cloudy acryl, or the like, and a transparent cover glass 6 are mounted sequentially thereon. A get subjected to electrophoresis and labeled with a fluorescent coloring matter is mounted on the transparent cover glass 6. A fluorescent image visualization unit 1 thus obtained is set in a slightly dark environment and light is emitted from the blue LED 2. The blue light is rendered nondirectional through the diffuser 5 and enters through the transparent cover glass 6 into the get where the fluorescent coloring matter in the sample is excited. The fluorescence can be observed with eyes and thereby an electrophoretic image can be visualized.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for visualizing a fluorescent image, and more particularly, to a fluorescent image visualizing apparatus capable of safely visualizing an electrophoretic image of a sample labeled with a fluorescent substance. It concerns the device.

[0002]

2. Description of the Related Art A fluorescence detection system using a fluorescent substance as a labeling substance is known. According to this fluorescence detection system, by reading a fluorescence image, it is possible to perform gene sequence, gene expression level, protein separation and identification, or molecular weight and property evaluation. D
After adding a fluorescent dye to the solution containing the NA fragment, a plurality of DNA fragments are subjected to electrophoresis on a gel support, or a plurality of DNA fragments are placed on a gel support containing a fluorescent dye.
The A fragment is electrophoresed, or a plurality of DNA fragments are electrophoresed on a gel support, and then the gel support is immersed in a solution containing a fluorescent dye. By labeling, exciting a fluorescent dye with excitation light, and detecting the generated fluorescence, an image is generated, and the distribution of DNA on the gel support is detected. After electrophoresis on a support, the DNA is denaturated and then
A probe prepared by transferring at least a portion of a denatured DNA fragment onto a transfer support such as nitrocellulose by Southern blotting, and labeling DNA or RNA complementary to the target DNA with a fluorescent dye D
An image is generated by hybridizing with an NA fragment, selectively labeling only a DNA fragment complementary to a probe DNA or a probe RNA, exciting a fluorescent dye with excitation light, and detecting generated fluorescence. Then, the distribution of the target DNA on the transcription support can be detected. Furthermore, by preparing a DNA probe complementary to the DNA containing the target gene labeled with a labeling substance,
The DNA is hybridized with the DNA on the transcription support, and the enzyme is
After binding with a complementary DNA labeled with a labeling substance, the fluorescent substance is brought into contact with a fluorescent substrate to convert the fluorescent substrate into a fluorescent substance that emits fluorescence, and the generated fluorescent substance is excited by excitation light to generate the fluorescent substance. By detecting the fluorescence, an image can be generated and the distribution of the target DNA on the transfer support can be detected. This fluorescence detection system
There is an advantage that a gene sequence or the like can be easily detected without using a radioactive substance.

[0003]

A sample labeled with such a fluorescent substance is electrophoresed on a gel, the obtained image is observed, and a portion where a specific target substance is distributed is cut out or sucked out. In some cases, the sample is taken out, further processed, and variously analyzed. In such a case,
Conventionally, a sample that has been electrophoresed with ultraviolet light having a wavelength of 365 nm or 315 nm is irradiated to visualize an electrophoretic image, visually observe, find a band portion in which a target substance is distributed, cut out with a scalpel, It was common to take it out by sucking it out.

[0004] However, such a method has a problem that the user is inevitably exposed to ultraviolet rays. Therefore, an object of the present invention is to provide a fluorescent image visualizing apparatus that can safely visualize an electrophoretic image of a sample labeled with a fluorescent substance.

[0005]

SUMMARY OF THE INVENTION The object of the present invention is as follows.
Blue LE with multiple blue LEDs (light emitting diodes)
This is achieved by a fluorescence image visualization device including a D array substrate and a diffusion member which is located on the blue LED array substrate, is labeled with a fluorescent substance, and can mount a gel containing a sample subjected to electrophoresis. According to the present invention, the fluorescent substance labeling the sample is excited by light having a wavelength of, for example, 450 nm emitted from the blue LED, and the fluorescent image can be visualized. No, it is safe to visualize the electrophoresis image of the sample labeled with the fluorescent substance, find the band where the target substance is distributed, cut out or suck out with a scalpel, take out, add further processing , It becomes possible to analyze the target substance.

[0006] The object of the present invention is also to provide a plurality of blue LEs.
A blue LED array substrate provided with D (light emitting diode); and a diffusion member located on the blue LED array substrate and capable of mounting an electrophoresis tank capable of housing a gel capable of electrophoresing a sample. Achieved by a fluorescence image visualizer. According to the present invention, on the spot, the sample is electrophoresed, the electrophoresis image of the sample is visualized, the band portion in which the target substance is distributed is found, cut out or sucked out with a scalpel, and taken out. In addition to the processing, the target substance can be analyzed.

[0007] In a preferred embodiment of the present invention, the fluorescent image visualizing device further comprises a CCD camera. According to a preferred embodiment of the present invention, an electrophoretic image of a sample is visualized on a display means such as a CRT by a cooled CCD camera, a band portion where a target substance is distributed is found, and cut out with a scalpel, It is possible to analyze the target substance by sucking out, taking out, and further processing. In the present invention, the fluorescent dye that can be used to label a sample is not particularly limited as long as it is a fluorescent dye that emits fluorescence when excited by a blue LED. For example, Fluorescein (CIN)
o. 45350), Fluorescein-X represented by the structural formula (1), YOYO-1 represented by the structural formula (2), and T represented by the structural formula (3)
OTO-1, YO-PRO-1 represented by structural formula (4), structural formula (5)
Cy-3 (registered trademark) represented by the formula: Ni represented by the structural formula (6):
le Red, BCECF represented by the structural formula (7), Cy-5 (registered trademark) represented by the structural formula (8), Eth represented by the structural formula (9)
idium Bromide, Texas Red represented by the structural formula (10), Propidium Iodide represented by the structural formula (11), POPO-3 represented by the structural formula (12), Rhodamine 6G (CI No. 45160),
SYBR Green (C ₂ H ₆ OS), Acridine Orange (CI No. 46
005), Quantum Red, R-Phycoerythrin, Red 613, R
ed 670, Fluor X, Fluorescein labeled amidite, FAM
, AttoPhos, Bodipy phosphatidylcholine, SNAFL, C
alcium Green, Fura Red, Fluo 3, AllPro, NBD phosp
hoethanolamine, Carboxyrhodamine (R6G), JOE, HEX
, Ethidium homodimer, Lissamine rhodamine B pepti
de, Allphycocyanin and the like.

[0008]

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[0009]

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[0010]

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[0011]

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[0012]

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[0013]

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[0014]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments according to the present invention will be described below in detail with reference to the accompanying drawings. FIG. 1 is a schematic exploded view of a fluorescent image visualizing device according to a preferred embodiment of the present invention, and FIG. 2 is a schematic perspective view thereof. In FIG. 1 and FIG.
A blue LED array substrate 3 having a plurality of blue LEDs 2, a band-pass filter 4 mounted on the blue LED array substrate 3, a diffusion plate 5 formed of opal glass, opaque acrylic, or the like; A transparent cover glass 6 mounted on the diffusion plate 5 is provided. On the transparent cover glass 6, a gel 7 containing a sample that has been electrophoresed and labeled with a fluorescent dye is placed. Here, the band pass filter 4 is provided to improve the contrast of the fluorescent image, and the transparent cover glass 6
This is provided to prevent the diffusion plate 5 from being damaged when the band portion of the target substance is cut off with a scalpel or the like.

The fluorescent image visualizing device 1 configured as described above is placed in a slightly dark environment, and the blue LED array substrate 3
When a plurality of blue LEDs 2 are turned on, a plurality of blue LEs
Blue light is emitted from D2, and the blue light transmitted through the band-pass filter 4 is converted to non-directional light by transmitting through the diffusion plate 5 and enters the gel 7 via the transparent cover glass 6. I do. After the gel 7 is subjected to electrophoresis,
A band 8 of the sample labeled with the fluorescent dye is formed, and the fluorescent dye is excited by light having a wavelength of, for example, 450 nm incident on the gel 7, and the band 8 emits fluorescence. This fluorescence is visually observable, and thus an electrophoretic image of the sample can be visualized. In this case, if the user wears sunglasses or the like that cuts light having the wavelength of the excitation light emitted from the blue LED 2, even if the amount of light emitted from the fluorescent dye is small,
Band 8 can be seen. According to this embodiment, the band 8 of the sample that has been safely electrophoresed on the gel 7 and labeled with a fluorescent dye can be visually observed, the band portion where the target substance is distributed is found, and cut out with a scalpel. It is possible to analyze the target substance by taking out, sucking out, taking out, and further processing.

FIG. 3 is a schematic perspective view of a fluorescent image visualizing apparatus according to another preferred embodiment of the present invention. As shown in FIG. 3, a fluorescent image visualizing device 1 according to another preferred embodiment of the present invention is mounted on a blue LED array substrate 3 having a plurality of blue LEDs 2 and a blue LED array substrate 3. The apparatus includes a bandpass filter 4, a diffusion plate 5, and a transparent cover glass 6 placed on the diffusion plate 5. In the present embodiment, on the transparent cover glass 6,
An electrophoresis tank 12 provided with electrodes 10 and 11 is placed, and a gel 7 is set in the electrophoresis tank 12. A sample labeled with a fluorescent dye can be electrophoresed on the gel 57. It is configured as follows.

The fluorescent image visualizing device 1 is placed in a slightly dark environment, and a voltage is applied to the gel 7 through the electrodes 10 and 11,
The sample is electrophoresed on the gel 7 in the electrophoresis tank 12.
After that, the plurality of blue LEDs 2 on the blue LED array substrate 3
Is turned on, the electrophoresis image of the sample is visualized and observed in exactly the same manner as in the above embodiment, a band portion in which the target substance is distributed is found, cut out or sucked out with a scalpel, and taken out. Process and analyze the target substance. According to the present embodiment, the sample can be electrophoresed on the spot, and the electrophoresis image of the sample can be visualized.

FIG. 4 is a schematic front view of an image generating apparatus including a fluorescent image visualizing apparatus according to another embodiment of the present invention. In FIG. 4, the image generation device includes an imaging device 20, a dark box 21 provided with a fluorescence image visualization device, and a personal computer 22. The personal computer 22 includes a CRT 23 and a keyboard 24. FIG. 5 shows an imaging apparatus 2 according to another embodiment of the present invention.
0 is a schematic longitudinal sectional view of FIG. As shown in FIG. 5, the imaging device 20 includes a CCD 25, a heat transfer plate 26 made of metal such as aluminum, a Peltier element 27 for cooling the CCD 25, and a shutter 28 disposed on the front of the CCD 25. And an A / D converter 2 for converting analog image data generated by the CCD 25 into digital image data.
9 and an image data buffer 30 for temporarily storing image data digitized by the A / D converter 29.
And a camera control circuit 31 for controlling the operation of the imaging device 20
And The opening formed between the imaging device 2 and the dark box 21 is closed by a glass plate 32.
Radiation fins 33 for dissipating the heat generated by the Peltier element 27 are formed around the periphery of the radiator 0 almost in the longitudinal direction. On the front surface of the glass plate 32 provided in the imaging device 20, an image
An intensifier 34 is provided, and a camera lens 35 is mounted on the front surface of the image intensifier 34.

FIG. 6 is a schematic vertical sectional view of the dark box 21. As shown in FIG. 6, for example, 45
A blue LED array substrate 43 having a plurality of blue LEDs that emit 0 nm excitation light, a bandpass filter 44 mounted on the blue LED array substrate 43, and a diffusion formed by opal glass, opaque acrylic, or the like. Board 4
5 and a transparent cover glass 46 placed on the diffusion plate 45
It has. On the transparent cover glass 46, the gel 4 containing the sample that has been electrophoresed and labeled with a fluorescent dye
7 is placed. A filter 36 that cuts light having a wavelength of excitation light emitted from the plurality of blue LEDs is attached to a front surface of the camera lens 35.

FIG. 7 is a block diagram around the personal computer 22. As shown in FIG. 7, the personal computer 22 includes a CPU 50 for controlling the exposure of the imaging device 20, and image data generated by the imaging device 1. Image data transfer means 51 for reading image data from the image data buffer 30, image processing means 53 for performing image processing on the image data read by the image data transfer means 51 and storing the image data in the image data storage means 52, and image data storage means 52 based on the image data stored in
Image display means 5 for displaying a visible image on the screen of RT23
4 is provided. The plurality of blue LEDs 42 are controlled by a light source control unit 55, and an instruction signal is input to the light source control unit 55 from the keyboard 24 via the CPU 50. The CPU 50
Various signals can be output to the camera control circuit 31 of the imaging device 20.

In this embodiment, when the operator inputs a lens focus adjustment signal via the keyboard 24, the CPU 50 outputs a lens focus adjustment mode control signal to the camera control circuit 31 to control the lens focus adjustment mode. Upon receiving the signal, the camera control circuit 31
Is configured to control a readout control circuit (not shown) so that the A / D converter 29 outputs image data accumulated in the form of electric charges by the CCD 25 every predetermined time. In reading the fluorescent image, first, the user places the gel 47 on the transparent cover glass 46 and operates the camera lens 16 to adjust the lens focus. When the lens focus adjustment is completed, the dark box 21 is closed. Thereafter, when the user inputs an exposure start signal to the keyboard 24, the light source control unit 55 turns on the plurality of blue LED 42 light sources,
Excitation light is emitted toward the gel 47. At the same time, the exposure start signal is input to the camera control circuit 31 of the imaging device 20 via the CPU 30, and the shutter 28 is opened by the camera control circuit 31 to start the exposure of the CCD 25.

After the gel 47 is electrophoresed, a band of a sample labeled with a fluorescent dye is formed, and when the excitation light is irradiated from the plurality of blue LEDs 42,
Fluorescence is emitted from the fluorescent dye. The fluorescent light emitted from the fluorescent dye enters the fluorescent screen of the image intensifier 34 via the filter 36 and the camera lens 35 to form an image. The CCD 25 receives the image light thus formed on the phosphor screen of the image intensifier 34 and accumulates the light in the form of electric charges. Filter 36
As a result, the light having the wavelength of the excitation light is cut off, so that only the fluorescent light emitted from the fluorescent dye
0 is received by the CCD 25. When the predetermined exposure time has elapsed, the CPU 50 outputs an exposure completion signal to the camera control circuit 31 of the imaging device 20. Camera control circuit 3
When an exposure completion signal is received from the CPU 50, the CCD 1
A / D 25 stores analog image data stored in the form of electric charges.
The data is transferred to the converter 29, digitized, and temporarily stored in the image data buffer 30. At the same time, CPU5
0 outputs a data transfer signal to the image data transfer means 51, causes the digital image data temporarily stored in the image data buffer 30 of the imaging device 20 to be read out, and causes the image processing means 53 to input the digital image data. The image processing unit 53 performs image processing on the image data input from the image data transfer unit 51 and stores the image data in the image data storage unit 52.

Thereafter, when the user inputs an image generation signal to the keyboard 24, the image display means 55 reads out the image data stored in the data storage means 53,
On the screen of the CRT 23, a fluorescent image composed of the band of the sample is displayed based on the read image data. According to this embodiment, the electrophoresis image of the sample is
Is visualized on the screen of the CRT 23 by using the imaging device 20 including the target, and a band portion where the target substance is distributed is found, cut out or sucked out with a scalpel, taken out, and further processed to analyze the target substance. It becomes possible to do. The present invention is not limited to the above embodiments, and various changes can be made within the scope of the invention described in the claims, which are also included in the scope of the present invention. Needless to say.

For example, in the embodiment shown in FIGS. 4 to 7, the electrophoresis image of the electrophoresed sample is visualized on the screen of the CRT 24 by the imaging device 20, but is not shown in FIG. As in the embodiment described above, the electrophoresis tank is placed on the transparent cover glass 46, the sample is electrophoresed on the gel set in the electrophoresis tank, and the electrophoresis image of the sample is visualized. Alternatively, a portion where the target substance is distributed may be found, cut out or sucked out, taken out, and further processed to analyze the target substance. Further, in the embodiment, the electrophoretic image of the sample is visualized, a portion where the target substance is distributed is found, cut out or sucked out, and taken out,
Although the system is configured to analyze the target substance by further processing, it may be sufficient to simply visualize the electrophoresis image of the sample.

Further, in the above embodiment, the band pass filters 4 and 44 are provided, but the band pass filter 4 is not necessarily required. In the above embodiment, the transparent cover glasses 6 and 46 are provided on the diffusion plates 5 and 45. However, when the diffusion plates 5 and 45 are made of a material that is not easily damaged, the transparent cover glasses 6 and 46 are provided. 4
6 need not be provided. Further, in the embodiments shown in FIGS. 4 to 7, the image intensifier 34 is provided, but the image intensifier 34 is not necessarily provided. Further, in the embodiment shown in FIGS. 4 to 7, the imaging device 20 includes a Peltier element 27 for cooling the CCD 25.
In addition, a radiating fin 33 for radiating heat generated by the Peltier element 27 is provided around the imaging device 20, but the fluorescent light emitted by the fluorescent dye that labeled the sample excited by the light emitted from the blue LEDs 2 and 42. Is not so weak light, it is not always necessary to provide the Peltier element 27 and the radiation fin 33.

[0026]

According to the present invention, it is possible to provide a fluorescent image visualizing apparatus capable of safely visualizing an electrophoretic image of a sample labeled with a fluorescent substance.

[Brief description of the drawings]

FIG. 1 is a schematic exploded view of a fluorescent image visualizing device according to a preferred embodiment of the present invention.

FIG. 2 is a schematic perspective view of a fluorescent image visualizing device according to a preferred embodiment of the present invention.

FIG. 3 is a schematic perspective view of a fluorescent image visualizing device according to another preferred embodiment of the present invention.

FIG. 4 is a schematic front view of an image generating apparatus including a fluorescent image visualizing apparatus according to another preferred embodiment of the present invention.

FIG. 5 is a schematic vertical sectional view of an imaging device according to another embodiment of the present invention.

FIG. 6 is a schematic vertical sectional view of a dark box.

FIG. 7 is a block diagram of the periphery of a personal computer.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Fluorescence image visualization apparatus 2, 42 Blue LED 3, 43 Blue LED array board 4, 44 Band pass filter 5, 45 Diffusion plate 6, 46 Transparent cover glass 7, 47 Gel 8 Band 10, 11 Electrode 12 Electrophoresis tank 20 Imaging Device 21 Dark Box 22 Personal Computer 23 CRT 24 Keyboard 25 CCD 26 Heat Transfer Plate 27 Peltier Element 28 Shutter 29 A / D Converter 30 Image Data Buffer 31 Camera Control Circuit 32 Glass Plate 33 Heat Dissipating Fin 34 Image Intensifier 35 Camera Lens 36 Filter 50 CPU 51 Image data transfer means 52 Image data storage means 53 Image processing means 54 Image display means 55 Light source control means

Claims (3)

[Claims] 1. A blue LED array substrate provided with a plurality of blue LEDs (light emitting diodes), and a gel, which is located on the blue LED array substrate, is labeled with a fluorescent substance and contains an electrophoresed sample, is placed on the substrate. A fluorescent image visualization device comprising a possible diffusion member. 2. A blue LED array substrate provided with a plurality of blue LEDs (light emitting diodes), and an electrophoresis tank which is located on the blue LED array substrate and which can accommodate a gel capable of electrophoresing a sample. A fluorescent image visualizing device, comprising a diffusion member that can be placed. 3. The fluorescence image visualizing device according to claim 1, further comprising a CCD camera.