CN211453364U - Multicolor fluorescence gene detection device - Google Patents
Multicolor fluorescence gene detection device Download PDFInfo
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- CN211453364U CN211453364U CN202020096664.3U CN202020096664U CN211453364U CN 211453364 U CN211453364 U CN 211453364U CN 202020096664 U CN202020096664 U CN 202020096664U CN 211453364 U CN211453364 U CN 211453364U
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Abstract
The utility model relates to the technical field of gene detection devices, in particular to a multicolor fluorescence gene detection device, which comprises a capillary electrophoresis gene analysis device, wherein the capillary electrophoresis gene analysis device comprises a third section of capillary, a first detection window and a second detection window are arranged on the third section of capillary, a first laser is arranged on one side of the first detection window, and the other side of the first detection window is connected with a CCD detector through an optical fiber; and a second laser is arranged on one side of the second detection window, and the other side of the second detection window is connected with the APD detector. The utility model solves the problem that the multicolor fluorogenic detection device in the prior art can only detect 6 to 8 colors; through designing first detection window and second detection window to realize twice detection, two detectors are made up and are can be detected 7 ~ 10 look fluorescence, thereby can satisfy the detection demand, can be used to the on-the-spot detection gene.
Description
Technical Field
The utility model relates to a gene detection device technical field especially relates to a polychrome fluorescence gene detection device.
Background
In the prior art, the gold standard method for judicial identification, criminal DNA identification of forensic doctors and clinical genetic variation detection is the capillary electrophoresis short-Segment (STR) analysis of nucleic acid, one of the key links of the method is to label primers of different targets with different dyes, then mix and amplify a plurality of PCR primers labeled with fluorescent dyes, denaturize the amplified product, put the denatured product into a capillary electrophoresis gene analyzer, and acquire the information of an original sample through the acquisition of fluorescent signals. The existing commercial capillary electrophoresis gene analyzer uses a 488nm laser or a 505nm laser as an excitation light source to excite a PCR product marked with fluorescence, and the marked fluorescent dye realizes 6-8 colors of fluorescence wavelength through energy transfer after modification.
In practical application, 30-60 fluorescent labeled STR loci need to be detected, so that detection of 6-8 colors cannot meet practical application.
SUMMERY OF THE UTILITY MODEL
In view of the above shortcomings of the prior art, an object of the present invention is to provide a multi-color fluorescent gene detection device, which is used for solving the problem that the multi-color fluorescent gene detection device can only detect 6-8 colors in the prior art. The utility model realizes twice detection by designing the first detection window and the second detection window, and the two detectors can detect 7-10 colors of fluorescence by combining, thereby meeting the detection requirement; the special light path formed by the two detectors can be used in a miniaturized gene detection device, so that the gene detection device is small in size and convenient to carry, can be used for detecting genes on site, and can be applied to identity identification by judicial expertise and DNA identification of criminals.
In order to achieve the above objects and other related objects, the present invention provides a multi-color fluorescent gene detecting device, comprising:
the sample cell is connected with the sample introduction end of the capillary electrophoresis gene analysis device through a first section of capillary, the sample outlet end of the capillary electrophoresis gene analysis device is connected with the buffer tank through a second section of capillary, the first section of capillary is connected with the negative end of the high-voltage electric field, and the second section of capillary is connected with the positive end of the high-voltage electric field; an electrophoresis gel injector is arranged on the second section of capillary tube;
the capillary electrophoresis gene analysis device comprises a third section of capillary, wherein the sample inlet end of the third section of capillary is communicated with the first section of capillary, and the sample outlet end of the third section of capillary is communicated with the second section of capillary; a first detection window and a second detection window are arranged on the third section of capillary tube, a first laser is arranged on one side of the first detection window, and the other side of the first detection window is connected with a CCD (charge coupled device) detector through an optical fiber; and one side of the second detection window is provided with a second laser, and the other side of the second detection window is connected with an APD (avalanche photo diode) detector.
The first detection window and the second detection window are designed, so that two times of detection are realized, and the two detectors are combined to detect 7-10 colors of fluorescence, so that the detection requirement can be met; the special light path formed by the two detectors can be used in a miniaturized gene detection device, so that the gene detection device is small in size and convenient to carry, can be used for detecting genes on site, and can be applied to identity identification by judicial expertise and DNA identification of criminals.
The nucleic acid sample of the first detection window is excited by the first laser (488nm or 505nm), the fluorescence signal is transmitted to the CCD multi-color spectral detector through the optical fiber, an angle of 90 degrees is formed between incidence and emission and receiving, and 6-8-color fluorescence detection can be realized.
The second laser (wavelength is larger than 650nm or smaller than 488nm) is used for marking different dyes according to actual needs, proper laser wavelength is selected to excite the nucleic acid sample of the second detection window, the fluorescent signal is transmitted to an APD detector consisting of optical filters with specific wavelengths through optical fibers, and the corresponding fluorescent signal is collected to realize the collection of additional STR locus information.
An APD detector (APD single-color detector capable of detecting 1-2 colors) and a CCD detector (CCD multi-color light-splitting detector with 6-8 colors).
In an embodiment of the present invention, the electrophoresis gel injector is installed between the junction of the second section of capillary and the high voltage electric field and the capillary electrophoresis gene analyzer.
In an embodiment of the present invention, the distance between the first detection window and the second detection window is 100 to 150 μm.
In an embodiment of the present invention, a distance between the first detection window and the second detection window is 150 μm.
The method is characterized in that a laser is added on the basis of an original 488nm or 505nm laser, the wavelength of the laser is not in the range of 488-650 nm, an APD detector (APD single-color detector capable of detecting 1-2 colors) and a capillary detection window are added at the same time, the detection window is arranged at the position 100-150 mu m (optimally 150 mu m) near the original excitation detection window, the migration rate of electrophoresis is 250 mu m per second, and an electrophoresis peak appears in 10 seconds, namely 2500 mu m, so the detection of the window does not influence the retention time of the electrophoresis peak.
In an embodiment of the present invention, a first focusing mirror is disposed between the first detection window and the first laser; and a second focusing mirror is arranged between the second detection window and the second laser.
In an embodiment of the present invention, the laser wavelength of the first laser is 488nm or 505 nm; the wavelength of the second laser is larger than 650nm or smaller than 488 nm.
In an embodiment of the present invention, the wavelength difference between the laser wavelength of the second laser and the wavelength of the optical filter of the APD detector is 20 to 30 nm.
The wavelength of a filter with a specific wavelength in the APD detector is determined by receiving fluorescence emission wavelength, the difference between the wavelength and the selected laser excitation wavelength is usually 20-30 nm, and the selection of the laser wavelength is determined by the excitation wavelength of a fluorescent dye.
As mentioned above, the utility model discloses a polychrome fluorescence gene testing device has following beneficial effect: the first detection window and the second detection window are designed, so that two times of detection are realized, and the two detectors are combined to detect 7-10 colors of fluorescence, so that the detection requirement can be met; the special light path formed by the two detectors can be used in a miniaturized gene detection device, so that the gene detection device is small in size and convenient to carry, can be used for detecting genes on site, and can be applied to identity identification by judicial expertise and DNA identification of criminals.
Drawings
FIG. 1 is a schematic view of an embodiment of the present invention showing an overall multi-color fluorescent gene detecting apparatus.
Fig. 2 is a schematic diagram of a capillary electrophoresis gene analyzer of a multi-color fluorescent gene detection device according to an embodiment of the present invention.
Description of the element reference numerals
1-a sample cell; 2-capillary electrophoresis gene analysis device, 201-third section of capillary, 202-first detection window, 203-second detection window, 204-first laser, 205-second laser, 206-first focusing mirror, 207-second focusing mirror, 208-CCD detector and 209-APD detector; 3-a first section of capillary; 4-a buffer tank; 5-a second section of capillary; 6-high voltage electric field; 7-electrophoresis gel syringe.
Detailed Description
The following description is provided for illustrative purposes, and other advantages and features of the present invention will become apparent to those skilled in the art from the following detailed description.
Please refer to fig. 1-2. It should be understood that the structure, ratio, size and the like shown in the drawings attached to the present specification are only used for matching with the content disclosed in the specification, so as to be known and read by those skilled in the art, and are not used for limiting the limit conditions that the present invention can be implemented, so that the present invention has no technical essential meaning, and any structure modification, ratio relationship change or size adjustment should still fall within the scope that the technical content disclosed in the present invention can cover without affecting the function that the present invention can produce and the purpose that the present invention can achieve. Meanwhile, the terms such as "upper", "lower", "left", "right", "middle" and "one" used in the present specification are for convenience of description, and are not intended to limit the scope of the present invention, and changes or adjustments of the relative relationship thereof may be made without substantial technical changes, and the present invention is also regarded as the scope of the present invention.
Referring to fig. 1, the present invention provides a multi-color fluorescent gene detection device, which includes:
referring to fig. 1, in a sample cell 1, a sample introduction end of the sample cell 1 is connected with a sample introduction end of a capillary electrophoresis gene analysis device 2 through a first section of capillary tube 3, a sample discharge end of the capillary electrophoresis gene analysis device 2 is connected with a buffer tank 4 through a second section of capillary tube 5, the first section of capillary tube 3 is connected with a negative end of a high-voltage electric field 6, and the second section of capillary tube 5 is connected with a positive end of the high-voltage electric field 6; the electrophoresis gel injector 7 is arranged between the connection part of the second section of capillary 5 and the high-voltage electric field 6 and the capillary electrophoresis gene analysis device 2;
referring to fig. 2, the capillary electrophoresis gene analyzer 2 includes a third section of capillary 201, a sample inlet of the third section of capillary 201 is connected to the first section of capillary 3, and a sample outlet of the third section of capillary 201 is connected to the second section of capillary 5; a first detection window 202 and a second detection window 203 are arranged on the third section of capillary 201, and the distance between the first detection window 202 and the second detection window 203 is 100-150 micrometers; a first laser 204 is arranged on one side of the first detection window 202, a first focusing mirror 206 is arranged between the first detection window 202 and the first laser 204, and the other side of the first detection window 202 is connected with a CCD detector 208 through an optical fiber; a second laser 205 is arranged on one side of the second detection window 203, a second focusing mirror 207 is arranged between the second detection window 203 and the second laser 205, and the other side of the second detection window 203 is connected with an APD detector 209;
the laser wavelength of the first laser 204 is 488nm or 505 nm; the wavelength of the second laser 205 is greater than 650nm or less than 488 nm; the difference between the laser wavelength of the second laser 205 and the wavelength of the optical filter of the APD detector 209 is 20-30 nm.
The working process is as follows: placing the nucleic acid fragment solution marked with the fluorescent dye into a sample cell 1, wherein a sample to be detected, namely nucleic acid, has negative charges, and a second section of capillary 5 is filled into electrophoresis gel through an injector before electrophoresis; the first section of capillary 3 sucks a nucleic acid sample into the capillary by applying a high-voltage electric field 6; under the high-voltage electric field 6, the nucleic acid is electrophoresed from the negative electrode to the positive electrode, the speed of the electrophoresis is inversely proportional to the molecular size of the nucleic acid, after a period of time of electrophoresis, the nucleic acids with different sizes are separated according to the molecular size to form specific zones, when the zones pass through the first detection window 202 and the second detection window 203, the marked fluorescent dye can emit specific fluorescent colors under the condition of laser excitation, and the condition of a detection sample can be known by collecting the fluorescent colors.
And (3) detection flow: nucleic acids with different fragment sizes can pass through the first detection window 202 and the second detection window 203 one by one in the electrophoresis process, when nucleic acid fragments with fluorescent dyes are electrophoresed to the first detection window 202, specific fluorescence can be emitted under the excitation of laser of the first laser 204, the specific fluorescence emitted by the dyes excited by the first laser 204 is transmitted to the CCD detector 208 through optical fibers and is detected and collected by the CCD, and 6-8 colors of fluorescence can be collected at the same time; when the nucleic acid with the fluorescent dye flows to the second detection window 203, the nucleic acid with the fluorescent dye emits specific fluorescence under the excitation of the laser of the second laser 205, the specific fluorescence emitted by the dye excited by the second laser 205 is transmitted to the APD detector 209 through the optical fiber, and is detected and collected by the APD, so that 1-2 color fluorescence can be collected simultaneously, and the two detectors can be combined to detect 7-10 color fluorescence to meet the detection of the gene locus.
To sum up, the utility model discloses a design first detection window 202 and second detection window 203 to realize twice detection, two detectors make up and to detect 7 ~ 10 look fluorescence, thereby can satisfy the detection demand. Therefore, the utility model effectively overcomes various defects in the prior art and has high industrial utilization value.
The above embodiments are merely illustrative of the principles and effects of the present invention, and are not to be construed as limiting the invention. Modifications and variations can be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which may be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (7)
1. A multicolor fluorogenic detection device, comprising:
the device comprises a sample cell (1), wherein the sample cell (1) is connected with a sample introduction end of a capillary electrophoresis gene analysis device (2) through a first section of capillary tube (3), a sample outlet end of the capillary electrophoresis gene analysis device (2) is connected with a buffer tank (4) through a second section of capillary tube (5), the first section of capillary tube (3) is connected with a negative end of a high-voltage electric field (6), and the second section of capillary tube (5) is connected with a positive end of the high-voltage electric field (6); an electrophoresis gel injector (7) is arranged on the second section of capillary (5);
the capillary electrophoresis gene analysis device (2) comprises a third section of capillary (201), the sample inlet end of the third section of capillary (201) is communicated with the first section of capillary (3), and the sample outlet end of the third section of capillary (201) is communicated with the second section of capillary (5); a first detection window (202) and a second detection window (203) are arranged on the third section of capillary (201), a first laser (204) is arranged on one side of the first detection window (202), and the other side of the first detection window (202) is connected with a CCD detector (208) through an optical fiber; and a second laser (205) is arranged on one side of the second detection window (203), and the other side of the second detection window (203) is connected with an APD detector (209).
2. The multicolor fluorogenic gene detection device according to claim 1, wherein: the electrophoresis gel injector (7) is arranged between the connection part of the second section of capillary (5) and the high-voltage electric field (6) and the capillary electrophoresis gene analysis device (2).
3. The multicolor fluorogenic gene detection device according to claim 1, wherein: the distance between the first detection window (202) and the second detection window (203) is 100-150 mu m.
4. The multicolor fluorogenic gene detection device according to claim 1 or 3, wherein: the distance between the first detection window (202) and the second detection window (203) is 150 μm.
5. The multicolor fluorogenic gene detection device according to claim 1 or 3, wherein: a first focusing mirror (206) is arranged between the first detection window (202) and the first laser (204); a second focusing mirror (207) is arranged between the second detection window (203) and the second laser (205).
6. The multicolor fluorogenic gene detection device according to claim 5, wherein: the laser wavelength of the first laser (204) is 488nm or 505 nm; the second laser (205) has a wavelength greater than 650nm or less than 488 nm.
7. The multicolor fluorogenic gene detection device according to claim 1, wherein: the difference between the laser wavelength of the second laser (205) and the wavelength of the optical filter of the APD detector (209) is 20-30 nm.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113125394A (en) * | 2020-01-16 | 2021-07-16 | 上海戈江仪器科技有限公司 | Multicolor fluorescence gene detection device |
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| CN113125394A (en) * | 2020-01-16 | 2021-07-16 | 上海戈江仪器科技有限公司 | Multicolor fluorescence gene detection device |
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