CN113092424A - Novel high-flux biochip analyzer - Google Patents
Novel high-flux biochip analyzer Download PDFInfo
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- CN113092424A CN113092424A CN202110291642.1A CN202110291642A CN113092424A CN 113092424 A CN113092424 A CN 113092424A CN 202110291642 A CN202110291642 A CN 202110291642A CN 113092424 A CN113092424 A CN 113092424A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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Abstract
The invention discloses a novel high-throughput biochip analyzer, and relates to the technical field of biochip analyzers. The invention comprises a chip bearing structure, an excitation light filter disc and a detection light filter disc which are arranged in a shell from bottom to top; an LED white light source is arranged on one side of the excitation filter assembly in the outer shell; white light emitted by the LED white light source is filtered by the excitation filter and then reflected and focused on the detection chip on the chip bearing structure; fluorescent signals generated by absorbing external energy by fluorescein molecules in PCR products in the micro-holes of the detection chip are acquired by a fluorescent detector after passing through a detection optical filter. The LED white light source provides a light source, the excitation filter disc and the detection light filter disc support the free combination of the 6-color excitation light channel and the 6-color detection light channel, multiple types of fluorescence can be detected, the detection range is wide, the resolution ratio is high, the accuracy is good, and the instrument performance is more stable; meanwhile, the driving device drives the tray frame to enter and exit the strip-shaped holes, so that the detection chip can be conveniently loaded and replaced.
Description
Technical Field
The invention belongs to the technical field of biochip analyzers, and particularly relates to a novel high-throughput biochip analyzer.
Background
As a common molecular biology experiment, the biochip is widely applied to a plurality of fields such as gene research, drug research, disease diagnosis, pathogenic microorganism detection and the like. The main principle is that the molecular hybridization principle is adopted, the fluorescence molecule mark is carried out on a sample to be detected, a biological analyzer is used for detecting a fluorescence signal in a reaction system, and after the fluorescence signal is recorded and analyzed, the result is used for qualitative, semi-quantitative or quantitative analysis of a target detection object.
At present, most of biochip analyzers are used for realizing dual-wavelength fluorescence detection by adopting full-mechanical two-dimensional scanning, a dual-light source and a dual-photodetector. The device comprises a double light source and a double photoelectric detector which are staggered according to a certain angle, wherein a laser beam emitted by the light source reaches a certain point on a biochip through a lens, a spectroscope and a prism, a fluorescent dye molecule on the position of the point is excited to emit fluorescence, the fluorescence passes through the prism, is reflected to a reflector by the spectroscope, and then is converged by a condenser to reach the photoelectric detector. The two-dimensional scanning of the chip is realized by driving the prism to move in the X direction by using a linear driver and driving the biochip to move in the Y direction by using a stepping motor. This biochip analyzer has the following disadvantages:
1) the scanning speed of the device is limited by the operating frequency of a linear driver, the analysis time of a biochip is long, the vibration is large, the scanning inertia is large, the reciprocating stroke is long, and the stroke of a corresponding prism is also long, so that the change of a focusing spot of a laser beam is large, and the resolution of an instrument is influenced;
2) the numerical aperture of the condenser is limited by a mechanical structure, so that the analysis sensitivity of the instrument is influenced;
3) the simultaneous scanning imaging of the dual-wavelength fluorescence is realized by adopting the dual light source and the dual detector, so that the structure of the whole instrument is complex, and the performance of the instrument is influenced due to crosstalk in the scanned image;
4) the method is limited by the problem of chip design, the sample detection flux and the number of detection sites are low, and the requirements of high-flux and rapid detection on clinic are difficult to adapt.
To solve the above problems, the present invention provides a novel high-throughput biochip analyzer.
Disclosure of Invention
The invention aims to provide a novel high-flux biochip analyzer, which provides a light source through an LED white light source, is provided with an excitation filter disc and a detection light filter disc, and supports the free combination of 6-color excitation light channels and 6-color detection light channels; the method realizes high-efficiency detection of multiple fluorescence and improves the detection range and resolution.
In order to solve the technical problems, the invention is realized by the following technical scheme:
the invention relates to a novel high-flux biochip analyzer, which comprises an outer shell, wherein a chip bearing structure, an excitation filter disc and a detection light filter disc are arranged in the outer shell from bottom to top; an LED white light source is arranged on one side of the excitation optical filter component in the shell;
the excitation filter disc comprises a first support ring and an excitation filter; six groups of excitation light filters are annularly and uniformly distributed on the peripheral side of the first supporting ring; the LED white light source corresponds to the excitation filter in position; the LED white light source emits white light, the white light is filtered by the excitation optical filter and then is reflected and focused on the detection chip on the chip bearing structure;
the detection light filter disc comprises a second support ring and a detection light filter; six groups of detection optical filters are annularly and uniformly arranged on the periphery of the second support ring; the second support ring and the second support ring are coaxial, and the detection optical filter corresponds to the excitation optical filter in the upper and lower positions;
a fluorescent detector is arranged at the top in the outer shell and corresponds to the detection optical filter in position; fluorescent signals generated by absorbing external energy by fluorescein molecules in PCR products in the micro-holes of the detection chip pass through the detection optical filter, are acquired by the fluorescent detector and are transmitted to the master control analyzer.
As a preferable technical solution, the first dichroic mirror is installed obliquely on the periphery of the first support ring and located inside the excitation filter; the LED white light source emits white light, the white light is filtered by the excitation optical filter and then reflected by the first dichroic mirror to irradiate the detection chip.
As a preferable technical solution, a second dichroic mirror is installed on the periphery of the second support ring below the detection filter; the first dichroic mirror and the second dichroic mirror which correspond to the positions are parallel; and the fluorescent signal is reflected by the first dichroic mirror and the second dichroic mirror and then irradiated onto the detection filter.
As a preferred technical solution, an amplifier is installed inside the outer shell and between the excitation filter disc and the chip carrying structure; the light beam reflected by the first dichroic mirror is amplified by an amplifier and then irradiates a detection chip.
As a preferred technical solution, the chip carrying structure includes a carrying seat and a cover plate; the detection chip is placed on the bearing seat; the cover plate covers and bears the weight of on the seat.
As a preferred technical scheme, a tray frame and a chip tray are movably arranged on the bearing seat; the chip tray is used for matching and mounting a detection chip; the chip tray is installed on the tray frame in a matching mode.
As a preferred technical scheme, a strip-shaped hole for the tray frame to enter and exit is formed in the side surface of the outer shell; the tray frame is driven by a driving device to enter and exit the strip-shaped holes; the driving device is electrically connected with the master control analyzer.
As a preferred technical scheme, a first servo motor is installed at the top in the outer shell; the first servo motor drives the excitation filter disc to rotate; the first servo motor is electrically connected with the main control analyzer.
As a preferred technical scheme, a second servo motor is installed on the inner side wall of the outer shell; the second servo motor drives the detection light filter disc to rotate; the first servo motor is electrically connected with the main control analyzer.
One aspect of the present invention has the following advantageous effects:
1. the invention provides a light source through the LED white light source, simultaneously excites the filter disc and detects the light filter disc to support the free combination of the 6-color excitation light channel and the 6-color detection light channel, detects multiple kinds of fluorescence, and has the advantages of wide detection range, high resolution, good accuracy and more stable instrument performance.
2. The driving device drives the tray frame to enter and exit the strip-shaped holes, so that the detection chip can be conveniently loaded and replaced; the detection system is as small as 33nL, and the analysis sensitivity is higher; meanwhile, scanning of more than two million data points can be carried out within 8 hours, and the detection flux is high.
Of course, it is not necessary for any product in which the invention is practiced to achieve all of the above-described advantages at the same time.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic diagram of the external structure of a novel high throughput biochip analyzer according to the present invention.
FIG. 2 is a schematic diagram of the structure of the internal components of a novel high throughput biochip analyzer according to the present invention.
In the drawings, the components represented by the respective reference numerals are listed below:
the detection device comprises a shell 1, a 2-LED white light source, a 31-first supporting ring, a 32-excitation optical filter, a 33-first dichroic mirror, a 41-detection chip, a 42-bearing seat, a 421-tray frame, a 422-chip tray, a 43-cover plate, a 51-second supporting ring, a 52-detection optical filter, a 53-second dichroic mirror, a 6-fluorescence detector and a 7-amplifier.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "open," "upper," "middle," "length," "inner," and the like are used in an orientation or positional relationship for convenience in describing the present invention and for simplicity of description, and do not indicate or imply that the referenced components or elements must have a particular orientation, be constructed and operated in a particular orientation, and therefore should not be construed as limiting the present invention.
Referring to fig. 1-2, the present invention is a novel high-throughput biochip analyzer, including an outer housing 1, a chip carrying structure, an excitation filter disk and a detection light filter disk are installed inside the outer housing 1 from bottom to top; an LED white light source 2 is arranged on one side of the excitation filter assembly in the outer shell 1; specifically, the LED white light source 2 adopts a white light semiconductor LED light source, has a wide spectral range, is used together with a matched Openarray chip, and can read fluorescence signals of 12288 targets in total from 4 chips in about 5 minutes; has the advantages of high flux, easy operation and high speed.
The excitation filter disk comprises a first support ring 31 and an excitation filter 32; six groups of excitation filters 32 are annularly and uniformly arranged on the peripheral side of the first support ring 31; the LED white light source 2 corresponds to the excitation filter 32 in position; the white light emitted by the LED white light source 2 is filtered by the excitation filter 32 and then reflected and focused on the detection chip 41 on the chip bearing structure; specifically, first dichroic mirror 33 is obliquely mounted on the inner side of excitation filter 32 on the peripheral side of first support ring 31; the white light emitted by the LED white light source 2 is filtered by the excitation filter 32 and then reflected by the first dichroic mirror 33 to irradiate the detection chip 41; further, an amplifier 7 is arranged between the excitation filter disc and the chip bearing structure inside the outer shell 1; the light beam reflected by the first dichroic mirror 33 is amplified by the amplifier 7 and then irradiates the detection chip 41, and the amplification of the amplifier 7 enlarges the area of the detection chip 41 irradiated by the light beam, thereby being beneficial to forming more effective fluorescent signals.
The detection light filter wheel comprises a second support ring 51 and a detection light filter 52; six groups of detection optical filters 52 are annularly and uniformly arranged on the peripheral side of the second support ring 51; the second support ring 51 and the second support ring 51 are coaxial, and the detection filter 52 corresponds to the excitation filter 32 in the upper and lower positions; specifically, a second dichroic mirror 53 is mounted on the periphery of the second support ring 51 below the detection filter 52; the first dichroic mirror 33 corresponding to the position is parallel to the second dichroic mirror 53; the fluorescent signal is reflected by the first dichroic mirror 33 and the second dichroic mirror 53 and then irradiated onto the detection filter 52; in fact, the fluorescence signal can be vertically irradiated on the detection filter 52 after being reflected by the first dichroic mirror 33 and the second dichroic mirror 53 which are parallel to each other, so as to be conveniently acquired by the fluorescence detector 6.
A fluorescent detector 6 is arranged at the top in the outer shell 1, and the position of the fluorescent detector 6 corresponds to that of the detection filter 52; fluorescent signals generated by absorbing external energy by fluorescein molecules in PCR products in the micro-holes of the detection chip 41 pass through the detection optical filter 52, are acquired by the fluorescent detector 6 and are transmitted to the master control analyzer; specifically, the fluorescence detector 6 is a high-definition CCD camera, and is configured to capture an image of the fluorescence signal irradiated on the detection filter 52 and transmit the image to the main control analyzer for further analysis processing.
As a more preferred embodiment, the chip carrying structure includes a carrying seat 42 and a cover plate 43; the detection chip 41 is placed on the bearing seat 42; the cover plate 43 is covered on the bearing seat 42; specifically, the cover plate 43 is provided with a plane light-transmitting mirror corresponding to the detection chip 41, and is used for light beams to pass through and irradiate on the detection chip 41; here, the cover plate 43 covers the detection chip 41 and the bearing seat 42, so as to avoid the influence on the light beam irradiation caused by the hot air flow generated by heating in the process of generating the fluorescent signal, and is convenient and practical.
In a more preferred embodiment, the carrier 42 is movably mounted with a tray frame 421 and a chip tray 422; the chip tray 422 is used for matching and mounting the detection chip 41; the chip tray 422 is matched and installed on the tray frame 421; in addition, the side surface of the outer shell 1 is provided with a strip-shaped hole 11 for the inlet and outlet of the tray frame 421; the tray frame 421 is driven by a driving device to enter and exit the strip-shaped holes 11; the driving device is electrically connected with the master control analyzer; in fact, a corresponding button is arranged outside the outer shell 1, and after the corresponding button is pressed down, the main control analyzer controls the driving device to drive the tray frame 421 to enter and exit the strip-shaped hole 11, so that the detection chip 41 is conveniently loaded and replaced; specifically, various detection chips 41 can be replaced conveniently, the detection system is as small as 33nL, and the analysis sensitivity is higher.
As a more preferable embodiment, a first servo motor is installed at the top in the outer shell 1; the first servo motor drives the excitation filter disc to rotate; the first servo motor is electrically connected with the master control analyzer; a second servo motor is arranged on the inner side wall of the outer shell 1; the second servo motor drives the detection light filter disc to rotate; the first servo motor is electrically connected with the master control analyzer; in fact, the first servo motor drives the excitation light filter disc to rotate, and the second servo motor drives the detection light filter disc to rotate, so that free combination of multiple lights is realized, free combination of 6-color excitation light channels and 6-color detection light channels is supported, up to 21 fluorescence lights are detected, the detection range is wide, the resolution ratio is high, the accuracy is good, and the instrument performance is more stable; in fact, scans of more than 2,000,000 data points can be made 8 hours a day, with high detected flux.
The working principle of the invention is as follows: mixing the probe marked with fluorescein with template DNA, high-temp. denaturation, low-temp. renaturation and proper-temp. extension chain polymerase reaction (PCR), cutting the probe complementary-paired with template DNA in the course of PCR amplification, and dissociating fluorescein in the reaction system. And uniformly injecting the PCR product subjected to the PCR reaction into micropores of the Openarray chip by using the cypress distribution principle, and placing the Openarray chip in a biochip analyzer. Excitation light generated by an LED white light source 2 of the biochip analyzer is filtered and reflected and then focused on a detection chip 41, and fluorescein molecules in PCR products in micropores of the chip become unstable excited states after absorbing external energy and then jump to a stable ground state, so that a fluorescence signal is generated. The generated specific fluorescent signal passes through the detection optical filter, is received by the fluorescent detector 6 and is stored. And analyzing the fluorescent signal so as to complete the detection and analysis of the detected object.
When the fluorescent detector is practically used, the LED white light source 2 provides a light source, the excitation filter disc and the detection light filter disc support the free combination of the 6-color excitation light channel and the 6-color detection light channel, 21 kinds of fluorescence can be detected, the detection range is wide, the resolution ratio is high, the accuracy is good, and the instrument performance is more stable. In addition, the driving device drives the tray frame 421 to enter and exit the strip-shaped holes 11, so that the detection chip 41 can be conveniently loaded and replaced; the detection system is as small as 33nL, and the analysis sensitivity is higher.
In the description herein, references to the description of "one embodiment," "an example," "a specific example" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (9)
1. A novel high-throughput biochip analyzer, comprising an outer housing (1), characterized in that:
a chip bearing structure, an excitation filter disc and a detection light filter disc are arranged in the outer shell (1) from bottom to top; an LED white light source (2) is arranged on one side of the excitation filter assembly in the shell (1);
the excitation filter disc comprises a first support ring (31) and an excitation filter (32); six groups of excitation filters (32) are annularly and uniformly arranged on the peripheral side of the first support ring (31);
the LED white light source (2) corresponds to the excitation filter (32) in position; the LED white light source (2) emits white light, the white light is filtered by the excitation filter (32) and then is reflected and focused on a detection chip (41) on the chip bearing structure;
the detection light filter disc comprises a second support ring (51) and a detection light filter (52); six groups of detection optical filters (52) are annularly and uniformly arranged on the peripheral side of the second support ring (51); the second support ring (51) and the second support ring (51) are coaxial, and the detection filter (52) corresponds to the excitation filter (32) in the vertical position;
a fluorescent detector (6) is arranged at the top in the outer shell (1), and the fluorescent detector (6) corresponds to the detection optical filter (52) in position;
fluorescent signals generated by absorbing external energy by fluorescein molecules in PCR products in the chip micropores of the detection chip (41) pass through the detection optical filter (52), are acquired by the fluorescent detector (6) and are transmitted to the master control analyzer.
2. The analyzer, according to claim 1, characterized in that the first dichroic mirror (33) is installed obliquely inside the excitation filter (32) around the first supporting ring (31); the LED white light source (2) emits white light, the white light is filtered by the excitation filter (32), and then is reflected by the first dichroic mirror (33) to irradiate the detection chip (41).
3. The new type of high throughput biochip analyzer according to claim 2, wherein the second dichroic mirror (53) is mounted around the second support ring (51) below the detection filter (52); the first dichroic mirror (33) and the second dichroic mirror (53) which correspond in position are parallel; the fluorescence signal is reflected by the first dichroic mirror (33) and the second dichroic mirror (53) and then irradiated onto the detection filter (52).
4. A new high throughput biochip analyzer according to claim 2 or 3, wherein an amplifier (7) is mounted inside the outer housing (1) between the excitation filter wheel and the chip carrying structure; the light reflected by the first dichroic mirror (33) is amplified by an amplifier (7) and then irradiates a detection chip (41).
5. The new high throughput biochip analyzer of claim 4, wherein the chip carrying structure comprises a carrying seat (42) and a cover plate (43); the detection chip (41) is placed on the bearing seat (42); the cover plate (43) is covered on the bearing seat (42).
6. The analyzer for biochip with high throughput as claimed in claim 5, wherein the tray holder (421) and the chip tray (422) are movably mounted on the carrier (42); the chip tray (422) is used for matching and mounting the detection chip (41); the chip tray (422) is matched and installed on the tray frame (421).
7. The analyzer is characterized in that the side of the outer casing (1) is provided with a strip-shaped hole (11) for the inlet and outlet of a tray rack (421); the tray frame (421) is driven by a driving device to enter and exit the strip-shaped holes (11); the driving device is electrically connected with the master control analyzer.
8. The analyzer, according to claim 1 or 7, characterized in that the first servo motor is installed on the top inside the outer casing (1); the first servo motor drives the excitation filter disc to rotate; the first servo motor is electrically connected with the main control analyzer.
9. The analyzer, according to claim 8, characterized in that a second servo motor is installed on the inner side wall of the outer casing (1); the second servo motor drives the detection light filter disc to rotate; the first servo motor is electrically connected with the main control analyzer.
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