CN212025347U

CN212025347U - Fluorescence detection device

Info

Publication number: CN212025347U
Application number: CN202020114029.3U
Authority: CN
Inventors: 吴保强; 马勇
Original assignee: Hangzhou Bio Gener Technology Co ltd
Current assignee: Hangzhou Bio Gener Technology Co ltd
Priority date: 2020-01-17
Filing date: 2020-01-17
Publication date: 2020-11-27
Anticipated expiration: 2030-01-17

Abstract

The utility model discloses a fluorescence detection device, including the inside base member of placing the sample test tube and the base that is provided with photoelectric detection system, be provided with temperature control system between base member and the base, photoelectric detection system arouses optical module and fluorescence detection module, photoelectric detection system is still including setting up the reciprocating type optical fiber scanning mechanism on the base, the base member sets up in reciprocating type optical fiber scanning mechanism one side, reciprocating type optical fiber scanning mechanism passes through optic fibre and arouses optical module and fluorescence detection module to be connected. The reciprocating type optical fiber scanning mechanism is driven to reciprocate through the motor, fluorescence of the detection reagent is scanned, the excitation light efficiency is favorably improved, higher fluorescence sensitivity and fluorescence intensity are obtained, the fluorescence signal-to-noise ratio is improved, the temperature uniformity of the module is improved, and the cost is reduced.

Description

Fluorescence detection device

Technical Field

The utility model belongs to the technical field of fluorescence detection technique and specifically relates to a fluorescence detection device is related to.

Background

Polymerase Chain Reaction (PCR) is a molecular biotechnology that amplifies target DNA millions of times in a short time by denaturation, renaturation and extension. The real-time fluorescence quantitative PCR is characterized in that a fluorescent group is added into a PCR reaction system, the whole PCR process is monitored in real time by utilizing fluorescence signal accumulation, and the concentration of an unknown template is quantitatively analyzed through a standard curve. At present, the industries such as agriculture and medical treatment are widely applied in a plurality of fields. The current fluorescence detection device can be divided into two detection modes, namely top detection and bottom scanning, for the test tube carrying the product to be detected. 1) The top detects, and this fluorescence detection device's major structure is including being provided with the metal module of test tube inserting groove, and the metal module bottom is confined, excites and detect the part and be located the metal module top. The advantage of this method is that the temperature of the test tube can be controlled by using a universal refrigerating plate or other types of heating plates and radiators; the disadvantage is that the top of the test tube is uneven, and evaporation and condensation can occur, resulting in poor fluorescence repeatability. 2) Bottom scanning, for example, a "fluorescence quantitative PCR detection system based on bottom scanning detection" disclosed in chinese patent document, whose publication number "CN 101328503", includes a temperature-changing metal module, a semiconductor refrigerator and a heat dissipation device, which are sequentially installed into an integral body from top to bottom, and a plurality of test tubes can be inserted into the temperature-changing metal module, the semiconductor refrigerator and the heat dissipation device. The method has the advantages that the position of the detection module is close to the position of the reagent stored in the test tube, the fluorescence signal is strong, and the detection sensitivity is high; the defects are that air convection can be formed up and down on the module, the stability and the uniformity of the metal module are influenced, the refrigerating fins or other types of heating fins and radiators need to be punched, the refrigerating fins or other types of heating fins and radiators with general sizes cannot be adopted, the processing and mounting difficulty is increased, the cost is increased, the bottoms of test tubes are not smooth generally, and the instability of signal acquisition can be caused. Therefore, a new fluorescence detection device is needed to solve the above problems.

Disclosure of Invention

The method aims at the following problems in the prior art: 1. the top detection type fluorescence detection device is far away from a sample substance to be detected, and is easy to interfere with a detection result by phenomena such as evaporation and condensation in a tube; 2. during bottom scanning, the fluorescence detection device cannot be matched with the temperature control device for use, so that a more accurate detection result cannot be obtained; the utility model provides a fluorescence detection device settles through the base member and carries the test tube of treating sample material, and reciprocating type optic fibre scanning mechanism that cooperation temperature control device and set up in the test tube below treats that sample material carries out the side scanning to treating in the test tube to this obtains the better and less fluorescence intensity signal of interference of uniformity.

In order to achieve the above purpose, the utility model adopts the following technical scheme:

the utility model provides a fluorescence detection device, includes that inside places the base member of sample test tube and the base that is provided with photoelectric detection system, photoelectric detection system is including arousing optical module and fluorescence detection module, photoelectric detection system is still including setting up the reciprocating type optical fiber scanning mechanism on the base, reciprocating type optical fiber scanning mechanism sets up in base member one side, it is provided with the inspection hole to correspond optical fiber scanning mechanism one side on the base member, reciprocating type optical fiber scanning mechanism passes through optic fibre and arouses optical module and fluorescence detection module and is connected. The excitation light module is used for emitting excitation light, so that fluorescent components in a sample substance to be detected absorb light energy and enter an excited state, and the fluorescence detection module is used for receiving a fluorescence signal, converting the fluorescence signal into an electric signal and analyzing PCR quantitative data. The vertical arrangement of test tube is inside the base member, and when reciprocating type optical fiber scanning mechanism scanned the test tube, the sample in the test tube was because of the even attached in test tube lateral wall of action of gravity, and the reciprocating type optical fiber scanning mechanism who is on a parallel with the test tube motion can carry out abundant scanning to the sample, and relies on smooth test tube lateral wall can avoid because of detection error and the repeatability that test tube body shape probably appears, effectively improves fluorescence detection's the degree of accuracy.

Preferably, a temperature control device is arranged between the base body and the base, and heat-conducting media are arranged between the temperature control device and the base body. The temperature control device is used for heating and refrigerating the substrate, and accelerating the denaturation-renaturation process of the sample by adjusting the temperature of the sample substance, so that the sample is fully paired with the fluorescent components, and the experimental purpose of obtaining sample component data by detecting the intensity of the fluorescent signal in a PCR quantitative experiment is achieved; the heat-conducting medium can buffer the temperature when the temperature control device heats the sample test tube, and can also play a role in accelerating heat dissipation in the cooling process.

Preferably, the bottom of the temperature control device is provided with a heat dissipation module, the heat dissipation module is provided with a plurality of heat dissipation fins, and the heat dissipation module is connected with the base. The heat dissipation module is used for assisting in dissipating redundant heat.

Preferably, the reciprocating optical fiber scanning mechanism comprises a scanning track arranged parallel to the base body, a motor and a synchronizing mechanism connected with the motor, wherein a signal interaction head is arranged on the synchronizing mechanism, and the signal interaction head reciprocates on the scanning track. Reciprocating type optical fiber scanning mechanism is used for inciting somebody to action and detecting its inside sample material along sample test tube lateral wall to export fluorescence signal through optic fibre, signal interaction head passes through the belt drive uniform motion on the scanning track under motor drive, improves the experimental result degree of accuracy.

Preferably, a placing groove is formed in the base body, the sample test tube is placed in the placing groove, the placing groove is parallel to the base, the detection hole is formed in the middle of the placing groove, the distance between the optical fiber and the detection hole can be adjusted to be minimum by the orifice of the detection hole towards the reciprocating type optical fiber scanning mechanism, the excitation light efficiency is favorably improved, higher fluorescence sensitivity and fluorescence signal intensity are obtained, and the stability is improved.

Preferably, the longitudinal section of the containing groove is in a conical structure, and the sample test tube is connected with the containing groove in an inserting and combining manner. The arrangement groove of back taper structure is used for cooperating the back taper test tube for test tube and base member fully laminate in the experimentation, can ensure going on of reaction step more rapidly high-efficient, also can ensure that photoelectric detection system can carry out abundant scanning to the in vitro sample under the testing process.

Preferably, the optical fiber is a Y-shaped structure including a branching end and a bundling end, the bundling end includes an excitation portion and a receiving portion, the branching end includes an input end communicated with the excitation portion and an output end communicated with the receiving portion, the excitation optical module is connected with the excitation portion through the input end, the fluorescence detection module is connected with the receiving portion through the output end, and the bundling end of the optical fiber is disposed on the signal interaction head. The Y-shaped optical fiber is connected with the excitation optical module and the fluorescence detection module at the branch ends respectively, and the beam collecting end of the Y-shaped optical fiber is connected with the signal interaction head, so that the hidden danger caused by mutual winding of the separated optical fibers during the movement of the signal limiting interaction head is avoided. The excitation portion is used for evenly shining the exciting light that the excitation optical module produced in the sample test tube, and correspondingly, the receiving unit then can be with the fluorescence intensity signal input fluorescence detection module after the excitation and convert the signal of telecommunication into, and then accomplish the collection of experimental data.

Therefore, the utility model discloses following beneficial effect has: (1) the test tube carrying the sample substance to be detected is arranged on the substrate, the motor drives the reciprocating type optical fiber scanning mechanism to reciprocate, and the fluorescence of the detection reagent is scanned, so that the excitation efficiency is improved, higher fluorescence sensitivity and fluorescence intensity are obtained, the fluorescence signal-to-noise ratio is improved, the temperature uniformity of the module is improved, and the cost is reduced; (2) the sample in the test tube is uniformly attached to the side wall of the test tube under the action of gravity, the reciprocating optical fiber scanning mechanism moving parallel to the test tube can fully scan the sample, and the smooth side wall of the test tube can avoid possible scanning errors caused by the shape of the test tube body, so that the accuracy of fluorescence detection is effectively improved; (3) the temperature control device is used for heating and cooling the sample test tube, and the denaturation-renaturation process of the sample is accelerated by adjusting the temperature of the sample substance, so that the sample test tube is fully paired with the fluorescent component, and the aim of obtaining sample component data by detecting the intensity of the fluorescent signal in a PCR quantitative experiment is fulfilled; (4) simple structure makes the device simple operation through the mutual first motion of motor drive signal, and the testing result of side formula testing mode is comparatively accurate.

Drawings

Fig. 1 is a schematic structural view (top view) of the present invention.

Fig. 2 is a vertical sectional view of the base body of fig. 1.

Fig. 3 is a schematic structural view of the optical fiber of fig. 1.

In the figure: 1. the device comprises a base body, 11, a placement groove, 12, a detection hole, 2, a photoelectric detection system, 21, an excitation optical module, 22, a fluorescence detection module, 3, a base, 4, a reciprocating type optical fiber scanning mechanism, 41, a scanning track, 42, a motor, 5, an optical fiber, 51, an input end, 52, an output end, 6, a temperature control device, 61, a heat conducting medium, 7, a heat dissipation module, 71, a heat dissipation sheet, 8 and a signal interaction head.

Detailed Description

The present invention will be further described with reference to the accompanying drawings and the detailed description.

Example 1

As shown in fig. 1, a fluorescence detection device, including the inside base member 1 of placing the sample test tube and the base 3 that is provided with photoelectric detection system 2, photoelectric detection system 2 includes exciting light module 21 and fluorescence detection module 22, photoelectric detection system 2 is still including reciprocating type optical fiber scanning mechanism 4 that sets up on base 3, reciprocating type optical fiber scanning mechanism 4 sets up in base member 1 one side, it is provided with inspection hole 12 to correspond optical fiber scanning mechanism one side on the base member, reciprocating type optical fiber scanning mechanism 4 is connected with exciting light module 21 and fluorescence detection module 22 through optic fibre 5. The excitation light module is used for emitting excitation light, so that fluorescent components in a sample substance to be detected absorb light energy and enter an excited state, and the fluorescence detection module is used for receiving a fluorescence signal, converting the fluorescence signal into an electric signal and analyzing PCR quantitative data. The vertical arrangement of test tube is inside the base member, and when reciprocating type optical fiber scanning mechanism scanned the test tube, the sample in the test tube was because of the even attached in test tube lateral wall of action of gravity, and the reciprocating type optical fiber scanning mechanism who is on a parallel with the test tube motion can carry out abundant scanning to the sample, and relies on smooth test tube lateral wall can avoid because of detection error and the repeatability that test tube body shape probably appears, effectively improves fluorescence detection's the degree of accuracy.

A temperature control device 6 is arranged between the base body and the base, and heat-conducting media 61 are arranged between the temperature control device and the base body. The temperature control device is used for heating and refrigerating the substrate, and the denaturation-renaturation process of the sample, namely the rapid amplification process of the fluorescent components, is accelerated by adjusting the temperature of the sample substances, so that the DNA and the fluorescent components are fully matched, and the experimental purpose of obtaining sample component data by detecting the intensity of a fluorescent signal in a PCR quantitative experiment is achieved; the heat-conducting medium can buffer the temperature when the temperature control device heats the sample test tube, and can also play a role in accelerating heat dissipation in the cooling process. In this embodiment, the heat conducting medium is an air layer, and an aluminum layer or a silver layer having a high heat conductivity may be used. The bottom of the temperature control device is provided with a heat dissipation module 7, the heat dissipation module is provided with a plurality of heat dissipation fins 71, and the heat dissipation module is connected with the base. The heat dissipation module further improves the renaturation process of the sample test tube and effectively accelerates the experiment progress. In this embodiment, the temperature control device employs a micro-heater, which denatures the sample at high temperature; adopt heat radiation module to adopt the gilled radiator of metal, carry out effective heat dissipation to the sample test tube through front and back formula ventilation structure.

The reciprocating optical fiber scanning mechanism comprises a scanning track 41, a motor 42 and a signal interaction head 8 connected with the motor, wherein the signal interaction head is sleeved on the scanning track and reciprocates on the scanning track. Reciprocating type optical fiber scanning mechanism is used for inciting somebody to action and detecting its inside sample material along sample test tube lateral wall to pass through optical fiber output with fluorescence signal, signal interaction head sliding connection is in scanning track and drive the uniform motion through the belt under motor drive, can carry out the abundant scanning of no dead angle formula to the sample test tube, improves the experiment result degree of accuracy. The top of the signal interaction head is provided with an excitation part and a receiving part, the excitation part and the receiving part are integrated into a whole, the excitation optical module is connected with the excitation part through an optical fiber, and the receiving part is connected with the fluorescence detection module through an optical fiber. The excitation part is used for uniformly irradiating the excitation light generated by the excitation light module on the sample test tube, correspondingly, the receiving part can input the excited fluorescence intensity signal into the fluorescence detection module, and then the collection of experimental data is completed.

As shown in fig. 2, a placing groove is vertically arranged in the base, the sample test tube is placed in the placing groove, the base is arranged in parallel with the placing groove, so that a detection hole with a diameter smaller than that of the placing groove is arranged at the bottom of the placing groove, the notch of the detection hole is arranged towards the reciprocating type optical fiber scanning mechanism, and in the embodiment, the center of the detection hole is 2-5mm higher than the bottom of the placing groove. The distance between the optical fiber and the detection hole can be adjusted to be minimum, so that the excitation light efficiency is improved, higher fluorescence sensitivity and fluorescence signal intensity are obtained, and the stability is improved. Be provided with arrangement groove 11 in the base member 1, the sample test tube sets up in arrangement groove 11, arrangement groove 11 sets up in a parallel with base 3, inspection hole 12 sets up in arrangement groove 11 middle part position under bias, the drill way orientation reciprocating type optical fiber scanning mechanism 4 setting of inspection hole 12. As shown in fig. 3, the optical fiber 5 has a Y-shaped structure including a bifurcated end and a bundled end, the bundled end includes an excitation portion and a receiving portion, the bifurcated end includes an input end 51 communicated with the excitation portion and an output end 52 communicated with the receiving portion, the excitation optical module 21 is connected with the excitation portion through the input end 51, the fluorescence detection module 22 is connected with the receiving portion through the output end 52, and the bundled end of the optical fiber 5 is disposed on the signal interaction head 8. The Y-shaped optical fiber is connected with the excitation optical module and the fluorescence detection module at the branch ends respectively, and the beam collecting end of the Y-shaped optical fiber is connected with the signal interaction head, so that the hidden danger caused by mutual winding of the separated optical fibers during the movement of the signal limiting interaction head is avoided. The longitudinal section of the containing groove is of a conical structure, and the sample test tube is connected with the containing groove in an inserting mode. The arrangement groove of back taper structure is used for cooperating the back taper test tube for test tube and base member fully laminate in the experimentation, can ensure going on of reaction step more rapidly high-efficient, also can ensure that photoelectric detection system can carry out abundant scanning to the in vitro sample under the testing process. More specifically, the excitation light module comprises a first lens assembly and an LED lamp, wherein an excitation light filter is arranged in the first lens assembly; the fluorescence detection module comprises a second lens assembly and a photoelectric detector, wherein a receiving optical filter is arranged in the second lens assembly, and the second lens assembly is arranged between the optical fiber and the photoelectric detector. The LED lamp is used for outputting optical signals, the first lens component and the optical fiber are matched to complete excitation of fluorescent components, the second lens component is used for converting the fluorescent signals of the excited fluorescent components into light rays which can be identified by the photoelectric detector, and the photoelectric detector converts the received optical signals into electric signals and outputs the fluorescent detection device disclosed by the invention to complete fluorescent detection.

Before the device is used, the sample substance in the test tube is firstly determined to be added with the auxiliary components of PCR experiment such as fluorescent component, random primer, Taq enzyme and the like, and then a PCR reaction system is formed. The fluorescent components used in the fluorescent quantitative PCR can be divided into two types: the fluorescent probe and the fluorescent dye are mainly SYBR fluorescent dye, excessive SYBR fluorescent dye is added into a PCR reaction system, the SYBR fluorescent dye is specifically doped into a DNA double chain and then emits a fluorescent signal, and SYBR dye molecules which are not doped into the DNA double chain cannot emit any fluorescent signal, so that the fluorescent signal is increased completely synchronously with the increase of a PCR product. Then the sample test tube is inserted into the placing groove of the base body, and the base body is fixed on the outer side of the base body through the screw, so that the sample test tube in the placing groove can be ensured to be in a position parallel to the photoelectric detection system; the formal experiment was started: a) denaturation: starting a temperature control device to heat the sample test tube at high temperature, controlling the temperature at 90-95 ℃, and breaking hydrogen bonds of a double-stranded DNA template in a sample substance under the action of heat to form single-stranded DNA; b) annealing: the temperature of the sample test tube is reduced to 45-60 ℃ by matching the heat dissipation module with the temperature control device, and the process aims to combine the fluorescent component with the DNA template to form a local double chain; c) extension: controlling the temperature to 72 ℃, using dNTP as raw material under the action of Taq enzyme, extending from the 5 'end → 3' end of the random primer, synthesizing a DNA chain complementary with the template, wherein SYBR fluorescent dye is specifically doped into the DNA double chain at the moment, and finishing the treatment of the sample substance. Three of the steps are cycles, each cycle doubling the DNA content. Starting to detect after the substance treatment is finished; the motor drive signal interaction head moves on a scanning track at a constant speed, firstly, an excitation ray is generated by an excitation light module, fluorescence components in a sample test tube are excited by an excitation part, so that the fluorescence components absorb light energy and are converted into an excited state, then, the excitation light module is closed, fluorescence signals formed by excitation are scanned and collected by a receiving part, and the fluorescence signals are transmitted to a signal detection module through optical fibers, so that the experiment is finished. In this embodiment, the excitation light module includes a plurality of lenses, an excitation filter and a monochromatic LED lamp, and the signal detection module includes a plurality of lenses, a detection filter, a detector and an optical circuit thereof. An excitation light source penetrates into the optical fiber through the primary lens, the excitation optical filter and the secondary lens, the test tube reagent is excited hole by hole through optical fiber conduction, reflected fluorescence penetrates into the optical fiber hole by hole and is conducted to the optical detection unit through the receiving optical fiber, the fluorescence sequentially penetrates through the primary lens, the receiving optical filter and the secondary lens of the optical detection unit and irradiates on the detector, and the detector converts the fluorescence into an electric signal for analysis and processing.

In addition to the above embodiments, the technical features of the present invention can be re-selected and combined within the scope of the present invention as disclosed in the claims and the specification to constitute new embodiments, which can be realized by those skilled in the art without any creative work, so that the embodiments of the present invention not described in detail should be regarded as specific embodiments of the present invention and are within the protection scope of the present invention.

Claims

1. The utility model provides a fluorescence detection device, characterized by, including inside base member (1) of placing the sample test tube and base (3) that are provided with photoelectric detection system (2), photoelectric detection system (2) are including exciting light module (21) and fluorescence detection module (22), photoelectric detection system (2) are still including reciprocating type fiber scanning mechanism (4) of setting on base (3), reciprocating type fiber scanning mechanism (4) set up in base member (1) one side, it is provided with inspection hole (12) to correspond fiber scanning mechanism one side on the base member, reciprocating type fiber scanning mechanism (4) are connected with exciting light module (21) and fluorescence detection module (22) through optical module (5).

2. The fluorescence detection device according to claim 1, wherein a temperature control device (6) is disposed between the base body (1) and the base (3), and a heat conducting medium (61) is disposed between the temperature control device (6) and the base body (1) and the base (3).

3. The fluorescence detection device according to claim 2, wherein a heat dissipation module (7) is arranged between the temperature control device (6) and the reciprocating fiber scanning mechanism (4), a plurality of heat dissipation fins (71) are arranged on the heat dissipation module (7), and the heat dissipation module (7) is connected with the base (3).

4. The fluorescence detecting device of claim 1, wherein the reciprocating fiber scanning mechanism comprises a scanning track parallel to the substrate, a motor, and a synchronizing mechanism connected to the motor, the synchronizing mechanism having a signal interacting head, the signal interacting head reciprocating on the scanning track.

5. The fluorescence detection device according to claim 4, wherein the optical fiber (5) has a Y-shaped structure including a branch end and a bundle end, the bundle end includes an excitation portion and a receiving portion, the branch end includes an input end (51) communicated with the excitation portion and an output end (52) communicated with the receiving portion, the excitation module (21) is connected with the excitation portion through the input end (51), the fluorescence detection module (22) is connected with the receiving portion through the output end (52), and the bundle end of the optical fiber (5) is disposed on the signal interaction head (8).

6. The fluorescence detection device according to claim 4, wherein the synchronous mechanism comprises a synchronous pulley arranged on the scanning track and a synchronous belt connected to the synchronous pulley, the signal interaction head (8) is connected to the synchronous belt, and the synchronous pulley is connected to the motor.

7. The fluorescence detection device according to claim 4, wherein an excitation portion (81) and a receiving portion (82) are arranged on the top of the signal interaction head (8), the excitation module (21) is connected with the excitation portion (81) through an optical fiber (5), and the receiving portion (82) is connected with the fluorescence detection module (22) through the optical fiber (5).

8. The fluorescence detection device according to any one of claims 1 to 7, wherein a placement groove (11) is provided in the base body (1), the placement groove (11) is arranged parallel to the base (3), the detection hole (12) is arranged in the middle of the placement groove (11), and the opening of the detection hole (12) is arranged towards the reciprocating type optical fiber scanning mechanism (4).