[go: up one dir, main page]

CN113969238A - A Portable Visual Imaging System for Fluorescence Detection of Gene Amplification - Google Patents

A Portable Visual Imaging System for Fluorescence Detection of Gene Amplification Download PDF

Info

Publication number
CN113969238A
CN113969238A CN202111344210.9A CN202111344210A CN113969238A CN 113969238 A CN113969238 A CN 113969238A CN 202111344210 A CN202111344210 A CN 202111344210A CN 113969238 A CN113969238 A CN 113969238A
Authority
CN
China
Prior art keywords
reaction
fluorescence detection
array
imaging system
sample
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202111344210.9A
Other languages
Chinese (zh)
Inventor
雷荣
吴品珊
张永江
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Chinese Academy of Inspection and Quarantine CAIQ
Original Assignee
Chinese Academy of Inspection and Quarantine CAIQ
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Chinese Academy of Inspection and Quarantine CAIQ filed Critical Chinese Academy of Inspection and Quarantine CAIQ
Priority to CN202111344210.9A priority Critical patent/CN113969238A/en
Publication of CN113969238A publication Critical patent/CN113969238A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844Nucleic acid amplification reactions
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Optics & Photonics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention relates to a portable visual imaging system for gene amplification fluorescence detection, which comprises an array reaction chip, a fluorescence detection system and a control monitoring terminal; the fluorescence detection system comprises imaging equipment, a mounting rack, an excitation light source and a controllable temperature table; the top surface of the mounting frame is provided with a window glass sheet, the excitation light source is arranged on one side of the mounting frame, a light filter II is arranged in front of the excitation light source, and a collimating lens is arranged in front of the light filter II; a dichroic mirror is arranged inside the mounting rack, the included angle between the dichroic mirror and the collimating lens is 45 degrees, the controllable temperature table is arranged at the bottom of the mounting rack, and the array type reaction chip is arranged above the controllable temperature table; the visualized imaging system integrates nucleic acid amplification and fluorescence detection, is portable and can be operated on site, so that the time and the detection cost of gene amplification fluorescence detection are greatly reduced, and manpower and material resources are effectively saved.

Description

Portable visual imaging system for gene amplification fluorescence detection
Technical Field
The invention belongs to detection equipment and a method, and particularly relates to a portable visual imaging system for gene amplification fluorescence detection.
Background
In the traditional germ detection, the fluorescence detection technology of the gene amplification product combines the advantages of gene specificity amplification, DNA hybridization specificity and spectrum sensitivity and specificity, directly detects the fluorescence of the gene amplification product, does not need post-treatment such as separation, derivation and the like or additional detection, and completely closes the tube operation. Fluorescent groups such as tetrachlorofluorescein (TET), carboxyfluorescein (FAM), hexachloro-6-methylfluorescein (HEX), carboxy-4 ',5' -dichloro-2 ',7' -dimethoxyfluorescein (JOE), carboxytetramethylrhodamine (TAMRA), and the like can be introduced by means of a probe, primer modification, or the like. A method for adding fluorescent groups into a PCR reaction system by using a Real-time fluorescent Quantitative Polymerase Chain Reaction (PCR) technology, utilizing fluorescent signal accumulation to monitor the whole PCR process in Real time, and finally carrying out Quantitative analysis on an unknown template through a standard curve. The fluorescence detector can directly detect the wavelength and intensity change of the fluorescence signal through the wall of the PCR tube. The fluorescence quantitative PCR method can perform full-closed automatic detection on target nucleic acid, but has expensive instruments, high detection cost and high requirement on operation sites, and limits the field application of nucleic acid detection.
In addition to PCR techniques that require thermal cycling, isothermal gene amplification techniques also incorporate fluorescent probes to monitor or detect the amplification of a gene or its products. The existing gene amplification fluorescence detection method mainly detects fluorescence signals in real time, has an exciting light detector, an emission light detector and a signal processor, needs complex signal processors and software, and cannot be carried and operated on site. If the amplification product is subjected to terminal detection, most of the conventional methods adopt an ultraviolet flashlight or a blue light gel cutting instrument for visual imaging detection, and a complete portable visual imaging system integrating gene amplification and fluorescence detection is not available, so that the rapid field detection of germs cannot be completed.
Disclosure of Invention
In order to make up for the defects of the prior art, the invention provides a portable visual imaging system for gene amplification fluorescence detection, which completely integrates nucleic acid amplification and fluorescence detection into a whole, is portable and can be operated on site, thereby greatly reducing the time and detection cost of gene amplification fluorescence detection and effectively saving manpower and material resources.
The invention aims to realize the technical scheme that a portable visual imaging system for gene amplification fluorescence detection comprises an array reaction chip, a fluorescence detection system and a control monitoring terminal; the technical key points are as follows: the fluorescence detection system comprises imaging equipment, a mounting rack, an excitation light source and a controllable temperature table;
a sample placing window is arranged at the lower part of one side of the mounting frame, a window glass sheet is arranged on the top surface of the mounting frame, the transmittance of the full-transparent window glass is more than 90%, and the thickness of the full-transparent window glass is 0.5-2 mm; an optical filter I is arranged below the full-transparent window glass sheet, and the wavelength of the optical filter I is 515-560 nm;
the excitation light source is arranged on one side of the mounting frame and comprises an LED surface array and an LED array controller, and the wavelength range of the LED array is 420-488 nm;
an optical filter II is arranged in front of the LED area array of the excitation light source, a collimating lens is arranged in front of the optical filter II, the collimating lens is a plano-convex lens, and the focal length of the collimating lens is equal to the distance from the collimating lens to the LED array;
a dichroic mirror is arranged inside the mounting rack, and the included angle between the dichroic mirror and the collimating lens is 45 degrees; the wavelength of the dichroic mirror is 495nm or 565 nm;
the controllable temperature table is arranged at the bottom of the mounting frame, and the array reaction chip is arranged above the controllable temperature table;
the array reaction chip consists of three layers, namely a heat-conducting bottom layer, a reaction middle layer and a light-transmitting upper cover from bottom to top in sequence, wherein the reaction middle layer is provided with a plurality of reaction holes, and the bottom layer is made of glass or organic glass; the material of the middle layer in the reaction is polystyrene and polydimethylsiloxane after silanization treatment; and a light-transmitting upper cover.
Furthermore, the controllable temperature table comprises a heating table, a temperature sensor, a heating element, a PID controller, a direct current power supply and a solid-state relay; the heating table is arranged above the heating element, the temperature sensor is inserted into the heating table and connected with the PID controller, and the solid-state relay is connected with the PID controller, the heating element and the direct-current power supply.
Further, the application method of the visual imaging system comprises the following steps:
(1) preparation of test samples
Amplifying the DNA to obtain a detection sample;
(2) sample application
Loading a detection sample into a reaction hole of an array reaction chip of a visual imaging system for gene amplification fluorescence detection;
(3) obtaining the detection result
The array reaction chip is placed on a controllable temperature table in a detection system through a sample window, the temperature of the controllable temperature table is preset to be 38 ℃, a fluorescence image of a sample in the reaction chip can be obtained during reaction, and green represents positive amplification.
Furthermore, the imaging system is composed of a camera and a lens, is positioned above the full transparent window glass sheet, enables the reaction chip to be positioned on the focal point of the lens by adjusting the focal length of the lens, and shoots the image of the detection fluorescent substance in the reaction hole of the reaction chip by the camera.
Furthermore, the control monitoring terminal is a computer or a movable terminal computer.
The invention has the beneficial effects that:
(1) the traditional PCR-gel electrophoresis detection needs additional electrophoresis detection on PCR products, needs long time and has the risk of sample adding errors or pollution. Real-time fluorescence PCR requires a complex data acquisition assembly, performs complex calculations on the emitted light, and has high instrument cost. By adopting the detection method, the amplification and the visual imaging can be carried out simultaneously, the time for detecting after the amplification is omitted, and the potential pollution possibly brought by absorbing the amplification product is avoided.
(2) The isothermal amplification technology adopted by the invention can be carried out at 38 ℃, the amplification condition can be simultaneously observed, and a group of samples can observe a strong enough fluorescence image of a positive sample in about 30 minutes.
Drawings
FIG. 1 is a schematic structural diagram of the present visualization imaging system;
FIG. 2 is a schematic diagram of a schematic structural diagram of the present visualization imaging system;
FIG. 3 is a schematic diagram of an array reaction chip;
FIG. 4 is a diagram of reactant addition;
FIG. 5 is a schematic diagram of a structure of an LED array;
FIG. 6 shows the results of isothermal amplification fluorescence detection of nucleic acids of Alternaria brasiliensis Schneid;
FIG. 7 is a schematic diagram of a temperature-controllable stage.
In fig. 1 to 7, specific names of the respective structures are: the device comprises an array type reaction chip 1, a heat conduction bottom layer 1-1, a reaction middle layer 1-2, a light transmission upper cover 1-3, a fluorescence detection system 2, a control monitoring terminal 3, an imaging device 4, a mounting rack 5, an excitation light source 6, an LED surface array 6-1, an LED array controller 6-2, a sample placing window 7, a window glass sheet 8, an optical filter I9, an optical filter II 10, a collimating lens 11, a dichroic mirror 12, a controllable temperature table 13, a heating table 13-1, a temperature sensor 13-2, a heating element 13-3, a PID controller 13-4, a direct current power supply 13-5 and a solid-state relay 13-6.
Detailed Description
The technical solutions of the present invention can be specifically described below with reference to the drawings and the embodiments, but the technical solutions of the present invention are not limited to the scope of the present invention.
Example 1A visual imaging System for fluorescence detection of Gene amplification
Referring to fig. 1-5, the specific structure of this embodiment is a visual imaging system for gene amplification fluorescence detection, which includes an array reaction chip 1, a fluorescence detection system 2, and a control monitoring terminal 3;
the fluorescence detection system 2 comprises an imaging device 4, a mounting rack 5, an excitation light source 6 and a temperature-controllable table 13;
a sample placing window 7 is arranged at the lower part of one side of the mounting frame 5, a window glass sheet 8 is arranged on the top surface of the mounting frame 5, the transmittance of the full-transparent window glass is more than 90%, the loss of a fluorescence signal is reduced, the thickness is 0.5-2mm, the full-transparent window glass has certain intensity and is used for protecting other internal optical devices, an optical filter I9 is arranged below the full-transparent window glass sheet 8, the parameter range of the optical filter I9 is 515-560nm, the optical filter I is a replaceable component and is replaced according to the emission wavelength of a detected fluorescent substance, and the optical filter I is 525/50 (green fluorescence) or 605/70 (red fluorescence) under general conditions;
the excitation light source 6 is arranged on one side of the mounting frame, the excitation light source 6 comprises an LED surface array 6-1 and an LED array controller 6-2, the wavelength range of the LED array is 420-488nm, and the wavelength of the LED array can be changed according to the excitation wavelength of the detected fluorescent substance, generally 470/40 (blue light excitation) and 545/25 (green light excitation); the size of the LED array is matched with that of the reaction chip, so that the light of the LED array can completely irradiate the reaction chip;
an optical filter II 10 is arranged in front of the LED surface array 6-1 of the excitation light source 6, a collimating lens 11 is arranged in front of the optical filter II 10, the collimating lens 11 is a plano-convex lens, and the focal length of the collimating lens 11 is equal to the distance from the collimating lens to the LED surface array 6-1;
a dichroic mirror 12 is arranged inside the mounting rack 5, and the included angle between the dichroic mirror 12 and the collimating lens 11 is 45 degrees; the wavelength of the dichroic mirror 12 is selected according to the detection fluorescent substance, and is generally 495nm (green fluorescence) or 565nm (red fluorescence).
The controllable temperature table 13 is arranged at the bottom of the mounting frame 5, the array type reaction chip 1 is arranged above the controllable temperature table 13, the size of the temperature control table is matched with that of the reaction chip, and the reaction chip is arranged on the surface of the controllable temperature table, so that the whole chip can be uniformly heated;
the temperature-controllable table 13 is composed of a heating table 13-1, a temperature sensor 13-2, a heating element 13-3, a PID controller 13-4, a direct-current power supply 13-5 and a solid-state relay 13-6. The connection relation is as shown in FIG. 7; the heating table 13-1 is arranged above the heating element 13-3, the temperature sensor 13-2 is inserted into the heating table 13-1, the temperature sensor 13-2 is connected with the PID controller 13-4, the direct current power supply 13-5 is connected with the heating element 13-3, and the solid state relay 13-6 is connected with the PID controller 13-4, the heating element 13-3 and the direct current power supply 13-5.
The heating stage 13-1 is made of a metal having good thermal conductivity, such as aluminum or copper. The temperature sensor is inserted into the heating stage 13-1 (the temperature sensor may be a thermocouple, a thermistor, a Resistance Temperature Detector (RTD), an IC temperature sensor, or the like); the heating element 13-3 is fixed at the bottom of the heating table (the heating element can be a film heater or a PTC heater, so as to realize uniform heating on the heating table). The PID controller 13-4, the solid state relay 13-6 and the dc power supply are all commercially available.
The working principle is as follows: the PID controller 13-4 is connected with the temperature sensor 13-2, and detects the temperature of the heating table 13-3 in real time through the temperature sensor 13-2; comparing the temperature of the heating table 13-1 with a set temperature, outputting a control signal through a PID algorithm, and controlling the solid-state relay 13-6; the PID controller 13-4 controls the connection or disconnection of the DC power supply 13-5 and the heating element 13-3 by controlling the solid-state relay. When the direct current power supply 13-5 is communicated with the heating element 13-3, the heating element 13-3 controls the temperature of the heating platform 13-1 to rise; when the DC power supply 13-5 is disconnected from the heating element 13-3, the heating element 13-3 does not heat the heating stage 13-1. In this way, the temperature of the heating stage 13-1 is controlled at the set temperature value.
The reaction chip 1 consists of three layers, namely a heat conduction bottom layer 1-1, a reaction middle layer 1-2 and a light-transmitting upper cover 1-3 in sequence from bottom to top, wherein the reaction middle layer 1-2 is provided with a plurality of reaction holes, and the heat conduction bottom layer 1-1 and the reaction middle layer 1-2 are integrally formed to form reaction holes, namely a sample cell; the bottom layer 1-1 is made of a material having an excellent heat conductive ability, such as glass or the like; the reaction layer 1-2 has good biocompatibility, and can prevent the nonspecific adsorption of biological substances in the sample on the wall of the sample cell, such as silanized polystyrene (PP), Polydimethylsiloxane (PDMS), etc.; the light-transmitting upper cover 1-3 has excellent light-transmitting performance, and can tightly cover the sample cell to prevent evaporation of the sample in the sample cell during the reaction process.
An imaging system: constitute by camera and camera lens, be located full transparent window glass piece top, through adjusting the camera lens focus, make reaction chip be located the camera lens focus, shoot the image of the detection fluorescent substance in the reaction chip reaction hole through the camera: the camera may be a CCD or CMOS camera. The camera is connected with the monitoring terminal through a data line and is used for controlling the camera and transmitting image data.
The control monitoring terminal 3 can be a computer or a movable terminal computer; the computer or the mobile terminal computer is provided with control software, and the control software comprises control software capable of setting and displaying the temperature of the temperature control console in real time; capable of controlling camera shooting parameters such as exposure time, shot image format, image storage location, shooting interval, etc.); the software can analyze the shot image, detect and identify fluorescent substances in the reaction holes through a fluorescence threshold value, quantify the average fluorescence intensity of the fluorescent substances, and generate a curve of the average fluorescence intensity-time; and calculating the content of the gene to be detected through fluorescence intensity.
The use process of the invention is as follows: loading the gene of a sample to be detected and an amplification reaction reagent containing fluorescence into a sample pool of the array reaction chip, and putting the sample pool into a detection system through a sample window; controlling the sample in the reaction chip to amplify (such as PCR or isothermal amplification method) by a controllable temperature table, wherein the amplification quantity of the sample gene is in direct proportion to the fluorescence of the sample; controlling a sample in the reaction chip to carry out gene amplification through a controllable temperature table; shooting fluorescence images of all samples in the reaction chip in real time or at an end point through a fluorescence detection module and an imaging system, and carrying out image analysis by controlling a monitoring terminal to obtain a sample gene amplification result; the fluorescence of the sample can also be observed by naked eyes, and qualitative analysis is carried out according to whether the sample has fluorescence or not.
Example 2 isothermal amplification fluorescent detection of Phosphaera brassicae
To be provided with
Figure BDA0003353374610000061
Taking 29.5 mu L of rehydration buffer solution carried by the kit, 4.8 mu L of primer and probe mixed solution (the ratio is 3.5: 3.5: 1), 2.0 mu L of DNA and 11.2 mu L of deionized water as an example, mixing uniformly, adding into lyophilized enzyme powder (A), (B), (C) and D) and (C)
Figure BDA0003353374610000062
Kit of parts) 0.2mL of TwistAmpExo reaction tube (1)
Figure BDA0003353374610000063
Kit self), adding 2.5 mu L of magnesium acetate solution (280mmol/L), mixing uniformly, loading into a sample cell of an array reaction chip, putting into a controllable temperature table with the preset temperature of 38 ℃ in a detection system through a sample window, observing a fluorescence image of a sample in the reaction chip while amplifying, shooting the fluorescence image by a mobile phone, and analyzing.
In the reaction system, the primer F, the primer R and the probe P are added into the reaction system in the form of working solution, and the concentration of the primer F, the primer R and the probe P in the working solution is 10 mu mol/L.
The results are shown in FIG. 6, where green represents positive amplification.
Primer F, primer R, fluorescent probe P references:
R.Lei,J.Kong,Y.H.Qiu,N.Z.Chen,S.F.Zhu,X.Y.Wang,P.S.Wu,Rapid detection of the pathogenic fungi causing blackleg of Brassica napus using a portable real-time fluorescence detector.Food Chem.288(2019)57-67.https://10.1016/j.foodchem.2019.02.089。
by adopting the structure of the imaging system, amplification and visual imaging can be carried out simultaneously, the time for detecting after amplification is saved, and potential pollution possibly brought by absorbing amplification products is avoided. Such as PCR-gel electrophoresis detection, requires additional electrophoresis detection of the PCR product, takes a long time, and has a risk of sample application errors or contamination. Real-time fluorescence PCR requires a complex data acquisition assembly, performs complex calculations on the emitted light, and has high instrument cost. The isothermal amplification technology adopted by the invention can be carried out at 38 ℃, the amplification condition can be observed at the same time, and a strong enough fluorescence image of a positive sample can be observed within about 30 minutes generally.
The principle of the invention is as follows:
(1) the fluorescence detection system comprises an excitation light source, an excitation filter, a collimating lens, a dichroic mirror, a sample placing window, an emission filter lens and a full-transparent window glass
(2) Excitation light source: the LED array is provided with a white light source, and the light intensity of the LED array can be adjusted by a controller. The size of the LED array can be selected based on the size of the sample being tested.
(3) The optical filter II in front of the LED array is an excitation optical filter: the desired excitation light filter is selected based on the excitation wavelength of the detected fluorescence, and the excitation light filter passes light of the particular wavelength of interest emitted by the LED array.
(4) A collimating lens: the exciting light is changed into parallel light (exciting light), and the intensity of the exciting light is uniform, and the device is composed of a plano-convex lens.
(5) A dichroic mirror: the exciting light is refracted and vertically irradiated on the sample, and simultaneously, the excited fluorescence of the sample is transmitted for detection or observation.
(6) The filter I below the window glass sheet is an emission filter, and the emission filter only passes through the emission wavelength after the fluorescent substance is excited, so that the interference of background light signals is reduced.
(7) Full transparent window glass sheet: over the bandpass filter, the optical elements in the protection system become contaminated.
(8) A temperature-controllable table: the reaction chip is fixed on the surface of a temperature table to control the problem of the sample on the chip. The controllable temperature table can be subjected to thermostatic control and programming control (temperature changes along with time) according to reaction needs.
(9) Array reaction chip: the reaction chip consists of three layers: the bottom layer and the middle layer are combined together to form a sample cell; the bottom layer is made of a material having excellent heat conductivity, such as glass or the like; the middle layer has good biocompatibility and can prevent the nonspecific adsorption of biological substances in a sample on the wall of the sample cell, such as polystyrene (PP), Polydimethylsiloxane (PDMS) and the like after silanization treatment; the upper layer has excellent light transmission performance, and can tightly cover the sample cell to prevent the evaporation of the sample in the sample cell during the reaction process.
(10) An imaging device: the fluorescence detection device is used for detecting and imaging the excited fluorescence of the sample, further acquiring a fluorescence image of the sample and analyzing the fluorescence intensity. The fluorescence change of the sample can be analyzed by controlling the imaging device to carry out continuous timing shooting, and the device is composed of a CCD or COMS imaging device. The imaging device can also be replaced by human eyes, and the fluorescence state of the sample can be visually observed.
(11) Controlling the monitoring terminal: can be composed of a computer and a movable terminal computer. And specific control software is installed for controlling the system LED laser light source light intensity, the temperature of the temperature control table, the collection and analysis of a sample fluorescence image, the storage, management and transmission of result data and the like.

Claims (5)

1. A portable visual imaging system for gene amplification fluorescence detection comprises an array reaction chip, a fluorescence detection system and a control monitoring terminal; the method is characterized in that: the fluorescence detection system comprises imaging equipment, a mounting rack, an excitation light source and a controllable temperature table;
a sample placing window is arranged at the lower part of one side of the mounting frame, a window glass sheet is arranged on the top surface of the mounting frame, the transmittance of the full-transparent window glass is more than 90%, and the thickness of the full-transparent window glass is 0.5-2 mm; an optical filter I is arranged below the full-transparent window glass sheet, and the wavelength of the optical filter I is 515-560 nm;
the excitation light source is arranged on one side of the mounting frame and comprises an LED surface array and an LED array controller, and the wavelength range of the LED array is 420-488 nm;
an optical filter II is arranged in front of the LED area array of the excitation light source, a collimating lens is arranged in front of the optical filter II, the collimating lens is a plano-convex lens, and the focal length of the collimating lens is equal to the distance from the collimating lens to the LED array;
a dichroic mirror is arranged inside the mounting rack, and the included angle between the dichroic mirror and the collimating lens is 45 degrees; the wavelength of the dichroic mirror is 495nm or 565 nm;
the controllable temperature table is arranged at the bottom of the mounting frame, and the array reaction chip is arranged above the controllable temperature table;
the array reaction chip consists of three layers, namely a heat-conducting bottom layer, a reaction middle layer and a light-transmitting upper cover from bottom to top in sequence, wherein the reaction middle layer is provided with a plurality of reaction holes, and the bottom layer is made of glass or organic glass; the material of the middle layer in the reaction is polystyrene and polydimethylsiloxane after silanization treatment; and a light-transmitting upper cover.
2. The portable visual imaging system for gene amplification fluorescence detection according to claim 1, wherein: the controllable temperature table comprises a heating table, a temperature sensor, a heating element, a PID controller, a direct current power supply and a solid-state relay; the heating table is arranged above the heating element, the temperature sensor is inserted into the heating table and connected with the PID controller, and the solid-state relay is connected with the PID controller, the heating element and the direct-current power supply.
3. The portable visual imaging system for gene amplification fluorescence detection according to claim 1, wherein: the application method of the visual imaging system comprises the following steps:
(1) preparation of test samples
Amplifying the DNA to obtain a detection sample;
(2) sample application
Loading a detection sample into a reaction hole of an array reaction chip of a visual imaging system for gene amplification fluorescence detection;
(3) obtaining the detection result
The array reaction chip is placed on a controllable temperature table in a detection system through a sample window, the temperature of the controllable temperature table is preset to be 38 ℃, a fluorescence image of a sample in the reaction chip can be obtained during reaction, and green represents positive amplification.
4. The portable visual imaging system for gene amplification fluorescence detection according to claim 1, wherein: the imaging device consists of a camera and a lens, is positioned above the full-transparent window glass sheet, enables the reaction chip to be positioned on the focal point of the lens by adjusting the focal length of the lens, and shoots an image of the detection fluorescent substance in the reaction hole of the reaction chip by the camera.
5. The portable visual imaging system for gene amplification fluorescence detection according to claim 1, wherein: the control monitoring terminal is a computer or a movable terminal computer.
CN202111344210.9A 2021-11-15 2021-11-15 A Portable Visual Imaging System for Fluorescence Detection of Gene Amplification Pending CN113969238A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202111344210.9A CN113969238A (en) 2021-11-15 2021-11-15 A Portable Visual Imaging System for Fluorescence Detection of Gene Amplification

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202111344210.9A CN113969238A (en) 2021-11-15 2021-11-15 A Portable Visual Imaging System for Fluorescence Detection of Gene Amplification

Publications (1)

Publication Number Publication Date
CN113969238A true CN113969238A (en) 2022-01-25

Family

ID=79589734

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202111344210.9A Pending CN113969238A (en) 2021-11-15 2021-11-15 A Portable Visual Imaging System for Fluorescence Detection of Gene Amplification

Country Status (1)

Country Link
CN (1) CN113969238A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114755211A (en) * 2022-04-24 2022-07-15 华南农业大学 Integrated visual fluorescent LAMP detection device and method
CN114860003A (en) * 2022-05-12 2022-08-05 中国科学院苏州生物医学工程技术研究所 PCR thermal cycle system control method, device, equipment and storage medium

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106140338A (en) * 2016-06-21 2016-11-23 中国科学院电子学研究所 Micro-fluidic chip system and apply the method that this system carries out isothermal duplication and detection
CN112525870A (en) * 2019-09-17 2021-03-19 北京达微生物科技有限公司 Large-area fluorescence imaging detection device
CN216808842U (en) * 2021-11-15 2022-06-24 中国检验检疫科学研究院 Portable visual imaging system for gene amplification fluorescence detection

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106140338A (en) * 2016-06-21 2016-11-23 中国科学院电子学研究所 Micro-fluidic chip system and apply the method that this system carries out isothermal duplication and detection
CN112525870A (en) * 2019-09-17 2021-03-19 北京达微生物科技有限公司 Large-area fluorescence imaging detection device
CN216808842U (en) * 2021-11-15 2022-06-24 中国检验检疫科学研究院 Portable visual imaging system for gene amplification fluorescence detection

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114755211A (en) * 2022-04-24 2022-07-15 华南农业大学 Integrated visual fluorescent LAMP detection device and method
CN114860003A (en) * 2022-05-12 2022-08-05 中国科学院苏州生物医学工程技术研究所 PCR thermal cycle system control method, device, equipment and storage medium

Similar Documents

Publication Publication Date Title
US10928321B2 (en) Portable device for detecting molecule(s)
KR102352586B1 (en) Pcr apparatus for real-time detecting multiplex fluorescent signals
KR101302353B1 (en) Automatic analysis device of molecular diagnosis for performing both real-time pcr detecting quantitatively gene and dna microarray for gene analysis
JPH07163397A (en) Simultaneous observation and analysis method for multiple amplification reactions
JP6158318B2 (en) Nucleic acid analyzer and nucleic acid analysis method using the same
US20170051335A1 (en) Apparatus and method for thermocyclic biochemical operations
CN113969238A (en) A Portable Visual Imaging System for Fluorescence Detection of Gene Amplification
KR102261902B1 (en) Real-time PCR Fluorescence detection device
CN102618439A (en) Deoxyribonucleic acid (DNA) fragment amplification and quantitative detection system based on closed reactors
CN104120077B (en) A kind of real-time fluorescence DNA cloning instrument
CN109811038B (en) Portable instrument for bottom-lighting type nucleic acid isothermal amplification detection
KR20190120947A (en) A device for real-time detecting nucleic acids amplification products
CN111592982A (en) Novel portable nucleic acid amplification analyzer
CN104120078A (en) Real-time fluorescence PCR (Polymerase Chain Reaction) amplification instrument
JP2003344290A (en) Fluorescence detector with temperature control
EP4182670A1 (en) An apparatus & method for processing and analysing one or more samples
CN216808842U (en) Portable visual imaging system for gene amplification fluorescence detection
CN210923478U (en) Real-time fluorescent quantitative PCR instrument for on-site rapid detection
CN109797097B (en) A top-illuminated nucleic acid isothermal amplification detection portable instrument
US7186989B2 (en) Low thermal mass fluorometer
CN111944678A (en) Portable nucleic acid colorimetric detection device and use method thereof
TW201339308A (en) Detection device for nucleic acid amplification
CN210215376U (en) Portable nucleic acid colorimetric detection device
US20220010369A1 (en) Method and apparatus for performing a real-time colorimetric nucleic acid amplification assay
Sun et al. Two colorimetric LAMP systems for nucleic acid-based diagnostics

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination