CN211374514U - Light-guide rod light-transmission lens-free illumination imaging device and micro-fluidic chip detection system - Google Patents
Light-guide rod light-transmission lens-free illumination imaging device and micro-fluidic chip detection system Download PDFInfo
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Abstract
The utility model provides a leaded light stick passes no lens illumination imaging device of light, including imaging element, light source, leaded light stick, excitation light filter, the even light of bottom reflection secret light section of thick bamboo and micro-fluidic chip, the light that the light source sent passes through earlier the transmission of leaded light stick passes through after the filtration of excitation light filter forms the quasi monochromatic light extension illuminating beam, some of quasi monochromatic light extension illuminating beam directly jets into micro-fluidic chip is right the imaging area of micro-fluidic chip throws light on, another part of quasi monochromatic light extension illuminating beam is in jet into under the reflection of the even light secret light section of thick bamboo wall of bottom reflection micro-fluidic chip is right the imaging area edge of micro-fluidic chip compensates the illumination. The utility model also provides a micro-fluidic chip detecting system. The utility model has the advantages that: the light guide rod is adopted to transmit light and illuminate without a lens, the structure is simple, the cost is low, and the fluorescence collection efficiency is high.
Description
Technical Field
The utility model relates to a biochip especially relates to a leaded light stick passes no lens illumination imaging device of light and micro-fluidic chip detecting system.
Background
Biochips are an advanced biomedical measurement method emerging at the end of the 20 th century, and realize rapid, parallel, efficient detection and analysis of hundreds of biomolecules and precise medical molecular diagnosis on one solid support. Biochips are mainly classified into microarray (micro array) chips and microfluidic (Microfluidics) chips. Microarray chips are also known as gene chips (Genechip) or DNA chips (DNA chip). The Micro-fluidic chip is also called Micro-Total Analysis System (μ TAS), which integrates the main functional units of sample preparation, biochemical reaction, result detection, etc. into one chip, realizes the integration, automation and microminiaturization from sample collection to result report, and can automatically complete the whole process of detection and Analysis. In practical application, the micro-fluidic chip mainly realizes the function of a micro total analysis system at present.
The fluorescence labeling detection method is a common method for biochip detection, and comprises confocal scanning and CCD imaging, wherein a light source is focused or collimated by a lens to illuminate a detected object (such as a DNA base), then the fluorescence labeling DNA base is utilized to generate emission fluorescence under the irradiation of exciting light with different wavelengths, and the emission fluorescence is filtered by a color filter and received by a photoelectric conversion detector to obtain information of mutual combination (hybridization) of interested biomolecules or change of the quantity of the biomolecules. By using the multicolor fluorescence labeling, multiple analyses can be simultaneously carried out on two or more biological samples in one analysis, and the accuracy of gene expression and mutation detection results is greatly improved. However, the method of focusing or collimating the light source based on the lens not only has a complicated structure and high cost, but also has a large number of lenses which cause a large attenuation of the light signal, thereby affecting the efficiency and detection sensitivity of the excitation light.
Disclosure of Invention
In order to solve the problems in the prior art, the utility model provides a leaded light stick passes no lens illumination imaging device of light and micro-fluidic chip detecting system.
The utility model provides a leaded light stick passes no lens illumination imaging device of light, including imaging element, light source, leaded light stick, excitation light filter, the even light of bottom reflection is tight a section of thick bamboo and micro-fluidic chip, the light that the light source sent passes through earlier the transmission of leaded light stick passes through after the filtration of excitation light filter forms the extension illuminating beam of quasi monochromatic light, some of the extension illuminating beam of quasi monochromatic light directly jets into micro-fluidic chip is right the imaging area of micro-fluidic chip throws light on, another part of the extension illuminating beam of quasi monochromatic light is in jet into under the reflection of the section of thick bamboo wall of the even light tight light section of thick bamboo of bottom reflection micro-fluidic chip is right the imaging area edge of micro-fluidic chip compensates the illumination, receives the fluorescence that the measured object produced through imaging element, realizes the fluorescence imaging to the measured object.
As a further improvement of the present invention, the light guide rod is seamlessly close to the light emitting surface of the light source, directly receiving the light emitted from the light source.
As a further improvement, be equipped with the aspherical mirror between light source, the leaded light stick, the light source pass through the aspherical mirror with the coupling of leaded light stick, the light that the light source produced passes through earlier behind the aspherical mirror spotlight, the coupling gets into the transmission of leaded light stick passes through arouse the light filter and filter, form the quasi monochromatic light extension illuminating beam.
As a further improvement, be equipped with the beam splitter between the even light tight light section of thick bamboo of excitation light filter, bottom reflection, the quasi monochromatic light extension illuminating beam who forms after the excitation light filter filters jets into the beam splitter, quilt the beam splitter reflection, partly directly jets into micro-fluidic chip, another part is in jet into under the reflection of the section of thick bamboo wall of the even light tight light section of thick bamboo of bottom reflection micro-fluidic chip.
As a further improvement, the light reflected by the beam splitter is perpendicularly injected into the micro-fluidic chip, the light reflected by the micro-fluidic chip is blocked by the dichroic gating function of the beam splitter, and cannot be reflected again to enter the imaging unit.
As a further improvement, the quasi-monochromatic light expanded illuminating light beam formed after the filter is excited is jetted into the micro-fluidic chip from the side direction.
As a further improvement, the light source is connected with the fin, the even light of imaging unit, bottom reflection is dense, and a light section of thick bamboo, the coaxial setting of micro-fluidic chip, the even light of bottom reflection is dense, and a light section of thick bamboo is located between imaging unit, the micro-fluidic chip, light source, leaded light stick, excitation light filter are located the side of micro-fluidic chip.
As a further improvement, the even light of bottom reflection is dense a light section of thick bamboo and is located directly over micro-fluidic chip, the upper portion part of the even light of bottom reflection is the light extinction area, the lower floor's part of the even light of bottom reflection is the reflecting area, another part of quasi monochromatic light extension illuminating beam is in jet into under the reflection of the reflecting area of the even light of bottom reflection is dense a light section of thick bamboo micro-fluidic chip.
The utility model also provides a micro-fluidic chip detecting system, include as in any one of above-mentioned leaded light stick biography light no lens illumination imaging device, the specific gene primer sequence that nucleic acid detection used is fixed respectively to micro-fluidic chip's reaction channel bottom, like the specific nucleic acid fragment sequence that is fit for detecting the nucleic acid isothermal amplification molecule diagnosis of yersinia lung sporophytes (Pneumocystis jiirovici):
primer No. 1-GTAGTGAAATACAAATCGGACT;
primer No. 2-CTGTTCTGGGCTGTTTCC;
primer No. 3-AGTGCTATACCTACTATTTTTAAGAGGAGGATATAGCTGGTTTTCTGC;
primer No. 4-TCGAGGGAGTATGAAAATATTTATCTCACCTTATCGCACATAGTCTGA;
primer No. 5-ATAAACAATTTGCCAAAACAATTTTC;
primer No. 6-GATATTTAATCTCAAAATAACTATTTCTTAA.
As a further improvement of the present invention, the micro-fluidic chip is embedded with a biomolecule detection probe for isothermal or temperature-varying amplification detection of nucleic acids, the imaging unit comprises a detector, an imaging lens and an emission filter, the imaging lens is located between the detector and the emission filter, the micro-fluidic chip detection system further comprises a heating film, a temperature control module, a motion control module and a computer display processor, the heating film is located under the micro-fluidic chip, the heating film is connected with the temperature control module, the micro-fluidic chip is connected with the motion control module, the temperature control module, the motion control module and the detector are respectively connected with the computer display processor, the computer display processor sends motion control signals to the motion control module, and the motion control module sends the motion control signals to the motion control module according to the received motion control signals, controlling the micro-fluidic chip to do one-dimensional or two-dimensional planar motion to realize detection and analysis of reaction channels in different areas on the micro-fluidic chip; the computer display processor transmits a temperature control signal to the temperature control module, and the temperature control module controls the temperature of the heating film according to the received temperature control signal to meet the requirement of the temperature control condition of nucleic acid amplification of the microfluidic chip; the nucleic acid amplification products of the reaction channels in different areas on the microfluidic chip generate fluorescent signals under the irradiation of exciting light of a light source, are filtered by the emission optical filter, are converged on the detector by the imaging lens, are input into the computer display processor after photoelectric conversion, and are further processed by digital signals of the computer display processor to display the nucleic acid amplification fluorescent signals in a digital or graphic mode, so that the visualization of the nucleic acid amplification signals of different reaction channels on the microfluidic chip is realized.
As a further improvement of the present invention, the biomolecule detection probe is preferably a nucleic acid detection molecular probe composed of 6 segments of primers, and the microfluidic chip 6 comprises more than 6 reaction channels.
The utility model has the advantages that: by adopting the scheme, the light guide rod is adopted for transmitting light and illuminating without a lens, the structure is simple, the cost is low, the fluorescence collection efficiency is higher, and the detection accuracy and sensitivity of the microfluidic chip are favorably improved.
Drawings
Fig. 1 is a schematic diagram of oblique incidence of the light-guiding rod light-transmitting lensless illumination imaging device of the present invention.
Fig. 2 is a schematic view of the coaxial normal incidence of the light-guiding rod light-transmitting lensless illumination imaging device of the present invention.
Fig. 3 is a schematic view of the illumination of the light-guiding rod light-transmitting lens-free illumination imaging device of the present invention.
Fig. 4 is a schematic view of the illumination of the light-guiding rod light-transmitting lens-free illumination imaging device of the present invention.
Fig. 5 is a schematic diagram of a bottom-reflection light-homogenizing light-tube of the light-guiding rod light-transmitting lens-less illumination imaging device of the present invention.
Detailed Description
The present invention will be further described with reference to the following description and embodiments.
As shown in FIG. 1, a light guide rod light transmission lens-free lighting imaging device comprises an imaging unit, a light source 2, a light guide rod 3, an excitation filter 4, a bottom reflection light-homogenizing light cylinder 5 and a microfluidic chip 6, the light emitted by the light source 2 is transmitted by the light guide rod 3 and filtered by the excitation filter 4, forming a quasi-monochromatic light expanded illuminating beam, a part of which is directly incident into the microfluidic chip 6, illuminating the imaging area of the microfluidic chip 6, wherein the other part of the quasi-monochromatic light expanded illuminating beam is emitted into the microfluidic chip 6 under the reflection of the cylinder wall of the light cylinder 5 with uniform and dense light reflected at the bottom, and the edge of the imaging area of the microfluidic chip 6 is subjected to compensation illumination, and fluorescence generated by the object to be detected is received through the imaging unit, so that fluorescence imaging of the object to be detected is realized.
As shown in fig. 3, the light guide rod 3 is seamlessly close to the light emitting surface of the light source 2, and directly receives the light emitted from the light source 2.
As shown in fig. 4, an aspheric mirror 15 is disposed between the light source 2 and the light guide rod 3, the light source 2 is coupled with the light guide rod 3 through the aspheric mirror 15, light generated by the light source 2 is condensed by the aspheric mirror 15, then coupled into the light guide rod 3 for transmission, and then filtered by the excitation filter 4 to form a quasi-monochromatic light expanded illumination beam.
As shown in fig. 2, a beam splitter 14 is disposed between the excitation filter 4 and the bottom reflection uniform light density optical tube 5, the quasi-monochromatic light expanded illumination light beam formed after being filtered by the excitation filter 4 is incident on the beam splitter 14, reflected by the beam splitter 14, and a part of the quasi-monochromatic light expanded illumination light beam is directly incident on the microfluidic chip 6, and another part of the quasi-monochromatic light expanded illumination light beam is incident on the microfluidic chip 6 under the reflection of the tube wall of the bottom reflection uniform light density optical tube 5.
As shown in fig. 2, the light reflected by the beam splitter 14 perpendicularly enters the microfluidic chip 6, and the light reflected by the microfluidic chip 6 is cut off by the dichroic gating function of the beam splitter 14 and does not pass through the beam splitter 14 to enter the imaging unit.
As shown in fig. 1, the quasi-monochromatic expanded illumination beam formed after filtering by the excitation filter 4 is emitted into the microfluidic chip 6 from the side.
As shown in fig. 1 and 2, the light source 2 is connected to a heat sink 1, the imaging unit, the bottom-reflection light-homogenizing light cylinder 5, and the microfluidic chip 6 are coaxially disposed, the bottom-reflection light-homogenizing light cylinder 5 is located between the imaging unit and the microfluidic chip 6, and the light source 2, the light guide rod 3, and the excitation filter 4 are located at a side of the microfluidic chip 6.
As shown in fig. 1 and 2, the bottom reflection dodging tube 5 is located right above the microfluidic chip 6, an upper layer portion of the bottom reflection dodging tube 5 is a light extinction area 51, a lower layer portion of the bottom reflection dodging tube 5 is a reflection area 52, and another portion of the quasi-monochromatic light expanded illumination beam is reflected by the reflection area 52 of the bottom reflection dodging tube 5 and enters the microfluidic chip 6.
As shown in fig. 1 and 2, a microfluidic chip detection system includes the light-guide rod light-transmission lens-free illumination imaging device as described in any one of the above.
Specific gene primer sequences for nucleic acid detection are respectively fixed at the bottoms of reaction channels of the microfluidic chip, and the specific gene primer sequences adopt specific nucleic acid fragment sequences suitable for detecting nucleic acid isothermal amplification molecules of yarrowia (Pneumocystis jiirovici):
primer No. 1-GTAGTGAAATACAAATCGGACT;
primer No. 2-CTGTTCTGGGCTGTTTCC;
primer No. 3-AGTGCTATACCTACTATTTTTAAGAGGAGGATATAGCTGGTTTTCTGC;
primer No. 4-TCGAGGGAGTATGAAAATATTTATCTCACCTTATCGCACATAGTCTGA;
primer No. 5-ATAAACAATTTGCCAAAACAATTTTC;
primer No. 6-GATATTTAATCTCAAAATAACTATTTCTTAA.
As shown in fig. 1 and 2, a biomolecule detection probe for isothermal or temperature-varying amplification detection of nucleic acid is embedded in the microfluidic chip 6, the biomolecule detection probe is preferably a molecular probe for nucleic acid detection composed of 6 segments of primers, the microfluidic chip 6 includes one or more reaction channels, the imaging unit includes a detector (CCD) 10, an imaging lens 9, and an emission filter 8, the imaging lens 9 is located between the detector 10 and the emission filter 8, the microfluidic chip detection system further includes a heating film 7, a temperature control module (HCP) 13, a motion control Module (MCP) 12, and a computer display processor (CCP) 11, the heating film 7 is located right below the microfluidic chip 6, the heating film 7 is connected to the temperature control module 13, and the microfluidic chip 6 is connected to the motion control module 12, the temperature control module 13, the motion control module 12 and the detector 10 are respectively connected with the computer display processor 11, the computer display processor 11 sends a motion control signal to the motion control module 12, and the motion control module 12 controls the microfluidic chip 6 to do one-dimensional or two-dimensional planar motion according to the received motion control signal, so as to realize detection and analysis of reaction channels in different areas on the microfluidic chip 6; the computer display processor 11 transmits a temperature control signal to the temperature control module 13, and the temperature control module 13 controls the temperature of the heating film 7 according to the received temperature control signal, so that the requirement of the nucleic acid amplification temperature control condition of the microfluidic chip 6 is met; the nucleic acid amplification products of the reaction channels in different areas on the microfluidic chip 6 generate fluorescent signals under the irradiation of the exciting light of the light source 2, are filtered by the emission optical filter 8, are converged on the detector 10 by the imaging lens 9, are input into the computer display processor 11 after photoelectric conversion, and are further processed by the computer display processor 11 in a digital or graphic manner, so that the nucleic acid amplification fluorescent signals of different reaction channels on the microfluidic chip are displayed in a digital or graphic manner, and the visualization of the nucleic acid amplification signals of different reaction channels on the microfluidic chip is realized.
As shown in fig. 1, the present invention provides a preferred lensless illumination scheme, i.e. an oblique incidence dark field illumination mode, which adopts a light guide rod 3 to be seamlessly close to the light emitting surface of a light source 2, directly receives the light emitted from the light source 2, and transmits the light to the vicinity of the illumination area through the light guide rod 3; then, expanding light at a certain divergence angle (10-45 degrees), illuminating the measured object from the side of the measured object to a dark field, and performing compensation illumination on the edge of an imaging area at a certain angle (10-45 degrees) by utilizing reflected residual light of the cylinder wall of the light cylinder 5 with uniform light and dense reflected at the bottom to improve the uniformity of the illumination of the side of the measured object to the dark field; and finally, receiving fluorescence generated by the measured object through the imaging unit to realize fluorescence imaging of the measured object, wherein peripheral residual light of the elliptical lighting light spot is reflected back through the bottom reflection uniform light-dense light cylinder 5 to supplement the edge of the lighting micro-fluidic chip 6, so that the uniformity of light irradiated on the whole surface of the micro-fluidic chip 6 is improved.
As shown in fig. 1, after the excitation light generated by the light source 2 is transmitted by the light guide rod 3 and filtered by the excitation filter 4, the light is obliquely incident on the microfluidic chip 6, the reflected light is reflected at the same angle on the other side of the vertical axis of the microfluidic chip 6 by taking the vertical axis of the microfluidic chip 6 as a symmetry axis, and the oblique incident angle is adjusted, so that the reflected light without the excitation light in a certain range right above the microfluidic chip 6 (in the vertical axis direction) can be realized, the influence of the reflection of the excitation light on a fluorescence signal can be effectively reduced, and the dark field illumination effect can be achieved. The nucleic acid amplification products of reaction channels in different areas on the microfluidic chip 6 generate fluorescent signals under the irradiation of exciting light of the light source 2, are filtered by the emission filter 8, are converged on the detector 10 by the imaging lens 9, are input into the computer display processor 11 after photoelectric conversion, and are further processed by the computer display processor 11 in a digital or graphic manner, so that the nucleic acid amplification fluorescent signals of different reaction channels on the microfluidic chip 6 are displayed in a digital or graphic manner, and the visualization of the nucleic acid amplification signals of different reaction channels on the microfluidic chip 6 is realized.
As shown in fig. 2, the utility model also provides another kind of preferred no lens illumination scheme, coaxial normal incidence illumination mode promptly, this scheme cross the even dense light section of thick bamboo 5 of bottom reflection and reflect the peripheral afterglow of oval illumination facula back, and 6 edges of supplementary illumination micro-fluidic chip improve the homogeneity of the whole surface illumination light of micro-fluidic chip 6.
As shown in fig. 2, after the excitation light generated by the light source 2 is transmitted by the light guide rod 3 and filtered by the excitation filter 4, the excitation light is reflected by the beam splitter 14, the normal incidence illuminates the microfluidic chip 6, the reflected light symmetrically returns along the vertical axis of the microfluidic chip 6, and is blocked by the dichroic gating function of the beam splitter 14, and does not enter the detector 10 through the beam splitter 14. The amplified nucleic acid products of the reaction channels in different areas on the microfluidic chip 6 generate fluorescent signals under the irradiation of exciting light of the light source 2, are transmitted by the beam splitter 14 and filtered by the emission filter 8, are converged on the detector 10 by the imaging lens 9, are input into the computer display processor 11 after photoelectric conversion, and are displayed in a digital or graphic manner through further digital signal processing of the computer display processor 11, so that the visualization of the amplified nucleic acid signals of different reaction channels on the microfluidic chip 6 is realized.
As shown in fig. 3, a preferred embodiment of the light-guiding rod for light-transmitting lensless illumination is: the light source 2 is directly coupled with the light guide rod 3, and exciting light generated by the light source 2 is transmitted by the light guide rod 3 and filtered by the exciting light filter 4 to form a relatively uniform quasi-monochromatic light expanded illuminating beam.
Another preferred embodiment of the light-conducting rod for light-conducting lensless illumination, as shown in fig. 4, is: the light source 2 is coupled with the light guide rod 3 through the aspherical mirror 15, exciting light generated by the light source 2 is condensed through the aspherical mirror 15, then is coupled into the light guide rod 3 for transmission, and is filtered through the exciting optical filter 4 to form a relatively uniform quasi-monochromatic light expanded illuminating beam.
As shown in fig. 5, in order to improve the excitation light irradiation uniformity of different regional reaction channels on the microfluidic chip 6, the utility model discloses be provided with a bottom reflection uniform light density optical cylinder 5, the first part extinction area 51 through the bottom reflection uniform light density optical cylinder 5 is handled, the stray light influence that the incident illumination light path produced can be effectively eliminated, and further through the reflex action of the lower half reflection area 52 of the bottom reflection uniform light density optical cylinder 5, the afterglow of the incident illumination light is reflected and folded at certain angle (10 ° -45 °), the marginal area of certain limit around the microfluidic chip 6 of supplementary illumination, can effectively improve the excitation light illumination uniformity of different regional reaction channels on the microfluidic chip 6, further improve the stable reliability that different reaction channels produced fluorescence signal on the microfluidic chip 6.
As shown in fig. 5, the bottom reflection dodging tube 5 is composed of an upper half portion light-eliminating region 51 and a lower half portion reflection region 52. The inner wall of the upper half light-eliminating area 51 is a concentric light-eliminating ring belt, and the surface treatment is carried out through a black oxidation process, so that the influence of stray light generated by an incident illumination light path can be effectively eliminated. The inner wall of the lower half reflection area 52 is a fan-shaped reflection area with a variable angle (10-45 degrees), a reflector can be adopted, or the lower half part is directly processed into a smooth inner wall with a certain reflection angle (10-45 degrees) by adopting a metal material, and bright and white surface treatment is carried out, so that the residual light of incident illumination light can be reflected and reflected at a certain angle (10-45 degrees), the edge area in a certain range around the microfluidic chip 6 is illuminated in a supplementary mode, the uniformity of the excitation light of the microfluidic chip 6 is improved, and the purpose of improving the stability of fluorescent signals generated by different reaction channels on the microfluidic chip 6 is achieved.
The utility model provides a pair of micro-fluidic chip detecting system, its testing process as follows:
1) respectively fixing corresponding molecular probes for nucleic acid detection at the bottom of a reaction channel of the microfluidic chip 6 according to a plurality of nucleic acid indexes to be detected;
the molecular probe for nucleic acid detection consists of 6 sections of primers, is fixed at the bottom of a reaction channel of the microfluidic chip 6, detects different nucleic acid indexes, and is designed into nucleic acid segments with different T, G, A, C base sequence sequences and lengths, thereby having the function of specific detection and identification of pathogenic bacteria, viruses, microorganisms, fungi and the like of different species;
the molecular probe for detecting nucleic acid can be embedded at the bottom of a reaction channel of the micro-fluidic chip MChip by using a low-melting-point biological meltable material, is heated by a heating film HF (hydrogen fluoride) and is melted at the temperature of 30-95 ℃ to release a primer, and is mixed with a sample to be detected and a reaction reagent to carry out nucleic acid amplification reaction.
The 6-segment primer of the molecular probe for detecting nucleic acid is suitable for the molecular diagnosis and analysis of nucleic acid isothermal amplification, and the sequence design result of a group of specific nucleic acid segments for detecting the yersinia sporogenes (Pneumocystis jiirovaci) is as follows:
primer No. 1-GTAGTGAAATACAAATCGGACT;
primer No. 2-CTGTTCTGGGCTGTTTCC;
primer No. 3-AGTGCTATACCTACTATTTTTAAGAGGAGGATATAGCTGGTTTTCTGC;
primer No. 4-TCGAGGGAGTATGAAAATATTTATCTCACCTTATCGCACATAGTCTGA;
primer No. 5-ATAAACAATTTGCCAAAACAATTTTC;
primer No. 6-GATATTTAATCTCAAAATAACTATTTCTTAA;
2) dissolving a nucleic acid sample to be analyzed into a reagent for nucleic acid detection, and injecting the reagent for nucleic acid detection dissolved with the nucleic acid sample into corresponding microfluidic channels from each sample inlet hole of the microfluidic chip 6 by using a liquid transfer device;
wherein the reagent for nucleic acid detection mainly comprises dNTPs, EvaGreen, DTT, BstDNA Polymerase Buffer, Tris-HCl (pH 8.8 at 25 ℃), MgSO4M-MLV reverse transcriptase, RNase Plus, BstDNA Polymerase and Betaine, can realize the molecular diagnosis application of nucleic acid samples on the microfluidic chip 6, and the reaction system of single index nucleic acid detection is less than or equal to10 mu L, the detection limit reaches 1000 nucleic acid molecule copies.
3) The computer display processor 11 controls the movement control module 12 to realize the detection of different areas of the microfluidic chip 6; meanwhile, the computer display processor 11 controls the heating film 7 through the temperature control module 13 to uniformly heat the microfluidic chip 6, and keeps the temperature of the microfluidic chip 6 to change within the temperature range of 30-95 ℃ according to practical application, so that primers of each reaction channel in the microfluidic chip 6 are released and mixed with a nucleic acid sample and a biological reagent, and a nucleic acid amplification reaction is carried out under the condition of temperature change or isothermal amplification, thereby realizing the molecular diagnosis of the nucleic acid sample;
4) the LED exciting light is guided into and irradiates the microfluidic chip 6, so that a nucleic acid sample in the microfluidic chip 6 generates fluorescence under the excitation of the exciting light, the fluorescence is filtered by the emission optical filter 8, the fluorescence is collected by the imaging lens 9 and is converged on the detector 10 to generate an analog signal, the generated analog signal is digitally converted by the detector 10 and then is sent to the computer display processor 11 to generate a real-time fluorescence detection signal, and a fluorescence detection signal curve is displayed by the display.
The molecular probe for nucleic acid detection is fixed at the bottom of the reaction channel on the microfluidic chip 6 by surface adsorption, or embedded and fixed at the bottom of the reaction channel of the microfluidic chip 6 by agarose or oligosaccharide or a biocompatible material with a melting point of 30-95 ℃.
The molecular probe for nucleic acid detection consists of 6 segments of primers, and is used for detecting different nucleic acid indexes, wherein the 6 segments of primers are designed into nucleic acid fragments with different A, T, G, C base sequence sequences and lengths; the reagent for nucleic acid detection mainly comprises dNTPs, EvaGreen, DTT, BstDNA Polymerase Buffer, Tris-HCl with pH 8.8 at 25 ℃, MgSO4, M-MLVrevererse transcriptase, RNase Plus, BstDNA Polymerase and Betaine, and a single index reaction system is less than or equal to 10 mu L.
The light guide rod 3 is preferably a glass rod, a plastic rod, a quartz fiber, or the like, and the light source 2 is preferably an LED, a tungsten halogen lamp, a laser, a fluorescent lamp, or the like.
The beam splitter 14 may be a dichroic plate with a certain reflectance and transmittance, a prism, or a dichroic mirror that is gated according to a certain wavelength range.
The heating film 7 can be resistance wire heating or semiconductor heating.
The bottom reflection uniform light-density light cylinder 5 adopts an up-down structure, the extinction area 51 on the upper layer is a black extinction ring, and the reflection area 52 on the lower layer is a mirror reflection layer with a certain angle, so that peripheral residual light of an illumination light spot can be reflected back to supplement the edge of the illumination micro-fluidic chip 6, and the uniformity of light irradiated on the whole surface of the micro-fluidic chip 6 is improved.
The utility model provides a pair of leaded light stick passes no lens illumination imaging device of light and micro-fluidic chip detecting system has following advantage:
(1) the utility model discloses owing to adopt the nucleic acid that comprises 6 sections primers to detect with molecular probe, to different nucleic acid indexes, 6 sections primers design into the nucleic acid fragment of different A, T, G, C base sequence order and length, consequently have the specificity detection recognition function to pathogenic bacteria, virus, microorganism, fungi of different species etc..
(2) The utility model discloses owing to adopt the nucleic acid to detect with reagent and the nucleic acid sample mixture that analyzes, can realize trace nucleic acid sample molecular diagnosis on micro-fluidic chip MC and use, the reaction system who realizes single index nucleic acid detection is less than or equal to 10 mu L, detects the limit and reaches within 1000 nucleic acid molecule copies.
(3) The utility model discloses owing to adopt the leaded light stick to pass the illumination of no lens of light, the system architecture becomes simple, and fluorescence collection efficiency is than higher.
(4) The utility model discloses owing to adopt the even light of bottom reflection to close a section of thick bamboo 5, stray light influence can be eliminated to upper black extinction ring, and the lower floor has certain angle (10-45) specular reflection layer, can reflect the peripheral afterglow of illumination facula back, and 6 edges of supplementary illumination micro-fluidic chip improve the homogeneity of the 6 whole surface illumination light of micro-fluidic chip.
The utility model provides a pair of no lens illumination imaging device of leaded light stick biography light and micro-fluidic chip detecting system can be used for fields such as clinical pathogenic bacteria molecular diagnosis, food inspection and quarantine, health and epidemic prevention, has great economic and social.
The foregoing is a more detailed description of the present invention, taken in conjunction with the specific preferred embodiments thereof, and it is not intended that the invention be limited to the specific embodiments shown and described. To the utility model belongs to the technical field of ordinary technical personnel, do not deviate from the utility model discloses under the prerequisite of design, can also make a plurality of simple deductions or replacement, all should regard as belonging to the utility model discloses a protection scope.
Claims (10)
1. The utility model provides a leaded light stick passes light no lens illumination image device which characterized in that: the device comprises an imaging unit, a light source, a light guide rod, an excitation light filter, a bottom reflection uniform-light-density light cylinder and a micro-fluidic chip, wherein light emitted by the light source is transmitted through the light guide rod and filtered through the excitation light filter to form a quasi-monochromatic light expanded illuminating beam, one part of the quasi-monochromatic light expanded illuminating beam directly enters the micro-fluidic chip to illuminate an imaging area of the micro-fluidic chip, the other part of the quasi-monochromatic light expanded illuminating beam enters the micro-fluidic chip under the reflection of the cylinder wall of the bottom reflection uniform-light-density light cylinder to perform compensation illumination on the edge of the imaging area of the micro-fluidic chip, and fluorescence generated by a measured object is received through the imaging unit to realize fluorescence imaging of the measured object.
2. The light-guiding rod light-transmitting lensless illumination imaging device of claim 1, wherein: the light guide rod is seamlessly close to the light emitting surface of the light source and directly receives light emitted from the light source.
3. The light-guiding rod light-transmitting lensless illumination imaging device of claim 1, wherein: an aspherical mirror is arranged between the light source and the light guide rod, the light source is coupled with the light guide rod through the aspherical mirror, light generated by the light source is condensed by the aspherical mirror, then is coupled to the light guide rod for transmission, and is filtered by the excitation filter to form a quasi-monochromatic light expanded illumination beam.
4. The light-guiding rod light-transmitting lensless illumination imaging device of claim 1, wherein: a beam splitter is arranged between the excitation optical filter and the bottom reflection uniform light density tube, the quasi monochromatic light expanded illumination light beam formed after the excitation optical filter is filtered is emitted into the beam splitter and is reflected by the beam splitter, one part of the quasi monochromatic light expanded illumination light beam is directly emitted into the microfluidic chip, and the other part of the quasi monochromatic light expanded illumination light beam is emitted into the microfluidic chip under the reflection of the tube wall of the bottom reflection uniform light density tube.
5. The light-guiding rod light-transmitting lensless illumination imaging device of claim 4, wherein: the light reflected by the beam splitter is vertically emitted into the microfluidic chip, and the light reflected by the microfluidic chip is cut off by the dichroic gating function of the beam splitter and cannot enter the imaging unit through the beam splitter.
6. The light-guiding rod light-transmitting lensless illumination imaging device of claim 1, wherein: and the quasi-monochromatic light expanded illuminating light beam formed after the filtering of the excitation filter is emitted into the microfluidic chip from the side direction.
7. The light-guiding rod light-transmitting lensless illumination imaging device of claim 1, wherein: the light source is connected with a radiating fin, the imaging unit, the bottom reflection light-homogenizing and light-concentrating light cylinder and the micro-fluidic chip are coaxially arranged, the bottom reflection light-homogenizing and light-concentrating light cylinder is located between the imaging unit and the micro-fluidic chip, and the light source, the light guide rod and the excitation filter are located on the side edge of the micro-fluidic chip.
8. The light-guiding rod light-transmitting lensless illumination imaging device of claim 1, wherein: the bottom reflection uniform light density light cylinder is positioned right above the microfluidic chip, the upper layer part of the bottom reflection uniform light density light cylinder is a light extinction area, the lower layer part of the bottom reflection uniform light density light cylinder is a reflection area, and the other part of the quasi-monochromatic light expanded illumination light beam is emitted into the microfluidic chip under the reflection of the reflection area of the bottom reflection uniform light density light cylinder.
9. A micro-fluidic chip detection system is characterized in that: the light-guide rod light-transmission lens-free illumination imaging device comprises the light-guide rod light-transmission lens-free illumination imaging device as claimed in any one of claims 1 to 8, wherein specific gene primer sequences for nucleic acid detection are respectively fixed at the bottoms of reaction channels of the microfluidic chip.
10. The microfluidic chip detection system according to claim 9, wherein: the micro-fluidic chip is embedded with a biomolecule detection probe for carrying out nucleic acid isothermal or temperature-varying amplification detection, the imaging unit comprises a detector, an imaging lens and an emission filter, the imaging lens is positioned between the detector and the emission filter, the micro-fluidic chip detection system further comprises a heating film, a temperature control module, a motion control module and a computer display processor, the heating film is positioned under the micro-fluidic chip and is connected with the temperature control module, the micro-fluidic chip is connected with the motion control module, the temperature control module, the motion control module and the detector are respectively connected with the computer display processor, the computer display processor sends a motion control signal to the motion control module, and the motion control module sends the motion control signal to the motion control module according to the received motion control signal, controlling the micro-fluidic chip to do one-dimensional or two-dimensional planar motion to realize detection and analysis of reaction channels in different areas on the micro-fluidic chip; the computer display processor transmits a temperature control signal to the temperature control module, and the temperature control module controls the temperature of the heating film according to the received temperature control signal to meet the requirement of the temperature control condition of nucleic acid amplification of the microfluidic chip; the nucleic acid amplification products of the reaction channels in different areas on the microfluidic chip generate fluorescent signals under the irradiation of exciting light of a light source, are filtered by the emission optical filter, are converged on the detector by the imaging lens, are input into the computer display processor after photoelectric conversion, and are further processed by digital signals of the computer display processor to display the nucleic acid amplification fluorescent signals in a digital or graphic mode, so that the visualization of the nucleic acid amplification signals of different reaction channels on the microfluidic chip is realized.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110132931A (en) * | 2019-06-17 | 2019-08-16 | 北京百康芯生物科技有限公司 | Lamp guide passes light without lens illumination imaging device and fluidic chip detecting system |
| CN112577932A (en) * | 2020-11-27 | 2021-03-30 | 苏州雅睿生物技术有限公司 | Single-light-source multi-sample fluorescence detection optical system and working method thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110132931A (en) * | 2019-06-17 | 2019-08-16 | 北京百康芯生物科技有限公司 | Lamp guide passes light without lens illumination imaging device and fluidic chip detecting system |
| CN112577932A (en) * | 2020-11-27 | 2021-03-30 | 苏州雅睿生物技术有限公司 | Single-light-source multi-sample fluorescence detection optical system and working method thereof |
| CN112577932B (en) * | 2020-11-27 | 2022-02-18 | 苏州雅睿生物技术股份有限公司 | Single-light-source multi-sample fluorescence detection optical system and working method thereof |
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