CN202492522U - Real-time fluorescent PCR (Polymerase Chain Reaction) working system of space-oriented micro-volume unit - Google Patents
Real-time fluorescent PCR (Polymerase Chain Reaction) working system of space-oriented micro-volume unit Download PDFInfo
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- CN202492522U CN202492522U CN2012200717805U CN201220071780U CN202492522U CN 202492522 U CN202492522 U CN 202492522U CN 2012200717805 U CN2012200717805 U CN 2012200717805U CN 201220071780 U CN201220071780 U CN 201220071780U CN 202492522 U CN202492522 U CN 202492522U
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Abstract
A real-time fluorescent PCR working system of a space-oriented micro-volume unit belongs to the detection fields of biology, analytical chemistry and medical science and comprises a ceramic refrigeration chip (1), an electrothermal film (2), a hard transparent plastic (3), a heat conduction channel (8), a temperature sensor (9), a rubber piston (10), an LED (4) and an excitation light filter (5), a silicon photocell (6) and a detection light filter (7) and further comprises an external microprocessor, a keyboard and a display screen, wherein the ceramic refrigeration chip (1), the electrothermal film (2), the hard transparent plastic (3), the heat conduction channel (8), the temperature sensor (9) and the rubber piston (10) forms a temperature circulation control device; the LED (4) and the excitation light filter (5) serve as an excitation light unit; the silicon photocell (6) and the detection light filter (7) serve as a fluorescence spectrum detector; the external microprocessor is used for controlling the ceramic refrigeration chip (1) to refrigerate and the electrothermal film to heat; and the keyboard and the display screen are used for inputting and output control parameters. The system conforms to the targets of miniature structure, function integration, light weight, small volume and full-automatic detection. The light excitation and the fluorescence detection are in zero-distance contact with an object to be detected and are unlimited by the force of gravity, so that the real-time fluorescent PCR working system of the space-oriented micro-volume unit is stable in detection results without being interfered and is more applicable to spatial working requirements.
Description
Technical field
The utility model relates to the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody, mainly is applicable to the real-time fluorescence PCR work of in the spatial dimension under the below-G conditions, using, and belongs to biology, analytical chemistry and medical science detection range.
Background technology
Along with reaching its maturity of China's manned spaceflight technology, the prolongation gradually in space mission cycle also can be increasingly high to the requirement of spacefarer's health care.Be in the mikrobe serious threat spacefarer's of non-monitor state health in the spacecraft.The correlated results of the medium-term and long-term flight of US and Russia shows that detected 234 kinds of mikrobes in the crew module, bacterium has 40 genus; 108 kinds; Kind surplus wherein pathogenic bacteria has 20, these pathogenic bacterias can cause the spacefarer to infect multiple disease, and even influence the smooth completion of aerial mission.In the long duration space flight, the growth of microorganism species and bacterial strain variation can influence the performance and the working environment of plant and instrument, to the normal operation generation harm of spacecraft and plant and instrument thereof.Simultaneously, carry the mikrobe that can under space environment, survive on the aerospacecraft, might take space station, martian surface to, can cause disadvantageous effect human research's space life form through the mankind or spacecraft.For the generation that prevents infection in the spaceflight, propagate and effectively treat, the mikrobe in must the monitor closely spacecraft.
Polymerase chain reaction (PCR) is a kind of nucleic acid amplification technologies in in-vitro simulated natural dna replication dna process; Extend (72 ℃) by elevated temperature heat sex change (about 94 ℃), low-temperature annealing renaturation (50～60 ℃) and thermophilic and form one-period; A circulation can make the DNA total amount double; Can be 1,000,000 times of the rapid and easy amplifications of the genetic material of trace; Re-use based on FRET (fluorescence resonance energy transmission technology) and carry out fluorescence and detect in real time,, can carry out qualitative and quantitative analysis the PCR product according to the information that fluorescence intensity and intensity change relatively.
Based on the real-time fluorescence quantitative PCR appearance of above-mentioned principle, can reach to mikrobe in the spacecraft of operation in the spatial dimension monitoring.The real-time quantitative PCR appearance adopts laser apparatus or fluorescence excitation light source to monitor in real time; Glitch-free fluorescence excitation is stablized in assurance; Forming optical system by a series of lens, filter and a dichroic mirror focuses on fluorescence excitation on the spectrograph; Spectrograph separates fluorescence with mode at interval according to the difference of wavelength; Get into charge coupled cell (CCD camera) or PM, the sequential detection application software is collected these fluorescent signals from charge coupled cell (CCD camera) or PM, data are analyzed.So far, of a great variety based on the commercial PCR appearance of above-mentioned reaction and detection principle.External brand mainly contains ABI, Roche, MJ, Bio-Rad, Stratagene, Corbtt, Cepheid etc., and rich day of domestic Hangzhou, Guangzhou reach peace gene and Shanghai Feng Ling etc.
Obviously this quasi-instrument has following shortcoming for the real-time fluorescence PCR work of carrying out the job space requirement:
1, bulky, be unfavorable for carrying.Because the volume of PM or charge coupled cell self is just very big, and be again that split is used, need supporting light path system, cause the bulky of whole fluorescence low-light spectrum detection system, be difficult to carry out again micro.
2, because optical excitation unit and light receiving unit use many optical fiber as optical transmission system, unavoidably in the optical fiber coupling process, there is the luminous energy loss when transmission, makes the raising of whole detection system sensitivity be severely limited.
3, because the light path that when exciting light conduction and reflected light collection, needs all kinds of optics and optical fiber to form is carried out light transmission, this has influenced spectral detection system stability in actual use.
4, owing to detect the metering system that does not reach the zero distance contact, make the low-light spectrum detect and receive non-detected object material,, cause measuring error as forming the interference of microchannel wall material.
The utility model content
The utility model relates to the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody, is mainly used under the spatial dimension below-G conditions, to use, to the detection of PCR reaction carrying out real-time fluorescence.Purpose is temperature cycle gear, exciting light conduction device, these three of fluorescence spectrum proofing units partly integrated; Realization is applicable to the portable minisize PCR fluorescence real-time detecting system of job space requirement, reaches that function is integrated, structure is miniature, the target of the little full-automatic detection of volume in light weight.
In order to achieve the above object, the utility model is taked following technical scheme:
The real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody, it includes the ceramic cooling piece 1 of forming the temperature cycle gear, Electric radiant Heating Film 2, hard transparent plastic 3, passage of heat 8, TP 9, rubber piston 10; As the unitary LED photodiode 4 of exciting light, exciting light spectral filter 5 as the silicon photocell 6 of fluorescence spectrum proofing unit, detects light spectral filter 7; Also include and be used to control ceramic cooling piece 1 refrigeration, the external microprocessor of Electric radiant Heating Film 2 heating, the import and export of keyboard and image display carry out to(for) controlled variable.
The real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody; 3 one-tenth tubulars of hard transparent plastic volume; Electric radiant Heating Film 2 is at the skin of hard transparent plastic volume 3, and the interior STH at the both ends up and down of hard transparent plastic volume 3 embeds exciting light spectral filter 5, is coated with LED photodiode 4 on the exciting light spectral filter 5; LED photodiode 4 is positioned at the outside of hard transparent plastic volume 3, and Electric radiant Heating Film 2 is in above-mentioned not perforate of position of opening; Hard transparent plastic is rolled up the inboard perforate in end, 3 front and back and is embedded detection light spectral filter 7, detects on the light spectral filter 7 and is coated with silicon photocell 6, and silicon photocell 6 is positioned at the outside of hard transparent plastic volume 3.Passage of heat 8 is left in the outside, two ends of Electric radiant Heating Film 2.To be spliced into lateral cross section be foursquare rectangular parallelepiped to pottery cooling piece 1, and above-mentioned hard transparent plastic volume 3 places the internal space of described rectangular parallelepiped tangent, and between ceramic cooling piece 1 and hard transparent plastic volume 3 the falling heat-transfer elargol.Above-mentioned transparent plastics is rolled up 3 two ends with piston 10 encapsulation, and embeds TP 9 on the piston 10 of a side therein.
Use the silica gel adhesion between Electric radiant Heating Film 2 and the hard transparent plastic volume 3.
The sheet number that is spliced into the ceramic cooling piece 1 of rectangular parallelepiped is 4.
Described piston 10 is a rubber piston.
Detecting light spectral filter 7 transmission peak wavelengths is the fluorescence of 525nm.
Described ceramic cooling piece size is 5mm * 5mm, and thickness is 1mm.
That described silicon photocell and TP are of a size of is long * widely be: 1mm * 1mm.
The utility model is that a non-electrician of the real-time fluorescence PCR work system of the long-pending unit of microbody makes the unit; Integrated all non-electric key elements; But need external microprocessor, keyboard and image display etc. during work, its working process is that ceramic cooling piece 1 rolls up 3 through the external microprocessor control and the feedback of TP 9 with hard transparent plastic; Being heated to 94 ℃ makes the genetic material in the P C R reaction solution high-temperature denatured; After be cooled to 55 ℃ of low temperature renaturation of withdrawing from a secret society or underworld gang, be warming up to 72 ℃ at last and make it thermophilic and extend these 3 temperature that circulate (30～40 cycles).The light that sends from LED photodiode 4 is simultaneously filtered at spectral filter 5, the 475nm excitation light irradiation object PCR to be detected reaction solution of selecting.Material to be detected goes out fluorescence by excitation, and after detecting light spectral filter 7, the fluorescence that leaches wavelength 525nm is received by silicon photocell 6, becomes electrical signal output.
The real-time fluorescence PCR work system of the long-pending unit of the microbody of the utility model compared with prior art has following remarkable advantages and beneficial effect:
The whole non-electric key element of real-time fluorescence PCR work that the utility model is integrated; Temperature circulatory system, reactive system and detection system; Wherein heating can reach 5mm * 5mm size with refrigeration plant, and cooling piece thickness can reach 1mm, and LED photodiode, silicon photocell and TP can reach 1mm * 1mm; Make a whole set of work system narrow down to 5mm * 5mm size, meet structure micro, the little full-automatic target that detects of integrated, the in light weight volume of function.Optical excitation can contact with thing zero distance to be detected with fluoroscopic examination, does not receive the gravity requirement, makes detected result stable, and is interference-free, more adapts to the space job requirement.
The utility model stands in the integrated aspect of real-time fluorescence PCR work, if the space-oriented need be accurate to automatic sampling in the sample introduction direction, could constitute the integrity of using.The hand sampling of the utility model can be accomplished by the cosmonaut in the space.
Description of drawings
The real-time fluorescence PCR work system of the long-pending unit of Fig. 1 microbody is along the longitudinal section view of fore-and-aft direction
The real-time fluorescence PCR work system of the long-pending unit of Fig. 2 microbody is along the longitudinal section view of left and right directions
The real-time fluorescence PCR work system overall schematic of the long-pending unit of Fig. 3 microbody
Among the figure: 1 ceramic cooling piece, 2 Electric radiant Heating Film, 3 hard transparent plastics, 4LED photodiode, 5 exciting light spectral filters, 6 silicon photocells, 7 detect light spectral filter, 8 passages of heat, 9 TPs, 10 pistons.
Specific embodiment
Specify present embodiment below in conjunction with accompanying drawing 1～3.
The structural representation of present embodiment such as Fig. 1,2 and 3.Fig. 1 is its longitudinal section view along fore-and-aft direction; Can only draw among the figure ceramic cooling piece 1, Electric radiant Heating Film 2, hard transparent plastic 3, as the unitary LED photodiode 4 of exciting light; Exciting light spectral filter 5, passage of heat 8 and TP 9 and rubber piston 10, the detecting unit side spectral filter 7 that can only draw.Said ceramic cooling piece 1 every be 5mm * 5mm size, 1mm thickness is spliced into the rectangular parallelepiped of 7mm * 5mm * 5mm size.LED photodiode 4 is formed the optical excitation unit with exciting light spectral filter 5, wherein as the LED photodiode 4 of excitation light source luminous peak wavelength be 475nm; The peak wavelength of exciting light spectral filter 5 transmissions also is 475nm; Silicon photocell 6, detection light spectral filter 7 are formed optical detecting units, and detecting light spectral filter 7 transmissive wavelength that measured matter produces is the fluorescence of 525nm, and the 525nm fluorescence that silicon photocell 6 will detect 7 transmissions of light spectral filter is converted into electrical signal; Passage of heat 8 is connected ceramic cooling piece 1 with hard transparent plastic 3, the falling heat-transfer elargol can make hard transparent plastic 3 internal reaction liquid better carry out temperature exchange in the middle of the passage of heat 8; Temperature variation in 9 pairs of PCR reaction process of TP feeds back to external microprocessor, is controlled the work of cooling piece 1 and Electric radiant Heating Film 2 again by microprocessor; 10 is rubber piston, with whole work system sealing.
The working process of the real-time fluorescence PCR work system of the long-pending unit of whole microbody is; Use needle tubing that the PCR reaction solution is injected by the piston place; Start work system; By three warm area times of keyboard input temp round-robin, microprocessor carries out the PCR reaction according to the feedback control cooling piece 1 and heating film 2 work of TP 9.The light that sends from LED photodiode 4 is simultaneously filtered at spectral filter 5, the 475nm excitation light irradiation object PCR to be detected reaction solution of selecting.Material to be detected goes out fluorescence by excitation, and after detecting light spectral filter 7, the fluorescence that leaches wavelength 525nm is received by silicon photocell 6, becomes electrical signal output.Fig. 2 is the longitudinal section view of this detection system along left and right directions; Can only be among the figure by ceramic cooling piece, heating film 2, hard transparent plastic 3, LED photodiode 4, exciting light spectral filter 5 and silicon photocell 6, detection light spectral filter 7 and a TP 9 and a side piston 10, heat transfer path 8 fails to draw.Form the optical excitation unit by exciting light spectral filter 5 and silicon photocell 6, form optical detecting unit with TP 9 by detecting light spectral filter 7; The purpose of using two groups of optical excitation unit and two groups of optical detecting units is further to improve detection sensitivity.The long-pending real-time fluorescence PCR work system of the microbody of the utility model has following characteristics: integrated whole key elements of real-time fluorescence PCR work system, and like temperature cycle and control, excitation light source, beam splitting system, light detection etc.Because the device that uses all is millimetre-sized, makes the characteristic dimension of total system narrow down to and have only 5mm * 5mm * the 7mm millimeter.
The long-pending real-time fluorescence PCR work system manufacture craft flow process of microbody is following: at first with 3 one-tenth tubulars of hard transparent plastic volume; Again Electric radiant Heating Film 2 usefulness silica gel are sticked in its skin; Embed LED photodiode 4 and exciting light spectral filter 5 in both sides up and down at heating film; Front and back embed silicon photocell 6 and detect light spectral filter 7, and passage of heat 8 is left in both sides.Once more, 1 four of ceramic cooling pieces are spliced into rectangular parallelepiped, the cylinder that the last step processes is wherein built-in, and at attaching space falling heat-transfer elargol not.Last two encapsulates with piston 10, and on the bottom piston, embeds TP 9.
Claims (8)
1. the real-time fluorescence PCR work system of the long-pending unit of a space-oriented microbody, it is characterized in that: it includes the ceramic cooling piece (1) of forming the temperature cycle gear, Electric radiant Heating Film (2); Hard transparent plastic (3); Passage of heat (8), TP (9), rubber piston (10); As the unitary LED photodiode of exciting light (4), exciting light spectral filter (5) as the silicon photocell (6) of fluorescence spectrum proofing unit, detects light spectral filter (7); Also include and be used to control ceramic cooling piece (1) refrigeration, the external microprocessor of Electric radiant Heating Film (2) heating, the import and export of keyboard and image display carry out to(for) controlled variable;
Hard transparent plastic volume (3) becomes tubular; Electric radiant Heating Film (2) is at the skin of hard transparent plastic volume (3); The interior STH of rolling up the both ends up and down of (3) at hard transparent plastic embeds exciting light spectral filter (5); Be coated with LED photodiode (4) on the exciting light spectral filter (5), LED photodiode (4) is positioned at the outside of hard transparent plastic volume (3), and Electric radiant Heating Film (2) is in above-mentioned not perforate of position of opening; The inboard perforate in end embeds and detects light spectral filter (7) before and after the hard transparent plastic volume (3), detects on the light spectral filter (7) and is coated with silicon photocell (6), and silicon photocell (6) is positioned at the outside of hard transparent plastic volume (3); Passage of heat (8) is left in the outside, two ends of Electric radiant Heating Film (2); To be spliced into lateral cross section be foursquare rectangular parallelepiped to pottery cooling piece (1), and above-mentioned hard transparent plastic volume (3) places the internal space of described rectangular parallelepiped tangent, and between ceramic cooling piece (1) and hard transparent plastic volume 3 the falling heat-transfer elargol; Above-mentioned transparent plastics volume (3) two ends are with piston (10) encapsulation, and the piston of a side (10) is gone up embedding TP (9) therein.
2. the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody according to claim 1 is characterized in that: use the silica gel adhesion between Electric radiant Heating Film (2) and the hard transparent plastic volume (3).
3. the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody according to claim 1, it is characterized in that: the sheet number that is spliced into the ceramic cooling piece (1) of rectangular parallelepiped is 4.
4. the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody according to claim 1, it is characterized in that: described piston (10) is a rubber piston.
5. the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody according to claim 1 is characterized in that: photodiode (4) luminous peak wavelength be 475nm; The peak wavelength of exciting light spectral filter (5) transmission also is 475nm.
6. the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody according to claim 1 is characterized in that: detecting light spectral filter (7) transmission peak wavelength is the fluorescence of 525nm.
7. the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody according to claim 1 is characterized in that: described ceramic cooling piece size is 5mm * 5mm, and thickness is 1mm.
8. the real-time fluorescence PCR work system of the long-pending unit of a kind of space-oriented microbody according to claim 1 is characterized in that: described silicon photocell and TP be of a size of grow * widely be: 1mm * 1mm.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012200717805U CN202492522U (en) | 2012-02-29 | 2012-02-29 | Real-time fluorescent PCR (Polymerase Chain Reaction) working system of space-oriented micro-volume unit |
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| CN2012200717805U CN202492522U (en) | 2012-02-29 | 2012-02-29 | Real-time fluorescent PCR (Polymerase Chain Reaction) working system of space-oriented micro-volume unit |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586098A (en) * | 2012-02-29 | 2012-07-18 | 北京工业大学 | Real-time fluorescent PCR (polymerase chain reaction) working system of space-oriented micro-volume unit |
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- 2012-02-29 CN CN2012200717805U patent/CN202492522U/en not_active Withdrawn - After Issue
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586098A (en) * | 2012-02-29 | 2012-07-18 | 北京工业大学 | Real-time fluorescent PCR (polymerase chain reaction) working system of space-oriented micro-volume unit |
| CN102586098B (en) * | 2012-02-29 | 2013-11-20 | 北京工业大学 | Real-time fluorescent PCR (polymerase chain reaction) working system of space-oriented micro-volume unit |
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| C14 | Grant of patent or utility model | ||
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| AV01 | Patent right actively abandoned |
Granted publication date: 20121017 Effective date of abandoning: 20131120 |
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| RGAV | Abandon patent right to avoid regrant |