CN102353659A - Detector for biochip fluorescent microspectrum and manufacture method thereof - Google Patents
Detector for biochip fluorescent microspectrum and manufacture method thereof Download PDFInfo
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- CN102353659A CN102353659A CN201110153526XA CN201110153526A CN102353659A CN 102353659 A CN102353659 A CN 102353659A CN 201110153526X A CN201110153526X A CN 201110153526XA CN 201110153526 A CN201110153526 A CN 201110153526A CN 102353659 A CN102353659 A CN 102353659A
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Abstract
A detector for biochip fluorescent microspectrum and a manufacture method thereof. (1) represents an upper cover chip, and (2) represents a lower loading chip; (11) represents a micro channel laden with biomicrofluidic to be detected; an upper side and a lower side of the micro channel (11) are equipped with optical excitation units (12) (13), and optical detection units (14) (15) are symmetrically distributed on the left and the right of the micro channel (11); light emitted from an excitation light source (3) is filtered by optical filters (4), and selected excitation light is focalized through optical microlenses (5) at tops of the excitation units and transmits into the micro channel (11) to irradiate the biomicrofluidic to be detected; the substance to be detected is excitated by the exciting light to radiate fluorescence, which is acquired by optical microlenses (7) tops of the detection units; fluorescence filtered through the optical filters is received by semiconductor photoelectric converters and changed into electric signals to be output. The invention combines all non-electrical factors of the spectrum detection; and the detector has a minimal structure to realize micromation of the chip detection apparatus.
Description
Technical field
The present invention relates to a kind of fluorescence biosensor chip low-light spectrum detection device, be mainly used in the detection of the microfluid fluorescence low-light spectrum in the biochip, belong to biology, analytical chemistry and medical science detection range.
Technical background
Biochip is a new and high technology that in life science, develops rapidly in recent years; It can realize all biological engineering analysis process and whole laboratory function on microminiaturized basis; To sample, dilute, add functions such as reagent, reaction, separation and detection is integrated in the chip; Thereby generically being called " chip lab ", its science and advance embody a concentrated reflection of miniature and integrated these two aspects of function of structure.Biochip technology has 4 bare bones: chip preparation, specimen preparation, biochemical reaction and input.Input is the important component part of biochip technology, mainly comprises signal generation, signal collection and transmission, signal Processing and discerns three parts.In various biochip signal detecting methods, fluorescence low-light spectrum detection method has advantages such as good reproducibility, selectivity are strong, highly sensitive, non-destructive detection, is one of present most widely used detection technique.Like the little detection of all kinds of biological medical science micro-fluidic chip, the little detection of all kinds of causal organism medical science microarray, the quantitative little detection of each collaurum demarcation property, the little detection of each solid state chemistry test paper qualitative, quantitative etc.
Employed excitation source is traditional encapsulated semiconductor luminescent device in the low-light spectrum detection technique at present, like semiconductor light-emitting-diode (LED) or semiconductor laser diode (LD).And fluorescence low-light spectrum detecting device all is directly to use traditional encapsulation photoelectric tube, like photomultiplier (PMT) or charge coupled cell (CCD)." laser-Induced Fluorescence Detection micro-fluidic chip Biochemical Analyzer " (analytical chemistry instrument device and experimental technique of Shanghai Spectrum Apparatus Co., Ltd. in 2008 and microanalysis device research institute of department of chemistry of Zhejiang University joint research and development for example; 36 (1): 127～131; 2008.1), " Portable Fluorescence Detection System Integrated with Disposable Microfluidic Chip " (nanometer technology and precision engineering of people such as Chen Xing; 7 (2): 127～131; 2009.3), 433～445,2009.3), " imaging of line sweep quasi confocal fluorescent " (optical precision engineering of people such as Yang Bin people such as Lin Ping Cheng " micro-fluidic chip Experiment of Analytical Chemistry chamber " (SCI, 30 (3):; 18 (5): 1028～1034,2010.5) etc.Obviously there is following shortcoming in general fluorescence biosensor chip low-light spectrum detection device at present:
1, because the volume of photomultiplier or charge coupled cell self is just very big, and is again that split is used, needs supporting light path device, cause the bulky of whole fluorescence low-light spectrum detection device, possibly not be embedded in the biochip.
2, because optical excitation unit and light receiving unit all do not have lens to focus on; Simultaneously use many optical fiber again as light transmitting device; Unavoidably in the optical fiber coupling process, there is the luminous energy loss, makes the raising of whole detection device sensitivity be severely limited.
3, because the light path that when exciting light conduction and reflected light collection, needs all kinds of optical device and optical fiber to form is carried out light transmission, this has influenced spectrum detection device 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.
Present fluorescence biosensor chip low-light spectrum detects present situation; Be in the peripherals of PC main frame size; Remove to detect the chip of thumbnail; The science advance of the scientific meaning of its instrument manufacturing and biochip theory runs in the opposite direction, and this situation has hindered the integrated raising of biochip widely, becomes the bottleneck of biochip development.People such as Bambang Kuswandi in 2007 summarize in " Optical sensing systems for micro-fluidic devices:A review " and have summarized the 106 piece documents of various countries biological chip micro fluid optical detection apparatus research field during last century the nineties to 2007 year; Point out research based on the microminiaturized integrated circuit of spectroscopy pick-up unit of MEMS (Micro Electro Mechanical System) Micrometer-Nanometer Processing Technology; Be the developing direction of biological chip micro fluid optical detection apparatus, the development volume is little of embedding one of chip and highly sensitive focus that has become the research of current various countries to the low-light spectrum detection device that can reach the biotechnology requirement.
Summary of the invention
The objective of the invention is to through a kind of fluorescence biosensor chip low-light spectrum detection device and method for making are provided, with signal generation, signal collection and transmission, signal Processing in the input and discern these three partly integrated, realize that chip is integrated.
The present invention adopts following technological means to realize:
A kind of fluorescence biosensor chip low-light spectrum detection device, 1 is the cover core sheet on top, 2 is the chip that carries of bottom; 11 for being filled with the microchannel of biological microfluid to be detected; 11 upper and lower sides has been provided with optical excitation unit 12,13 in the microchannel, and 11 left-right symmetric is distributed with optical detecting unit 14,15 in the microchannel; The optical excitation unit is a tubulose, by excitation source 3, exciting light optical filter 4, excite the unit optical microlens 5 and with whole excite the unit around the multilayer film 6 that are surrounded form; Optical detecting unit is a tubulose, by the optical microlens 7 of detecting unit, detect light optical filter 8, electrooptical device 9 and the multilayer film 10 that are surrounded around the whole detecting unit are formed; The light that sends from excitation source 3 is filtered at optical filter 4, and the exciting light of selecting focuses on through the optical microlens 5 that excites the top, unit, propagates into 11 li of microchannels, shines object organisms microfluid to be detected; Material to be detected goes out fluorescence by excitation, and the optical microlens 7 at top, unit to be detected is gathered, and behind optical filter 8, the fluorescence that leaches is received by semiconductor light power conversion device 9, becomes electric signal output.
Aforesaid semiconductor light power conversion device 9 is photodiode or silicon blue streak battery.
The transmission peak value wavelength of aforesaid exciting light is 525nm.
A kind of method for making of fluorescence biosensor chip low-light spectrum detection device may further comprise the steps: make the optical excitation unit, excitation source 3 and exciting light optical filter 4 usefulness multilayer film 6 are surrounded as tubulose; On optical filter, make micro optical lens 5 with the formed in situ legal system then, the glue of ultraviolet optics curing soon drips from certain altitude and vertically discharges, and drops onto on the optical filter 4; And from top to bottom to diffusion trickling all around; In time drop is shone, make its curing, form micro optical lens with Ultra-Violet Laser;
Make optical detecting unit, at first electrooptical device 9 is surrounded as tubulose with detecting light optical filter 8 usefulness multilayer film 10, on optical filter, make micro optical lens then with the formed in situ legal system; On upper cap chip 1, make a call to a hole, the optical excitation unit is placed in one, lenticule 7 down;
Carry on the chip 2 in the bottom and to go out the microchannel 11 as biological microfluid to be detected with laser ablation, at the bottom and two side perforatings of microchannel, optical excitation unit 13 is placed in the base apertures then, lenticule up; Optical detecting unit 14 and 15 is placed respectively in the hole of both sides, the lenticule of optical detecting unit 14 towards a left side, carries chip adhesive together with upper cap chip and bottom towards the lenticule of right, optical detecting unit 15, and four lenticules all have the microchannel of determinand towards stream.
Fluorescence low-light spectrum detection device of the present invention compared with prior art has following remarkable advantages and beneficial effect:
The whole non-electric key element of spectral detection that the present invention is integrated is spared bundle, light collection, light detection etc. like gathering, transmission, the light of excitation source, light-dividing device, light.Owing to replaced the photomultiplier PMT or the charge coupled cell CCD that can't embed chip, make the characteristic dimension of whole device narrow down to and have only the millimeter magnitude, even can also be more further micro to micron dimension.
Description of drawings
Fig. 1 is the longitudinal section view of fluorescence low-light spectrum detection device along fore-and-aft direction;
Fig. 2 is the longitudinal section view of fluorescence low-light spectrum detection device along left and right directions;
Fig. 3 is the technological process of fluorescence low-light spectrum detection device;
Fig. 4 is the integral core chip architecture.
Among the figure: 1 is the cover core sheet, and 2 for carrying chip, and 3 is excitation source, and 4 is the exciting light optical filter; 5 for exciting the optical microlens of unit, and 6 for exciting the multilayer film of unit, and 7 is the optical microlens of detecting unit; 8 for detecting the light optical filter, and 9 is electrooptical device, and 10 is the multilayer film of detecting unit; 11 is the microchannel, and 12,13 are the optical excitation unit, and 14,15 is optical detecting unit.
Embodiment
Specify present embodiment below in conjunction with accompanying drawing 1～4.
The structural representation of present embodiment such as Fig. 1 and 2.Fig. 1 is its longitudinal section view along fore-and-aft direction, by two optical excitation unit up and down, about the canaliculus of two optical detecting units and central authorities form.1,2 are respectively the cover core sheet 1 on top and year chip 2 of bottom among the figure, use organic glass (PMMA) to make, the about thick 2mm of upper cap chip, and the about thick 3.5mm of chip is carried in the bottom.3,4,5,6 form the optical excitation unit, and wherein 3 is excitation source, can use semiconductor light-emitting-diode (LED) or semiconductor laser diode (LD), luminous peak wavelength be 475nm; 4 is the exciting light optical filter, and the peak wavelength of transmission also is 475nm; 5 for exciting the optical microlens of unit, with luminous the focusing on the tested biological microfluid of excitation source; 6 is multilayer film, with whole excite the unit around be surrounded, form tubulose, play to the high reflection of light wave in managing with to managing the effect that outer light wave intercepts fully.7,8,9,10 form optical detecting units, and wherein 7 is the optical microlens of detecting unit, are that the fluorescence of 525nm focuses on to wavelength that tested biological microfluid produces; 8 for detecting the light optical filter, and the transmission peak value wavelength is 525nm; 9 is electrooptical device, can be photodiode (PIN) or silicon blue streak battery, and the peak wavelength of opto-electronic conversion is 525nm; 10 is multilayer film, with being surrounded around the whole detecting unit, forms tubulose, and role is the same.11 microchannels that are square for the cross section, sectional area is approximately 0.8mm * 0.8mm, and the inside is full of biological microfluid to be detected.Generally speaking, the photosensitive area of photoelectricity testing part is bigger than the light-emitting area of excitation source, so the diameter of optical detecting unit than optical excitation unit more greatly.The course of work of whole low-light spectrum detection device is; The light that sends from excitation source 3 is filtered at optical filter 4; The 475nm exciting light of selecting focuses on through the optical microlens 5 that excites the top, unit, propagates into 11 li of microchannels, shines object organisms microfluid to be detected.Material to be detected goes out fluorescence by excitation, and the optical microlens 7 at top, unit to be detected is gathered, and behind optical filter 8, the fluorescence that leaches 525nm is received by semiconductor light power conversion device 9, becomes electric signal output.
Fig. 2 is the longitudinal section view of this pick-up unit along left and right directions, and the lenticule 7 in chip 2, microchannel 11 and optical excitation unit 3,4,5,6 and the optical detecting unit is carried in the bottom of can only drawing among the figure.And the lenticule 5 in chip 2, microchannel 11 and optical detecting unit 7,8,9,10 and the optical excitation unit is carried in the bottom of can only drawing on the view in transverse section of pick-up unit.The purpose of using two groups of optical excitation unit and two groups of optical detecting units is further to improve detection sensitivity.
Fluorescence low-light spectrum detection device of the present invention has following characteristics: the integrated whole non-electric key element of spectral detection, and like the even bundle of gathering, transmission, light of excitation source, light-dividing device, light, light collection, light detection etc.Owing to replaced the photomultiplier PMT or the charge coupled cell CCD that can't embed chip, make the characteristic dimension of whole device narrow down to and have only the millimeter magnitude, even can also be more further micro to micron dimension.
The manufacture craft flow process of fluorescence biosensor chip low-light spectrum detection device is as shown in Figure 3.Make optical excitation unit and optical detecting unit respectively, the manufacturing process of optical excitation unit is at first excitation source and exciting light optical filter to be surrounded as tubulose with multilayer film; On optical filter, make micro optical lens with the formed in situ legal system then, the glue of ultraviolet optics curing soon drips from certain altitude and vertically discharges, and drops onto on the optical filter; And from top to bottom to diffusion trickling all around; In time drop is shone, make its curing, form micro optical lens with Ultra-Violet Laser.The method for making of optical detecting unit is similar with the optical excitation unit, at first electrooptical device is surrounded as tubulose with detecting the light optical filter with multilayer film, on optical filter, makes micro optical lens with the formed in situ legal system then.On upper cap chip 1, make a call to a hole, optical excitation unit 12 is placed in one, lenticule down, and is as shown in Figure 4.On lower chips 2, go out square trench 11 with laser ablation, as the microchannel of biological microfluid to be detected, make a call to three holes in the bottom and the both sides of microchannel then, optical excitation unit 13 is placed in the base apertures, lenticule up.Optical detecting unit 14 and 15 is placed respectively in the hole of both sides, the lenticule of optical detecting unit 14 towards the lenticule of right, optical detecting unit 15 towards a left side, as shown in Figure 4.At last chip 2 is carried in upper cap chip 1 and bottom and be bonded together, just be assembled into fluorescence biosensor chip low-light spectrum detection device, this moment, 4 lenticules all were the microchannels that determinand is arranged towards stream.
Claims (5)
1. fluorescence biosensor chip low-light spectrum detection device, it is characterized in that: (1) is the cover core sheet on top, (2) are the chip that carries of bottom; (11) for being filled with the microchannel of biological microfluid to be detected; Upper and lower sides in the microchannel is provided with optical excitation unit (12) (13), and the left-right symmetric of (11) is distributed with optical detecting unit (14) (15) in the microchannel;
Described optical excitation unit is a tubulose, by excitation source (3), exciting light optical filter (4), excite the unit optical microlens (5) and with whole excite the unit around the multilayer film (6) that are surrounded form;
Described optical detecting unit is a tubulose, by the optical microlens (7) of detecting unit, detect light optical filter (8), electrooptical device (9) and the multilayer film (10) that are surrounded around the whole detecting unit are formed;
The light that sends from excitation source (3) is filtered at optical filter (4), and the exciting light of selecting focuses on through the optical microlens (5) that excites the top, unit, propagates into lining, microchannel (11), shines object organisms microfluid to be detected; Material to be detected goes out fluorescence by excitation, and the optical microlens (7) at top, unit to be detected is gathered, and behind optical filter (8), the fluorescence that leaches is received by semiconductor light power conversion device (9), becomes electric signal output.
2. fluorescence biosensor chip low-light spectrum detection device according to claim 1 is characterized in that: described semiconductor light power conversion device (9) is photodiode or silicon blue streak battery.
3. fluorescence biosensor chip low-light spectrum detection device according to claim 1 is characterized in that: the transmission peak value wavelength of described exciting light is 525nm.
4. the method for making of a kind of fluorescence biosensor chip low-light spectrum detection device according to claim 4 is characterized in that: described microchannel (11) are square trench.
5. the method for making of a fluorescence biosensor chip low-light spectrum detection device is characterized in that may further comprise the steps:
Make the optical excitation unit, excitation source (3) and exciting light optical filter (4) are surrounded as tubulose with multilayer film (6), on optical filter, make micro optical lens (5) then with the formed in situ legal system; The glue of ultraviolet optics curing soon drips from certain altitude and vertically discharges; Drop onto on the optical filter (4), and, in time drop is shone with Ultra-Violet Laser from top to bottom to diffusion trickling all around; Make its curing, form micro optical lens;
Make optical detecting unit, at first electrooptical device (9) is surrounded as tubulose with detecting light optical filter (8) with multilayer film (10), on optical filter, make micro optical lens then with the formed in situ legal system; Upward make a call to a hole at upper cap chip (1), optical excitation unit (12) are placed in one, lenticule (7) down;
Carry chip (2) in the bottom and upward go out the microchannel (11) as biological microfluid to be detected with laser ablation, at the bottom and two side perforatings of microchannel, optical excitation unit (13) are placed in the base apertures then, lenticule (7) up; Optical detecting unit (14) and (15) are placed respectively in the hole of both sides; The lenticule (7) of optical detecting unit (14) is towards lenticule (a 7) court left side right, optical detecting unit (15); Upper cap chip and bottom are carried chip adhesive together, and four lenticules all have the microchannel of determinand towards stream.
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| CN102998234A (en) * | 2012-12-14 | 2013-03-27 | 江苏苏净集团有限公司 | Micro liquid grain counter chip |
| CN106979940A (en) * | 2017-04-17 | 2017-07-25 | 浙江大学 | A kind of device and method for carrying out nucleic acid amplification detection |
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| CN108913565A (en) * | 2018-08-01 | 2018-11-30 | 苏州索真生物技术有限公司 | Multichannel micro-fluidic Systems for optical inspection |
| CN110672008A (en) * | 2019-09-30 | 2020-01-10 | 苏州索真生物技术有限公司 | Micro-channel positioning structure of silicon photocell and positioning method based on structure |
| CN110770552A (en) * | 2017-06-14 | 2020-02-07 | 陈敬红 | High-sensitivity optical detection system |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102998234A (en) * | 2012-12-14 | 2013-03-27 | 江苏苏净集团有限公司 | Micro liquid grain counter chip |
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| CN110770552A (en) * | 2017-06-14 | 2020-02-07 | 陈敬红 | High-sensitivity optical detection system |
| CN110770552B (en) * | 2017-06-14 | 2023-09-12 | 陈敬红 | High-sensitivity optical detection system |
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| CN108913565A (en) * | 2018-08-01 | 2018-11-30 | 苏州索真生物技术有限公司 | Multichannel micro-fluidic Systems for optical inspection |
| CN108913565B (en) * | 2018-08-01 | 2021-07-02 | 苏州索真生物技术有限公司 | Multi-channel micro-fluidic optical detection system |
| CN110672008A (en) * | 2019-09-30 | 2020-01-10 | 苏州索真生物技术有限公司 | Micro-channel positioning structure of silicon photocell and positioning method based on structure |
| CN110672008B (en) * | 2019-09-30 | 2021-02-05 | 苏州索真生物技术有限公司 | Micro-channel positioning structure of silicon photocell and positioning method based on structure |
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