CN201014990Y - Laser-induced fluorescence confocal scanning device - Google Patents
Laser-induced fluorescence confocal scanning device Download PDFInfo
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- CN201014990Y CN201014990Y CNU2007201035183U CN200720103518U CN201014990Y CN 201014990 Y CN201014990 Y CN 201014990Y CN U2007201035183 U CNU2007201035183 U CN U2007201035183U CN 200720103518 U CN200720103518 U CN 200720103518U CN 201014990 Y CN201014990 Y CN 201014990Y
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Abstract
The utility model relates to a confocal laser-induced fluorescence scan device, which belongs to the biochip detection technology. As to the prior devices with similar functions, they cannot collect and use the fluorescence efficiently, and they have great difficulty in controlling the focus during scanning. The utility model adopts a full-reflection mirror with a hole in the center to separate the incoming laser and the reflected induced-fluorescence. When the laser goes through the hole, the energy of the laser almost totally focuses on the biochip with very little loss. When collecting the fluorescence, the larger the angle of the fluorescence collection is, the more efficient the laser shoots and the fluorescence collects. With the help of the computing-control system, the pinholes of the light and the detector can be more confocal, which means only once adjustment of the focus is necessary before scan.
Description
Technical field
The utility model is a kind of laser-induction fluorescence co-focusing scan device, and laser-induction fluorescence co-focusing scan device and method belong to the biological chip testing technology field.
Background technology
Biochip is being widely used in life science, medical research and application as a kind of emerging high-tech product, and the notion of biochip stems from computer chip.The biochip of narrow sense is meant the microarray of bioactivators such as the high-density DNA that is coated on solid phase carrier such as silicon chip, glass, plastics and the nylon membrane etc., protein, cell, mainly comprises cDNA microarray, oligonucleotide microarray and protein microarray.These microarraies are to be fixed on the solid phase carrier in an orderly manner with the form of dot matrix by bioactivator to form.Carry out biochemical reaction under certain conditions, reaction result shows with chemiluminescence method, enzyme linked immunosorbent assay, isotope method, carries out data acquisition with optical instruments such as scanners again, carries out data analysis by special computer software at last.For the broad sense biochip, except above-mentioned passive type micro-array chip, also comprise utilize photoetching technique and micro-processing technology make up on the solid substrate surface microfluid analysis unit and system with realize to biomolecule carry out fast, the slim device of miniature solid of large information capacity parallel processing and analysis.Comprise nucleic acid amplification chip, capillary array electrophoresis chip, active electromagnetism biochip etc.
Biochip test aspect major technology means have the confocal scanning method and based on the detection method of CCD (Charge-coupled device).Because it is highly sensitive that the confocal scanning method has, what most micro-array biochip scanner adopted all is the confocal scanning principle.
The classical bio-chip test device general using half-reflecting half mirror based on laser confocal scanning carries out beam split to the laser of incident and the fluorescence that derives, as shown in Figure 1.The laser that laser instrument 1 sends becomes pointolite after through first lens 2 and first pin hole 3, through second lens, 4 collimations, collimated light through a half-reflecting half mirror 6 after, converge on its back focal plane through the 3rd lens 8 again, induce that biochip 10 sends fluorescence on this lens focal plane, fluorescence is through the 3rd lens 8 back collimations, project on the half-reflecting half mirror 6, the fluorescence that half-reflecting half mirror 6 will throw on it reflexes on the 4th lens 12, the 4th lens 12 will reflect the back focus that fluorescence on it converges to the 4th lens 12, place a pin hole 13 on this focus, be close to pin hole and place a photodetector 14, photodetector is sent into computing machine 15 after the fluorescence signal that receives is converted into digital signal.In order to obtain the good fluorescence signal of concentration of energy signal to noise ratio (S/N ratio), must focusing, classical focusing is to utilize the imaging eyepiece to observe the whether blur-free imaging of hot spot, manual focusing.
Utilize half-reflecting half mirror that the fluorescence that laser and induced with laser come out is carried out beam split in the said apparatus, the collection of fluorescence and utilization ratio are not high, the laser of incident is in the energy loss of minute light time nearly 50%, and the fluorescence that the while lens are collected also has nearly 50% energy loss.If the multichannel different wavelength of laser, induce the fluorescence of different wave length, phosphor collection and utilization ratio will be lower; The pin hole of the pin hole of light source and detector is strict simultaneously keeps copolymerization Jiao to concern, the focusing control in the scanning process is difficulty very.
The utility model content
The purpose of this utility model is to provide a kind of bio-chip test device, and one of purpose is exactly to improve the light path of fluoroscopic examination, and the lens that are all-trans of punching improve phosphor collection efficient as light-splitting device in the middle of utilizing.Two of purpose is the burnt change that proposes of copolymerization, biochip focal depth range in the focal plane and up and down during gated sweep, by before scanning along the mode of two diagonal line prescans of rectangle of biochip, the position of regulating camera lens makes the live part of whole biochip drop on the focal plane of camera lens in pre-scanning process and up and down in the focal depth range.
A kind of method of the scanning biochip based on laser-induction fluorescence co-focusing scan device may further comprise the steps:
1) opens laser instrument 1 preheating;
2) laser that sends of laser instrument 1 is through by first lens 2, first pin hole 3, the beam-expanding system that second lens 4 are formed expands that to restraint into diameter be 1 millimeter light beam 5, this light beam 5 passes through from the light hole of the completely reflecting mirror 7 of centre punching, vertical irradiation to the 3rd lens 8 on, the 3rd lens 8 converge to laser on the biochip 10 at back focal plane place, the part laser that converges on the biochip 10 sees through biochip 10, reflection through catoptron 16, obtained by detection system based on four-quadrant photo detector, entering computing machine 15 focuses, on lens 8 focuses and in the focal depth range, such second pin hole 13 becomes the burnt relation of copolymerization with first pin hole 3 to biochip 10 all the time approx all the time in the assurance scanning process.Therefore must utilize 9 pairs of biochip focusing of focus control, the focusing process is: the laser of part vertical incidence is from biochip 10 transmissions, through the focusing optical system, after being lens 17 and cylindrical mirror 18, image in four-quadrant photo detector 19, suitably select the parameter of lens and cylindrical mirror, can obtain certain focusing range and certain focusing accuracy.The light signal that four-quadrant photo detector 19 is accepted is converted into electric signal, through filtering and processing and amplifying, be transferred to 15 li in computing machine, through calculating generation differential wave and preset value relatively, produce the mechanical driving device motion that the drive motor is linked to each other with drive motor by driven by motor, lens 8 along the translation up and down of optical axis direction, arrive the target focal plane position that requires with mechanical driving device up to object lens.To biochip diagonal line scanning, the pointwise focusing makes the focal depth range of whole biochip 10 at lens 8.
3) converge to that a certain spot sends fluorescence 11 on the biological sample on another part induced with laser biochip 10 on the biochip 10, the part of the fluorescence that sends is through the 3rd lens 8, become collimated light and project reflection on the completely reflecting mirror 7, on the 4th lens 12 that propagate into, converge on the pin hole 13 of back focal plane of the 4th lens 12, photodetector 14 receives fluorescence, and with the fluorescence signal conversion electric signal of this point, digitizing enters computing machine 15;
4) mobile biochip 10, the whole biochip 10 of LASER SPECKLE pointwise two-dimensional scan that lens 8 are converged.
Advantage of the present utility model is:
1. the completely reflecting mirror of punching is to the laser of incident with induce the fluorescence of reflection to carry out beam split in the middle of adopting, laser beam is during through the aperture, energy loss is little, almost all laser energy can shine on the biochip, and when collecting fluorescence, the phosphor collection angle is big, and the efficient height not only improves laser incident efficient but also improved the collection efficiency of fluorescence, can reach 80-90%.
2. utilize computer control, make the pin hole of light source and the pin hole of detector keep approximate copolymerization Jiao relation, only need the preceding focusing of scanning once, focusing control is simple and convenient.
Description of drawings
The structural representation of the confocal scanning bio-chip test device of the single channel incident light of Fig. 1 classics;
Fig. 2 is the synoptic diagram of confocal scanning bio-chip test device of the present utility model;
Fig. 3 is the biochip synoptic diagram among the embodiment of the present utility model
Fig. 4 embodiment scan mode of the present utility model.
Embodiment:
Below in conjunction with accompanying drawing, in detail the utility model is described.
Embodiment:
It is 1 millimeter light beam 5 that laser that laser instrument 1 sends is restrainted into diameter through the beam-expanding system expansion of being made up of first lens 2, first pin hole 3, second lens 4, this light beam is slightly larger than the light hole process of 1 millimeter completely reflecting mirror 7 from the middle aperture of punching, vertical irradiation to the 3rd lens 8 on, the 3rd lens 8 converge to laser on the biochip 10 at back focal plane place.
Part laser converges on the biochip 10 and sees through biochip, reflection through catoptron 16, obtained by detection system based on four-quadrant photo detector, entering computing machine 15 focuses, on lens 8 focuses and in the focal depth range, such second pin hole 13 becomes the burnt relation of copolymerization with first pin hole 3 to biochip 10 all the time approx all the time in the assurance scanning process.Utilize 9 pairs of biochip focusing of focus control, in optical system shown in Figure 2 and focus control shown in Figure 3, the focusing process is: the laser of part vertical incidence is from biochip 10 transmissions, through the focusing optical system, after being lens 17 and cylindrical mirror 18, image in four-quadrant photo detector 19, suitably select the parameter of lens and cylindrical mirror, can obtain certain focusing range and certain focusing accuracy.The light signal that four-quadrant photo detector 19 is accepted is converted into electric signal, through filtering and processing and amplifying, be transferred to 15 li in computing machine, through calculating generation differential wave and preset value relatively, produce the drive motor, by the mechanical driving device motion that driven by motor links to each other with drive motor, lens 8 along the translation up and down of optical axis direction, arrive the target focal plane position that requires with mechanical driving device up to object lens.To biochip diagonal line scanning, the pointwise focusing makes whole biochip 10 at the focal depth range of lens 8, thinks defective otherwise abandon this biochip, finishes focusing.
Another part laser converges to the biological sample on the biochip 10 of inducing on the biochip 10, send fluorescence, part fluorescence 11 is through the 3rd lens 8, become collimated light and project reflection on the completely reflecting mirror 7, on the 4th lens 12 that propagate into, converge on the pin hole 13 of back focal plane of the 4th lens 12, photodetector 14 next-door neighbour's pin holes 14 receive fluorescence, fluorescence signal is transformed electric signal, and digitizing enters computing machine 15.So just survey the fluorescence light intensity that obtains a point on the biochip, move biochip by control by linear electric motors and stepper motor and the two-dimensional stage that driven thereof, here do not draw, two-dimensional scan obtains the fluorescence light intensity of two-dimensional lattice on the whole biochip, the quantity of biochemical reactant is proportional on fluorescence intensity and the biochip, can calculate the quantity of biochemical reactant on the biochip according to fluorescence intensity, biochip is seen Fig. 3, interval 150 nanometer point samples in 60 millimeters * 20 the scope on 75 millimeters * 25 millimeters microslide, about diameter 150 nanometers of point sample, the mode of two-dimensional scan is seen Fig. 4; Among Fig. 4 along short-axis direction promptly 25 millimeters directions move, simultaneously along major axis 75 millimetres of movement, circulation is carried out, and scans whole biochip.
Flow process: beginning, open laser instrument 1 preheating 10 minutes; Detection is adjusted lens 8 based on the probe value of four-quadrant photo detector system 17 by electromechanical assembly 9, makes biochip 10 upper surfaces be in the back focal plane of lens 8, and is in the focal depth range all the time in scanning process; Synchronous scanning biochip and fluorescence detection, i.e. data acquisition; Computing machine 15 data processing and demonstration.
Claims (2)
1. laser-induction fluorescence co-focusing scan device, comprise laser instrument (1), first lens (2), first pin hole (3), second lens (4), the 3rd lens (8), biochip (10), the 4th lens (12), second pin hole (13), photodetector (14), computing machine (15) successively, it is characterized in that:
The completely reflecting mirror (7) of punching in the middle of between second lens (4) and the 3rd lens (8), being provided with, place catoptron (16) successively and based on the detection system of four-quadrant photo detector in biochip (10) one sides, should be made up of lens (17), cylindrical mirror (18), four-quadrant photo detector (19) based on the detection system of four-quadrant photo detector, four-quadrant photo detector (19) is connected with computing machine (15);
The 3rd lens (8) link to each other with focus control (9), this focus control (9) by drive motor with form with the mechanical driving device of the drive motor that links to each other, focus control (9) is connected with computing machine (15).
2. laser-induction fluorescence co-focusing scan device according to claim 1 is characterized in that: laser instrument (1) is connected with computing machine (15).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNU2007201035183U CN201014990Y (en) | 2007-02-08 | 2007-02-08 | Laser-induced fluorescence confocal scanning device |
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| Application Number | Priority Date | Filing Date | Title |
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| CNU2007201035183U CN201014990Y (en) | 2007-02-08 | 2007-02-08 | Laser-induced fluorescence confocal scanning device |
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| CN201014990Y true CN201014990Y (en) | 2008-01-30 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101013136B (en) * | 2007-02-08 | 2011-07-20 | 北京工业大学 | Laser-induction fluorescence co-focusing scanning device and method |
| CN102519913A (en) * | 2011-12-22 | 2012-06-27 | 中国科学院理化技术研究所 | Polarizing modulation type laser cofocal surface plasma resonance apparatus |
| CN103528610A (en) * | 2013-10-30 | 2014-01-22 | 苏州晋翌生物医学仪器有限公司 | Reflective photoelectric sensing device |
-
2007
- 2007-02-08 CN CNU2007201035183U patent/CN201014990Y/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101013136B (en) * | 2007-02-08 | 2011-07-20 | 北京工业大学 | Laser-induction fluorescence co-focusing scanning device and method |
| CN102519913A (en) * | 2011-12-22 | 2012-06-27 | 中国科学院理化技术研究所 | Polarizing modulation type laser cofocal surface plasma resonance apparatus |
| CN102519913B (en) * | 2011-12-22 | 2013-05-08 | 中国科学院理化技术研究所 | Polarizing modulation type laser cofocal surface plasma resonance apparatus |
| CN103528610A (en) * | 2013-10-30 | 2014-01-22 | 苏州晋翌生物医学仪器有限公司 | Reflective photoelectric sensing device |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080130 Termination date: 20120208 |