CN103676122A - Small fluorescent/bright field optical imaging system, optical imaging method and purpose thereof - Google Patents
Small fluorescent/bright field optical imaging system, optical imaging method and purpose thereof Download PDFInfo
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- CN103676122A CN103676122A CN201310251546.XA CN201310251546A CN103676122A CN 103676122 A CN103676122 A CN 103676122A CN 201310251546 A CN201310251546 A CN 201310251546A CN 103676122 A CN103676122 A CN 103676122A
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Abstract
The invention provides a small fluorescent/bright field optical imaging system, an optical imaging method and the application of the system in the aspects of a biological experiment, medical examination, archaeology, geological exploration and the like. The small fluorescent/bright field optical imaging system is characterized in that the system comprises an imaging unit which comprises a CMOS module which is connected with an external controller and does not comprise an eye lens, a focusing unit, an achromatization unit, a filter unit, a light excitation unit which comprises an LED module (8) as a light source, and an object lens unit. The dimension of the optical imaging system is in a range of 1 cubic centimeter to 1*103 cubic centimeters, and the weight of the system is in a range of 1 gram to 1*103 grams.
Description
Technical field
The present invention relates to a kind of small fluorescent/light field optical imaging system, more particularly, the present invention relates to a kind of remarkable physical dimension that reduced and (be less than 10
3cubic centimetre) and weight (be less than 10
3gram) small fluorescent/light field optical imaging system, utilize this system fluorescence/light field optical imaging method, with and in the purposes of the aspects such as biological experiment.
Background technology
Fluorescence/light field optical imaging system is for observing and study the optical precision instrument of object fine structure, being widely used in the field of each subject, and the exploration of microworld and theoretical research are played an important role.For example, epifluorescence microscope optical imaging system is wherein the important research means of biology and medical science, in the researchs such as cell biology, Neuscience, be widely used, along with emerging in multitude of fluorescence molecule label in biotechnology, this kind equipment is showing important effect day by day aspect observation of cell activity.
Traditional fluorescence/light field optical imaging system is a kind of large scale and expensive equipment, and it is a large-scale instrument that takies desktop normally.In Neuscience and medical research, fluorescence/light field optical imaging system is widely applied in living animal, but because the size of legacy system is large (generally 10
4more than cubic centimetre), weight is heavier (generally 10
4more than gram), be difficult to realize free moving animals (such as mouse, its head size is 10 cubic centimetres of magnitudes, body wt is 10 grams of magnitudes) carry out the movable imaging research of cerebral nerve, this has restricted scientist greatly to the central issue of the bio-science possibility that " the nerve ring roadbed plinth of behavior " furtherd investigate.Movable formation method (the function nuclear-magnetism of existing live body cerebral nerve, PET, CT etc.) substantially all need that animal is carried out to physics and fix, this not only makes a lot of animal behaviors (such as mutual social interactions between animal, reproductive behavior, the behavior etc. of going after profits and advoiding disadvantages) cannot study; And whether the data that animal obtains under bound condition can reflect real cerebral nerve mechanics, it is also a considerable problem.Therefore, little, the lightweight fluorescence/light field of a kind of physical dimension optical imaging system can be observed the cerebral nerve activity of free moving animals scientist, thereby advance greatly bio-science, especially the development of Neuscience and cognitive science, has great meaning to research " the nerve ring roadbed plinth of behavior " this important scientific problems.
The structure of existing fluorescence/light field optical imaging system possesses conventionally: image formation unit, and it comprises monocular tube eyepiece or binocular eyepiece and is connected to the CMOS module on external unit (such as controller and computing machine etc.) by data line; For regulating the focus unit of focal plane; Achromatism unit; Filter unit (falling to penetrating fluorescence filter unit), it mainly comprises the first optical filter, the second optical filter, becomes the dichronic mirror of miter angle with level; Exciting light unit, comprises collector lens and as the mercury lamp of light source; Object lens, comprises many group object lens and Nosepiece.Because above-mentioned part count is various, also comprise various adjusting knobs, optical fiber etc., and conventionally adopt mercury lamp or high-intensity halogen lamps as light source, need to use independent power supply, be therefore difficult to miniaturization.
Modern micro-integrated technology is an important technical progress, utilizes this technology easily to obtain cheap product, good effect and the application of novelty.The influence power of modern micro-integrated technology has appeared at a plurality of research fields, comprises communication, calculating and genomics etc.But current present situation is, still do not develop a kind of size little, lightweight, be applicable to small fluorescent/light field optical imaging system that the cerebral nerve activity of freely movable animal is observed.
Summary of the invention
Fundamental purpose of the present invention is to provide little, the lightweight small fluorescent/light field of a kind of volume optical imaging system, thereby serve, the weight and volume of imaging system is had to the scientific researches such as biological experiment (as the brain cell of experiments of measuring animal changes), medical inspection (as minute lesion in body), archaeology (observation cultural artifact surface slight crack), geologic prospecting (as jade mineral are invaded the degree of soaking) of strict restriction and industrial use and the purposes among the people of the small-sized imaging system of various needs.
For above-mentioned technical matters of the prior art, the present inventors use micro-integrated technology and in conjunction with technology such as Neuscience, micro-optic and semiconductor photoelectricities, have developed a kind of remarkable fluorescence/light field optical imaging system that has reduced system physical dimension and weight.
Small fluorescent/light field optical imaging system of the present invention is characterised in that to possess:
Image-generating unit, comprises receiving end (18) and is fixed on the CMOS module (1) on receiving end (18), this CMOS module (1) is connected with peripheral control unit by wireless transmission or data line, does not comprise eyepiece in described image-generating unit;
Focus unit, comprises setting nut (3) and screw rod (17), and screw rod (17) is connected by screw thread with setting nut (3); Receiving end (18) is installed in rotation on setting nut (3);
Achromatism unit, comprises achromat body (16) and is embedded in the inner achromat (4) of achromat body (16); Achromat body (16) is fixedly connected on screw rod (17);
Filter unit, comprises mirror body (14), is embedded in the inner inner support (15) of mirror body (14), embeds optical filter (5) in the upper groove of inner support (15), embeds exciter filter (10) in the side groove of inner support (15), to be fixed on inner support (15) inner and become the dichronic mirror (13) of miter angle with level; Mirror body (14) is fixedly connected on achromat body (16);
Exciting light unit, comprises collector lens body (7), is embedded in collector lens body (7) inner collector lens (9), LED fixing body (6) and is fixed on the LED module (8) as light source on LED fixing body (6); The two ends of collector lens body (7) are fixedly connected with mirror body (14) with LED fixing body (6) respectively;
Object lens, comprises object lens body (12) and is embedded in the inner object lens (11) of object lens body (12), object lens body (12) is fixedly connected on mirror body (14).
Preferably, on the outer peripheral face of setting nut (3), be formed with circular groove, by two annulus that fixedly semicircular ring (2A), (2B) are spliced into, embedded in this circular groove and can freely be rotated, receiving end (18) is bolted at described two fixedly on semicircular ring (2A), (2B).
Preferably, be fixedly connected be bolted, riveted joint or mechanical stamping is connected and fixed.
Preferably on object lens body (12), be provided with four small boss, form and to coordinate with mirror body (14) medial surface, (15 are clipped in the middle and are fixed by described four small boss and achromat body (16) to be embedded in the inner support of mirror body (14) inside.
Preferably in focus unit, do not comprise screw rod (17), in achromat body (16) periphery, be formed with screw thread, directly by screw thread, be connected and fixed with setting nut (3).
The physical dimension of preferred described optical imaging system is 1～1 * 10
3the scope of cubic centimetre, and weight is 1～1 * 10
3gram scope.
More preferably the physical dimension of described optical imaging system is 5～1 * 10
2the scope of cubic centimetre, and weight is 5～1 * 10
2gram scope.
Preferred described small fluorescent/light field optical imaging system also possesses built-in lithium battery, and for replacing external power source, driving LED module (8) is luminous.
The present invention also provides a kind of fluorescence/light field optical imaging method, it is characterized in that: use above-mentioned small fluorescent/light field optical imaging system to observe the cell of moving animals, possess following steps:
Make the rear utilizing emitted light of LED module (8) energising, light is converged to nearly directional light by collector lens (9); It is upper that nearly directional light after this convergence is irradiated to exciter filter (10), the light transmission exciter filter (10) of particular range of wavelengths, and the remaining light optical filter (10) that is excited stops; It is upper that the illumination of particular range of wavelengths is mapped to dichronic mirror (13), and the transmission range that is shorter than dichronic mirror due to wavelength is reflected on object lens (11); The light reflexing on object lens (11) is irradiated to and is observed on cell by object lens (11); Intracellular fluorescent material is subject to exciting light and irradiates and the longer fluorescence of generation wavelength, this fluorescence is irradiated on dichronic mirror (13) by the convergence of object lens (11), because glimmering light wavelength reaches the transmission range of dichronic mirror, thereby be irradiated on optical filter (5) through dichronic mirror (13), optical filter (5) filters to light, only by the fluorescence of particular range of wavelengths; It is upper that illumination after optical filtering is mapped to achromat (16), by achromat (16), again can coalescence be irradiated to CMOS module (1) above, directly gathered image data generating by wireless transmission or data line by peripheral control unit.
The present invention also provides the application of above-mentioned small fluorescent/light field optical imaging system at aspects such as biological experiment, medical inspection, archaeology, geologic prospectings.
With respect to prior art, to be that volume is little (be less than 10 to the advantage of small fluorescent/light field optical imaging system of the present invention
3cubic centimetre), quality gently (is less than 10
3gram), compare at volume with currently marketed similar commercial product and all reduced qualitatively by least 1 order of magnitude, go for the cerebral nerve activity of freely movable animal to observe, and make system cost have significant decline.
The present invention will greatly advance the development in a plurality of fields, for example medical inspection, archaeology, geologic prospecting, examination of water, bio-science etc., especially the development of Neuscience and cognitive science, has great meaning to research " the nerve ring roadbed plinth of behavior " this important scientific problems.
Accompanying drawing explanation
Fig. 1 is the sectional view of small fluorescent/light field optical imaging system of the present invention.
Fig. 2 is the three-dimensional exploded view of small fluorescent/light field optical imaging system of the present invention.
Fig. 3 means the stereographic map of the overall appearance after small fluorescent/light field optical imaging system of the present invention has assembled.
Fig. 4 is the figure that schematically represents the light path of small fluorescent/light field optical imaging system of the present invention.
Fig. 5 means the photo of the physical dimension of small fluorescent/light field optical imaging system of the present invention.
Symbol description:
1.CMOS module, 2A, 2B. be semicircular ring, 3. setting nut, 4. achromat, 5. optical filter, 6.LED fixing body, 7. collector lens body, 8.LED module, 9. collector lens, 10. exciter filter, 11. object lens, 12. object lens bodies, 13. dichronic mirrors, 14. mirror bodies, 15. inner supports, 16. achromat bodies, 17. screw rods, 18. receiving ends fixedly.
Embodiment
The technology such as micro-integrated technology, micro-optic, semiconductor photoelectricity have been merged in the present invention, small fluorescent/light field optical imaging system of take carries out observation and analysis as imaging platform, use micromechanics, electronics, optical imagery and image processing techniques to object, has realized the small-sized and lightweight of fluorescence/light field optical imaging system.
Below in conjunction with accompanying drawing, small fluorescent/light field optical imaging system of the present invention is described in further detail.
Fig. 1 is the sectional view of small fluorescent/light field optical imaging system of the present invention.Fig. 2 is the three-dimensional exploded view of small fluorescent/light field optical imaging system of the present invention.
As depicted in figs. 1 and 2, in small fluorescent/light field optical imaging system of the present invention, image-generating unit comprises receiving end (18) and is fixed on the CMOS module (1) on receiving end (18), this CMOS module (1) is connected with peripheral control unit (not shown) by wireless transmission or data line, it is characterized in that, in described image-generating unit, do not comprise eyepiece.
For the ease of direct observation, in current fluorescence/light field optical imaging system, conventionally possess monocular tube eyepiece or binocular eyepiece, so volume is larger.But in the present invention, due to the small-sized object that turns to, therefore can omit eyepiece, only by CMOS module, data-signal is transferred to peripheral control unit, by computing machine, process and obtain image.
Focus unit of the present invention comprises setting nut (3) and screw rod (17), and screw rod (17) is connected by screw thread with setting nut (3); Receiving end (18) is installed in rotation on setting nut (3).
Because CMOS module is connected on external unit (controller etc.) by data line, when needs regulate focal plane and rotate setting nut (3), if receiving end (18) is fixedly connected with setting nut (3), can produce the situation that data line etc. also rotates thereupon.Therefore, preferably receiving end (18) is bolted at two fixedly on semicircular ring (2A), (2B), and be formed with circular groove on the outer peripheral face of setting nut (3), to be embedded in the circular groove of setting nut (3) by two annulus that fixedly semicircular ring (2A), (2B) are spliced into, because annulus can freely rotate, therefore, even if rotated setting nut (3), also can suitably adjust the position of receiving end (18), it is not rotated thereupon.
In addition, for further miniaturization, more preferably in focus unit, do not comprise screw rod (17), can be formed with screw thread in achromat body (16) periphery, achromat body (16) and setting nut (3) are directly connected and fixed by screw thread.
Achromatism of the present invention unit comprises achromat body (16) and is embedded in the inner achromat (4) of achromat body (16), and achromat body (16) is fixedly connected on screw rod (17).
Filter of the present invention unit comprises mirror body (14), be embedded in the inner inner support (15) of mirror body (14), embed optical filter (5) in the upper groove of inner support (15), embed exciter filter (10) in the side groove of inner support (15), to be fixed on inner support (15) inner and become the dichronic mirror (13) of miter angle with level; Mirror body (14) is fixedly connected on achromat body (16);
Exciting light of the present invention unit comprises collector lens body (7), is embedded in collector lens body (7) inner collector lens (9), LED fixing body (6) and is fixed on the LED module (8) as light source on LED fixing body (6); The two ends of collector lens body (7) are fixedly connected with mirror body (14) with LED fixing body (6) respectively.
Conventionally, fluorescence/light field optical imaging system adopts mercury lamp or high-intensity halogen lamps as light source, and itself volume is larger, and need to use independent power supply, therefore aspect miniaturization, has very large restriction.The present invention replaces conventional light source by employing LED module (8), thereby has obtained obvious progress aspect miniaturization.
In addition, LED module can be connected with external power source by shared power lead (not shown) with CMOS module.In the present invention, for further miniaturization, built-in lithium battery can also be set in exciting light unit, coin shape lithium battery as very little in volume, for replacing external power source, driving LED module (8) is luminous.
LED module (8) is used as to light source, although slightly low with conventional light source phase specific luminance, still can obtain enough in the present invention image clearly.
Object lens of the present invention comprises object lens body (12) and is embedded in the inner object lens (11) of object lens body (12), and object lens body (12) is fixedly connected on mirror body (14).
Four small boss are preferably set on object lens body (12), form and coordinate with mirror body (14) medial surface, to reach the object being mounted with.Inner support (15) is embedded in mirror body (14) inside, and is clipped in the middle and is fixed by four small boss of object lens body (12) and achromat body (16).By four small boss are set on object lens body (12), can reduce the area that coordinates of object lens body (12) and inner support (15), to reach the object that improves the precision coordinating, can alleviate the weight of object lens body (12), to reach the light weight requirement of optical imaging system simultaneously.
Common fluorescence/light field optical imaging system adopts the different object lens of many group multiplying powers, and disposes Nosepiece, so volume is larger, is difficult to miniaturization.The present invention, with the small-sized object that turns to, has therefore only adopted one group of object lens, has omitted Nosepiece.When needs different multiplying, can adopt the small fluorescent/light field optical imaging system of the present invention that possesses different object lens.
In being fixedly connected with between above-mentioned each parts, adopted the technology of micromechanics, described in be fixedly connected be bolted, riveted joint or mechanical stamping is connected and fixed.For example, three bolts hole that achromat body (16) is corresponding with screw rod (17) carry out the heart to coordinate, and are connected and fixed by bolt.The external cylindrical surface of achromat (4) carries out the heart to coordinate with the inner cylinder face of achromat body (16).Two bolts hole that object lens body (12) is corresponding with mirror body (14) carry out the heart to coordinate, and are connected and fixed by bolt or mechanical stamping.Object lens (11) external cylindrical surface carries out the heart to coordinate with the inner cylinder face of object lens body (12).The external cylindrical surface of the outside outshot of collector lens body (7) coordinates to reach the effect to the heart with the circular hole of mirror body (14) sidepiece, and is connected and fixed by bolt or mechanical stamping.The external cylindrical surface of collector lens (9) carries out the heart to coordinate with the inner cylinder face of collector lens body (7).The inner cylinder face of LED fixing body (6) carries out the heart to coordinate with the external cylindrical surface of collector lens body (7), two bolts hole on LED fixing body (6) carry out the heart to coordinate with two bolts hole on collector lens body (7) simultaneously, and connect by bolt or mechanical stamping.
Fig. 3 means the stereographic map of the overall appearance after small fluorescent/light field optical imaging system of the present invention has assembled.As can be seen from the figure, small fluorescent/light field optical imaging system profile compact of the present invention, its physical dimension is 1～1 * 10
3the scope of cubic centimetre, and weight is 1～1 * 10
3gram scope.For example, for the ease of meiofauna (mouse) is carried out to biological study, more preferably the physical dimension of described optical imaging system is 5～1 * 10
2the scope of cubic centimetre, and weight is 5～1 * 10
2gram scope.
Below, the process of using small fluorescent/light field optical imaging system of the present invention to observe the cell of moving animals is briefly described.
As shown in Figure 4, fluorescence/light field optical imaging method of the present invention possesses following steps:
By external power source or built-in power, make the rear utilizing emitted light of LED module (8) energising, light is converged to nearly directional light by collector lens (9); It is upper that nearly directional light after this convergence is irradiated to exciter filter (10), the light transmission exciter filter (10) of particular range of wavelengths, and the remaining light optical filter (10) that is excited stops; It is upper that the illumination of particular range of wavelengths is mapped to dichronic mirror (13), and the transmission range that is shorter than dichronic mirror due to wavelength is reflected on object lens (11); The light reflexing on object lens (11) is irradiated to and is observed on cell by object lens (11); Intracellular fluorescent material is subject to exciting light and irradiates and the longer fluorescence of generation wavelength, this fluorescence is irradiated on dichronic mirror (13) by the convergence of object lens (11), because glimmering light wavelength reaches the transmission range of dichronic mirror, thereby be irradiated on optical filter (5) through dichronic mirror (13), optical filter (5) filters to light, only by the fluorescence of particular range of wavelengths; It is upper that illumination after optical filtering is mapped to achromat (16), by achromat (16), again can coalescence be irradiated to CMOS module (1) above, directly gathered image data generating by wireless transmission or data line by peripheral control unit.
Computing machine, from controller reads image data, can carry software by native system and process, and comprises brightness, contrast adjustment; Image size adjustment; Mark chi; Export as single image; Export as the common image processing functions such as dynamic film, and comprise elementary time series analysis function.
Therefore, small fluorescent/light field optical imaging system of the present invention has great importance for biological experiment, medical inspection aspect, particularly the cranial nerve research for moving animals.
When adopting outside bright field light source, also suitably changing behind the filter unit of small fluorescent/light field optical imaging system of the present invention, just can be used as light field optical imaging system and use.Because its volume is little, lightweight, at aspects such as archaeology, geologic prospectings, also there is important application.
Fig. 5 means the photo of an example of small fluorescent/light field optical imaging system of the present invention and the relative size of 1 yuan of coin.Can find out, the optical texture physical dimension of small fluorescent/light field optical imaging system of the present invention can reach the degree of approximately 2 cubic centimetres, and weight is only approximately 1.6 grams.
The optimal way of more than enumerating and concrete example are only for technical scheme of the present invention and inventive concept are done to indicative explaination but not limit the scope of the invention.All those skilled in the art on the basis of the application's inventive concept in conjunction with prior art, by the available other technologies scheme of logic analysis, reasoning or limited experiment, within also should being considered to drop on protection scope of the present invention.
Claims (10)
1. small fluorescent/light field optical imaging system, is characterized in that possessing:
Image-generating unit, comprises receiving end (18) and is fixed on the CMOS module (1) on receiving end (18), this CMOS module (1) is connected with peripheral control unit by wireless transmission or data line, does not comprise eyepiece in described image-generating unit;
Focus unit, comprises setting nut (3) and screw rod (17), and screw rod (17) is connected by screw thread with setting nut (3); Receiving end (18) is installed in rotation on setting nut (3);
Achromatism unit, comprises achromat body (16) and is embedded in the inner achromat (4) of achromat body (16); Achromat body (16) is fixedly connected on screw rod (17);
Filter unit, comprises mirror body (14), is embedded in the inner inner support (15) of mirror body (14), embeds optical filter (5) in the upper groove of inner support (15), embeds exciter filter (10) in the side groove of inner support (15), to be fixed on inner support (15) inner and become the dichronic mirror (13) of miter angle with level; Mirror body (14) is fixedly connected on achromat body (16);
Exciting light unit, comprises collector lens body (7), is embedded in collector lens body (7) inner collector lens (9), LED fixing body (6) and is fixed on the LED module (8) as light source on LED fixing body (6); The two ends of collector lens body (7) are fixedly connected with mirror body (14) with LED fixing body (6) respectively;
Object lens, comprises object lens body (12) and is embedded in the inner object lens (11) of object lens body (12), object lens body (12) is fixedly connected on mirror body (14).
2. small fluorescent/light field optical imaging system according to claim 1, it is characterized in that: on the outer peripheral face of setting nut (3), be formed with circular groove, by two annulus that fixedly semicircular ring (2A), (2B) are spliced into, embedded in this circular groove and can freely be rotated, receiving end (18) is bolted at described two fixedly on semicircular ring (2A), (2B).
3. small fluorescent/light field optical imaging system according to claim 1 and 2, is characterized in that: described in be fixedly connected be bolted, riveted joint or mechanical stamping is connected and fixed.
4. according to the small fluorescent/light field optical imaging system described in any one in claim 1～3, it is characterized in that: on object lens body (12), be provided with four small boss, form and to coordinate with mirror body (14) medial surface, the inner support (15) that is embedded in mirror body (14) inside is clipped in the middle and is fixed by described four small boss and achromat body (16).
5. according to the small fluorescent/light field optical imaging system described in any one in claim 1～4, it is characterized in that: in focus unit, do not comprise screw rod (17), achromat body (16) periphery is formed with screw thread, directly by screw thread, is connected and fixed with setting nut (3).
6. according to the small fluorescent/light field optical imaging system described in any one in claim 1～5, it is characterized in that: the physical dimension of described optical imaging system is 1～1 * 10
3the scope of cubic centimetre, and weight is 1～1 * 10
3gram scope.
7. small fluorescent/light field optical imaging system according to claim 6, is characterized in that: the physical dimension of described optical imaging system is 5～1 * 10
2the scope of cubic centimetre, and weight is 5～1 * 10
2gram scope.
8. according to the small fluorescent/light field optical imaging system described in any one in claim 1～7, it is characterized in that: described small fluorescent/light field optical imaging system also possesses built-in lithium battery, and for replacing external power source, driving LED module (8) is luminous.
9. fluorescence/light field optical imaging method, is characterized in that: right to use requires the small fluorescent/light field optical imaging system described in any one in 1～8 to observe the cell of moving animals, possesses following steps:
Make the rear utilizing emitted light of LED module (8) energising, light is converged to nearly directional light by collector lens (9); It is upper that nearly directional light after this convergence is irradiated to exciter filter (10), the light transmission exciter filter (10) of particular range of wavelengths, and the remaining light optical filter (10) that is excited stops; It is upper that the illumination of particular range of wavelengths is mapped to dichronic mirror (13), and the transmission range that is shorter than dichronic mirror due to wavelength is reflected on object lens (11); The light reflexing on object lens (11) is irradiated to and is observed on cell by object lens (11); Intracellular fluorescent material is subject to exciting light and irradiates and the longer fluorescence of generation wavelength, this fluorescence is irradiated on dichronic mirror (13) by the convergence of object lens (11), because glimmering light wavelength reaches the transmission range of dichronic mirror, thereby be irradiated on optical filter (5) through dichronic mirror (13), optical filter (5) filters to light, only by the fluorescence of particular range of wavelengths; It is upper that illumination after optical filtering is mapped to achromat (16), by achromat (16), again can coalescence be irradiated to CMOS module (1) above, directly gathered image data generating by wireless transmission or data line by peripheral control unit.
10. the application of the small fluorescent/light field optical imaging system described in any one aspect biological experiment, medical inspection, archaeology, geologic prospecting in claim 1～8.
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