CN104887318A - Mini-type confocal microscope - Google Patents
Mini-type confocal microscope Download PDFInfo
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- CN104887318A CN104887318A CN201510220771.6A CN201510220771A CN104887318A CN 104887318 A CN104887318 A CN 104887318A CN 201510220771 A CN201510220771 A CN 201510220771A CN 104887318 A CN104887318 A CN 104887318A
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Abstract
The invention discloses a mini-type confocal microscope. The mini-type confocal microscope is characterized in that a scanning mirror, a relay lens group and a self-focusing lens are sequentially arranged in a lighting light path; a light filter group, an imaging lens, an aperture or slit and a photoelectric detector are sequentially arranged in an imaging light path; the scanning mirror is used for adjusting the directions of both lighting light and emitting light; the relay lens group enables the scanning mirror and the self-focusing lens to be in pupil match; the self-focusing lens is used for gathering the lighting light on a sample to be imaged and collecting fluorescent light emitted by the sample; the imaging lens is used for focusing fluorescent light of the emitting light; and the aperture or slit is formed on the focal plane of the imaging lens. By utilizing the confocal principle, optical chromatography is realized in the mini-type fluorescent microimaging device, interferences caused by afocal signals are inhibited, and both the contrast ratio and the signal-to-noise ratio are increased.
Description
Technical field
The invention belongs to fluorescence imaging field, more specifically, relate to the burnt mini microscope of a kind of copolymerization.
Background technology
To vivo biological tissue imaging, it is an important means in life science field.And wherein to the realtime imaging of live body brain cell, become the important means contacted of zoologizeing between behavior and cerebral neuron inside neuroscience, in experimentation, give animal stimulation makes animal make behavior, and to observe in animal brain district specific region neuron produce reaction, can set up like this animal behavior and its related Neurons distribute between contact.
By measuring in active procedure, the action potential change of animal's nerve cells, therewith whether behavior is relevant to detect this neurocyte.The optical signal excited due to electric indicator is too weak, and poor selectivity is unsatisfactory in the effect of clinical trial, therefore research worker can use the change that calcon-carboxylic acid carrys out indirect inspection action potential usually.Calcium ion is maintain intraor extracellular electrochemical gradient in intracellular effect, intracellular second message,second messenger, action potential change frequency is relevant with calcium ion concentration change, when neurocyte activates the change of generation action potential in action process, the increase of intracellular calcium concentration can be caused, now calcium ion is combined with calcon-carboxylic acid and produces fluorescin, and fluorescence excitation under the stimulation of extraneous illumination, just can detect fluorescence signal by imaging device.
At neuroscience field, studying that this animal behavior and functional neurosurgery unit associates is method mainly through live body realtime imaging.At present, a kind of method utilizing miniature fluorescence microscopy device animal to be carried out to live body realtime imaging has been developed in the industry.Miniature fluorescence microscopy device volume is little, and mirror body can be fixed on mouse head, and its weight can not have influence on the activity of mice., there is afocal signal disturbing during imaging of samples to three dimensional structure, be lowered into image contrast and signal to noise ratio, time serious, even can cause the erroneous judgement of signal in the wide field optical imagery that existing miniature fluorescent microscopic imaging device adopts.
Summary of the invention
For above defect or the Improvement requirement of prior art, the invention provides a kind of Laser Scanning Confocal Microscope, its object is to improve image contrast and signal to noise ratio, solve existing miniature fluorescence microscope thus and there is afocal signal disturbing, cause the technical problem of the erroneous judgement of signal.
For achieving the above object, according to one aspect of the present invention, provide the burnt mini microscope of a kind of copolymerization, illumination path is disposed with scanning mirror, relay lens group and GRIN Lens; Imaging optical path is disposed with filter set, imaging len, aperture or slit and photodetector;
Described scanning mirror, for adjusting illumination light and radiative direction;
Described relay lens group, makes scanning mirror mate with GRIN Lens pupil;
Described GRIN Lens, for gather illumination light on sample to be imaged and to collect the fluorescence of described electromagnetic radiation;
Described imaging len, for by radiative fluorescent foci;
Described aperture or slit, be arranged on the focal plane of described imaging len;
During work, the illumination light through collimation reflects on the scan mirror through filter set, is radiated in GRIN Lens, focuses on sample to be imaged, fluorescence excitation by GRIN Lens after reflection by relay lens group; The fluorescence of described electromagnetic radiation is by being collected by GRIN Lens, reverse along illumination light light path, through relay lens group and scanning mirror, is incident upon in filter set, is focused on after transmission by imaging len, collected by aperture or slit by photodetector.
Preferably, the burnt mini microscope of described copolymerization, its imaging optical path is provided with aperture.
Preferably, the burnt mini microscope of described copolymerization, its photodetector is miniature photomultiplier.
Preferably, the burnt mini microscope of described copolymerization, its scanning mirror is Scan mirror, preferred MEMS scanning mirror.
Preferably, the burnt mini microscope of described copolymerization, it also comprises illuminating spotlight mirror, is arranged between light source and filter set, for illumination light being collimated in a scanning direction, for turning round symmetrical lens.
Preferably, the burnt mini microscope of described copolymerization, its imaging optical path is provided with slit.
Preferably, the burnt mini microscope of described copolymerization, its photodetector photosurface overlaps with described imaging len focal plane, is photodiode array.
Preferably, the burnt mini microscope of described copolymerization, its photodiode array is linear array CMOS.
Preferably, the burnt mini microscope of described copolymerization, its scanning mirror is miniscanning mirror, preferred MEMS scanning mirror or miniature polygonal rotating mirror.
Preferably, the burnt mini microscope of described copolymerization, it also comprises illuminating spotlight mirror, is arranged between light source and filter set, for illumination light being collimated in a scanning direction, is the battery of lens of the post lens composition that pair of orthogonal is arranged.
In general, the above technical scheme conceived by the present invention compared with prior art, can obtain following beneficial effect:
The present invention utilizes common focusing principle, achieves optical chromatography, inhibits afocal signal disturbing, improve contrast and signal to noise ratio in miniature fluorescent microscopic imaging device.
Accompanying drawing explanation
Fig. 1 is the light channel structure figure of embodiment 1;
Fig. 2 is the light channel structure figure of embodiment 2;
Fig. 3 is the light channel structure figure of embodiment 3;
Fig. 4 is that embodiment 3 is perpendicular to scanning direction illumination path structure;
Fig. 5 is that embodiment 3 is perpendicular to scanning direction imaging optical path structure.
In all of the figs, identical Reference numeral is used for representing identical element or structure, wherein: 1 is scanning mirror, 2 is relay lens, and 3 is GRIN Lens, and 4 is optical filter, 5 is imaging len, 6 is aperture, and 7 is miniature photomultiplier, and 8 is light source, 9 is illuminating spotlight mirror, 10 is dichroic mirror, and 11 is linear array CMOS photodetector, and 12 is post lens.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.In addition, if below in described each embodiment of the present invention involved technical characteristic do not form conflict each other and just can mutually combine.
The burnt mini microscope of copolymerization provided by the invention, illumination path is disposed with scanning mirror 1, relay lens group 2 and self-focusing object lens 3; Imaging optical path is disposed with filter set, imaging len 5, aperture 6 or slit and photodetector; Preferably, the illuminating spotlight mirror 9 be arranged between light source 8 and filter set is also comprised.
Described scanning mirror 1, for adjusting illumination light and radiative direction;
Described relay lens group 2, makes scanning mirror 1 mate with self-focusing object lens 3 pupil;
Described self-focusing object lens 3, for gather illumination light on sample to be imaged and to collect the fluorescence of described electromagnetic radiation;
Described imaging len 5, for by radiative fluorescent foci;
Described aperture 6 or slit, be arranged on the focal plane of described imaging len 5;
Described illuminating spotlight mirror 9, for collimating illumination light in a scanning direction.
During work, the illumination light through collimation is reflected on scanning mirror 1 through filter set, is radiated on self-focusing object lens 3, focuses on sample to be imaged, fluorescence excitation by self-focusing object lens 3 after reflection by relay lens group 2; The fluorescence of described electromagnetic radiation is by being collected by self-focusing object lens 3, reverse along illumination light light path, through relay lens group 2 and scanning mirror 1, is incident upon in filter set, is focused on after transmission by imaging len 5, collected by aperture 6 or slit by photodetector.
When described imaging optical path is arranged be aperture 6 time: described photodetector is miniature photomultiplier 7; Described scanning mirror 1 is two and scans mirror, preferred MEMS scanning mirror; Described illuminating spotlight mirror 9 is the lens of revolution symmetry.
When described imaging optical path is arranged be slit time: described photodetector photosurface overlaps with described imaging len 5 focal plane, is photodiode array, be preferably linear array CMOS photodetector 11; Described scanning mirror 1 is miniscanning mirror, preferred MEMS scanning mirror or miniature polygonal rotating mirror; The battery of lens of the post lens composition that described illuminating spotlight mirror 9 is arranged for pair of orthogonal.
Be below embodiment:
Embodiment 1
The burnt mini microscope of a kind of copolymerization, as shown in Figure 1, illumination path is disposed with scanning mirror 1, relay lens group 2 and self-focusing object lens 3; Imaging optical path is disposed with filter set, imaging len 5, aperture 6 and miniature photomultiplier 7; Also comprise the illuminating spotlight mirror 9 be arranged between light source 8 and filter set.
Described filter set comprises optical filter 4 and dichroic mirror 10, for adjusting light path and optical filtering.
Described scanning mirror 1, for adjusting illumination light and radiative direction, is MEMS scanning mirror;
Described relay lens group 2, by the mirror image of scanning mirror 1 on the entrance pupil of self-focusing object lens 3, makes scanning mirror 1 mate with self-focusing object lens 3 pupil, is pair of alignment aspheric surface plus lens, and the focal plane of the two overlaps;
Described self-focusing object lens 3, for illumination light to be gathered on sample to be imaged and to collect the fluorescence of described electromagnetic radiation, described self-focusing object lens 3 length is 0.23 pitch, and equivalent focal length is about 0.6mm;
Described imaging len 5, for the fluorescent foci by electromagnetic radiation, focal length is about 6mm;
Described aperture 6, is arranged on the focal plane of described imaging len 5;
Described illuminating spotlight mirror 9, for illumination light being collimated in scanning plane, be aspheric surface plus lens, focal length is about 3mm;
Described photodetector is miniature photomultiplier 7, is arranged on aperture 6 rear.
During work, the illumination light through collimation is reflected on scanning mirror 1 through filter set, is radiated on self-focusing object lens 3, focuses on sample to be imaged, fluorescence excitation by self-focusing object lens 3 after reflection by relay lens group 2; The fluorescence of described electromagnetic radiation is by being collected by self-focusing object lens 3, reverse along illumination light light path, through relay lens group 2 and scanning mirror 1, is incident upon in filter set, is focused on after transmission by imaging len 5, collected by aperture 6 by miniature photomultiplier 7.When scanning mirror 1 rotates, its object space beam direction changes thereupon, the focus position corresponded on self-focusing object lens 3 focal plane also changes thereupon, a point on the corresponding object plane of each attitude of scanning mirror 1, and the output of miniature photomultiplier 7 can reflect the intensity of the fluorescence that this point sends.Scanning mirror 1 rotates according to certain rules and makes the focus point on self-focusing object lens 3 focal plane sweep complete object plane, and the output of miniature photomultiplier 7 is combined into a face according to same rule with mosaic formation, can obtain the picture of object plane.
Embodiment 2
The burnt mini microscope of a kind of copolymerization, as shown in Figure 2, illumination path is disposed with scanning mirror 1, relay lens group 2 and self-focusing object lens 3; Imaging optical path is disposed with filter set, imaging len 5, aperture 6 and photodetector; Also comprise the illuminating spotlight mirror 9 be arranged between light source 8 and filter set.
Described filter set comprises optical filter 4 and dichroic mirror 10, for adjusting light path and optical filtering.
Described scanning mirror 1, for adjusting illumination light and radiative direction, is miniature polygonal rotating mirror;
Described relay lens group 2, by the mirror image of scanning mirror 1 on the entrance pupil of self-focusing object lens 3, makes scanning mirror 1 mate with self-focusing object lens 3 pupil, is pair of alignment aspheric surface plus lens, and the focal plane of the two overlaps;
Described self-focusing object lens 3, for illumination light to be gathered on sample to be imaged and to collect the fluorescence of described electromagnetic radiation, described self-focusing object lens 3 length is 0.23 pitch, and equivalent focal length is about 0.6mm;
Described imaging len 5, for the fluorescent foci by electromagnetic radiation;
Described aperture 6, is arranged on the focal plane of described imaging len 5;
Described illuminating spotlight mirror 9, for illumination light being collimated in a scanning direction, is aspheric collimation lens;
Described photodetector is miniature photomultiplier 7, is arranged on aperture 6 rear.
During work, the illumination light through collimation is reflected on scanning mirror 1 through filter set, is radiated on self-focusing object lens 3, focuses on sample to be imaged, fluorescence excitation by self-focusing object lens 3 after reflection by relay lens group 2; The fluorescence of described electromagnetic radiation is by being collected by self-focusing object lens 3, reverse along illumination light light path, through relay lens group 2 and scanning mirror 1, is incident upon in filter set, is focused on after transmission by imaging len 5, collected by aperture 6 by miniature photomultiplier 7.When scanning mirror 1 rotates, its object space beam direction changes thereupon, the focus position corresponded on self-focusing object lens 3 focal plane also changes thereupon, a point on the corresponding object plane of each attitude of scanning mirror 1, and the output of miniature photomultiplier 7 can reflect the intensity of the fluorescence that this point sends.Scanning mirror 1 rotates according to certain rules and makes the focus point on self-focusing object lens 3 focal plane sweep complete object plane, and the output of miniature photomultiplier 7 is combined into a face according to same rule with mosaic formation, can obtain the picture of object plane.
Embodiment 3
The burnt mini microscope of a kind of copolymerization, as shown in Figure 3, illumination path is disposed with scanning mirror 1, relay lens group 2 and self-focusing object lens 3; Imaging optical path is disposed with filter set, imaging len 5, slit and photodetector; Also comprise the illuminating spotlight mirror 9 be arranged between light source 8 and filter set.Fig. 3 is along scanning direction light channel structure; Fig. 4 is perpendicular to scanning direction illumination path structure; Fig. 5 is perpendicular to scanning direction imaging optical path structure.
Described filter set comprises optical filter 4 and dichroic mirror 10, for adjusting light path and optical filtering.
Described scanning mirror 1, for adjusting illumination light and radiative direction, is MEMS scanning mirror;
Described relay lens group 2, makes scanning mirror 1 mate with self-focusing object lens 3 pupil, is pair of alignment aspheric surface plus lens, and the focal plane of the two overlaps;
Described self-focusing object lens 3, for illumination light to be gathered on sample to be imaged and to collect the fluorescence of described electromagnetic radiation, described self-focusing object lens 3 length is 0.23 pitch, and equivalent focal length is about 0.6mm;
Described imaging len 5, for the fluorescent foci by electromagnetic radiation;
Described slit, is arranged on the focal plane of described imaging len 5;
Described illuminating spotlight mirror 9, be the battery of lens of the post lens composition that pair of orthogonal is arranged, the post lens placed along scanning direction are used for illumination light to collimate in a scanning direction, and the post lens focal plane placed perpendicular to scanning direction is positioned near described scanning mirror 1 reflecting surface;
Described photodetector is linear array CMOS photodetector 11, is close to slit and places, and ensures that the distance of the photosurface of slit and linear array CMOS photodetector 11 to imaging len 5 focal plane is in focal depth range.
During work, the illumination light through collimation is reflected on scanning mirror 1 through filter set, is radiated on self-focusing object lens 3, focuses on sample to be imaged by self-focusing object lens 3 after reflection by relay lens group 2, forms linear light speckle, fluorescence excitation; The fluorescence of described electromagnetic radiation is by being collected by self-focusing object lens 3, reverse along illumination light light path, through relay lens group 2 and scanning mirror 1, is incident upon in filter set, is focused on after transmission by imaging len 5, collected by slit by linear array CMOS photodetector 11.The output of linear array CMOS photodetector 11 is the picture of thing on linear light speckle, and scanning mirror 1 rotates then linear light speckle mobile formation line sweep thereupon, and linear array CMOS is stitched together according to each two field picture that sequential exports and is the picture of object plane.
Those skilled in the art will readily understand; the foregoing is only preferred embodiment of the present invention; not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (10)
1. the burnt mini microscope of copolymerization, is characterized in that, illumination path is disposed with scanning mirror, relay lens group and GRIN Lens; Imaging optical path is disposed with filter set, imaging len, aperture or slit and photodetector;
Described scanning mirror, for adjusting illumination light and radiative direction;
Described relay lens group, makes scanning mirror mate with GRIN Lens pupil;
Described GRIN Lens, for gather illumination light on sample to be imaged and to collect the fluorescence of described electromagnetic radiation;
Described imaging len, for by radiative fluorescent foci;
Described aperture or slit, be arranged on the focal plane of described imaging len;
During work, the illumination light through collimation reflects on the scan mirror through filter set, is radiated in GRIN Lens, focuses on sample to be imaged, fluorescence excitation by GRIN Lens after reflection by relay lens group; The fluorescence of described electromagnetic radiation is by being collected by GRIN Lens, reverse along illumination light light path, through relay lens group and scanning mirror, is incident upon in filter set, is focused on after transmission by imaging len, collected by aperture or slit by photodetector.
2. the burnt mini microscope of copolymerization as claimed in claim 1, is characterized in that, described imaging optical path is provided with aperture.
3. focus on mini microscope as claimed in claim 2, it is characterized in that, described photodetector is miniature photomultiplier.
4. focus on mini microscope as claimed in claim 2, it is characterized in that, described scanning mirror is Scan mirror, preferred MEMS scanning mirror.
5. the burnt mini microscope of copolymerization as claimed in claim 2, is characterized in that, also comprise illuminating spotlight mirror, be arranged between light source and filter set, for illumination light being collimated in a scanning direction, for turning round symmetrical lens.
6. the burnt mini microscope of copolymerization as claimed in claim 1, is characterized in that, described imaging optical path is provided with slit.
7. focus on mini microscope as claimed in claim 6, it is characterized in that, described photodetector photosurface overlaps with described imaging len focal plane, is photodiode array.
8. focus on mini microscope as claimed in claim 7, it is characterized in that, described photodiode array is linear array CMOS.
9. focus on mini microscope as claimed in claim 6, it is characterized in that, described scanning mirror is miniscanning mirror, preferred MEMS scanning mirror or miniature polygonal rotating mirror.
10. focusing on mini microscope as claimed in claim 6, it is characterized in that, also comprise illuminating spotlight mirror, be arranged between light source and filter set, for illumination light being collimated in a scanning direction, is the battery of lens of the post lens composition that pair of orthogonal is arranged.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510220771.6A CN104887318A (en) | 2015-05-04 | 2015-05-04 | Mini-type confocal microscope |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510220771.6A CN104887318A (en) | 2015-05-04 | 2015-05-04 | Mini-type confocal microscope |
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| CN104887318A true CN104887318A (en) | 2015-09-09 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107966799A (en) * | 2017-12-27 | 2018-04-27 | 南方医科大学 | A kind of miniature mating plate microscope of wear-type |
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| CN103776769A (en) * | 2012-10-19 | 2014-05-07 | 赖博 | Confocal optical scanner |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070255143A1 (en) * | 2006-04-27 | 2007-11-01 | Olympus Corporation | Imaging apparatus |
| US7460248B2 (en) * | 2006-05-15 | 2008-12-02 | Carestream Health, Inc. | Tissue imaging system |
| US20120242992A1 (en) * | 2011-03-21 | 2012-09-27 | University Of Central Florida Research Foundation Inc. | Gradient index (grin)-based absorption spectroscopy apparatus, method, and applications |
| CN202204470U (en) * | 2011-07-06 | 2012-04-25 | 黄书伟 | Balanced detection confocal microscope imaging system |
| CN103776769A (en) * | 2012-10-19 | 2014-05-07 | 赖博 | Confocal optical scanner |
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| CN107966799A (en) * | 2017-12-27 | 2018-04-27 | 南方医科大学 | A kind of miniature mating plate microscope of wear-type |
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