CN104865687B - A kind of autohemagglutination focus objective lens and miniature fluorescent microscopic imaging device - Google Patents
A kind of autohemagglutination focus objective lens and miniature fluorescent microscopic imaging device Download PDFInfo
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- CN104865687B CN104865687B CN201510220640.8A CN201510220640A CN104865687B CN 104865687 B CN104865687 B CN 104865687B CN 201510220640 A CN201510220640 A CN 201510220640A CN 104865687 B CN104865687 B CN 104865687B
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/006—Optical details of the image generation focusing arrangements; selection of the plane to be imaged
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/02—Objectives
- G02B21/025—Objectives with variable magnification
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/368—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements details of associated display arrangements, e.g. mounting of LCD monitor
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Abstract
The invention discloses a kind of autohemagglutination focus objective lens and miniature fluorescent microscopic imaging device.The autohemagglutination focus objective lens, including GRIN Lens and the glass bar of end face polishing, the end face of glass bar of the end face polishing are connected with the image space end face of the GRIN Lens.The fluorescent microscopic imaging device, including excitation source, condenser lens, dichroscope, the autohemagglutination focus objective lens, knot are as lens and face battle array photodetector；The excitation source sends exciting light line focus lens focus, transmits on the dichroscope；The dichroscope is by the excitation light reflection of line focus, by the lengthening object lens transmission on sample；The fluorescence that sample excitation goes out, is collected by the lengthening object lens, through being transmitted on the dichroscope, is then transmitted and is imaged on the battle array photodetector of the face as lens focus through the knot.Autohemagglutination focus objective lens provided by the invention and miniature fluoroscopic imaging device it is small, from heavy and light, be adapted to living imaging.
Description
Technical field
The invention belongs to fluorescent microscopic imaging field, more particularly, to a kind of autohemagglutination focus objective lens and miniature fluorescence microscopy
Imaging device.
Background technology
Vivo biological tissue is imaged, is an important means in life science field.It is and wherein thin to live body brain
The real time imagery of born of the same parents, has become the important hand contacted between research animal behavior and cerebral neuron inside Neuscience
Section, one stimulation of animal is given in experimentation makes animal make behavior, and observes specific region neuron in animal brain area
Reaction is produced, can so set up the contact between animal behavior neuron distribution associated therewith.
By measuring in active procedure, the action potential change of animal's nerve cells, whether to detect the nerve cell
It is related with this behavior.Since the optical signal of electric indicator excitation is too weak, poor selectivity is unsatisfactory in the effect of clinical test,
Therefore usually researcher can use the change that calcon-carboxylic acid measures action potential indirectly.The effect of calcium ion in the cell is dimension
Intraor extracellular electrochemical gradient is held, intracellular second messenger, action potential change frequency and calcium ion concentration change are related, when
Nerve cell can cause the increase of intracellular calcium concentration, at this time when activation during behavior produces action potential change
Calcium ion is combined with calcon-carboxylic acid produces fluorescin, and fluorescence is excited under the stimulation of extraneous illumination, can be filled by being imaged
Put and detect fluorescence signal.
At present in neuroscience field, studying this animal behavior, with functional neurosurgery member to associate owner real-time by live body
The method of imaging.In order to realize real-time living imaging, fiber optic bundle two-photon excitation imaging mode can be used, fiber optic bundle is passed through
A kind of miniature head-mounted device insertion needs the area of biological tissue observed, and exciting light is passed in detector by optical fiber；In addition
Fiber optic bundle can also be replaced with miniature object lens, by the way that signal is passed to detector after specific light path imaging.In experimentation into
The part being fixed on as device on living animal must be small, light-weight, could not influence the freely activity of animal, exclude it
He disturbs factor.Same technological means, can also come to carry out its hetero-organization of biological living real time imagery, such as observation disease
Become tissue regions, vascular flow, internal organs etc..
However, existing living body fluorescent imaging device, volume is larger, and dead weight is heavier, for the physiology of somatoscopy object
Activity hinders substantially, and therefore, one side fluorescence imaging result is undesirable, and another aspect imaging results are not enough close in free activity
Under physiological conditions.
The content of the invention
For the disadvantages described above or Improvement requirement of the prior art, the present invention provides a kind of autohemagglutination focus objective lens and miniature fluorescence
Microscopic imaging device, its object is to lengthen autohemagglutination focus objective lens fluoroscopic imaging device volume by one kind to be substantially reduced, subtracts
Light dead-weight, is imaged easy to living body fluorescent, thus solves the technology that existing living body fluorescent imaging device volume is larger, dead weight is heavier
Problem.
To achieve the above object, one side according to the invention, there is provided one kind lengthens autohemagglutination focus objective lens, including autohemagglutination
Focus lens and the glass bar of end face polishing, the image space end of the end face and the GRIN Lens of the glass bar of the end face polishing
Face connects.
Preferably, the lengthening autohemagglutination focus objective lens, its glass bar are truncated cone-shaped, its object space end face and the GRIN Lens
Image space end face be engaged, its image space end face diameter D >=d+Ltan θ, wherein d are GRIN Lens diameter, and L is glass bar length
Degree, θ is field angle.
Preferably, the lengthening autohemagglutination focus objective lens, its glass bar is cylindrical, its diameter of section D >=d+Ltan θ, wherein d
For GRIN Lens diameter, L is glass bar length, and θ is field angle.
Other side according to the invention, there is provided a kind of fluorescent microscopic imaging device, it includes excitation source, focuses on
Lens, dichroscope, the autohemagglutination focus objective lens as described in claims 1 to 3 any one, knot are as lens and face battle array photoelectricity spy
Survey device；The excitation source sends exciting light line focus lens focus, transmits on the dichroscope；The dichroscope will
The excitation light reflection of line focus, by the lengthening object lens transmission on sample；The fluorescence that sample excitation goes out, by the lengthening thing
Mirror is collected, and through being transmitted on the dichroscope, is then transmitted through the knot as lens focus on the battle array photodetector of the face
Imaging.
Preferably, the microscopic imaging device, it further includes shell, and the shell sees the autohemagglutination focus objective lens and live body
Object is examined to fix.
Preferably, the microscopic imaging device, its shell include fixing piece and movable part；The fixing piece and self-focus lens
Head fixed-link, for self-focusing camera lens and somatoscopy object to be fixed；The movable part, is equipped with excitation source, focuses on
Lens, dichroscope, knot are as lens and face battle array photodetector；The fixing piece and movable part are detachably connected.
Preferably, the microscopic imaging device, its shell assembling knot are set as the position of lens knead dough battle array photodetector
There is length adjustment device, for adjusting knot as the distance between lens knead dough battle array photodetector.
Preferably, the microscopic imaging device, its length adjustment device are screw feeding adjusting device.
Preferably, the microscopic imaging device, its excitation source are LED light source or optical fiber-coupled laser.
Preferably, the microscopic imaging device, its face its data cable of battle array photodetector use anti-interference stable multiple twin
Line.
In general, by the contemplated above technical scheme of the present invention compared with prior art, it can obtain down and show
Beneficial effect：
(1) autohemagglutination focus objective lens provided by the invention, compared to existing microscopic imaging device, can significantly reduce micro-imaging
Device volume and weight, microscopic imaging device are miniaturized, portability；
(2) autohemagglutination focus objective lens provided by the invention, compared to common autohemagglutination focus objective lens, total length is longer, can more go deep into
Living tissue, applicability is stronger, can observe superficial structure and observe the tissue of deeper；
(3) autohemagglutination focus objective lens provided by the invention, light path is passed to using glass bar, avoids picture caused by long GRIN Lens
Difference-product tires out, optical resolution higher, and imaging effect is more preferable；It is greatly reduced at the same time relative to long GRIN Lens, production cost；
(4) autohemagglutination focus objective lens provided by the invention, using truncated cone-shaped glass bar so that it is inserted into the surface light of living tissue
Sliding, living tissue damage is small, and observation result is closer in lossless case；
(5) microscopic imaging device provided by the invention, using the autohemagglutination focus objective lens, has the advantages that miniature, portable；
(6) microscopic imaging device provided by the invention, can be fixed on somatoscopy object, reduce to somatoscopy object
Influence, can also use softer photodetector its data cable, further reduce the influence to the free activity of live body, it is convenient compared with
Continuously observe for a long time and result is closer to live body free state；
(7) microscopic imaging device provided by the invention, can use detachable structure, when not observing conveniently, allow and observe object
Freely activity, reduces the pain of observation object；
(8) microscopic imaging device provided by the invention, is tied as the distance between lens knead dough battle array photodetector is adjustable, side
Just focus；In addition coordinate spring to carry out length adjustment using screw feeding adjusting device, avoid face battle array photodetector opposite
Rotated in mirror body, cause data cable to knot.
Brief description of the drawings
Fig. 1 is lengthening GRIN Lens structure diagram provided by the invention；
Fig. 2 is lengthening GRIN Lens light path schematic diagram provided by the invention；
Fig. 3 is miniature silver-colored light microscopic imaging device index path provided by the invention；
Fig. 4 is shell mechanism schematic diagram provided by the invention；
Fig. 5 is the lengthening GRIN Lens structure diagram of embodiment 1；
Fig. 6 is the lengthening GRIN Lens structure diagram of embodiment 2；
Fig. 7 is the structure diagram of embodiment 3；
Fig. 8 is the structure diagram of embodiment 4.
In all of the figs, identical reference numeral is used for representing identical element or structure, wherein：01 is saturating for self-focusing
Mirror, 02 is glass bar, and 11 be excitation source, and 12 be condenser lens, and 13 be exciter filter, and 14 be dichroscope, and 15 be lengthening
Autohemagglutination focus objective lens, 16 be sample, and 17 be transmitting optical filter, and 18 is tie as lens, and 19 be face battle array photodetector, and 21 fill for shell
Position with face battle array photodetector, 22 tie as the position of lens for shell assembling, and 23 be spring, and 24 be screw, and 31 be shell,
32 be fixing piece, and 33 be movable part.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to the accompanying drawings and embodiments, it is right
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.As long as in addition, technical characteristic involved in each embodiment of invention described below
Not forming conflict each other can be mutually combined.
Lengthening autohemagglutination focus objective lens provided by the invention, as shown in Figure 1, the glass polished including GRIN Lens 01 and end face
Rod 02, the end face of the glass bar 02 of the end face polishing are connected with the image space end face of the GRIN Lens 01.The glass
02 preferred truncated cone-shaped of rod or cylinder.Its index path is as shown in Figure 2.
If the glass bar 02 is truncated cone-shaped, its object space end face is engaged with the image space end face of the GRIN Lens 01,
Its image space end face diameter D >=d+Ltan θ, wherein d are 01 diameter of GRIN Lens, and L is 02 length of glass bar, and θ is field angle.Circle
The structure of platform shape, can seamlessly transit GRIN Lens 01, and formation is needle-shaped, be easy to be pierced into somatoscopy object tissue, and injury is small,
It is small to In vivo effects.
If the glass bar 02 is cylinder, its diameter of section D >=d+Ltan θ, wherein d are 01 diameter of GRIN Lens, L
For 02 length of glass bar, θ is field angle.The structure of cylinder, it is relatively low to process simple cost, can be saturating in glass bar 02 and self-focusing
01 junction of mirror, it is excessive to form the gradient using adhesive.
Wherein D >=d+Ltan θ in order to ensure from after GRIN Lens 01 pupil be emitted light beam light will not be blocked by glass bar 02.
Fluorescent microscopic imaging device provided by the invention, as shown in figure 3, including excitation source 11, condenser lens 12, two to
Look mirror 14, lengthening autohemagglutination focus objective lens 15 provided by the invention, knot are as lens 18 and face battle array photodetector 19；The excitation
Light source 11 sends exciting light line focus lens 12 and focuses on, and transmits on the dichroscope 14；The dichroscope 14 will be through poly-
Burnt excitation light reflection, by the lengthening object lens transmission on sample 16；The fluorescence that sample 16 inspires, by the lengthening thing
Mirror is collected, and through being transmitted on the dichroscope 14, is then focused on through the knot as lens 18, is transmitted in face battle array photodetection
It is imaged on device 19.Preferably, the fluorescent microscopic imaging device further includes exciting light optical filter and transmitting optical filter 17, described to swash
Hair optical filter 13 be arranged in the light path between excitation source 11 and dichroscope 14, it is described transmitting optical filter 17 be arranged on two to
In light path between 14 knead dough battle array photodetector 19 of Look mirror.
The microscopic imaging device, further includes shell 31, and the shell 31 is by the autohemagglutination focus objective lens and somatoscopy
Object is fixed.Preferably, the shell 31 includes fixing piece 32 and movable part 33；The fixing piece 32 is fixed with self-focusing camera lens
Link, for self-focusing camera lens and somatoscopy object to be fixed；The movable part 33, is equipped with excitation source 11, focuses on thoroughly
Mirror 12, dichroscope 14, knot are as lens 18 and face battle array photodetector 19；The fixing piece 32 and movable part 33 are detachable
Connection.Preferably, between the position 21,22 as 18 knead dough battle array photodetector 19 of lens is tied in the assembling of shell 31, it is provided with
Length adjustment device, as shown in figure 4, for adjusting knot as the distance between 18 knead dough battle array photodetector 19 of lens.The length
It is preferably 24 feeding adjusting device of screw to spend regulating device.When adjusting image definition, it is necessary to adjust lens barrel and shank up and down
Relative position, with change knot as the distance between 18 knead dough battle array photodetector 19 of lens.The focusing structure that the present invention realizes
It is preferred that complete to adjust by 24 screw of screw.As shown in figure 4, in the case where spring 23 makes nut and lens barrel wall is close to, rotation
Turn screw to move back and forth, the mobile band moving lens barrel of screw is translatable back and forth relative to shank realizes focusing.The device advantage is to adjust
Translation focusing is realized during Jiao, it is entirely avoided face battle array photodetector 19 is rotated relative to mirror body, causes data cable to be beaten
The problem of knot.
The excitation source 11 is LED light source or optical fiber-coupled laser.
The face battle array photodetector 19 uses the miniature CMOS camera of mobile phone, its data cable uses anti-interference stable
Twisted-pair feeder.
It is embodiment below：
Embodiment 1
One kind lengthens autohemagglutination focus objective lens, as shown in figure 5, the glass bar 02 polished including GRIN Lens 01 and end face, institute
The end face for stating the glass bar 02 of end face polishing is connected with the image space end face of the GRIN Lens 01.
The glass bar 02 is truncated cone-shaped, its object space end face is engaged with the image space end face of the GRIN Lens 01, its
Image space end face diameter D=1mm, 01 diameter d=0.5mm of GRIN Lens, 02 length L=3mm of glass bar.The structure of truncated cone-shaped,
GRIN Lens 01 can be seamlessly transitted, formation is needle-shaped, is easy to be pierced into somatoscopy object tissue, injury is small, to In vivo effects
It is small.
Embodiment 2
One kind lengthens autohemagglutination focus objective lens, as shown in fig. 6, the glass bar 02 polished including GRIN Lens 01 and end face, institute
The end face for stating the glass bar 02 of end face polishing is connected with the image space end face of the GRIN Lens 01.
The glass bar 02 is cylindrical, its diameter of section diameter D=1mm, 01 diameter d=0.5mm of GRIN Lens, glass
02 length L=3mm of glass rod.The structure of cylinder, it is relatively low to process simple cost, can be connected in glass bar 02 and GRIN Lens 01
Place, it is excessive to form the gradient using adhesive.
Embodiment 3
A kind of fluorescent microscopic imaging device, as shown in fig. 7, comprises excitation source 11, condenser lens 12, dichroscope 14,
The lengthening autohemagglutination focus objective lens 15 of the offer of embodiment 1, tie as lens 18, face battle array photodetector 19, exciting light optical filter and transmitting
Optical filter 17；The excitation source 11 sends exciting light line focus lens 12 and focuses on, and transmits on the dichroscope 14；It is described
Dichroscope 14 is by the excitation light reflection of line focus, by the lengthening object lens transmission on sample 16；What sample 16 inspired
Fluorescence, is collected by the lengthening object lens, through being transmitted on the dichroscope 14, is then focused on as lens 18, is transmitted through the knot
It is imaged on the battle array photodetector 19 of the face.The exciter filter 13 is arranged between excitation source 11 and dichroscope 14
Light path on, it is described transmitting optical filter 17 be arranged in the light path between 14 knead dough battle array photodetector 19 of dichroscope.Its light
Road figure is as shown in Figure 3.
The microscopic imaging device, further includes shell 31, as shown in figure 4, the assembling knot of the shell 31 is as lens knead dough
The position of battle array photodetector 19, is provided with 24 feeding adjusting device of screw, for adjusting knot as lens knead dough battle array photodetection
The distance between device 19.
The excitation source 11 is optical fiber-coupled laser.
The face battle array photodetector 19 uses the miniature CMOS camera of mobile phone, its data cable uses anti-interference stable
Twisted-pair feeder.
Embodiment 4
A kind of fluorescent microscopic imaging device, as shown in figure 8, including excitation source 11, condenser lens 12, dichroscope 14,
The lengthening autohemagglutination focus objective lens 15 of the offer of embodiment 2, tie as lens 18, face battle array photodetector 19, exciting light optical filter and transmitting
Optical filter 17；The excitation source 11 sends exciting light line focus lens 12 and focuses on, and transmits on the dichroscope 14；It is described
Dichroscope 14 is by the excitation light reflection of line focus, by the lengthening object lens transmission on sample 16；What sample 16 inspired
Fluorescence, is collected by the lengthening object lens, through being transmitted on the dichroscope 14, is then focused on as lens 18, is transmitted through the knot
It is imaged on the battle array photodetector 19 of the face.The exciter filter 13 is arranged between excitation source 11 and dichroscope 14
Light path on, it is described transmitting optical filter 17 be arranged in the light path between 14 knead dough battle array photodetector 19 of dichroscope.Its light
Road figure is as shown in Figure 3.
The microscopic imaging device, further includes shell 31, as shown in figure 4, the shell 31 is by the autohemagglutination focus objective lens
Fixed with somatoscopy object.The shell 31 includes fixing piece 32 and movable part 33；The fixing piece 32 and self-focusing camera lens
Fixed-link, for self-focusing camera lens and somatoscopy object to be fixed；The movable part 33, is equipped with excitation source 11, gathers
Focus lens 12, dichroscope 14, knot are as lens 18, face battle array photodetector 19, exciting light optical filter and transmitting optical filter 17；Institute
State fixing piece 32 and movable part 33 is detachably connected.The assembling of shell 31 knot is as the portion of 18 knead dough battle array photodetector 19 of lens
Position, is provided with 24 feeding adjusting device of screw, for adjusting knot as the distance between 18 knead dough battle array photodetector 19 of lens.
The excitation source 11 is optical fiber-coupled laser.
The face battle array photodetector 19 uses the miniature CMOS camera of mobile phone, its data cable uses anti-interference stable
Twisted-pair feeder.
As it will be easily appreciated by one skilled in the art that the foregoing is merely illustrative of the preferred embodiments of the present invention, not to
The limitation present invention, all any modification, equivalent and improvement made within the spirit and principles of the invention etc., should all include
Within protection scope of the present invention.
Claims (8)
1. one kind lengthens autohemagglutination focus objective lens, it is characterised in that the glass bar (02) polished including GRIN Lens (01) and end face,
The end face of the glass bar (02) of the end face polishing is connected with the image space end face of the GRIN Lens (01)；The glass bar
(02) it is truncated cone-shaped or cylinder, is that homogeneous refractive index is distributed；
When the glass bar (02) is truncated cone-shaped, its object space end face and the image space end face of the GRIN Lens (01) match
Close, its image space end face diameter D >=d+Ltan θ, wherein d be GRIN Lens (01) diameter, and L is glass bar (02) length, θ for regarding
Rink corner；
When the glass bar (02) is cylinder, its diameter of section D >=d+Ltan θ, wherein d are straight for GRIN Lens (01)
Footpath, L are glass bar (02) length, and θ is field angle.
2. a kind of fluorescent microscopic imaging device, it is characterised in that including excitation source (11), condenser lens (12), dichroscope
(14), autohemagglutination focus objective lens as claimed in claim 1, knot are as lens (18) and face battle array photodetector (19)；The excitation
Light source (11) sends exciting light line focus lens (12) focusing, transmits on the dichroscope (14)；The dichroscope
(14) by the excitation light reflection of line focus, transmitted by the autohemagglutination focus objective lens on sample (16)；Sample (16) inspires
Fluorescence, is collected by the autohemagglutination focus objective lens, through being transmitted on the dichroscope (14), then through the knot as lens (18) are poly-
Jiao, transmits and is imaged on face battle array photodetector (19).
3. microscopic imaging device as claimed in claim 2, it is characterised in that further include shell, the shell is by the autohemagglutination
Focus objective lens are fixed with somatoscopy object.
4. microscopic imaging device as claimed in claim 3, it is characterised in that the shell includes fixing piece (32) and movable part
(33)；The fixing piece (32) is fixedly connected with autohemagglutination focus objective lens, for autohemagglutination focus objective lens and somatoscopy object to be fixed；Institute
Movable part (33) is stated, is equipped with excitation source (11), condenser lens (12), dichroscope (14), knot as lens (18) and face
Battle array photodetector (19)；The fixing piece (32) and movable part (33) are detachably connected.
5. microscopic imaging device as claimed in claim 4, it is characterised in that the shell assembling knot is as lens (18) knead dough battle array
The position of photodetector (19), is provided with length adjustment device, for adjusting knot as lens (18) knead dough battle array photodetector
The distance between (19).
6. microscopic imaging device as claimed in claim 5, it is characterised in that the length adjustment device is screw feeding adjusting
Device.
7. microscopic imaging device as claimed in claim 2, it is characterised in that the excitation source (11) is LED light source or light
Fine coupled laser.
8. microscopic imaging device as claimed in claim 2, it is characterised in that the data cable of the face battle array photodetector (19)
Using anti-interference stable twisted-pair feeder.
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| CN110699241A (en) * | 2019-10-28 | 2020-01-17 | 军事科学院系统工程研究院卫勤保障技术研究所 | Automatic disinfection effect rapid evaluation device and method |
| CN110974173A (en) * | 2019-12-17 | 2020-04-10 | 北京脑科学与类脑研究中心 | Fluorescence imaging system for experimental animals |
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| CN203364759U (en) * | 2013-07-22 | 2013-12-25 | 杭州精飞光学仪器制造有限公司 | Reading microscope |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109828363A (en) * | 2019-01-02 | 2019-05-31 | 浙江大学 | A kind of experimental animal wearable minisize imaging in vivo system |
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