CN106691386A - Detection device of living body fluorescence signal and method - Google Patents
Detection device of living body fluorescence signal and method Download PDFInfo
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- CN106691386A CN106691386A CN201611272914.9A CN201611272914A CN106691386A CN 106691386 A CN106691386 A CN 106691386A CN 201611272914 A CN201611272914 A CN 201611272914A CN 106691386 A CN106691386 A CN 106691386A
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- optical fiber
- fluorescence signal
- detection means
- exciting light
- detection device
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0071—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0082—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes
- A61B5/0084—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes for introduction into the body, e.g. by catheters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
- A61B5/6846—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive
- A61B5/6847—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive mounted on an invasive device
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
- A61B5/6846—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive
- A61B5/6867—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive specially adapted to be attached or implanted in a specific body part
- A61B5/6868—Brain
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
- A61B5/6846—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive
- A61B5/6879—Means for maintaining contact with the body
- A61B5/688—Means for maintaining contact with the body using adhesives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B2503/00—Evaluating a particular growth phase or type of persons or animals
- A61B2503/40—Animals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B2503/00—Evaluating a particular growth phase or type of persons or animals
- A61B2503/42—Evaluating a particular growth phase or type of persons or animals for laboratory research
Abstract
The invention discloses a detection device of a living body fluorescence signal and a method. The detection device of the living body fluorescence signal comprises two parts of an exciting light source and the detection device. Optical fibers inserted into the animal brain tissue are formed by tightly combining two same optical fibers, and the tail ends are aligned. One of two optical fibers inserted into the tissue is used for transmitting the exciting light, and the other end is connected with a laser; the other optical fiber is used for transmitting the fluorescence signal, and the other end thereof is connected with the detection device after connecting to a filtering device. the exciting light is projected to the animal brain tissue by one optical fiber so as to generate the fluorescence by exciting; and the exciting light is transmitted out through the other optical fiber, and sent into the detection device after filtering a background light signal by the filtering device, so that the fluorescence signal can be detected. The detection device of the living body fluorescence signal is capable of monitoring the virus expression situation of the monkey brain in real time by the fluorescence signal, improving the experiment efficiency, and saving labor source and material resources. In addition, the device has the advantages of simple operation and cheap cost, and the damage to the animal tissue is small.
Description
Technical field
The invention belongs to the central nervous system vivo fluorescence signal record in biological visual light imaging technical field, tool
Body is related to the detection means and method of a kind of vivo fluorescence signal.
Background technology
With the rise of gene technology, photogene, retrovirus spike, gene knockout and gene therapy etc. are a series of new
Emerging technology plays important role in the research of Neuscience.The realization of all these technologies will be relied on and noted to target organ
Different vector virus are penetrated to realize, and these viral success transfection aim cells often turn into decision experimental implementation success or failure
It is crucial.Generally we detect whether virus has transfected aim cell with fluorescence signal, if the normal work in experimental animal body
Make and the scope of infected tissue has much.Generally need to sacrifice animal in conventional fluorescence signal detection to verify acquisition group
Sample is knitted, afterwards by a series of fixation, dehydration and section, the histotomy slide that will finally make is placed in fluorescence
Observed one by one under microscope.So not only waste time and energy but also costly (when the high zooperies such as macaque are carried out), especially
It often requires that researcher continues to observe the behavior of same experimental animal after virus transfection operation is completed in neural research field
Learn or other measurement indexes, or researcher generally require observation be a certain process consecutive variations situation.In the past, studied
Personnel typically carry out same treatment by a collection of animal, sacrifice animal to obtain sample the different times of observation are needed.
Not only cost is huge for such method, takes time and effort, and the factor of individual difference between individuality is not fee from again.Can not thus carry
Whether preceding execution animal detection virus transfection succeeds, and can only continue to complete row detection again after all experiments.To so waste a large amount of
Increase workload, reduce conventional efficient.
In the middle of actual process of scientific research, particularly in the middle of primate experiment, valuable primate resource
Determine that this is the work for being difficult to complete.In the transgenic experiments with virus as carrier, how fluorescence is observed in live body
Signal, and then the expression of expression and transgene of the virus in experimental animal body is observed, for transgenic protocol
Quality control, improve later experiments success rate have great meaning.
The content of the invention
In view of the shortcomings of the prior art, the invention provides it is a kind of can in the device of live body Continuous Observation fluorescence signal and
Method, the detection means and method of particularly a kind of vivo fluorescence signal.
Technical scheme：The detection means and method of a kind of vivo fluorescence signal, including exciting light sources and inspection
Survey device two parts.Wherein, the optical fiber of insertion animal brain by two same fibres and it is tight together, it is ensured that end is concordant.
Insert an optical fiber in two optical fiber of tissue to be used for transmitting exciting light, its other end connecting laser；Another optical fiber is used
To transmit fluorescence signal, detection means is reconnected after its other end connection filtering apparatus.Exciting light is thrown by wherein one optical fiber
Animal brain is injected, the fluorescence of generation is excited；Spread out of by another optical fiber, by filtering apparatus wiping out background optical signal
Enter detection means afterwards, just can detect fluorescence signal.
The present invention has following distinguishing feature：
1. observation fluorescence signal need not sacrifice experimental animal.The light source part of the device and fluorescence signal record part are two
The individual separate and adjustable self-contained unit of difference.Portability and flexibility using the device, researcher only need to be using suitable
When method will need the tissue exposure of observation first, it is possible to carry out the record of fluorescence signal in any place, and without sacrificial
Domestic animal animal makes histotomy.
2. detection depth is unrestricted.The different depth that present apparatus system can be expressed according to FLuorescent selects the insertion of optical fiber
Depth, so as to realize that other different depth fluorescence are recorded in addition to cortex to animal brain.It is different additionally by selection
The optical fiber of diameter, the scope that light source can be controlled to irradiate, it is possible thereby to realize the fluorescence signal record of full brain scope.
3. cost is less than most of existing fluorescence signal tape deck.The device mainly comprising exciting light sources,
Optical filter, optical fiber, and a spectrometer.
Brief description of the drawings
Fig. 1 is the principle schematic of the detection means of vivo fluorescence signal of the present invention；
Fig. 2 is the optical fiber structure schematic diagram using present invention insertion animal brain.
Specific embodiment
As shown in figure 1, the detection means of vivo fluorescence signal described in the present embodiment, including light source and detection means two
Point.Light source is a LED light source for power stability (generating laser), can stablize the blue light that release main peak is 470.By increasing
One light-intensity test instrument, can detect excitating light strength.The optical fiber (insertion optical fiber) of animal brain is inserted by two same fibres
And tightly together, it is ensured that end is concordant.Wherein one one end connection LED light source of optical fiber, other end connection insertion optical fiber is (as schemed
2) and by exciting light project in tissue；Another one end connection insertion optical fiber of optical fiber, other end connection filtering apparatus, tissue
The green fluorescence of generation is spread out of and by after filtering apparatus wiping out background signal by the optical fiber, then enters spectrometer through optical fiber.
Installed and the supporting software of spectrometer in computer end, you can by the sample detecting of software to fluorescence signal.
As Fig. 2 be insert optical fiber structural representation figure, insert optical fiber end by two micron optical fibers 13 of core diameter 200,
14 is tight and formed together, and its end is concordant.In two optical fiber, wherein one is used for projecting exciting light, another is used for passing
Go out the fluorescence (reception light) produced after exciting.Shown in label 15 it is effectively exciting and range of receiving for two optical fiber in figure, leads to
Crossing change optical fiber core diameter can make this this encloses expansion.Can using conical hose (not shown) by two optical fiber and together,
Glue is scribbled on sebific duct and optical fiber and plays fixation.In manufacturing process, optical fiber and together can also be made using metal sleeve,
And fixation is played at fibre cladding.Fiber stub (joint) is used for connecting two optical fiber in connection figure 1.Optical fiber insert
Enter part for exposed end, it is different according to insertion brain tissue depth, can change fiber lengths and lengthen or contract when optical fiber is made
Short exposed end.
In specific implementation process, in the optical fiber insertion Shu Nao linked groups that will be made.The optical fiber being inserted and fixed, can
To fix optical fiber with denture fixing device cement.Fiber stub (joint) is connected by metal sleeve with two fibre-optical splices in Fig. 1.Should
Optical fiber is disposable insertion, measurement can be repeated several times in different time.When implementing the method on the animals such as macaque, with mouse brain class
Seemingly, simply the exposed end length of optical fiber, or the optical fiber for changing different-diameter are inserted in corresponding adjustment.
The present invention passes through LED light source (laser), optical fiber, the combination of optical filter, there is provided one kind can be simply in live body
The approach of upper detection fluorescence signal, and animal tissue is caused to damage small, repeat measurement.For the quality handle of transgenic protocol
Control, the success rate for improving later experiments has great meaning.
The above is the preferred embodiment of the present invention, it is noted that for those skilled in the art
For, under the premise without departing from the principles of the invention, some improvement and modification can also be made, these are improved and modification should also be regarded
It is protection scope of the present invention.
Claims (8)
1. a kind of detection means of vivo fluorescence signal, it is characterised in that：Including exciting light sources and detection means two parts, its
In, insert the optical fiber of animal brain by two optical fiber and it is tight together, end is concordant；In two optical fiber of insertion tissue one
Root optical fiber is used for transmitting exciting light, its other end connecting laser；Another optical fiber is used for transmitting fluorescence signal, and its other end connects
Detection means is reconnected after connecing filtering apparatus.
2. the detection means of vivo fluorescence signal according to claim 1, it is characterised in that：The insertion animal brain
Optical fiber connector is tight by two micron optical fibers of core diameter 200 and is formed together, and its end is concordant.
3. the detection means of vivo fluorescence signal according to claim 2, it is characterised in that：The insertion animal brain
In two optical fiber, wherein one is used for projecting exciting light, another be used for spreading out of excite after the fluorescence that produces.
4. according to Claims 2 or 3 vivo fluorescence signal detection means, it is characterised in that：Two optical fiber passes through
Conical hose or metal sleeve are tight and together.
5. the detection means of vivo fluorescence signal according to claim 4, it is characterised in that：The insertion portion of the optical fiber is
Exposed end, it is different according to insertion brain tissue depth, can change fiber lengths and be lengthened or shortened exposed end when optical fiber is made
End.
6. a kind of detection method of vivo fluorescence signal, it is characterised in that：By two optical fiber and tight insertion animal brain group together
Knit, exciting light is projected into animal brain by wherein one optical fiber, excites the fluorescence of generation；Passed by another optical fiber
Go out, by entering detection means after filtering apparatus wiping out background optical signal, you can detect fluorescence signal.
7. the detection method of vivo fluorescence signal according to claim 6, it is characterised in that：In two optical fiber, one
One end connection LED light source of optical fiber, other end connection insertion optical fiber is simultaneously projected in tissue exciting light；Another the one of optical fiber
End connection insertion optical fiber, other end connection filtering apparatus organizes the green fluorescence for producing to be spread out of by the optical fiber and pass through optical filtering
After device wiping out background signal, then enter spectrometer through optical fiber.
8. the detection method of vivo fluorescence signal according to claim 7, it is characterised in that：Installed and the light in computer end
The supporting software of spectrometer, you can by the sample detecting of software to fluorescence signal.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611272914.9A CN106691386A (en) | 2016-12-31 | 2016-12-31 | Detection device of living body fluorescence signal and method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201611272914.9A CN106691386A (en) | 2016-12-31 | 2016-12-31 | Detection device of living body fluorescence signal and method |
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| CN106691386A true CN106691386A (en) | 2017-05-24 |
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| CN201611272914.9A Pending CN106691386A (en) | 2016-12-31 | 2016-12-31 | Detection device of living body fluorescence signal and method |
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| CN85100424A (en) * | 1985-04-01 | 1985-10-10 | 上海医疗器械研究所 | Natural fluorscence diagnosis instrument used for diagnosising malignant tumour |
| CN101178460A (en) * | 2006-11-06 | 2008-05-14 | 株式会社藤仓 | Multi-core optical fiber |
| CN101612035A (en) * | 2009-07-17 | 2009-12-30 | 清华大学 | Minimally invasive multiple channel in vivo fluorescence signal real-time detection system and method |
| CN101919686A (en) * | 2010-09-21 | 2010-12-22 | 华中科技大学 | Multi-mode imaging system for observing cerebral cortex functions of moving animals |
| CN201859120U (en) * | 2010-09-17 | 2011-06-08 | 福建师范大学 | Human tissue autofluorescence multi-wavelength detector |
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| CN206330887U (en) * | 2016-12-31 | 2017-07-14 | 中国科学院昆明动物研究所 | A kind of detection means of vivo fluorescence signal |
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Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN85100424A (en) * | 1985-04-01 | 1985-10-10 | 上海医疗器械研究所 | Natural fluorscence diagnosis instrument used for diagnosising malignant tumour |
| CN101178460A (en) * | 2006-11-06 | 2008-05-14 | 株式会社藤仓 | Multi-core optical fiber |
| US8475506B1 (en) * | 2007-08-13 | 2013-07-02 | Lockheed Martin Corporation | VCSEL array stimulator apparatus and method for light stimulation of bodily tissues |
| CN101612035A (en) * | 2009-07-17 | 2009-12-30 | 清华大学 | Minimally invasive multiple channel in vivo fluorescence signal real-time detection system and method |
| CN201859120U (en) * | 2010-09-17 | 2011-06-08 | 福建师范大学 | Human tissue autofluorescence multi-wavelength detector |
| CN101919686A (en) * | 2010-09-21 | 2010-12-22 | 华中科技大学 | Multi-mode imaging system for observing cerebral cortex functions of moving animals |
| CN202168817U (en) * | 2011-08-01 | 2012-03-21 | 上海亿长实业有限公司 | Medical 3D (three-dimensional) endoscope system |
| CN104755007A (en) * | 2012-10-25 | 2015-07-01 | 奥林巴斯株式会社 | Insertion system, insertion support device, insertion support method and program |
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