CN107228847A - The micro-fluidic radiation injury monitoring device and method of collection capture culture detection integration - Google Patents
The micro-fluidic radiation injury monitoring device and method of collection capture culture detection integration Download PDFInfo
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- CN107228847A CN107228847A CN201710353411.2A CN201710353411A CN107228847A CN 107228847 A CN107228847 A CN 107228847A CN 201710353411 A CN201710353411 A CN 201710353411A CN 107228847 A CN107228847 A CN 107228847A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
Abstract
The invention discloses a kind of micro-fluidic radiation injury monitoring device and method for collecting capture culture detection integration, described device includes micro-fluidic chip component, excitation source component, light field enhancing component, micro-imaging component, microcontroller, electronic moving assembly, vavuum pump and host computer processing assembly.The present invention is under space radiation environment, Caenorhabditis elegans is cultivated in micro-fluidic chip and the life state of Caenorhabditis elegans is monitored in real time, its degree damaged by space radiation is assessed by Caenorhabditis elegans motion feature and internal feature, space radiation degree of injury of the biologist according to suffered by Caenorhabditis elegans carries out space radiation biology research, and how solution defends spacefarer by the health problem of space radiation.The present invention is after sample inlet adds Caenorhabditis elegans, and subsequent step is all to carry out intelligent completion by controller control, and the present invention has the advantages that small volume, lightweight, cost are low, amount of samples is few, be easy to carry.
Description
Technical field
It is particularly a kind of to collect the micro-fluidic of capture culture detection integration the present invention relates to space radiation damage monitoring technology
Radiation injury monitoring device and method
Background technology
Space environment is with the compound extreme environment such as complicated radiation and microgravity, due to consideration that following Chinese Space
Standing needs by long-term in-orbit spacefarer's performance of work task, and following moonfall again and explores the Mars, and whether spacefarer
Can bear the radiation from space environment, how early warning, how to defend, be always a restriction manned astro-engineering on space flight
The problem of member's health protection.Therefore, the problem of space radiation damage monitoring has become primary in space radiation biology research.
Because real space flight test chance is considerably less, cycle length, restrictive factor it is also many, therefore selection is suitable
Biologic material is even more important, and Caenorhabditis elegans is then a kind of space radiation damage monitoring most classical model organism material.
And space radiation damage monitoring equipment is mainly biomicroscope.
Biomicroscope is advanced biological study technology, and it can the intuitive and accurate shape for observing Caenorhabditis elegans
State structure, while the fluorescent characteristics that Caenorhabditis elegans is sent can also be observed.But, biomicroscope huge structure, structure
Complicated, expensive, function is single, can not field monitoring.
The content of the invention
In order to solve above mentioned problem present in space radiation damage monitoring, the present invention will design a kind of small volume, structure
Simply, the micro-fluidic radiation injury prison of collection capture culture detection integration cheap, multiple functional, available for field monitoring
Survey device and method.
To achieve these goals, technical scheme is as follows：
The micro-fluidic radiation injury monitoring device of collection capture culture detection integration, including micro-fluidic chip component, excite
Light source assembly, light field enhancing component, micro-imaging component, microcontroller, electronic moving assembly, vavuum pump and host computer treatment group
Part；
The microcontroller by data wire respectively with host computer processing assembly, excitation source component, light field strengthen component,
Electronic moving assembly and vavuum pump connection；The host computer processing assembly is connected by data wire with micro-imaging component；It is described
Vavuum pump is connected by pipeline with micro-fluidic chip component.
The micro-imaging component includes fluorescence microscopy object lens, spectroscope, green color filter, collector lens group and shooting
Head, the collector lens group includes planoconvex spotlight and biconvex lens；The fluorescence microscopy object lens by light path successively with spectroscope,
Green color filter, collector lens group and camera connection；The camera is connected by bus with host computer processing assembly；
The micro-fluidic chip component is made up of six identical micro-fluidic chips, and the micro-fluidic chip is by substrate and admittedly
The coating composition on substrate is scheduled on, the substrate is sheet glass, and the coating is dimethyl silicone polymer coating, the coating table
Face is provided with sample inlet, culture tank, nutrient solution entrance, side suction inlet, gate and waste liquid outlet；Sample inlet is used to add beautiful hidden
Rhabditida；Culture tank is connected by passage I and passage II with sample inlet, for cultivating Caenorhabditis elegans；Nutrient solution entrance
It is connected by passage V and passage VI with culture tank, for adding nutrient solution；Side suction inlet is connected by passage VII with culture tank, is used
Caenorhabditis elegans is fixed in producing negative pressure；Gate is used to make culture tank with extraneous air completely cut off；Waste liquid outlet passes through passage III
It is connected with passage IV with culture tank, for excluding waste liquid；II, the passage allow single Caenorhabditis elegans by and limit
Caenorhabditis elegans escapes from culture tank；The passage IV and passage VI escape from culture tank for limiting Caenorhabditis elegans；
The excitation source component is used to excite Caenorhabditis elegans to make it produce fluorescence,
The light field enhancing component is used to strengthen the light intensity in Caenorhabditis elegans visual field, improves Caenorhabditis elegans light field
Contrast when video and bright field image acquisition；
The micro-imaging component is used for light field video, bright field image and the fluoroscopic image for obtaining Caenorhabditis elegans；
The microcontroller be used for control excitation source component, light field enhancing component, micro-imaging component, electronic mobile group
Part and vavuum pump；
The electronic moving assembly is used to drive micro-imaging component to move left and right；
The vavuum pump is used to make the side suction inlet of micro-fluidic chip to produce negative pressure；
The host computer processing assembly is used for light field video, bright field image and the fluorescence for storing and handling Caenorhabditis elegans
Image；The motion feature of Caenorhabditis elegans is obtained by light field video and bright field image, the motion feature includes beautiful hidden
Rhabditida motion frequency and motion amplitude；The internal feature of Caenorhabditis elegans, the internal feature are obtained by fluoroscopic image
Including Caenorhabditis elegans luminescence sites number and total fluorescence intensity.
The micro-fluidic radiation injury monitoring method of collection capture culture detection integration, comprises the following steps：
A, the sample inlet that Caenorhabditis elegans is added to micro-fluidic chip, make beautiful hidden bar line by syringe control
Worm enters in culture tank, incubation from sample inlet by passage I, passage II regularly replaces nutrient solution.
B, by microprocessor control micro-imaging component successively in each micro-fluidic chip in micro-fluidic chip component
Caenorhabditis elegans carry out data acquisition；Data acquisition flow includes successively：Light field video acquisition, bright field image collection, it is glimmering
Light image is gathered.
C, host computer processing assembly are analyzed the Caenorhabditis elegans data obtained in step B, assess beautiful with this
Hidden rhabditida radiation injury degree, specific appraisal procedure is as follows：
C1, the radiation injury degree by motion frequency assessment Caenorhabditis elegans；Motion frequency is smaller to show beautiful hidden
Rhabditida raying degree of injury is more serious.
C2, the radiation injury degree by motion amplitude assessment Caenorhabditis elegans；Motion amplitude is smaller to show beautiful hidden
Rhabditida raying degree of injury is more serious.
C3, the radiation injury degree by luminescence sites number assessment Caenorhabditis elegans, luminescence sites number are more, table
The degree of bright Caenorhabditis elegans raying damage is more serious；
C4, the radiation injury degree by total fluorescence intensity assessment Caenorhabditis elegans, fluorescence intensity shows more by force beautiful
Hidden rhabditida raying degree of injury is more serious.
C5, four index comprehensives assessments to more than, finally draw Caenorhabditis elegans raying degree of injury.
The operation principle of the present invention is as follows：
Under space radiation environment, Caenorhabditis elegans is cultivated in micro-fluidic chip and Caenorhabditis elegans is monitored in real time
Life state, its degree damaged by space radiation is assessed by Caenorhabditis elegans motion feature and internal feature, transport
Dynamic feature refers to Caenorhabditis elegans motion frequency and motion amplitude；Internal feature refers to Caenorhabditis elegans luminescence sites number and total
Fluorescence intensity.Finally, the space radiation degree of injury that biologist can be according to suffered by Caenorhabditis elegans carries out space radiation life
Thing is studied, and how solution is vigilant, and how to defend spacefarer by the health problem of space radiation.
Compared with prior art, the invention has the advantages that：
1st, the present invention is after sample inlet adds Caenorhabditis elegans, and subsequent step is all to carry out intelligence by controller control
Energyization is completed, therefore, simple to operate.
2nd, due to the microfluidic platform of the invention for using micro-fluidic chip component as space radiation damage monitoring, and related inspection
Measurement equipment can also use the structure type of small volume, therefore, and the present invention has that small volume, lightweight, cost be low, amount of samples
Less, the advantages of being easy to carry.
3rd, the present invention is realized surveys integration to Caenorhabditis elegans capture, culture and monitoring, is the space spoke of complete set
Penetrate damage monitoring device and method.
Brief description of the drawings
The shared width of accompanying drawing 3 of the present invention, wherein：
Fig. 1 is monitoring device schematic diagram of the invention.
Fig. 2 is micro-imaging assembly principle figure of the invention.
Fig. 3 is microfluidic chip structure schematic diagram of the invention.
In figure：1st, host computer processing assembly, 2, microcontroller, 3, excitation source component, 4, electronic moving assembly, 5, micro-
Image-forming assembly, 6, micro-fluidic chip component, 7, vavuum pump, 8, light field enhancing component, 9, bus, 10, camera, 11, optically focused it is saturating
Microscope group, 12, green color filter, 13, spectroscope, 14, fluorescence microscopy object lens, 15, sample inlet, 16, culture tank, 17, nutrient solution
Entrance, 18, side suction inlet, 19, gate, 20, waste liquid outlet, 21, passage I, 22, passage II, 23, passage III, 24, passage IV, 25,
Passage V, 26, passage VI, 27, passage VII, 28, syringe.
Embodiment
The present invention is further described through below in conjunction with the accompanying drawings.As Figure 1-3, collection capture culture detection integration
Micro-fluidic radiation injury monitoring device, including micro-fluidic chip component 6, excitation source component 3, light field enhancing component 8, micro-
Image-forming assembly 5, microcontroller 2, electronic moving assembly 4, vavuum pump 7 and host computer processing assembly 1；
The microcontroller 2 by data wire respectively with host computer processing assembly 1, excitation source component 3, light field enhancing group
Part 8, electronic moving assembly 4 and vavuum pump 7 are connected；The host computer processing assembly 1 is connected by data wire with micro-imaging component 5
Connect；The vavuum pump 7 is connected by pipeline with micro-fluidic chip component 6；
The micro-imaging component 5 includes fluorescence microscopy object lens 14, spectroscope 13, green glow optical filter 12, collector lens group
11 and camera 10, the collector lens group 11 includes planoconvex spotlight and biconvex lens；The fluorescence microscopy object lens 14 pass through light
Road is connected with spectroscope 13, green color filter 12, collector lens group 11 and camera 10 successively；The camera 10 passes through bus
9 are connected with host computer processing assembly 1；
The micro-fluidic chip component 6 is made up of six identical micro-fluidic chips, the micro-fluidic chip by substrate and
The coating composition on substrate is fixed on, the substrate is sheet glass, and the coating is dimethyl silicone polymer coating, the coating
Surface is provided with sample inlet 15, culture tank 16, nutrient solution entrance 17, side suction inlet 18, gate 19 and sample export 20；Sample inlet
15 are used to add Caenorhabditis elegans；Culture tank 16 is connected by passage I 21 and passage II 22 with sample inlet 15, for cultivating
Caenorhabditis elegans；Nutrient solution entrance 17 is connected by passage V 25 and passage VI 26 with culture tank 16, for adding nutrient solution；
Side suction inlet 18 is connected by passage VII 27 with culture tank 16, and Caenorhabditis elegans is fixed for producing negative pressure；Gate 19 is used to make
Culture tank 16 completely cuts off with extraneous air；Waste liquid outlet 20 is connected by passage III 23 and passage IV 24 with culture tank 16, for arranging
Except waste liquid；II 22, the passage allow single Caenorhabditis elegans by and limit Caenorhabditis elegans and escape from culture tank 16；
The passage IV 24 and passage VI 26 escape from culture tank 16 for limiting Caenorhabditis elegans；
The excitation source component 3 is used to excite Caenorhabditis elegans to make it produce fluorescence,
The light field enhancing component 8 is used to strengthen the light intensity in Caenorhabditis elegans visual field, improves Caenorhabditis elegans bright
Contrast when field video and bright field image acquisition；
The micro-imaging component 5 is used for light field video, bright field image and the fluoroscopic image for obtaining Caenorhabditis elegans；
The microcontroller 2 is used to control excitation source component 3, light field enhancing component 8, micro-imaging component 5, electric moving
Dynamic component 4 and vavuum pump 7；
The electronic moving assembly 4 is used to drive micro-imaging component 5 to move left and right；
The vavuum pump 7 is used to make the side suction inlet 18 of micro-fluidic chip to produce negative pressure；
The host computer processing assembly 1 is used to storing and handle the light field video of Caenorhabditis elegans, bright field image and glimmering
Light image；The motion feature of Caenorhabditis elegans is obtained by light field video and bright field image, the motion feature includes beautiful
Hidden rhabditida motion frequency and motion amplitude；The internal feature of Caenorhabditis elegans is obtained by fluoroscopic image, it is described internal special
Levy including Caenorhabditis elegans luminescence sites number and total fluorescence intensity.
The micro-fluidic radiation injury monitoring method of collection capture culture detection integration, comprises the following steps：
A, the sample inlet 15 that Caenorhabditis elegans is added to micro-fluidic chip, are made beautiful hidden by the control of syringe 28
Rhabditida enters in culture tank 16, incubation from sample inlet 15 by passage I 21, passage II 22 regularly replaces nutrient solution；
B, by microcontroller 2 control micro-imaging component 5 successively to each micro-fluidic chip in micro-fluidic chip component 6
In Caenorhabditis elegans carry out data acquisition；Data acquisition flow includes successively：Light field video acquisition, bright field image collection,
Fluoroscopic image is gathered；
C, host computer processing assembly 1 are analyzed the Caenorhabditis elegans data obtained in step B, assess beautiful with this
Hidden rhabditida radiation injury degree, specific appraisal procedure is as follows：
C1, the radiation injury degree by motion frequency assessment Caenorhabditis elegans；Motion frequency is smaller to show beautiful hidden
Rhabditida raying degree of injury is more serious；
C2, the radiation injury degree by motion amplitude assessment Caenorhabditis elegans；Motion amplitude is smaller to show beautiful hidden
Rhabditida raying degree of injury is more serious；
C3, the radiation injury degree by luminescence sites number assessment Caenorhabditis elegans, luminescence sites number are more, table
The degree of bright Caenorhabditis elegans raying damage is more serious；
C4, the radiation injury degree by total fluorescence intensity assessment Caenorhabditis elegans, fluorescence intensity shows more by force beautiful
Hidden rhabditida raying degree of injury is more serious；
C5, four index comprehensives assessments to more than, finally draw Caenorhabditis elegans raying degree of injury.
The groundwork flow of the present invention is as follows：
When gathering light field video and bright field image, Caenorhabditis elegans freely grows in culture tank 16, exciting light
Source component 3 is closed, and light field enhancing component 8 is opened；When gathering fluoroscopic image, excitation source component 3 is opened, light field enhancing component
8 close, while vavuum pump 7 is opened, produce negative pressure, the cell wall for Caenorhabditis elegans to be fixed on to culture tank 16；
When gathering light field video and bright field image, workflow is：Light field enhancing component 8 irradiate from below → it is beautiful hidden
Rhabditida amplifies through microcobjective → and collector lens group 11 focuses on → carrying out data acquisition by camera 10 → and is transferred to host computer
Processing assembly 1 is stored and handled；
When gathering fluoroscopic image, workflow is：Excitation source component 3 irradiated → reflected through spectroscope 13 from side →
Excite Caenorhabditis elegans produce fluorescence data that fluorescence → Caenorhabditis elegans produces amplify through fluorescence microscopy object lens 14 → it is green
Colo(u)r filter 12 filters out veiling glare data → collector lens group 11 and focused on, and → carrying out data acquisition by camera 10 → is transferred to upper
Machine processing assembly 1 is stored and handled；
The present invention is not limited to the present embodiment, any equivalent concepts in the technical scope of present disclosure or changes
Become, be classified as protection scope of the present invention.
Claims (2)
1. the micro-fluidic radiation injury monitoring device of collection capture culture detection integration, it is characterised in that：Including micro-fluidic chip
Component (6), excitation source component (3), light field enhancing component (8), micro-imaging component (5), microcontroller (2), electronic movement
Component (4), vavuum pump (7) and host computer processing assembly (1)；
The microcontroller (2) is strengthened with host computer processing assembly (1), excitation source component (3), light field respectively by data wire
Component (8), electronic moving assembly (4) and vavuum pump (7) connection；The host computer processing assembly (1) by data wire with it is micro-
Image-forming assembly (5) is connected；The vavuum pump (7) is connected by pipeline with micro-fluidic chip component (6)；
It is saturating that the micro-imaging component (5) includes fluorescence microscopy object lens (14), spectroscope (13), green color filter (12), optically focused
Microscope group (11) and camera (10), the collector lens group (11) include planoconvex spotlight and biconvex lens；The fluorescence microscopy thing
Mirror (14) is connected with spectroscope (13), green color filter (12), collector lens group (11) and camera (10) successively by light path；
The camera (10) is connected by bus (9) with host computer processing assembly (1)；
The micro-fluidic chip component (6) is made up of six identical micro-fluidic chips, and the micro-fluidic chip is by substrate and admittedly
The coating composition on substrate is scheduled on, the substrate is sheet glass, and the coating is dimethyl silicone polymer coating, the coating table
Face is provided with sample inlet (15), culture tank (16), nutrient solution entrance (17), side suction inlet (18), gate (19) and waste liquid outlet
(20)；Sample inlet (15) is used to add Caenorhabditis elegans；Culture tank (16) passes through passage I (21) and passage II (22) and sample
Product entrance (15) is connected, for cultivating Caenorhabditis elegans；Nutrient solution entrance (17) passes through passage V (25) and passage VI (26)
It is connected with culture tank (16), for adding nutrient solution；Side suction inlet (18) is connected by passage VII (27) with culture tank (16), is used for
Produce negative pressure and fix Caenorhabditis elegans；Gate (19) is used to make culture tank (16) with extraneous air completely cut off；Waste liquid outlet (20)
It is connected by passage III (23) and passage IV (24) with culture tank (16), for excluding waste liquid；The passage II (22) only allows
Single Caenorhabditis elegans by and limit Caenorhabditis elegans and escape from culture tank (16)；The passage IV (24) and passage VI
(26) culture tank (16) is escaped from for limiting Caenorhabditis elegans；
The excitation source component (3) is used to excite Caenorhabditis elegans to make it produce fluorescence,
The light field enhancing component (8) is used to strengthen the light intensity in Caenorhabditis elegans visual field, improves Caenorhabditis elegans light field
Contrast when video and bright field image acquisition；
The micro-imaging component (5) is used for light field video, bright field image and the fluoroscopic image for obtaining Caenorhabditis elegans；
The microcontroller (2) is used to control excitation source component (3), light field enhancing component (8), micro-imaging component (5), electricity
Dynamic moving assembly (4) and vavuum pump (7)；
The electronic moving assembly (4) is used to drive micro-imaging component (5) to move left and right；
The vavuum pump (7) is used to make the side suction inlet (18) of micro-fluidic chip to produce negative pressure；
The host computer processing assembly (1) is used for light field video, bright field image and the fluorescence for storing and handling Caenorhabditis elegans
Image；The motion feature of Caenorhabditis elegans is obtained by light field video and bright field image, the motion feature includes beautiful hidden
Rhabditida motion frequency and motion amplitude；The internal feature of Caenorhabditis elegans, the internal feature are obtained by fluoroscopic image
Including Caenorhabditis elegans luminescence sites number and total fluorescence intensity.
2. the micro-fluidic radiation injury monitoring method of collection capture culture detection integration, it is characterised in that：Comprise the following steps：
A, the sample inlet (15) that Caenorhabditis elegans is added to micro-fluidic chip, are made beautiful hidden by syringe (28) control
Rhabditida enters in culture tank (16), incubation from sample inlet (15) by passage I (21), passage II (22) to be regularly replaced
Nutrient solution；
B, by microcontroller (2) control micro-imaging component (5) successively to each micro-fluidic core in micro-fluidic chip component (6)
Caenorhabditis elegans in piece carries out data acquisition；Data acquisition flow includes successively：Light field video acquisition, bright field image are adopted
Collection, fluoroscopic image collection；
C, host computer processing assembly (1) are analyzed the Caenorhabditis elegans data obtained in step B, assess beautiful hidden with this
Rhabditida radiation injury degree, specific appraisal procedure is as follows：
C1, the radiation injury degree by motion frequency assessment Caenorhabditis elegans；Motion frequency is smaller to show beautiful hidden bar line
Worm raying degree of injury is more serious；
C2, the radiation injury degree by motion amplitude assessment Caenorhabditis elegans；Motion amplitude is smaller to show beautiful hidden bar line
Worm raying degree of injury is more serious；
C3, the radiation injury degree by luminescence sites number assessment Caenorhabditis elegans, luminescence sites number are more, show show
The degree of beautiful hidden rhabditida raying damage is more serious；
C4, the radiation injury degree by total fluorescence intensity assessment Caenorhabditis elegans, fluorescence intensity show more by force beautiful hidden bar
Nematode raying degree of injury is more serious；
C5, four index comprehensives assessments to more than, finally draw Caenorhabditis elegans raying degree of injury.
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| CN109187463A (en) * | 2018-08-31 | 2019-01-11 | 哈尔滨工业大学 | Micro-fluidic type radiation injury biological dose analysis and detection device and its determination method based on photobacteria |
| CN112763468A (en) * | 2020-12-28 | 2021-05-07 | 南通大学 | Micro-fluidic chip nematode adsorption and fixation device and use method thereof |
| CN113340921A (en) * | 2021-02-01 | 2021-09-03 | 中国人民解放军军事科学院军事医学研究院 | Quantitative evaluation method and evaluation system for electromagnetic radiation damage and electromagnetic radiation biological effect |
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| CN109187463A (en) * | 2018-08-31 | 2019-01-11 | 哈尔滨工业大学 | Micro-fluidic type radiation injury biological dose analysis and detection device and its determination method based on photobacteria |
| CN112763468A (en) * | 2020-12-28 | 2021-05-07 | 南通大学 | Micro-fluidic chip nematode adsorption and fixation device and use method thereof |
| CN113340921A (en) * | 2021-02-01 | 2021-09-03 | 中国人民解放军军事科学院军事医学研究院 | Quantitative evaluation method and evaluation system for electromagnetic radiation damage and electromagnetic radiation biological effect |
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