CN106052866A - Biophoton spectrum detection system and method - Google Patents
Biophoton spectrum detection system and method Download PDFInfo
- Publication number
- CN106052866A CN106052866A CN201610310424.7A CN201610310424A CN106052866A CN 106052866 A CN106052866 A CN 106052866A CN 201610310424 A CN201610310424 A CN 201610310424A CN 106052866 A CN106052866 A CN 106052866A
- Authority
- CN
- China
- Prior art keywords
- bio
- photon
- detection system
- sample
- detection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 52
- 238000001228 spectrum Methods 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 9
- 238000003384 imaging method Methods 0.000 claims abstract description 32
- 238000002474 experimental method Methods 0.000 claims abstract description 6
- 230000003287 optical effect Effects 0.000 claims abstract description 6
- 238000004020 luminiscence type Methods 0.000 claims abstract description 5
- 230000003595 spectral effect Effects 0.000 claims description 31
- 238000007405 data analysis Methods 0.000 claims description 7
- 239000002826 coolant Substances 0.000 claims description 6
- 238000009826 distribution Methods 0.000 claims description 5
- 238000007639 printing Methods 0.000 claims description 5
- 101100117236 Drosophila melanogaster speck gene Proteins 0.000 claims description 3
- 238000004364 calculation method Methods 0.000 claims description 3
- 239000003086 colorant Substances 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 238000007689 inspection Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 230000000007 visual effect Effects 0.000 claims description 3
- 239000006185 dispersion Substances 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 3
- 238000004458 analytical method Methods 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract 1
- 210000004556 brain Anatomy 0.000 description 14
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 229930195712 glutamate Natural products 0.000 description 5
- 230000006698 induction Effects 0.000 description 5
- 239000000463 material Substances 0.000 description 4
- 230000010412 perfusion Effects 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000005855 radiation Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 101700004678 SLIT3 Proteins 0.000 description 2
- 102100027339 Slit homolog 3 protein Human genes 0.000 description 2
- 238000012271 agricultural production Methods 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- AAOVKJBEBIDNHE-UHFFFAOYSA-N diazepam Chemical compound N=1CC(=O)N(C)C2=CC=C(Cl)C=C2C=1C1=CC=CC=C1 AAOVKJBEBIDNHE-UHFFFAOYSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 238000005086 pumping Methods 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- 241000931526 Acer campestre Species 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102100025490 Slit homolog 1 protein Human genes 0.000 description 1
- 101710123186 Slit homolog 1 protein Proteins 0.000 description 1
- 238000004378 air conditioning Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- JJWKPURADFRFRB-UHFFFAOYSA-N carbonyl sulfide Chemical compound O=C=S JJWKPURADFRFRB-UHFFFAOYSA-N 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000012809 cooling fluid Substances 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 230000007102 metabolic function Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008054 signal transmission Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/2803—Investigating the spectrum using photoelectric array detector
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/02—Details
- G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
- G01J3/0243—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows having a through-hole enabling the optical element to fulfil an additional optical function, e.g. a mirror or grating having a throughhole for a light collecting or light injecting optical fiber
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/2823—Imaging spectrometer
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/255—Details, e.g. use of specially adapted sources, lighting or optical systems
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
-
- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/27—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using photo-electric detection ; circuits for computing concentration
- G01N21/274—Calibration, base line adjustment, drift correction
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J2003/006—Fundamentals or review articles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/2803—Investigating the spectrum using photoelectric array detector
- G01J2003/282—Modified CCD or like
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/06—Illumination; Optics
- G01N2201/068—Optics, miscellaneous
Abstract
The present invention provides a biophoton spectrum detection system and method. The biophoton spectrum detection system is characterized by adding a sub-photon system composed of slits and a grating on the basis of an ultraweak biophoton imaging system. The grating is an optical element composed of a lot of parallel slits of equal widths and equal intervals, and enables the light to generate chromatic dispersion by utilizing the multi-slit diffraction principle. According to the multi-slit diffraction principle, the ultraweak biophotons emitted out by a to-be-detected sample form a line light source after passing the slits, the line light source irradiates the grating and then is diffracted by the multiple slits to form the bright fringes which are arranged at equal intervals, and the distance is relevant with the wavelength of the biophotons. An imaging subsystem is used for the image recording and analysis. The experiment detection proves that the biophoton spectrum detection system of the present invention is high in sensitivity, can detect the spectrums of the ultraweak luminescence of the biologies, at the same time, has the characteristics of being simple and convenient in operation, good in expansibility and abundant in function, and can be widely used in the fields, such as the life science research, the medical clinical examination, the agriculture production, the food safety, the environment protection, etc.
Description
Technical field
The present invention relates to one and utilize superweak biophoton emission signal, to biological (individual, tissue, cell and molecule etc.)
Carry out bio-photon spectral detection, it is achieved bio-photon light spectrum image-forming, belong to biological and biomedical technology imaging field.This skill
Art is equally applicable to other field needs long-time imaging to carry out the field of low light level spectrum analysis.
Background technology:
Bio-photon is the abbreviation of Bio-photon emission, in itself, does not has difference with the light of usual indication, and spectral coverage is
200-800nm, is prevalent in the various life entity such as microorganism, plant and animal.Bio-photon activity can be the most anti-
Reflect organic physiological status.
From the viewpoint of molecular physics, bio-photon can be understood as biomolecule from upper state to lower state transition
During the energy that discharges.Biosystem has metabolic function, and it is a typical open systems, with the external world
Environment also exists eternal material, energy, the exchange of information.The extraneous biosystem that constantly pumping dissipates, is allowed to be in one
Plant away from thermally equilibrated state.High energy living matter molecule is unstable, it is necessary to lower state transition, radiate birth in the process
Object light, and the molecule returning to lower state transits to upper state under external influence.Extraneous pumping occurs together mutually with photon radiation
Raw, reach a dynamic equilibrium.
Because the motion conditions of the atomic interior electronics of different living matters is different, so they bio-photons of giving off
Also different.These bio-photons after dispersion system (such as prism, grating) light splitting, the monochromatic light opened by dispersion by wavelength (or
Frequency) size and be arranged in order the pattern of formation, be referred to as spectrum.Luminous and the light absorbing situation of research different material, has
Important theoretical and practical significance.Therefore, the spectral characteristic of bio-photon contributes to us and furthers investigate, and can extensively apply
In fields such as life science, clinical medicine inspection, agricultural production, food safety and environmental conservation.
In the twenties in 20th century, Soviet Union biologist Gurwitsch uses bioprobe bio-photon to be detected first
Radiation.But this bioprobe be not the most given sufficient attention, and because bio-photon intensity is the most weak, it is subject to
The limitation of technology at that time, does not has sensitiveer bio-photon detection means occur.Until nineteen fifty-five, the Talents employing such as Colli is worked as
Time the photomultiplier tube (PMT) just invented at first the bio-photon of plant is directly detected.Along with electronics communicates with information
Developing rapidly of technology, photon detector is also being continuously updated the replacement, occurs in that photoconductive detector, optical transistor subsequently
With charge coupled cell (CCD).Wherein, electron multiplication-charge coupled cell (EM-CCD) has quantum efficiency height, signal to noise ratio
High, volume is little and the advantage of life-span length, is widely used in bio-photon detection field.
Construct a set of bio-photon imaging system (UBIS) with EM-CCD for core, can detect that bio-photon is not
With time and strength information spatially.But, in order to the character of bio-photon is carried out deeper into research, in addition it is also necessary to a kind of
New technology and method detect the spectral characteristic of bio-photon.
Summary of the invention:
The present invention, on the basis of the bio-photon imaging system (UBIS) with EM-CCD as core, adds by slit and light
Grid composition light splitting subsystem, the bio-photon spectral detection system after improvement can detect delicately bio-photon time and
Spatial information, the intensity of bio-photon and spectral characteristic.
Bio-photon spectral detection system, is made up of light splitting subsystem and imaging subsystems two parts, imaging subsystems by
Shade, high printing opacity camera lens, EMCCD image device and controller thereof, computer and coolant recirculation pump composition, shade connects
In between the camera lens and sample stage of EMCCD image device, for shielding the impact on imaging of the luminescence generated by light in camera bellows, high transmissive mirror
Head is for imaging, and coolant recirculation pump is used for ensureing that EMCCD image device works at low ambient temperatures, it is characterised in that: described
Light splitting subsystem is made up of slit and grating, and slit is placed on above sample, makes the bio-photon of sample become through after slit
The bright fringe equidistantly arranged, imaging subsystems is formed through too much seam diffraction after one line source, and this line source irradiation grating
Record bio-photon image analytical calculation go out bio-photon spectrum.Each striped distance is relevant to the wavelength of bio-photon.Light splitting
Subsystem makes bio-photon form the most different bright fringes according to different wavelength.
Described grating is the optical element being made up of some wide equally spaced parallel slits.
The detection method of the bio-photon spectral detection system described in utilization, it is characterised in that comprise the steps: that (1) is examined
Survey and prepare and imaging subsystems preheating；(2) laser calibration；(3) LED light source reliability demonstration；(4) sample detection；(5) at image
Reason；(6) data analysis.
Step (2) laser calibration is particularly as follows: use the laser of at least 3 kinds of different wave lengths to enter bio-photon detecting system
Rower is fixed, and, the LASER Light Source of every kind of wavelength needs and to carry out respectively under bio-photon intensity under normal light intensities respectively
Demarcate.
LED light source reliability demonstration particularly as follows: complete laser to bio-photon detecting system demarcate after, keep experiment
In the case of condition is consistent with laser calibration, use bio-photon detecting system detection blueness, green, yellow and red four
The normal intensity of kind of LED modules with different colors light source and the spectrum of bio-photon intensity, that verifies this bio-photon detecting system can
By property.
Sample detection and image procossing are particularly as follows: be placed on ready detection sample in the middle of visual field, and ensure
Slit location level, after detection obtains the bio-photon spectrum picture of sample, is first to be converted to by bio-photon spectrum picture
TIF form, after matlab algorithm removes speck, extracts the gray value on each pixel of bio-photon spectrum picture,
And be saved in Microsoft Excel, then the gray value of every a line is added and obtains gray value summation distribution curve.
Data analysis is particularly as follows: pass through computer Andor Solis (Solis according to bio-photon spectrum picture
Version 4.27.30001.0, Belfast., Northern Ireland) software determines zero order fringe and the limit of one-level striped
The pixel position at edge place, the data then in conjunction with gray value summation distribution curve confirm, obtain zero order fringe and one-level
Spacing △ Lmin, △ Lc, △ Lmax of striped.Wherein, two edges centrally through striped of striped determine, such as Fig. 2 c institute
Show.△ Lmin is the one-level striped nearest edge distance to zero order fringe center, and △ Lc is that one-level fringe center is to zero order fringe
The distance at center, △ Lmax is the one-level striped distal-most edge distance to zero order fringe center.Distance is represented by pixel difference.Finally
Linear equation according to laser calibration matching calculates the bio-photon spectral region of this sample.
During the bio-photon spectral detection of the present invention, sample, light splitting subsystem and part imaging subsystems device need
Be positioned in the camera bellows being made up of outer layer plate internal layer stereotype, to completely cut off ambient and cosmic ray, make imaging process from
The impact of ambient light.
The technical characterstic of the present invention:
The invention provides a kind of bio-photon spectral detection system, have the following characteristics that
(1) the bio-photon light that bio-photon imaging system (UBIS) with EM-CCD as core and light splitting subsystem are constituted
Spectrum detecting system, it is possible to achieve the spectral characteristic detection to the radiation of organism Ultra-weak Luminescence；
(2) highly sensitive: with X-10375 type EM-CCD of ANDOR company of Britain for photon imaging device, and to coordinate camera bellows
Deng relevant element, good signal to noise ratio can be reached, and detected sample directly contacts with light splitting subsystem, decrease life
Loss during thing photon detection, thus sensitiveer bio-photon signal detected；
(3) easy and simple to handle: according to stitching diffraction principle more, simple with the light splitting subsystem principle that slit and grating form, behaviour
Facilitate；
(4) favorable expandability: in this system, each device is separate, and it is convenient to change, and selects various, has well extension
Property and upgrading potential；
(5) feature richness: can adjust different experiment parameters according to testing conditions, can be widely applied to life sciences and grinds
Study carefully, clinical medicine inspection, agricultural production, bio-photon spectral detection in the field such as food safety and environmental conservation.
Accompanying drawing explanation
Fig. 1 is the bio-photon spectral detection system structural representation of the present invention；
Fig. 2 a is the laser calibration spectrogram of the present invention；
Fig. 2 b is the LED light source detection spectrogram of the present invention；
Fig. 2 c is the spectral image analysis schematic diagram of the present invention；
Fig. 2 d is the matched curve figure of the normal intensity laser calibration of the present invention；
Fig. 2 e is the matched curve figure of the bio-photon strength laser demarcation of the present invention；
Fig. 2 f is the LED light source spectral detection comparison diagram of the present invention；
Fig. 3 a is the 532nm laser light spectrogram of the present invention；
Fig. 3 b is the gray value summation scatter chart of the 532nm laser light spectrogram of the present invention；
Fig. 4 a is the bio-photon spectrogram of the mouse brain slices of the present invention；
Fig. 4 b is the Spectroscopic analysis results figure of the mouse brain slices of the present invention.
Detailed description of the invention
In order to be more fully understood that the present invention, it is further elucidated with the particular content of the present invention, operation stream below in conjunction with embodiment
Journey and actual performance.But present disclosure is not limited solely to the following examples, those skilled in the art can be to this
Bright being used for various modifications, upgrade and expansion etc., these equivalent form of values are equally within the scope of claims listed by the application limit.
As it is shown in figure 1, the bio-photon spectral detection system of the present invention, by light splitting subsystem and imaging subsystems 1 two
Being grouped into, imaging subsystems is followed by shade, high printing opacity camera lens, EMCCD image device and controller, computer and coolant
Ring pump forms, and shade is connected between camera lens and the sample stage of EMCCD image device, for shielding the luminescence generated by light pair in camera bellows
The impact of imaging, high printing opacity camera lens is used for imaging, and coolant recirculation pump is used for ensureing EMCCD image device work at low ambient temperatures
Make, it is characterised in that: described light splitting subsystem is made up of slit 3 and grating 2, and slit 3 is placed on above sample, makes the life of sample
Formed through too much seam diffraction after becoming a line source, and this line source irradiation grating 2 after thing transmission of photons slit and equidistantly arrange
The bright fringe of row, imaging subsystems 1 records bio-photon image and analytical calculation goes out bio-photon spectrum.Described grating be by
The optical element that some wide equally spaced parallel slits are constituted.
Correlational study is pointed out, and there may be bio-photon activity and have the merit of signal transmission in central nervous system
Energy.Based on this, the present inventor utilizes bio-photon spectral detection system of the present invention, with the X-10375 type of ANDOR company
EM-CCD is photon imaging device, and the GT13-12 type transmission grating 2 of Thorlabs company and slit 1 form light splitting subsystem, just
Step detects and have studied the bio-photon spectrum of mouse brain slices glutamate induction.
Embodiment: the bio-photon spectrum of mouse brain slices glutamate induction
One, detection material: kunming mice of growing up, from Hubei Province's laboratory animal Public service center.
Two, detecting step:
(1) detection prepares: before starting detection, first has to open air-conditioning, makes laboratory room temperature maintain about 25 DEG C,
Then use ultra-pure water to clean brain sheet perfusion system, and open EM-CCD and low-temperature cooling fluid circulating pump, make EM-CCD be in-95
DEG C working environment in preheat.
(2) laser calibration: use the laser of 3 kinds of different wave lengths of 405nm, 532nm and 650nm to come bio-photon and detect
System is demarcated.And, LASER Light Source needs and to demarcate under bio-photon intensity under normal intensity respectively.
As depicted in figure 2, upper figure is the laser spectrum under normal intensity to calibration result, and figure below is for being attenuated to bio-photon
Laser spectrum under intensity.It can be seen that spectrum picture includes a zero order fringe (the brightest middle striped) and two
Individual one-level striped (striped that both sides are the brightest).Along with the increase of optical maser wavelength, two one-level stripeds are relative to zero order fringe
Present significantly away from trend, and under varying strength, the spectrogram result of laser calibration is consistent.
(3) LED light source reliability demonstration: complete laser to bio-photon detecting system demarcate after, keep other experiment
In the case of consistent, use the normal of this system detection blueness, green, yellow and red four kinds of LED modules with different colors light sources
Intensity and the spectrum of bio-photon intensity, as shown in figure 2f, to verify the reliability of this bio-photon detecting system.
The spectral results of LED light source is as shown in accompanying drawing 2b, and upper figure is the LED light source spectrum under normal intensity, and figure below is for subtracting
Weak LED light source spectrum under bio-photon intensity.
(4) preparation of material is detected: its full brain, after sacrificed by decapitation, is taken out rapidly, is placed on 0～4 DEG C by mice
In ACSF, then cut the sagittal brain sections of 1～2mm thickness with vibratome.
(5) perfusion system is opened: the mouse brain being immersed in 0～4 DEG C of ACSF cut into slices and place at room temperature 1h, then,
Brain sections is transferred in perfusion system.Perfusate is contained in a vial, and is connected with the mixing of 95%O2+5%CO2
Gas.Perfusate is filled in camera bellows equipped with in the perfusion groove of brain sheet by peristaltic pump, and flow velocity is 5ml/min.
(6) start to detect imaging: be first the EM-CCD location photograph that shoots brain sheet under normal lighting conditions, be then shut off
Camera bellows, after lucifuge 30 minutes is to avoid the interference of ambient light, adds glutamic acid and starts realtime imaging, and time of exposure is 60s, holds
Continuous 120min.
(7) bio-photon spectral detection: add light splitting subsystem and adjust position, mainly keeps slit level.Then
Continuing realtime imaging, time of exposure is 1500s, glutamic acid carries out in imaging process eluting and agent-feeding treatment again, shoots
The biology of phase (50min), eluting phase (25min) and dosing phase (25min) again is maintained during the biophoton emission of glutamate induction
Photon spectrum image, and carry out overlap-add procedure.
(8) image procossing: after obtaining the spectrum picture of brain sheet biophoton emission of glutamate induction, with accompanying drawing 3a's
Image procossing is carried out as a example by 532nm laser spectrum image.First it is that this spectrum picture is converted to TIF form, through matlab
After algorithm removes speck, extract the gray value on each pixel of this image, and be saved in Microsoft Excel, so
After the gray value of every a line be added obtain gray value summation distribution curve.The scatterplot drawn is as shown in fig. 3b.
(9) data analysis: according to images above and data, we can determine whether zero order fringe as referring to fig. 2 c and
The lower edges of level striped.First computer Andor Solis (Solis version 4.27.30001.0, Belfast.,
Northern Ireland) software determines the pixel at place, image border by vision, divides then in conjunction with gray value summation
The data of cloth curve confirm, obtain spacing △ Lmin, △ Lc, △ Lmax of zero order fringe and one-level striped.The edge of striped
It is generally selected that pixel that neighbor grayscale value summation difference is maximum.
By above analysis method, we just can calculate △ Lmin, △ Lc, △ Lmax, and the wavelength of laser is
Know, this makes it possible to the equation of linear regression obtaining fringe spacing △ L with wavelength X, and carry out correlation test.Such as Fig. 2 d, figure
Shown in 2e, be respectively under normal intensity and bio-photon intensity under laser calibration matched curve.
Zero order fringe and the edge of one-level striped is determined by Visual Observations Observations and data analysis.Finally calculate between striped
Away from the wavelength with bio-photon.
Three, testing result:
In the case of ensureing that other experiment conditions are consistent, calibrated bio-photon spectral detection system is used to detect
The bio-photon spectrum of the mouse brain slices of 50mM glutamate induction, result is as shown in accompanying drawing 4a, it can be seen that significantly from figure
Article one, zero order fringe and two one-level stripeds.According to previously described data analysing method, by data analysis, measure striped
The bio-photon spectral region of these brain sheets just can be calculated, as depicted in fig. 4b after spacing △ Lmin, △ Lc, △ Lmax.
The above, be only to illustrate that the present invention's is embodied as case, and be not limited to the present invention can
Practical range, such as those skilled in the art completed under without departing from the spirit indicated by the present invention and principle all
Equivalence improves, upgrade and expansion etc. changes or modifies, and must be covered by the scope of the claims in the present invention.
Claims (7)
1. bio-photon spectral detection system, is made up of light splitting subsystem and imaging subsystems two parts, and imaging subsystems is by hiding
Light shield, high printing opacity camera lens, EMCCD image device and controller thereof, computer and coolant recirculation pump composition, shade is connected to
Between the camera lens of EMCCD image device and sample stage, for shielding the impact on imaging of the luminescence generated by light in camera bellows, high printing opacity camera lens
For imaging, coolant recirculation pump is used for ensureing that EMCCD image device works at low ambient temperatures, it is characterised in that: described point
Photonic system is made up of slit and grating, and slit is placed on above sample, makes the bio-photon of sample become one through after slit
Forming, through too much seam diffraction, the bright fringe equidistantly arranged after individual line source, and this line source irradiation grating, imaging subsystems is remembered
Record bio-photon image analytical calculation go out bio-photon spectrum.
Bio-photon spectral detection system the most according to claim 1, it is characterised in that: described grating is by some etc.
The optical element that wide equally spaced parallel slits is constituted.
3. utilize the detection method of bio-photon spectral detection system described in claim 1 or 2, it is characterised in that include as follows
Step: (1) detection prepares and imaging subsystems preheating；(2) laser calibration；(3) LED light source reliability demonstration；(4) sample inspection
Survey；(5) image procossing；(6) data analysis.
The detection method of bio-photon spectral detection system the most according to claim 3, it is characterised in that: step (2) swashs
Light is demarcated particularly as follows: use the laser of at least 3 kinds of different wave lengths to demarcate bio-photon detecting system, and, every kind
The LASER Light Source of wavelength needs and to demarcate respectively under bio-photon intensity under normal light intensities respectively.
The detection method of bio-photon spectral detection system the most according to claim 3, it is characterised in that: LED light source can
By property checking particularly as follows: complete after bio-photon detecting system demarcated by laser, keeping experiment condition and laser calibration bar
In the case of part is consistent, use bio-photon detecting system detection blueness, green, yellow and red four kinds of LED modules with different colors light
The normal intensity in source and the spectrum of bio-photon intensity, verify the reliability of this bio-photon detecting system.
The detection method of bio-photon spectral detection system the most according to claim 3, it is characterised in that sample detection and
Image procossing is particularly as follows: be placed on ready detection sample in the middle of visual field, and ensures slit location level, and detection obtains
After obtaining the bio-photon spectrum picture of sample, first it is that bio-photon spectrum picture is converted to TIF form, calculates through matlab
After method removes speck, extract the gray value on each pixel of bio-photon spectrum picture, and be saved in Microsoft
In Excel, then the gray value of every a line is added and obtains gray value summation distribution curve.
The detection method of bio-photon spectral detection system the most according to claim 6, it is characterised in that: data analysis has
Body is: determine zero order fringe and the limit of one-level striped according to bio-photon spectrum picture by computer Andor Solis software
The pixel position at edge place, the data then in conjunction with gray value summation distribution curve confirm, obtain zero order fringe and one-level
Spacing △ Lmin, △ Lc, △ Lmax of striped.Wherein, two edges centrally through striped of striped determine, △ Lmin is
The nearest edge of one-level striped is to the distance at zero order fringe center, and △ Lc is the one-level fringe center distance to zero order fringe center,
△ Lmax is the one-level striped distal-most edge distance to zero order fringe center, and distance is represented by pixel difference, finally according to laser mark
The linear equation determining matching calculates the bio-photon spectral region of this sample.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610310424.7A CN106052866A (en) | 2016-05-11 | 2016-05-11 | Biophoton spectrum detection system and method |
| PCT/CN2017/080342 WO2017193759A1 (en) | 2016-05-11 | 2017-04-13 | Bio-photon spectrum detection system and detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610310424.7A CN106052866A (en) | 2016-05-11 | 2016-05-11 | Biophoton spectrum detection system and method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106052866A true CN106052866A (en) | 2016-10-26 |
Family
ID=57177664
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610310424.7A Pending CN106052866A (en) | 2016-05-11 | 2016-05-11 | Biophoton spectrum detection system and method |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN106052866A (en) |
| WO (1) | WO2017193759A1 (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017193759A1 (en) * | 2016-05-11 | 2017-11-16 | 中南民族大学 | Bio-photon spectrum detection system and detection method |
| CN108078542A (en) * | 2017-11-10 | 2018-05-29 | 上海康奥实业发展有限公司 | High-precision wide visual field imaging spectrometer apparatus |
| CN108226014A (en) * | 2017-12-25 | 2018-06-29 | 中南民族大学 | A kind of red blood cell bio-photon detection method |
| CN108414476A (en) * | 2018-03-14 | 2018-08-17 | 中南民族大学 | A method of freshness of meat is detected by superweak bio-photon imaging system |
| CN108693183A (en) * | 2018-03-27 | 2018-10-23 | 中南民族大学 | The method for detecting cosmetic composition sensitization by superweak bio-photon imaging system |
| CN109593718A (en) * | 2018-10-16 | 2019-04-09 | 中子康(武汉)医药科技有限公司 | The application of biomolecule quantum energy common technology is reduced using bio-molecular interaction |
| CN111551253A (en) * | 2020-05-25 | 2020-08-18 | 重庆冠雁科技有限公司 | Calibration method for ensuring consistency of spectrum modules |
| CN117686008A (en) * | 2024-02-01 | 2024-03-12 | 广东海洋大学 | Fiber Bragg grating signal demodulation system and method based on image processing |
| CN117686008B (en) * | 2024-02-01 | 2024-04-26 | 广东海洋大学 | Fiber Bragg grating signal demodulation system and method based on image processing |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110850069A (en) * | 2019-10-24 | 2020-02-28 | 中南民族大学 | Detection kit for biological photon radiation of gastrointestinal tract tissue and application |
| CN112733604B (en) * | 2020-12-08 | 2022-11-22 | 黑龙江省爱格机械产品检测有限公司 | Cooling liquid impurity detection platform and method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5329353A (en) * | 1991-02-07 | 1994-07-12 | Research Development Corp. Of Japan | High sensitive multi-wavelength spectral analyzer |
| CN102706837A (en) * | 2012-03-29 | 2012-10-03 | 中南民族大学 | Method and system for identifying swill-cooked dirty oil by photoinduced ultraweak luminescence imaging technology |
| CN103536277A (en) * | 2013-10-30 | 2014-01-29 | 中南民族大学 | Automated ultra-weak light imaging system and operation method |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0558216A1 (en) * | 1992-02-24 | 1993-09-01 | Thermo Jarrell Ash Corporation | Spectrometry |
| CN1289918A (en) * | 2000-10-08 | 2001-04-04 | 暨南大学 | Method and device for multi-channel spectrophotometric detection of rapid microscope |
| CN102334978A (en) * | 2011-07-08 | 2012-02-01 | 山东省医药生物技术研究中心 | Device and method for imaging superweak photons radiated by human body by utilizing charge coupled device (CCD) camera |
| CN103115680B (en) * | 2013-01-24 | 2014-11-12 | 中国科学院空间科学与应用研究中心 | Super sensitive spectrograph and spectrum detection method |
| CN104515754B (en) * | 2014-11-14 | 2018-03-06 | 中国科学院上海光学精密机械研究所 | Laser plasma spectrometry device |
| CN205049240U (en) * | 2015-09-30 | 2016-02-24 | 中国农业科学院农业资源与农业区划研究所 | Hyperspectral imager system |
| CN106052866A (en) * | 2016-05-11 | 2016-10-26 | 中南民族大学 | Biophoton spectrum detection system and method |
-
2016
- 2016-05-11 CN CN201610310424.7A patent/CN106052866A/en active Pending
-
2017
- 2017-04-13 WO PCT/CN2017/080342 patent/WO2017193759A1/en active Application Filing
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5329353A (en) * | 1991-02-07 | 1994-07-12 | Research Development Corp. Of Japan | High sensitive multi-wavelength spectral analyzer |
| CN102706837A (en) * | 2012-03-29 | 2012-10-03 | 中南民族大学 | Method and system for identifying swill-cooked dirty oil by photoinduced ultraweak luminescence imaging technology |
| CN103536277A (en) * | 2013-10-30 | 2014-01-29 | 中南民族大学 | Automated ultra-weak light imaging system and operation method |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017193759A1 (en) * | 2016-05-11 | 2017-11-16 | 中南民族大学 | Bio-photon spectrum detection system and detection method |
| CN108078542A (en) * | 2017-11-10 | 2018-05-29 | 上海康奥实业发展有限公司 | High-precision wide visual field imaging spectrometer apparatus |
| CN108226014A (en) * | 2017-12-25 | 2018-06-29 | 中南民族大学 | A kind of red blood cell bio-photon detection method |
| CN108414476A (en) * | 2018-03-14 | 2018-08-17 | 中南民族大学 | A method of freshness of meat is detected by superweak bio-photon imaging system |
| CN108693183A (en) * | 2018-03-27 | 2018-10-23 | 中南民族大学 | The method for detecting cosmetic composition sensitization by superweak bio-photon imaging system |
| CN109593718A (en) * | 2018-10-16 | 2019-04-09 | 中子康(武汉)医药科技有限公司 | The application of biomolecule quantum energy common technology is reduced using bio-molecular interaction |
| CN109593718B (en) * | 2018-10-16 | 2022-03-29 | 中子康(武汉)医药科技有限公司 | Application of technology for reducing quantum energy common property of biomolecules by utilizing interaction of biomolecules |
| CN111551253A (en) * | 2020-05-25 | 2020-08-18 | 重庆冠雁科技有限公司 | Calibration method for ensuring consistency of spectrum modules |
| CN117686008A (en) * | 2024-02-01 | 2024-03-12 | 广东海洋大学 | Fiber Bragg grating signal demodulation system and method based on image processing |
| CN117686008B (en) * | 2024-02-01 | 2024-04-26 | 广东海洋大学 | Fiber Bragg grating signal demodulation system and method based on image processing |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2017193759A1 (en) | 2017-11-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106052866A (en) | Biophoton spectrum detection system and method | |
| Xu et al. | In-vivo fluorescence imaging with a multivariate curve resolution spectral unmixing technique | |
| Mansfield et al. | Autofluorescence removal, multiplexing, and automated analysis methods for in-vivo fluorescence imaging | |
| Dubach et al. | In vivo imaging of specific drug–target binding at subcellular resolution | |
| JP6235886B2 (en) | Biological tissue image reconstruction method and apparatus, and image display apparatus using the biological tissue image | |
| CN106092994B (en) | A kind of micro-array chip fluorescence detection method of great power LED | |
| US20180042483A1 (en) | Systems and methods for hyperspectral imaging | |
| Qin et al. | New optical molecular imaging systems | |
| Erkkilä et al. | Widefield fluorescence lifetime imaging of protoporphyrin IX for fluorescence‐guided neurosurgery: An ex vivo feasibility study | |
| Lilo et al. | Revising Fourier-transform infrared (FT-IR) and Raman spectroscopy towards brain cancer detection | |
| Radaelli et al. | μMAPPS: a novel phasor approach to second harmonic analysis for in vitro-in vivo investigation of collagen microstructure | |
| Mohsin et al. | Probing the intracellular refractive index and molecular interaction of gold nanoparticles in HeLa cells using single particle spectroscopy | |
| US8254650B2 (en) | System and method for contrast enhancement of time-resolved fluorescence images | |
| Zhang et al. | Tracking the formation and degradation of fatty-acid-accumulated mitochondria using label-free chemical imaging | |
| CN113330298A (en) | System, method and apparatus for correcting transmission deviation of interference filter due to incident angle | |
| Zhang et al. | Hot-band-absorption-induced anti-Stokes fluorescence of aggregation-induced emission dots and the influence on the nonlinear optical effect | |
| Yin et al. | A dynamic system with digital lock-in-photon-counting for pharmacokinetic diffuse fluorescence tomography | |
| JP2022122181A (en) | Correction parameter setting method and data correction method | |
| WO2016203617A1 (en) | Light emission observation method | |
| WO2018230318A1 (en) | Scale, image capturing device, image capturing system, kit, and method for adjusting image capturing device | |
| Schlothauer et al. | Spatially resolved singlet oxygen detection and imaging | |
| Maklygina et al. | Phototherapy of Brain Tumours Using a Fibre Optic Neurosystem | |
| Odion et al. | Direct detection of nanostar probes through a monkey skull using inverse surface-enhanced spatially offset Raman spectroscopy (SESORS) | |
| Talbot et al. | A multispectral FLIM tomograph for in-vivo imaging of skin cancer | |
| Stewart | Fluorophore studies & instrumentation development for tumour margin estimation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161026 |
|
| RJ01 | Rejection of invention patent application after publication |