CN106073701A - A kind of tissue infrared non-intuitive microscopic imaging device and method - Google Patents
A kind of tissue infrared non-intuitive microscopic imaging device and method Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
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Abstract
The invention discloses a kind of tissue infrared non-intuitive microscopic imaging device.This device includes computer, motor driver, infrared light supply controller, infrared CMOS camera, micromachine, wave plate, infrared annular LED light source and analyzer.Method is: human tissue specimens to be measured is positioned over the object stage of stereomicroscope, open infrared annular LED light source the sample on object stage is illuminated, simultaneously by computer drives motor driver and infrared CMOS camera, control micromachine rotation and figure adopted by infrared CMOS camera, wherein micromachine control wave plate periodically rotates, often rotate fixed angle infrared CMOS camera and once adopted figure by analyzer, multiple repairing weld obtains several figures with different polarization information, the figure with different polarization information that CMOS camera is gathered inputs computer and processes, thus obtain the non-intuitive image of human tissue specimens to be measured.The present invention has the advantage that image taking speed is fast, low cost, resolution are high.
Description
Technical field
The invention belongs to optics medical imaging technology field, particularly a kind of tissue infrared non-intuitive micro-imaging
Device and method.
Background introduction
For modern medicine iconography, the most conventional has CT (Computed Tomography), PET (Positron
Emission tomography), B ultrasonic and MRI (Magnetic Resonance Imaging) etc..But above-mentioned side
Method has their defect, and what CT utilized is high frequency high-energy ray, and tissue can cause irreversible destruction；PET sweeps
Retouch then along with substantial amounts of radiation dose, even carcinogenic risk；B ultrasonic is the ultrasound wave utilized, and its resolution is the lowest, is easily generated leakage
Examine, and anemia of pregnant woman looks into excessively B ultrasonic and may easily cause fetal anomaly；The diagnosis rate of MRI is the highest, uncomfortable for anemia of pregnant woman and critical patient
With, the time of inspection is the most long.
In order to solve problem above, people start to explore this lossless tissue detection method of optical imagery.90 years
In generation, starts people and has developed a series of advanced optical image-taking method: such as time resolved transmission projects tomography ultrashort laser
View finding method that pulse combines with time gate, two-photon fluorescence imaging, photon density wave method, space low-frequency light cofocus scanning
Transmission imaging method and optical coherence tomography (OCT) etc..But due to the complexity of organizational structure, optical image-taking method is the most very
Immature.Comprehensive Assessment in terms of investigation depth, resolution and practicality etc., current OCT technology is the most rising one
Kind, but OCT has preferable result of detection only for the relatively simple eyes of organizational structure, for complex organization's imaging at present
There is the problems such as speed is low compared with the resolution of slow, cost height, imaging.
Summary of the invention
It is an object of the invention to provide the tissue that a kind of image taking speed is fast, low cost, resolution are high infrared non-straight
See microscopic imaging device and method.
The technical solution realizing the object of the invention is: a kind of tissue infrared non-intuitive microscopic imaging device, bag
Including computer, motor driver, infrared light supply controller and stereomicroscope module, wherein stereomicroscope module includes infrared
CMOS camera, micromachine, wave plate, infrared annular LED light source and analyzer, infrared annular LED light source is arranged at stereoscopic micro-
Between object stage and the object lens of mirror, arranging a wave plate between object lens and the infrared CMOS camera of stereomicroscope, this wave plate leads to
Cross minitype motor driving to rotate, between wave plate and infrared CMOS camera, analyzer is set；Computer passes through motor driver
Accessing micromachine, infrared light supply controller is connected with infrared annular LED light source, and infrared CMOS camera is connected with computer；
Human tissue specimens to be measured is positioned over the object stage of stereomicroscope, opens infrared annular LED light source to loading
Sample on platform is illuminated, and simultaneously by computer drives motor driver and infrared CMOS camera, controls micromachine and rotates
Adopting figure with infrared CMOS camera, wherein micromachine control wave plate periodically rotates, and often rotates fixed angle infrared
Figure once adopted by CMOS camera by analyzer, and multiple repairing weld obtains several figures with different polarization information, CMOS camera
The figure with different polarization information gathered inputs computer and processes, thus obtains the non-straight of human tissue specimens to be measured
See image.
Preferably, described infrared LED annular light source uses LED array illumination, and light emission direction and light source axle center angle are 30
～45 degree, wave band is near infrared wavelength region.
Preferably, described infrared LED annular light source 7 is 75 apart from the distance of the sample stage upper surface of stereomicroscope
～85mm.
A kind of tissue micro-one-tenth of infrared non-intuitive based on described tissue infrared non-intuitive microscopic imaging device
Image space method, step is as follows:
Step 1, the ethanol utilizing volumetric concentration to be 70% is applied to the surface of human tissue specimens, then sample is placed in body
On the object stage of stereomicroscope；
Step 2, opens infrared light supply controller and lights infrared annular LED light source, utilizes stereoptics microscope and infrared
CMOS camera obtains a width light intensity picture, as not processed original image and input computer；
Step 3, controls wave plate by micromachine and periodically rotates, and by infrared CMOS collected by camera figure
Picture, it is thus achieved that several plot of light intensity also input computer；
Step 4, computer determines phase contrast and the azimuth of plot of light intensity according to the plot of light intensity of input；
Step 5, utilizes step 4 gained phase contrast, azimuthal value to form gray level image respectively, every in described gray level image
The gray value of point represents phase contrast or azimuthal size, different colours carries out assignment and forms false color image, be adjusted to picture
Contrast obtain the phase contrast non-intuitive image of human tissue specimens to be measured, azimuth non-intuitive image；
Step 6, according to phase contrast, azimuth, determines Stokes parameter by Mueller matrix and carries out Stokes parameter
Non-intuitive imaging.
Further, according to phase contrast, azimuth described in step 6, determine that Stokes parameter is gone forward side by side by Mueller matrix
Row Stokes parameter non-intuitive imaging, the formula used is:
In formula, IdpBeing the average intensity information of pixel place every width figure, δ is phase contrast,It is azimuth, S0、S1、S2、
S3It is four parameters of Stokes.
Compared with prior art, its remarkable advantage is the present invention: (1) image taking speed is fast: use optical imaging method, becomes
The time of picture is almost zero, and main time is used in the image obtained under various polarization states, but time-consuming also within 5 minutes；(2) knot
Structure is simple and visual field is bigger, it is not necessary to carry out point-to-point scanning, low cost；(3) use non-intuitive light wave parametric imaging, utilize
Polarization parameter imaging, by calculating multiple image, reduces weight to point spread function, thus has got around diffraction limit, it is thus achieved that high
The false color image of resolution.
Accompanying drawing explanation
The structural representation of Tu1Shi the present inventor soma infrared non-intuitive microscopic imaging device.
Fig. 2 is the simple microscope direct imaging design sketch of step epidermis sample in embodiment of the present invention.
Fig. 3 is the non-intuitive micro-imaging design sketch of step epidermis sample in embodiment of the present invention.
Detailed description of the invention
Below in conjunction with the accompanying drawings and the present invention is described in further detail by specific embodiment.
The present invention utilizes non-intuitive light wave Parameter Map to carry out human tissue structure analysis, and wherein non-intuitive light wave Parameter Map is
Refer to utilize the phase contrast of imaging, phase angle, Stokes parameter to carry out imaging.Tissue infrared non-intuitive microscopic imaging device
Advantage be the tissue of anisotropic is had ultra high sensitivity, thus strengthen optical microphotograph by measuring matching
The resolution of system.This device is mainly transformed, so greatly reducing the design of device according to the microscope that a Daepori is logical
Difficulty.
Specifically, the present inventor soma infrared non-intuitive microscopic imaging device is as it is shown in figure 1, include computer 1, electricity
Machine driver 2, infrared light supply controller 3 and stereomicroscope module, wherein stereomicroscope module includes infrared CMOS camera
4, micromachine 5, wave plate 6, infrared annular LED light source 7 and analyzer 8, infrared annular LED light source 7 is arranged at stereomicroscope
Object stage and object lens between, between object lens and the infrared CMOS camera 4 of stereomicroscope, a wave plate 6 is set, this wave plate 6 leads to
Cross micromachine 5 drive to rotate, between wave plate 6 and infrared CMOS camera 4, analyzer 8 is set；Computer 1 is driven by motor
Dynamic device 2 accesses micromachine 5, and infrared light supply controller 3 is connected with infrared annular LED light source 7, infrared CMOS camera 4 and calculating
Machine 1 connects；
Human tissue specimens to be measured is positioned over the object stage of stereomicroscope, opens infrared annular LED light source 7 to loading
Sample on platform is illuminated, and simultaneously by computer 1 drive motor driver 2 and infrared CMOS camera 4, controls micromachine 5
Rotating and figure adopted by infrared CMOS camera 4, wherein micromachine 5 controls wave plate 6 and periodically rotates, and often rotates fixed angle
Infrared CMOS camera 4 once adopts figure by analyzer 8, and multiple repairing weld obtains several figures with different polarization information,
The figure with different polarization information that CMOS camera 4 is gathered inputs computer 1 and processes, thus obtains tissue to be measured
The non-intuitive image of sample.
Described infrared LED annular light source 7 uses LED array illumination, and light emission direction and light source axle center angle are 30～45 degree,
Wave band is near infrared wavelength region.For lighting source, the wavelength visible ray less than 600nm, the absorption of light can be due to blood red egg
In vain, melanin and the impact of other pigment and raise；At ultraviolet band, can rise due to protein, the strong absorption of nucleic acid again
High；Water in infrared portion, tissue absorbs in the highest flight.The most commonly called is 650-for " treatment window " i.e. wavelength
In the range of 950nm, for most of soft tissues, light absorbs relatively low, and scattering is relatively strong, thus has stronger scattering
Light reflects or transmits from tissue becomes the light that can be detected.
Described infrared LED annular light source 7 is 75～85mm apart from the distance of the sample stage upper surface of stereomicroscope.
The ultimate principle of non-intuitive algorithm is: utilizes the polarization information of light to carry out imaging, enters for anisotropic sample
Row imaging, by utilizing the physical model of linear birefrigence, it is thus achieved that the intensity signal under several different polarization states, by light intensity table
Reach formula, by utilizing the method calculating Fourier space coefficient, it is thus achieved that required parameter, thus the parameter required for obtaining
Figure.
The present invention infrared non-intuitive of tissue based on power described tissue infrared non-intuitive microscopic imaging device shows
Micro-formation method, step is as follows:
Step 1, the ethanol utilizing volumetric concentration to be 70% is applied to the surface of human tissue specimens, then sample is placed in body
On the object stage of stereomicroscope.
Step 2, opens infrared light supply controller 3 and lights infrared annular LED light source 7, utilizes stereoptics microscope and red
Outer CMOS camera 4 obtains a width light intensity picture, as not processed original image and input computer 1.
Step 3, controls wave plate 6 by micromachine 5 and periodically rotates, and is adopted by infrared CMOS camera 4
Collection image, it is thus achieved that several plot of light intensity also input computer 1；
Described micromachine 5 controls wave plate 6 and carries out Periodic Rotating, it is possible to obtain the image under out of phase delay,
Enough intensity signals are provided for non-intuitive light wave parametric imaging.
Step 4, computer 1 determines phase contrast and the azimuth of plot of light intensity according to the plot of light intensity of input, specific as follows:
Light intensity function I (ω) is obtained according to Jones matrix,
I (ω)=a1+a2sin(2α)+a3sin(4α)+b1cos(4α)
Wherein a1, a2, a3, b1By the electric field component E of light intensityx,EyRepresent, by the value matching sides of obtaining organizing light intensity function more
Parallactic angleIt is the angle that wave plate rotates with phase contrast δ, α.
Step 5, utilizes step 4 gained phase contrast, azimuthal value to form gray level image respectively, every in described gray level image
The gray value of point represents phase contrast or azimuthal size, different colours carries out assignment and forms false color image, be adjusted to picture
Contrast obtain the phase contrast non-intuitive image of human tissue specimens to be measured, azimuth non-intuitive image.
Step 6, according to phase contrast, azimuth, determines Stokes parameter by Mueller matrix and carries out Stokes parameter
Non-intuitive imaging, the formula used is:
In formula, IdpBeing the average intensity information of pixel place every width figure, δ is phase contrast,It is azimuth (slow axis and x
The angle in direction), S0、S1、S2、S3It is four parameters of Stokes.
The present invention is to utilize infrared polarization parameter to carry out imaging of tissue.First with Polarization Modulation, obtain several single polarizations
Image under state, therefrom carries out parameter extraction, is obtained by each parameter value and carries out the image of imaging.With directly utilize far field
Light intensity carries out imaging and compares, and polarization parameter is the sensitiveest for the change of object structures anisotropic, and what is more important is led to
Cross and utilize root-mean-square matching, filter out the degree of fitting pixel higher than 95% so that PSF narrowed width, thus can break through
The diffraction limit of optical imagery, substantially increases the resolution of imaging.Therefore, the invention provides a kind of new modern medicine shadow
As the method learned, the method, based on non-intuitive Parametric imaging techniques, breaches diffraction limit, it is thus achieved that high-definition picture.
Composite geophysical methods and the light source used, infrared light supply makes to detect certain degree of depth, but infrared imaging
Resolution relatively low, and polarization parameter imaging compensate for the shortcoming of infrared light supply just, improves the resolution of imaging, and should
The advantage of method is to be made without scanning, is greatly saved imaging efficiency and cost.
To sum up, the present invention mainly has three advantages: image taking speed is fast, low cost, the resolution height of imaging.
Embodiment 1
This invention device is discussed in detail below in conjunction with the accompanying drawings and realizes sample is carried out the step of light wave parametric imaging.
(1) combine accompanying drawing and this invention device be discussed in detail:
In conjunction with Fig. 1, the present invention utilizes non-intuitive light wave Parameter Map to carry out the device of human tissue structure analysis, including calculating
Machine 1, motor driver 2, infrared light supply controller 3 and stereomicroscope module, wherein stereomicroscope module includes infrared
CMOS camera 4, micromachine 5, wave plate 6, infrared annular LED light source 7 and analyzer 8, infrared annular LED light source 7 is arranged at body
Between object stage and the object lens of stereomicroscope, between object lens and the infrared CMOS camera 4 of stereomicroscope, a wave plate 6 is set,
This wave plate 6 is driven by micromachine 5 and rotates, and arranges analyzer 8 between wave plate 6 and infrared CMOS camera 4；Computer 1
Accessing micromachine 5 by motor driver 2, infrared light supply controller 3 is connected with infrared annular LED light source 7, infrared CMOS phase
Machine 4 is connected with computer 1.
The epidermis sample of the foot of human body to be measured is positioned on the object stage of Stereo microscope, opens infrared annular LED
Light source 7 is illuminated, simultaneously by computer 1 drive motor driver 2 and infrared CMOS camera 4, by the most integrated good algorithm
Software, controls micromachine 5 and rotates and figure adopted by infrared CMOS camera 4, and wherein micromachine 5 controls wave plate 6 and carries out periodically
Rotate, utilize stereomicroscope module imaging to carry out adopting figure, it is thus achieved that several are with the figure of different polarization information, carry out calculating acquisition
Preferably non-intuitive light wave parametric image.Infrared annular LED light source 7 uses the LED array illumination, annular array outside dimension to be
120mm, light emission direction and light source axle center angle are 45 degree, and wave band is the illumination of near-infrared 940nm black light, and wherein annular is passed through
Infrared light supply controller 3 controls, and infrared light supply controller 3 can realize light intensity consecutive variations, uses short-circuit protection, output voltage
24V, output is 25W, single channel.Infrared annular LED light source 7 apart from the distance of sample stage upper surface be 75～
85mm.Micromachine 5 controls wave plate 6 and carries out Periodic Rotating, it is possible to obtain the image under out of phase delay, for non-straight sightseeing
Wave parameter imaging provides enough intensity signals.Algorithm integration software in computer 1, can be entered obtaining several plot of light intensity
The calculating of row non-intuitive algorithm, final acquisition various parametric images, namely non-intuitive light wave parametric image.
(2) specifically comprising the following steps that of the light wave parameter imaging method to this device is realized
Step 1, utilizes the Ethanol Treatment human tissue specimens of 70%, increases the transmitance of infrared light, sample is placed in body
On the object stage of stereomicroscope.
Step 2, opens infrared light supply controller 3 and lights the infrared annular LED light source 7 of infrared light 940nm, utilize body
Optometry microscope and infrared CMOS camera 4, it is thus achieved that the light intensity picture that a width visual field is bigger, as not processed original graph
Picture；
Step 3, controls wave plate 6 by micromachine 5 and periodically rotates, and is entered by infrared CMOS camera 4
Row takes figure, it is thus achieved that several plot of light intensity；
Step 4, by the image obtained by step 3, is carried out non-intuitive micro-imaging by computer 1 by integrated good software
Algorithm computing, thus obtain the phase contrast of plot of light intensity, the parameter information such as azimuth.
Step 5, utilize the value of each parameter to form a width gray level image, wherein the gray value parameter value of every
Size, it is also possible to different colours carries out assignment, is formed false color image, can also be adjusted to the contrast of picture by algorithm,
Obtain the phase contrast non-intuitive image of higher contrast, azimuth non-intuitive image；
Step 6, according to phase contrast, azimuth, determines Stokes parameter by Mueller matrix and carries out Stokes parameter
Non-intuitive imaging.
The image obtained is processed, it is possible to obtain the image that resolution is higher.Mainly have employed the non-straight of parameter
See algorithm, by the weight reducing of point spread function, thus broken the limit of resolution, thus obtain higher resolution and become
Picture.The ultimate principle of non-intuitive algorithm carries out imaging mainly by the polarization information of light, carries out for anisotropic sample
Imaging, by utilizing the physical model of linear birefrigence, it is thus achieved that the intensity signal under several different polarization states, by light strongly expressed
Formula, can be by utilizing the method calculating Fourier space coefficient, it is thus achieved that the parameter that we are required, thus obtains required ginseng
Number figure.
Except the phase difference that can will calculate, the parameter such as polarization azimuth carries out imaging, it is also possible to utilizes and passes through
The Stokes parameter that Mueller matrix calculus goes out carries out parametric imaging, is shown below:
I in formuladpThe average intensity information of the pixel place every width figure referred to, δ is phase contrast,Azimuth (slow axis and x
The angle in direction).
To sum up, the present invention, by the transformation at common Stereo microscope, adds the core of non-intuitive light wave parametric imaging
Parts, by the plot of light intensity under repetitive measurement different polarization states, Inversion Calculation obtains the resolution ratio more preferable effect of former plot of light intensity
Figure.Simultaneously because non-intuitive algorithm is the most sensitive for scattered light, under infrared " medical treatment window ", it is possible to realize certain depth
Imaging.Relative to traditional optical imaging method, there is the advantage that resolution is high, compared to traditional medical imaging apparatus tool
There is the advantage of low cost.
Claims (5)
1. a tissue infrared non-intuitive microscopic imaging device, it is characterised in that include computer (1), motor driver
(2), infrared light supply controller (3) and stereomicroscope module, wherein stereomicroscope module include infrared CMOS camera (4),
Micromachine (5), wave plate (6), infrared annular LED light source (7) and analyzer (8), infrared annular LED light source (7) is arranged at body
Between object stage and the object lens of stereomicroscope, between object lens and the infrared CMOS camera (4) of stereomicroscope, a wave plate is set
(6), this wave plate (6) is driven by micromachine (5) and rotates, and arranges analyzing between wave plate (6) and infrared CMOS camera (4)
Device (8)；Computer (1) accesses micromachine (5), infrared light supply controller (3) and infrared annular by motor driver (2)
LED light source (7) connects, and infrared CMOS camera (4) is connected with computer (1)；
Human tissue specimens to be measured is positioned over the object stage of stereomicroscope, opens infrared annular LED light source (7) to object stage
On sample be illuminated, simultaneously by computer (1) drive motor driver (2) and infrared CMOS camera (4), control micro electric
Machine (5) rotates and infrared CMOS camera (4) adopts figure, and wherein micromachine (5) control wave plate (6) periodically rotates, often
Rotating fixed angle infrared CMOS camera (4) and once adopt figure by analyzer (8), multiple repairing weld obtains several with difference
The figure of polarization information, the figure with different polarization information that CMOS camera (4) is gathered inputs computer (1) and processes, from
And obtain the non-intuitive image of human tissue specimens to be measured.
Tissue the most according to claim 1 infrared non-intuitive microscopic imaging device, it is characterised in that described infrared
LED annular light source (7) uses LED array illumination, and light emission direction and light source axle center angle are 30～45 degree, and wave band is near-infrared ripple
Long scope.
Tissue the most according to claim 1 infrared non-intuitive microscopic imaging device, it is characterised in that described infrared
The distance of the sample stage upper surface of LED annular light source (7) distance stereomicroscope is 75～85mm.
4. the infrared non-intuitive of tissue based on the infrared non-intuitive microscopic imaging device of tissue described in claim 1
Micro imaging method, it is characterised in that step is as follows:
Step 1, the ethanol utilizing volumetric concentration to be 70% is applied to the surface of human tissue specimens, is then placed in by sample stereoscopic aobvious
On the object stage of micro mirror；
Step 2, opens infrared light supply controller (3) and lights infrared annular LED light source (7), utilizes stereoptics microscope and red
Outer CMOS camera (4) obtains a width light intensity picture, as not processed original image and input computer (1)；
Step 3, controls wave plate (6) by micromachine (5) and periodically rotates, and by infrared CMOS camera (4)
Gather image, it is thus achieved that several plot of light intensity also input computer (1)；
Step 4, computer (1) determines phase contrast and the azimuth of plot of light intensity according to the plot of light intensity of input；
Step 5, utilizes step 4 gained phase contrast, azimuthal value to form gray level image respectively, in described gray level image every
Gray value represents phase contrast or azimuthal size, different colours carries out assignment and forms false color image, be adjusted to the right of picture
The phase contrast non-intuitive image of human tissue specimens to be measured, azimuth non-intuitive image is obtained than degree；
Step 6, according to phase contrast, azimuth, determines Stokes parameter by Mueller matrix and to carry out Stokes parameter non-straight
See imaging.
Tissue the most according to claim 4 infrared non-intuitive micro imaging method, it is characterised in that described in step 6
According to phase contrast, azimuth, determine Stokes parameter by Mueller matrix and carry out Stokes parameter non-intuitive imaging, institute
The formula used is:
In formula, IdpBeing the average intensity information of pixel place every width figure, δ is phase contrast,It is azimuth, S0、S1、S2、S3It is
Four parameters of Stokes.
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| CN (1) | CN106073701A (en) |
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| CN107102430A (en) * | 2017-06-01 | 2017-08-29 | 暨南大学 | Arc light source scanning lighting device and method for Fourier's lamination micro-imaging |
| CN107290279A (en) * | 2017-06-30 | 2017-10-24 | 湖北器长光电股份有限公司 | A kind of gene molecule fluorescence non-intuitive microscopic imaging device and method |
| CN110672530A (en) * | 2019-09-16 | 2020-01-10 | 南京理工大学 | Hyperspectrum-combined non-visual biological tissue imaging detection device and method |
| CN111579075A (en) * | 2020-05-25 | 2020-08-25 | 中国人民解放军国防科技大学 | Fast detection method for light wave polarization state based on Fourier analysis |
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| CN101441170A (en) * | 2008-12-22 | 2009-05-27 | 浙江大学 | Image polarization spectrum microscope |
| CN103477205A (en) * | 2011-01-31 | 2013-12-25 | 穆斯泰克计算服务有限公司 | Optical polarimetric imaging |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107102430A (en) * | 2017-06-01 | 2017-08-29 | 暨南大学 | Arc light source scanning lighting device and method for Fourier's lamination micro-imaging |
| CN107290279A (en) * | 2017-06-30 | 2017-10-24 | 湖北器长光电股份有限公司 | A kind of gene molecule fluorescence non-intuitive microscopic imaging device and method |
| CN110672530A (en) * | 2019-09-16 | 2020-01-10 | 南京理工大学 | Hyperspectrum-combined non-visual biological tissue imaging detection device and method |
| CN111579075A (en) * | 2020-05-25 | 2020-08-25 | 中国人民解放军国防科技大学 | Fast detection method for light wave polarization state based on Fourier analysis |
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