CN106204486B - A kind of micro- cell imaging device and method of near-infrared fluorescent based on Hadamard transform - Google Patents
A kind of micro- cell imaging device and method of near-infrared fluorescent based on Hadamard transform Download PDFInfo
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Abstract
The present invention relates to a kind of micro- cell imaging apparatus and method of near-infrared fluorescent based on Hadamard transform, including near-infrared fluorescent excitation light path, preposition imaging optical path and Path of Convergent Rays.Near-infrared fluorescent sample is excited to generate near-infrared fluorescent micro-image；Digital Micromirror Device encode along time shaft and carries out single-point imaging by near-infrared single-element detector to near-infrared fluorescent micro-image；The reconstruct of near-infrared fluorescent micro-image is realized using Hadamard transform theory.Advantage of the invention is that replacing expensive near-infrared planar array detector imaging device without increasing mechanical scan arrangement, the complexity and cost of near-infrared fluorescent micro imaging system being greatly reduced.
Description
Technical field
The invention belongs to near-infrared fluorescent micro-imaging fields, and in particular to a kind of near-infrared based on Hadamard transform is glimmering
The micro- cell imaging device and method of light.
Background technique
The space fluorescent intensity distribution that Induced Fluorescence Microscopy is issued by detection fluorescent samples, to realize fluorescence sample
The detection of product internal structural information, it is with a wide range of applications in the fields such as the detection such as biomedicine, chemical analysis.Phase
For fluorescence emission spectrum visible region fluorescent samples, the transmitting fluorescence bands of near-infrared fluorescent sample Δ λ=800~
1400nm.The corresponding sample autofluorescence light intensity of wave band Δ λ is weaker, can interfere to improve imaging letter to avoid background fluorescence
Make an uproar than and detection sensitivity, while near-infrared fluorescent has stronger penetration capacity, small to the damage of sample, therefore near-infrared
Induced Fluorescence Microscopy is one of most promising technology in current Induced Fluorescence Microscopy.
What the photosurface of moment detector mostly used is silicon matrix material, and spectral response range is mostly in UV, visible light light wave
Section, is not particularly suited for near-infrared fluorescent micro-imaging.Therefore, it is necessary to use near-infrared planar array detector to carry out near-infrared fluorescent
Imaging.But near-infrared planar array detector has the following problems at present:
(1) since near-infrared planar array detector development in China's is started late, domestic production technique and technology still prematurity；
(2) near-infrared planar array detector is related to sensitive national defense and military fields, and foreign countries have limit purchase policy；
(3) cost of near-infrared planar array detector is sufficiently expensive, and it is tens of thousands of or even up to a million to reach ten；
(4) there are non-uniformities for near-infrared planar array detector, and there are intrinsic spatial noises, cause distortion dry image
It disturbs.
The above problem strongly limits the further development and application of domestic near-infrared fluorescent microtechnic.
Summary of the invention
The present invention proposes a kind of based on Adama change for the problems of above-mentioned near-infrared fluorescent micro-imaging technique
The micro- single-element detector imaging device of the near-infrared fluorescent changed and method.The present invention is reducing near-infrared fluorescence imaging installation cost
While cost, near-infrared fluorescent micro-imaging signal-to-noise ratio is improved, there is wide application value.
The invention is realized by the following technical scheme:
A kind of micro- cell imaging device of near-infrared fluorescent based on Hadamard transform, including near-infrared fluorescent exciting light
Road, preposition imaging optical path and Path of Convergent Rays, near-infrared fluorescent excitation light path by excitation light source, near-infrared fluorescent microscope and
Near-infrared fluorescent sample is constituted；The preposition imaging optical path includes the first lens group, Digital Micromirror Device；The Path of Convergent Rays packet
Include the second lens group and near-infrared single-element detector.
Further, the micro- cell imaging device of a kind of near-infrared fluorescent based on Hadamard transform, including near-infrared
Fluorescent exciting road, preposition imaging optical path and Path of Convergent Rays, near-infrared fluorescent excitation light path is by excitation light source, near-infrared fluorescent
Microscope and near-infrared fluorescent sample are constituted；The preposition imaging optical path includes the first lens group, Digital Micromirror Device；It is described
Path of Convergent Rays includes the second lens group and near-infrared single-element detector；Wherein the excitation light source uses 660nm LED light source.
Further, the micro- cell imaging device of a kind of near-infrared fluorescent based on Hadamard transform, including near-infrared
Fluorescent exciting road, preposition imaging optical path and Path of Convergent Rays, near-infrared fluorescent excitation light path is by excitation light source, near-infrared fluorescent
Microscope and near-infrared fluorescent sample are constituted；The preposition imaging optical path includes the first lens group, Digital Micromirror Device；It is described
Path of Convergent Rays includes the second lens group and near-infrared single-element detector；Imaging device further includes reflecting mirror and computer, reflecting mirror
Connect the near-infrared fluorescent excitation light path and the preposition imaging optical path, computer connects the Digital Micromirror Device and described
Near-infrared single-element detector.
The application also provides a kind of micro- list of the near-infrared fluorescent based on Hadamard transform for above-mentioned imaging device
First imaging method, it is characterised in that: the following steps are included:
1) excitation light source excites near-infrared fluorescent sample, generates near-infrared fluorescent image F；
2) near-infrared fluorescent image F is focused in Digital Micromirror Device and is imaged through the first lens group:
In above formula, the element in matrix S respectively corresponds the rollover states of the miniature mirror surface in Digital Micromirror Device, value
Respectively 0 or 1, numerical value 0 indicates that miniature mirror surface nearly IR fluorescence is reflected on baffle, and numerical value 1 indicates miniature mirror surface nearly
IR fluorescence is reflected on detector and receives；P indicates the total pixel size of near-infrared fluorescent image, P=p1×p2；p1And p2Respectively
For horizontal, vertical direction pixel total size；
3) according to Hadamard transform theory, every a line of matrix S in step 2) is carried out by near-infrared fluorescent image size
It rearranges, generates two-dimensional modulation image sequence S1,S2,…,SP:
4) Digital Micromirror Device is according to two-dimensional modulation image sequence S1,S2,…,SP, along time shaft control micromirror in face of close
IR fluorescence image is encoded；
5) near-infrared fluorescent image after Digital Micromirror Device coding focuses on the detection of near-infrared unit through the second lens group
On device:
In above formula, f1,f2,…,fPFor the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,yPFor near-infrared
Single-element detector is in different moments collected one point data；
6) computer uses Adama inversion according to near-infrared single-element detector in different moments collected one point data
It changes and near-infrared fluorescent image F is reconstructed:
In above formula, subscript " -1 " is indicated to matrix inversion.
Further, the excitation light source is LED light source, and the near-infrared fluorescent sample is single-walled carbon nanotube label
Sample, the wave band of near-infrared fluorescent image F are 800~1600nm.
Further, in step 2), p1=400, p2=300, then P=120000；In step 3), generates two dimension and adjust
Imaged sequence S1,S2,…,S120000:
Digital Micromirror Device is according to two-dimensional modulation image sequence S in step 4)1,S2,…,S120000, controlled along time shaft micro-
Type mirror surface encodes near-infrared fluorescent image；
In step 5), Digital Micromirror Device coding after near-infrared fluorescent image, focus on near-infrared through the second lens group
On single-element detector:
In above formula, f1,f2,…,f120000For the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,y120000
It is near-infrared single-element detector in different moments collected one point data；
In step 6), computer, in different moments collected one point data, uses Ah according to near-infrared single-element detector
Near-infrared fluorescent image F is reconstructed up to Ma inverse transformation:
Compared with existing near-infrared fluorescent micro-imaging technique, advantage of the invention is embodied in:
(1) Hadamard transform is introduced into near-infrared fluorescent micro-imaging, is detected by matrix transformation method from unit
Near-infrared fluorescence imaging is reconstructed in device.Compared with traditional single-element detector imaging technique, the present invention is without increasing mechanical scanning
Structure can realize the micro- whole audience imaging of near-infrared, significantly reduce the complexity of imaging system structure；
(2) near infrared imaging spatial resolution of the invention depends on the miniature mirror surface number of Digital Micromirror Device, and digital
The cost of micro mirror element is well below near-infrared planar array detector, therefore present invention considerably reduces near-infrared micro imaging systems
Cost, so that the practicability of near-infrared micro-imaging technique is greatly improved.
Detailed description of the invention
To describe the technical solutions in the embodiments of the present invention more clearly, make required in being described below to embodiment
Attached drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the invention, for
For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings other
Attached drawing.
The micro- cell imaging structure drawing of device of near-infrared fluorescent of the Fig. 1 based on Hadamard transform；
Fig. 2 single-walled carbon nanotube marks sample reconstruct to restore image.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with attached drawing to embodiment party of the present invention
Formula is described in further detail.
Embodiment one
Referring to attached drawing 1, a kind of micro- cell imaging device of near-infrared fluorescent based on Hadamard transform, including near-infrared are glimmering
Phot-luminescence road, preposition imaging optical path and Path of Convergent Rays, the near-infrared fluorescent excitation light path are glimmering by excitation light source, near-infrared
Light microscope and near-infrared fluorescent sample are constituted；The preposition imaging optical path includes the first lens group, Digital Micromirror Device；Institute
Stating Path of Convergent Rays includes the second lens group and near-infrared single-element detector.
Embodiment two
The present embodiment is the further improvement made on the basis of example 1, and referring to attached drawing 1, imaging device includes close
IR fluorescence excitation light path, preposition imaging optical path and Path of Convergent Rays, the near-infrared fluorescent excitation light path is by excitation light source, close
IR fluorescence microscope and near-infrared fluorescent sample are constituted；The preposition imaging optical path includes the first lens group, digital micro-mirror
Device；The Path of Convergent Rays includes the second lens group and near-infrared single-element detector；Wherein, the excitation light source uses 660nm
LED light source.
Embodiment three
The present embodiment is the further improvement made on the basis of example 1, and referring to attached drawing 1, imaging device includes close
IR fluorescence excitation light path, preposition imaging optical path and Path of Convergent Rays, the near-infrared fluorescent excitation light path is by excitation light source, close
IR fluorescence microscope and near-infrared fluorescent sample are constituted；The preposition imaging optical path includes the first lens group, digital micro-mirror
Device；The Path of Convergent Rays include the second lens group and near-infrared single-element detector, wherein imaging device further include reflecting mirror and
Computer, reflecting mirror connect the near-infrared fluorescent excitation light path and the preposition imaging optical path, and computer connects the number
Micro mirror element and the near-infrared single-element detector.
Example IV
The present embodiment is related to a kind of micro- cell imaging method of the near-infrared fluorescent based on Hadamard transform comprising following
Several steps:
1) excitation light source excites near-infrared fluorescent sample, generates near-infrared fluorescent image F；
2) near-infrared fluorescent image F is focused in Digital Micromirror Device and is imaged through the first lens group:
In above formula, the element in matrix S respectively corresponds the rollover states of the miniature mirror surface in Digital Micromirror Device, value
Respectively 0 or 1, numerical value 0 indicates that miniature mirror surface nearly IR fluorescence is reflected on baffle, and numerical value 1 indicates miniature mirror surface nearly
IR fluorescence is reflected on detector and receives；P indicates the total pixel size of near-infrared fluorescent image, P=p1×p2；p1And p2Respectively
For horizontal, vertical direction pixel total size；
3) according to Hadamard transform theory, every a line of matrix S in step 2) is carried out by near-infrared fluorescent image size
It rearranges, generates two-dimensional modulation image sequence S1,S2,…,SP:
4) Digital Micromirror Device is according to two-dimensional modulation image sequence S1,S2,…,SP, along time shaft control micromirror in face of close
IR fluorescence image is encoded；
5) near-infrared fluorescent image after Digital Micromirror Device coding focuses on the detection of near-infrared unit through the second lens group
On device:
In above formula, f1,f2,…,fPFor the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,yPFor near-infrared
Single-element detector is in different moments collected one point data；
6) computer uses Adama inversion according to near-infrared single-element detector in different moments collected one point data
It changes and near-infrared fluorescent image F is reconstructed:
In above formula, subscript " -1 " is indicated to matrix inversion.
Embodiment five
The present embodiment is the further improvement made on the basis of example IV, wherein the excitation light source is LED light
Source, the near-infrared fluorescent sample are that single-walled carbon nanotube marks sample, the wave band of near-infrared fluorescent image F is 800~
1600nm。
A kind of micro- cell imaging method of near-infrared fluorescent based on Hadamard transform comprising following steps:
1) excitation LED light source excitation single-walled carbon nanotube marks sample, and it is glimmering to generate the near-infrared that wave band is 800~1600nm
Light image F；
2) near-infrared fluorescent image F is focused in Digital Micromirror Device and is imaged through the first lens group:
In above formula, the element in matrix S respectively corresponds the rollover states of the miniature mirror surface in Digital Micromirror Device, value
Respectively 0 or 1, numerical value 0 indicates that miniature mirror surface nearly IR fluorescence is reflected on baffle, and numerical value 1 indicates miniature mirror surface nearly
IR fluorescence is reflected on detector and receives；P indicates the total pixel size of near-infrared fluorescent image, P=p1×p2；p1And p2Respectively
For horizontal, vertical direction pixel total size；
3) according to Hadamard transform theory, every a line of matrix S in step 2) is carried out by near-infrared fluorescent image size
It rearranges, generates two-dimensional modulation image sequence S1,S2,…,SP:
4) Digital Micromirror Device is according to two-dimensional modulation image sequence S1,S2,…,SP, along time shaft control micromirror in face of close
IR fluorescence image is encoded；
5) near-infrared fluorescent image after Digital Micromirror Device coding focuses on the detection of near-infrared unit through the second lens group
On device:
In above formula, f1,f2,…,fPFor the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,yPFor near-infrared
Single-element detector is in different moments collected one point data；
6) computer uses Adama inversion according to near-infrared single-element detector in different moments collected one point data
It changes and near-infrared fluorescent image F is reconstructed:
In above formula, subscript " -1 " is indicated to matrix inversion.
The single-walled carbon nanotube label sample reconstructed image obtained using this method is referring to attached drawing 2.
Embodiment six
The present embodiment is the further improvement made on the basis of embodiment five, in this example, p1=400, p2=300, in
It is P=120000, the specific steps of imaging method are as follows:
1) excitation LED light source excitation single-walled carbon nanotube marks sample, and it is glimmering to generate the near-infrared that wave band is 800~1600nm
Light image F；
2) near-infrared fluorescent image is focused in Digital Micromirror Device and is imaged through the first lens group:
In above formula, the element in matrix S respectively corresponds the miniature mirror surface in Digital Micromirror Device, value be respectively 0 or
1；Numerical value 0 indicates that miniature mirror surface nearly IR fluorescence is reflected on baffle, and numerical value 1 indicates that miniature mirror surface nearly IR fluorescence is anti-
It is mapped on detector and receives；P indicates the total pixel size of near-infrared fluorescent image, P=p1×p2；p1And p2It is respectively horizontal, vertical
Direction pixel total size；In this example, p1=400, p2=300, then P=120000；
3) according to Hadamard transform theory, every a line of matrix S in step 2) is carried out by near-infrared fluorescent image size
It rearranges, generates two-dimensional modulation image sequence S1,S2,…,S120000:
4) Digital Micromirror Device is according to two-dimensional modulation image sequence S1,S2,…,S120000, miniature mirror surface is controlled along time shaft
Near-infrared fluorescent image is encoded；
5) near-infrared fluorescent image after Digital Micromirror Device coding focuses on the detection of near-infrared unit through the second lens group
On device:
In above formula, f1,f2,…,f120000For the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,y120000
It is near-infrared single-element detector in different moments collected one point data；
6) computer uses Adama inversion according to near-infrared single-element detector in different moments collected one point data
It changes and near-infrared fluorescent image F is reconstructed:
In above formula, subscript " -1 " is indicated to matrix inversion.Sample weight is marked using the single-walled carbon nanotube that this method obtains
Composition picture is referring to attached drawing 2.
By the various embodiments described above it is found that compared with existing near-infrared fluorescent micro-imaging technique, advantage body of the invention
It is present:
(1) Hadamard transform is introduced into near-infrared fluorescent micro-imaging, is detected by matrix transformation method from unit
Near-infrared fluorescence imaging is reconstructed in device.Compared with traditional single-element detector imaging technique, the present invention is without increasing mechanical scanning
Structure can realize the micro- whole audience imaging of near-infrared, significantly reduce the complexity of imaging system structure；
(2) near infrared imaging spatial resolution of the invention depends on the miniature mirror surface number of Digital Micromirror Device, and digital
The cost of micro mirror element is well below near-infrared planar array detector, therefore present invention considerably reduces near-infrared micro imaging systems
Cost, so that the practicability of near-infrared micro-imaging technique is greatly improved.
Obviously, the above embodiment of the present invention be only to clearly illustrate example of the present invention, and not be pair
The restriction of embodiments of the present invention.For those of ordinary skill in the art, may be used also on the basis of the above description
To make other variations or changes in different ways.There is no necessity and possibility to exhaust all the enbodiments.And these
Belong to connotation changes and variations that derived from of the invention to still fall in protection scope of the present invention.
Claims (6)
1. a kind of micro- cell imaging device of near-infrared fluorescent based on Hadamard transform, including near-infrared fluorescent excitation light path,
Preposition imaging optical path and Path of Convergent Rays, it is characterised in that: near-infrared fluorescent excitation light path is shown by excitation light source, near-infrared fluorescent
Micro mirror and near-infrared fluorescent sample are constituted；The preposition imaging optical path includes the first lens group, Digital Micromirror Device；The meeting
Optically focused road includes the second lens group and near-infrared single-element detector；
The micro- cell imaging method of the near-infrared fluorescent based on Hadamard transform of the imaging device the following steps are included:
1) excitation light source excites near-infrared fluorescent sample, generates near-infrared fluorescent image F；
2) near-infrared fluorescent image F is focused in Digital Micromirror Device and is imaged through the first lens group:
In above formula, the element in matrix S respectively corresponds the rollover states of the miniature mirror surface in Digital Micromirror Device, value difference
It is 0 or 1, numerical value 0 indicates that miniature mirror surface nearly IR fluorescence is reflected on baffle, and numerical value 1 indicates that miniature mirror surface is nearly infrared
It is received on fluorescent reflection to detector；P indicates the total pixel size of near-infrared fluorescent image, P=p1×p2；p1And p2Respectively water
Flat, vertical direction pixel total size；
3) according to Hadamard transform theory, every a line of matrix S in step 2) is carried out again by near-infrared fluorescent image size
Arrangement generates two-dimensional modulation image sequence S1,S2,…,SP；
4) Digital Micromirror Device is according to two-dimensional modulation image sequence S1,S2,…,SP, near-infrared is faced along time shaft control micromirror
Fluorescent image is encoded；
5) near-infrared fluorescent image after Digital Micromirror Device coding, focuses on near-infrared single-element detector through the second lens group
It is upper:
In above formula, f1,f2,…,fPFor the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,yPFor near-infrared unit
Detector is in different moments collected one point data；
6) computer uses Adama inversion swap-in according to near-infrared single-element detector in different moments collected one point data
Row reconstruct near-infrared fluorescent image F:
In above formula, subscript " -1 " is indicated to matrix inversion.
2. the micro- cell imaging device of near-infrared fluorescent as described in claim 1 based on Hadamard transform, it is characterised in that:
The excitation light source uses 660nm LED light source.
3. the micro- cell imaging device of near-infrared fluorescent as described in claim 1 based on Hadamard transform, it is characterised in that:
It further include reflecting mirror and computer, reflecting mirror connects the near-infrared fluorescent excitation light path and the preposition imaging optical path, calculates
Machine connects the Digital Micromirror Device and the near-infrared single-element detector.
4. a kind of near-infrared based on Hadamard transform for imaging device as claimed any one in claims 1 to 3 is glimmering
The micro- cell imaging method of light, it is characterised in that: the following steps are included:
1) excitation light source excites near-infrared fluorescent sample, generates near-infrared fluorescent image F；
2) near-infrared fluorescent image F is focused in Digital Micromirror Device and is imaged through the first lens group:
In above formula, the element in matrix S respectively corresponds the rollover states of the miniature mirror surface in Digital Micromirror Device, value difference
It is 0 or 1, numerical value 0 indicates that miniature mirror surface nearly IR fluorescence is reflected on baffle, and numerical value 1 indicates that miniature mirror surface is nearly infrared
It is received on fluorescent reflection to detector；P indicates the total pixel size of near-infrared fluorescent image, P=p1×p2；p1And p2Respectively water
Flat, vertical direction pixel total size；
3) according to Hadamard transform theory, every a line of matrix S in step 2) is carried out again by near-infrared fluorescent image size
Arrangement generates two-dimensional modulation image sequence S1,S2,…,SP；
4) Digital Micromirror Device is according to two-dimensional modulation image sequence S1,S2,…,SP, near-infrared is faced along time shaft control micromirror
Fluorescent image is encoded；
5) near-infrared fluorescent image after Digital Micromirror Device coding, focuses on near-infrared single-element detector through the second lens group
It is upper:
In above formula, f1,f2,…,fPFor the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,yPFor near-infrared unit
Detector is in different moments collected one point data；
6) computer uses Adama inversion swap-in according to near-infrared single-element detector in different moments collected one point data
Row reconstruct near-infrared fluorescent image F:
In above formula, subscript " -1 " is indicated to matrix inversion.
5. the micro- cell imaging method of near-infrared fluorescent as claimed in claim 4 based on Hadamard transform, it is characterised in that:
The excitation light source is LED light source, and the near-infrared fluorescent sample is that single-walled carbon nanotube marks sample, near-infrared fluorescent image
The wave band of F is 800~1600nm.
6. the micro- cell imaging method of near-infrared fluorescent as claimed in claim 4 based on Hadamard transform, it is characterised in that:
In step 2), p1=400, p2=300, then P=120000；
In step 3), two-dimensional modulation image sequence S is generated1,S2,…,S120000:
Digital Micromirror Device is according to two-dimensional modulation image sequence S in step 4)1,S2,…,S120000, micromirror is controlled along time shaft
It is encoded in face of near-infrared fluorescent image；
In step 5), Digital Micromirror Device coding after near-infrared fluorescent image, focus on near-infrared unit through the second lens group
On detector:
In above formula, f1,f2,…,f120000For the data that near-infrared fluorescent image F boil down to is one-dimensional；y1,y2,…,y120000It is close red
Outer single-element detector is in different moments collected one point data；
In step 6), computer, in different moments collected one point data, uses Adama according to near-infrared single-element detector
Near-infrared fluorescent image F is reconstructed in inverse transformation:
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610541848.4A CN106204486B (en) | 2016-07-08 | 2016-07-08 | A kind of micro- cell imaging device and method of near-infrared fluorescent based on Hadamard transform |
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| CN201610541848.4A CN106204486B (en) | 2016-07-08 | 2016-07-08 | A kind of micro- cell imaging device and method of near-infrared fluorescent based on Hadamard transform |
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