CN103837461B - A kind of gray scale photographic head and there is the cell comprehensive analysis device of high efficiency illumination - Google Patents
A kind of gray scale photographic head and there is the cell comprehensive analysis device of high efficiency illumination Download PDFInfo
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- CN103837461B CN103837461B CN201410020477.6A CN201410020477A CN103837461B CN 103837461 B CN103837461 B CN 103837461B CN 201410020477 A CN201410020477 A CN 201410020477A CN 103837461 B CN103837461 B CN 103837461B
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Abstract
A kind of attachment typed gray scale photographic head for micrometron of the present invention, including light inlet, light-emitting window, and 45 ° of band reduction of fractions to a common denominator light color separation films, lens devices, the gray level image sensors being coaxially disposed, light is injected by light inlet, two-way light is formed through 45 ° of band reduction of fractions to a common denominator light color separation films, it is divided into two-way according to different wavelength, the light of the most specific band-pass wavelength is carried out imaging by reflected illumination to gray level image sensor, the light of the wavelength in addition to specific band-pass wavelength, then keep original direction, light-emitting window penetrate.A kind of cell comprehensive analysis device with high efficiency illumination, including micrometron, colour imagery shot, gray scale photographic head, image analysis system；Gray scale photographic head, as adnexa, is arranged between the eyepiece of micrometron and the light path of object lens, and colour imagery shot is arranged on above eyepiece；Micrometron, colour imagery shot, gray scale photographic head are connected with image analysis system respectively, carry out data collection and analysis.
Description
Technical field
The present invention relates to a kind of cell analysis apparatus, more particularly, it relates to a kind of attachment typed ash for micrometron
Degree photographic head, and the cell comprehensive analysis device with high efficiency illumination of described gray scale photographic head is installed.
Background technology
Analysis to cell at present mainly includes qualitative and quantitative two kinds of methods.Method for qualitative analysis, is generally based on cellular morphology
Credit is analysed, i.e. by cell being carried out qualitative point by observation through the morphological feature of hyperchromatic exfoliative cyte under the microscope
Analysis, including the depth of cytoplasm Yu nuclear targeting, volume size, chromosome thickness in nucleus, Chromosomal arrangement and carefully
Karyon and cytoplasmic ratio, according to the whether canceration of the micro-judgment cell of observer, this kind of observation procedure subjective factors is too strong,
Relying on experience and the skill of doctor, misdiagnosis rate is higher, accuracy rate typically only about 60%.
Quantitative analysis method, it is simply that intracellular chemical composition is carried out quantitative analysis, such as hereditary material DNA (deoxyribonucleic acid)
(DNA).The essence of cell carcinogenesis is the rapid infinite multiplication of cell and transfer, when biological cell generation canceration, and nucleus
Interior DNA content can increase, and can combine, dyestuff or developer when cell nuclear dna with cell nuclear dna specificity
When content increases, DNA binding dye or developer increase accordingly, and its optical density (OD) increases.Then by measuring microscope image
Optical density (OD), nuclear DNA quantitative analysis can be carried out, by comparing with normal cell, find early carcinomatous change
Cell.
Although DNA quantitative analysis method can provide more objectively data, but the method is analyzed just for nucleus,
Information is not comprehensive, and in the neoplastic cell nuclei of early stage patient, DNA content increase is not notable, needs the morphology of comprehensive cell to believe
Breath, to provide analysis result the most accurately.
Chinese invention patent application 200810070940.2 discloses a kind of method for integrative detection of cells, can carry out cell shape simultaneously
State credit analysis and DNA detection by quantitative, the advantage combining two kinds of methods, improve the accuracy rate analyzed.
Foregoing invention application only discloses method, does not provide the device of implementation method, can obtain the colour of the same visual field simultaneously
Image and the gray level image of specific wavelength, wherein coloured image is in order to cellular morphology analysis, and gray level image is in order to nucleus DNA
Quantitative analysis.
Chinese invention patent application 201110071790.6 discloses a kind of full-automatic double-ccd photosensitive element digital microscope, simultaneously
Colored ccd photo-sensitive cell and black and white ccd photo-sensitive cell is utilized to carry out specimen image information collecting, to obtain the multi-focal-plane of specimen
Hologram three-dimensional image, the diagosis knot of the nuclear integrated optical density information output specimen simultaneously gathered according to black and white ccd photo-sensitive cell
Opinion, final output specimen picture and text report.The another present invention uses fluctuating plate device, moves horizontally clamping device and be automatically performed specimen and carry
The replacing of slide, then to using three-dimensional frame that microscope slide is carried out three-dimensional localization, in order to colored ccd photo-sensitive cell and black
The white ccd photo-sensitive cell image information collecting to specimen.
The technical scheme of foregoing invention application is directly respectively mounted a colour imagery shot on the binocular of viewing head and a black and white is taken the photograph
As head, owing to the binocular of viewing head is the light splitting of 50:50, the brightness of image that each photographic head receives have lost half, for
Black and white photographic head, reception is the image of narrow-band spectrum, and required time of exposure requires longer, adds light splitting loss, and it exposes
Light time lengthening one times, as used " scan mode of exposure in motion " that scanning speed will be made to reduce half.As kept former
The time of exposure come, to be at least enhanced about more than once brightness of illumination, need to use the light source of more high brightness, and redesign efficiently
Rate optical system, it will be greatly improved cost；Further, since binocular is assembled with photographic head, just without visualization function,
It is not easy to doctor carry out rechecking under mirror；And eyepiece typically has diopter to regulate, it is rotating, is easily caused two in use and takes the photograph
As head not parfocalization or visual field are inconsistent；Simultaneously because be mounted directly two photographic head and corresponding mechanical-optical setup observing head,
Its weight is relatively big, also can affect the stability of microscopic structure.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, it is provided that a kind of illumination efficiency is higher, more stable reliably, it is easy to
The colour of operation and black white image acquisition device, retain visualization function simultaneously, to ensure that the high efficiency that has rechecked under mirror is shone
Bright cell comprehensive analysis device, and the attachment typed gray scale photographic head for micrometron as digital middleware.
Technical scheme is as follows:
A kind of attachment typed gray scale photographic head for micrometron, including light inlet, light-emitting window, and be coaxially disposed
45 ° of band reduction of fractions to a common denominator light color separation films, lens devices, gray level image sensors, light is injected by light inlet, divides through 45 ° of band reduction of fractions to a common denominator light
Color chips formed two-way light, be divided into two-way according to different wavelength, the light of the most specific band-pass wavelength by reflected illumination to gray-scale map
As sensor carries out imaging, the light of the wavelength in addition to specific band-pass wavelength, then keep original direction, light-emitting window penetrate.
As preferably, the band-pass wavelength of 45 ° of band reduction of fractions to a common denominator light color separation film reflections matches with nuclear colouring method.
As preferably, 45 ° of band reduction of fractions to a common denominator light color separation films, reflection wavelength is the light of 585-595nm, transmission peak wavelength be 400-585nm,
The light of 595-700nm.
As preferably, the wavelength of the light of 45 ° of band reduction of fractions to a common denominator light color separation film foveal reflexs is 590nm, carries a width of 10nm.
A kind of cell comprehensive analysis device with high efficiency illumination, including micrometron, colour imagery shot, above-mentioned gray scale
Photographic head, image analysis system；Gray scale photographic head, as adnexa, is arranged between the eyepiece of micrometron and the light path of object lens,
Colour imagery shot is arranged on above eyepiece；Micrometron, colour imagery shot, gray scale photographic head are respectively with image analysis system even
Connect, carry out data collection and analysis.
As preferably, the communication interface being connected by differing of colour imagery shot, gray scale photographic head and image analysis system.
As preferably, thing side's sampling resolution that colour imagery shot is corresponding with gray scale photographic head is identical.
As preferably, the field of view center that colour imagery shot is corresponding with gray scale photographic head overlaps.
As preferably, image analysis system gathers coloured image and the gray level image of gray scale photographic head of colour imagery shot simultaneously, enters
After row space registration, with coloured image and gray level image, same cell is carried out qualitative analysis and quantitative analysis respectively, in conjunction with fixed
Property analysis and quantitative analysis draw analysis result.
Beneficial effects of the present invention is as follows:
The present invention provide a kind of can carry out morphocytology analysis and nucleus DNA detection by quantitative simultaneously there is high efficiency illumination
Cell comprehensive analysis device, utilize device of the present invention can gather coloured image needed for qualitative analysis and quantitatively point simultaneously
Gray level image needed for analysis, improves cell divide and the accuracy of analysis.
Present invention also offers a kind of attachment typed gray scale photographic head for micrometron, can be on the basis of micrometron
On realize the collection of gray level image.Gray scale photographic head of the present invention and cell comprehensive analysis device, remain microscope original
Visual function, meet under mirror the needs of check；Gray scale photographic head, as digital intermediate, uses attachment version design, it is easy to
Install, need not after installation adjust；Maintain original microstructural stability.
Accompanying drawing explanation
Fig. 1 is the structural representation of cell comprehensive analysis device；
Fig. 2 is the structural representation of micrometron；
Fig. 3 is the sectional view of gray scale photographic head；
Fig. 4 is in the present embodiment, the characteristic curve of the light of 45 ° of band reduction of fractions to a common denominator light color separation film reflections；
In figure: 100 is to observe head, and 101 is eyepiece, and 102 is object lens, and 103 is object stage, and 104 is main body, and 105 is the
Three, 200 is gray scale photographic head, and 201 is light inlet, and 202 is light-emitting window, and 203 is 45 ° of band reduction of fractions to a common denominator light color separation films, 204
Being lens devices, 205 is gray level image sensor, and 300 is colour imagery shot, and 400 is image analysis system.
Detailed description of the invention
Below in conjunction with drawings and Examples, the present invention is described in further detail.
As it is shown in figure 1, a kind of cell comprehensive analysis device, including micrometron as shown in Figure 2, colour imagery shot 300,
Gray scale photographic head 200, image analysis system 400；Gray scale photographic head 200, as adnexa, is arranged on the eyepiece of micrometron
Between 101 and the light path of object lens 102, in the present embodiment, gray scale photographic head 200 is arranged on the viewing head of automatic digit microscope
Between portion 100 and microscope body 104.Colour imagery shot 300 is arranged on above eyepiece 101, and in the present embodiment, colour is taken the photograph
As 300 are arranged on micrometron and observe on the 3rd 105 of head 100.Micrometron, colour imagery shot 300,
Gray scale photographic head 200 is connected with image analysis system 400 respectively, carries out data collection and analysis.Described micrometron,
Optical system is infinity chromatic aberration correction CCIS structure, and observing head 100 is three mesh, and object stage 103 is electronic, Ke Yixuan
BA610Mot with Motic company.
Gray scale photographic head 200 is designed as the form of microscopical adnexa, and one of the present invention is used for micrometron
Attachment typed gray scale photographic head 200 as it is shown on figure 3, include light inlet 201, light-emitting window 202, and be coaxially disposed
45 ° of band reduction of fractions to a common denominator light color separation films 203, lens devices 204, gray level image sensors 205, light injected by light inlet 201, warp
45 ° of band reduction of fractions to a common denominator light color separation films 203 form two-way light, are divided into two-way according to different wavelength, the light of the most specific band-pass wavelength
Imaging is carried out to gray level image sensor 205 by reflected illumination, the light of the wavelength in addition to specific band-pass wavelength, then keep original
Direction, light-emitting window 202 penetrate, enter the observation head 100 of described micrometron, the 3rd 105 can be arrived
Colour imagery shot 300.45 ° of band reduction of fractions to a common denominator light color separation films 203 are arranged at the forward position of gray scale photographic head 200, and it is distinguished up and down
Just to light-emitting window 202, light inlet 201.Lens devices 204 is for being converged to picture to the light path entering gray scale photographic head 200；
Gray level image sensor 205 is for receiving the imaging after lens devices 204 is assembled.
The light of the colour imagery shot 300 for entering observe head 100 105, owing to only having the light of the narrowest spectrum to be reflected,
Major part wavelength light still enters colour imagery shot 300, so color intensity is held essentially constant, close to 201110071790.6
Disclose the twice of the technical scheme of a kind of full-automatic double-ccd photosensitive element digital microscope；And for entering Lycoperdon polymorphum Vitt photographic head 200
Light, owing to there is no the light splitting of prism, disclose a kind of full-automatic double ccd photo-sensitive cell numerals close to 201110071790.6 and show
The twice of the technical scheme of micro mirror.
Optical filter 203 center matches with nuclear colouring method through wavelength, bandwidth, i.e. the spectral characteristic of optical filter 203
Determined by nuclear colouring method, when nucleus being carried out Feulgen dyeing, 45 ° of band reduction of fractions to a common denominator light color separation films 203, reflection
Wavelength is the light of 585-595nm, and transmission peak wavelength is the light of 400-585nm, 595-700nm.In the present embodiment, 45 ° of bands are logical
The wavelength of the light of color-division sheet 203 foveal reflex is 590nm, carries a width of 10nm, as shown in Figure 4.
Speed is reduced, colour imagery shot 300, ash in order to avoid influencing each other when same type communication interface transmits data at the same time
Degree photographic head 200 and the communication interface being connected by differing of image analysis system 400.Such as colour imagery shot 300 is optional
Using Moticam Pro285E, be 1394 interfaces, gray scale photographic head 200 selects USB interface, or other communication interfaces.
Thing side's sampling resolution that colour imagery shot 300 is corresponding with gray scale photographic head 200 is identical.Colour imagery shot 300 and gray scale
The field of view center of photographic head 200 correspondence overlaps.
Image analysis system 400 gathers coloured image and the gray level image of gray scale photographic head 200 of colour imagery shot 300 simultaneously,
After carrying out spatial registration, it is ensured that be that same cell is carried out quantitative analysis and analyzes qualitatively simultaneously.Image analysis system 400
With coloured image and gray level image, same cell is carried out qualitative analysis and quantitative analysis respectively, in conjunction with qualitative analysis and quantitatively divide
Analysis draws analysis result.
Described qualitative analysis is to be analyzed based on coloured image, including the classification of cell, and cytoplasmic color and size, carefully
Chromosome thickness in karyon, Chromosomal arrangement and nucleus and cytoplasmic ratio.Described quantitative analysis is based on gray-scale map
As nucleus being carried out DNA quantitative analysis, including nucleus gray scale, integral optical density information.
When specifically applying, first section is carried out dyeing process, including nucleus being carried out Feulgen dyeing and cytoplasm being entered
Row EA50 or eosin stains.
After twice dyeing, section is placed on the object stage 103 of micrometron, image analysis system 400 controls
Object stage 103 moves to interested position and focuses, and controls colour imagery shot 300 simultaneously and gray scale photographic head 200 carries out image
Obtain, obtain coloured image and the gray level image of 590nm wavelength of same area-of-interest respectively, two width images are carried out space
Registration.
It is analyzed processing to image by image analysis system 400: carry out according to the morphological characteristic parameter of cell with coloured image
Classification, and analyze cytoplasmic color and size, chromosome thickness in nucleus, Chromosomal arrangement and nucleus and cytoplasm
Ratio etc.；With gray level image, homocellular nucleus is carried out DNA quantitative analysis, including nucleus gray scale, integral light
Density information etc.；Finally according to spatial registration information, the comprehensively morphology letter of the nuclear DNA content of certain cell, cell
Breath provides comprehensive analysis report.
Above-described embodiment is intended merely to the present invention is described, and is not used as limitation of the invention.As long as the skill according to the present invention
Art essence, be changed above-described embodiment, modification etc. all will fall in the range of the claim of the present invention.
Claims (7)
1. the attachment typed gray scale photographic head for micrometron, it is characterised in that include light inlet, light-emitting window,
And 45 ° of band reduction of fractions to a common denominator light color separation films, lens devices, the gray level image sensors being coaxially disposed, light injected by light inlet, warp
45 ° of band reduction of fractions to a common denominator light color separation films form two-way light, are divided into two-way according to different wavelength, and the light of the most specific band-pass wavelength is by instead
Penetrate and expose to gray level image sensor and carry out imaging, the light of the wavelength in addition to specific band-pass wavelength, then keep original direction,
Penetrated by light-emitting window；The band-pass wavelength of 45 ° of band reduction of fractions to a common denominator light color separation film reflections matches with nuclear colouring method.
Attachment typed gray scale photographic head for micrometron the most according to claim 1, it is characterised in that 45 °
Band reduction of fractions to a common denominator light color separation film, reflection wavelength is the light of 585-595nm, and transmission peak wavelength is the light of 400-585nm, 595-700nm.
Attachment typed gray scale photographic head for micrometron the most according to claim 1, it is characterised in that 45 °
The wavelength of the light with reduction of fractions to a common denominator light color separation film foveal reflex is 590nm, carries a width of 10nm.
4. a cell comprehensive analysis device with high efficiency illumination, it is characterised in that include micrometron, colored shooting
Head, the gray scale photographic head as described in any one of claim 1 to claim 3, image analysis system；Gray scale photographic head conduct
Adnexa, is arranged between the eyepiece of micrometron and the light path of object lens, and colour imagery shot is arranged on above eyepiece；The most micro-
Mirror, colour imagery shot, gray scale photographic head are connected with image analysis system respectively, carry out data collection and analysis；Graphical analysis system
System gathers coloured image and the gray level image of gray scale photographic head of colour imagery shot simultaneously, after carrying out spatial registration, respectively with colour
Image and gray level image carry out qualitative analysis and quantitative analysis to same cell, draw analysis in conjunction with qualitative analysis and quantitative analysis
Result.
The cell comprehensive analysis device with high efficiency illumination the most according to claim 4, it is characterised in that colored shooting
The communication interface being connected by differing of head, gray scale photographic head and image analysis system.
The cell comprehensive analysis device with high efficiency illumination the most according to claim 4, it is characterised in that colored shooting
Thing side's sampling resolution that head is corresponding with gray scale photographic head is identical.
The cell comprehensive analysis device with high efficiency illumination the most according to claim 4, it is characterised in that colored shooting
The field of view center that head is corresponding with gray scale photographic head overlaps.
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| CN201410020477.6A CN103837461B (en) | 2014-01-16 | 2014-01-16 | A kind of gray scale photographic head and there is the cell comprehensive analysis device of high efficiency illumination |
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| EP4194801A1 (en) | 2015-02-24 | 2023-06-14 | The University of Tokyo | Dynamic high-speed high-sensitivity imaging device and imaging method |
| US11098275B2 (en) | 2015-10-28 | 2021-08-24 | The University Of Tokyo | Analysis device |
| CN106442528A (en) * | 2016-09-09 | 2017-02-22 | 上海新纤仪器有限公司 | Dual camera microscope testing device and testing method for contents of fiber components |
| EP3807005B1 (en) | 2018-06-13 | 2023-10-25 | ThinkCyte K.K. | Methods and systems for cytometry |
| CN112987279A (en) * | 2021-02-10 | 2021-06-18 | 光速视觉（北京）科技有限公司 | Telescope, and electronic eyepiece and eyepiece adapter for telescope |
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