CN103837461A - Gray scale camera and cell comprehensive analysis device with high-efficiency illumination - Google Patents
Gray scale camera and cell comprehensive analysis device with high-efficiency illumination Download PDFInfo
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- CN103837461A CN103837461A CN201410020477.6A CN201410020477A CN103837461A CN 103837461 A CN103837461 A CN 103837461A CN 201410020477 A CN201410020477 A CN 201410020477A CN 103837461 A CN103837461 A CN 103837461A
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Abstract
The invention relates to a gray scale camera used for an automatic microscope in an accessory manner, which comprises a light inlet and a light outlet, and also comprises a 45-degree bandpass light-splitting color separation film, a lens device, and a grey-scale image sensor arranged coaxially; light enters the light inlet, and is splitted into two ways by the 45-degree bandpass light-splitting color separation film according to different wavelengths; light with a specific bandpass wavelength is reflected to the grey-scale image sensor for imaging, and light with a wavelength other than the specific bandpass wavelength has its original direction maintained and exits the light outlet. A cell comprehensive analysis device with high-efficiency illumination comprises an automatic microscope, a color camera, a gray scale camera, and an image analysis system; the gray scale camera is used as an accessory, and is mounted between optical paths of an ocular lens and an object lens of the automatic microscope; the color camera is disposed above the ocular lens; the automatic microscope, the color camera, and the gray scale camera are respectively connected with the image analysis system for data acquisition and analysis.
Description
Technical field
The present invention relates to a kind of cell analysis apparatus, more particularly, relate to a kind of attachment typed gray scale camera for micrometron, and the cell comprehensive analysis device with high efficiency illumination of described gray scale camera is installed.
Background technology
At present the analysis of cell is mainly comprised to two kinds of methods of quantitative and qualitative analysis.Method for qualitative analysis, generally based on cellular morphology Epidemiological Analysis, by cell being carried out to qualitative analysis by observing through the morphological feature of hyperchromatic cast-off cells under the microscope, comprise the depth of cytoplasm and nucleus dyeing, volume size, chromosome thickness, Chromosomal arrangement and nucleus and cytoplasmic ratio in nucleus, according to the whether canceration of observer's micro-judgment cell, this kind of observation procedure subjective factor is too strong, rely on doctor's experience and skill, misdiagnosis rate is higher, and accuracy rate generally only has 60% left and right.
Quantitative analysis method, carries out quantitative test to intracellular chemical composition exactly, such as inhereditary material-DNA (deoxyribonucleic acid) (DNA).The essence of cell carcinogenesis is the rapid infinite multiplication of cell and transfer, in the time of biological cell generation canceration, endonuclear DNA content can increase, dyestuff or developer can be combined with cell nuclear dna specificity, in the time that in nucleus, DNA content increases, DNA binding dye or developer are corresponding to be increased, and its optical density (OD) increases.Then by the optical density (OD) of measuring microscope image, can carry out nuclear DNA quantitative test,, by comparing with normal cell, find the cell of early carcinomatous change.
Although DNA quantitative analysis method can provide comparatively objectively data, but the method is just analyzed for nucleus, and information is not comprehensive, and in the neoplastic cell nuclei of early stage patient, DNA content increase is not remarkable, need the morphologic information of comprehensive cell, so that analysis result more to be accurately provided.
Chinese invention patent application 200810070940.2 discloses a kind of method for integrative detection of cells, can carry out cellular morphology Epidemiological Analysis and DNA simultaneously and quantitatively detect, and combines the advantage of two kinds of methods, improves the accuracy rate of analyzing.
Foregoing invention application only discloses method, and the device of implementation method is not provided, and can obtain the coloured image of the same visual field and the gray level image of specific wavelength simultaneously, and wherein coloured image is in order to cellular morphology analysis, and gray level image is in order to nucleus DNA quantitative test.
Chinese invention patent application 201110071790.6 discloses a kind of full-automatic double-ccd photosensitive element digital microscope, utilize colored ccd photo-sensitive cell and black and white ccd photo-sensitive cell to carry out sample image information collecting simultaneously, to obtain multifocal hologram three-dimensional image of sample, simultaneously according to the nuclear integrated optical density information output sample of black and white ccd photo-sensitive cell collection read sheet conclusion, finally export sample picture and text report.Another the present invention adopts fluctuating plate device, move horizontally clamping device completes the replacing of sample microslide automatically, then to adopting three-dimensional frame to carry out three-dimensional localization to microslide, so that colored ccd photo-sensitive cell and the image information collecting of black and white ccd photo-sensitive cell to sample.
The technical scheme of foregoing invention application is directly installed respectively a colour imagery shot and a black and white camera on the binocular of viewing head, because the binocular of viewing head is the light splitting of 50:50, the brightness of image that each camera receives has been lost half, for black and white camera, what receive is the image of narrow-band spectrum, and the required time shutter requires longer, adds a point light loss, its time shutter extends one times, as adopted " scan mode of exposing in motion " will make sweep velocity reduce half.As kept the original time shutter, at least brightness of illumination to be enhanced about more than once, need adopt the more light source of high brightness, and redesign high-level efficiency optical system, will greatly raise the cost; In addition, because binocular has all been installed camera, just there is no visualization function, be not easy to doctor and carry out rechecking under mirror; And eyepiece generally have diopter regulate, be rotating, easily cause in use two cameras not parfocalization or visual field inconsistent; Due to two cameras and corresponding mechanical-optical setup being directly installed at observation head, its weight is larger, also can affect the stability of microscopic structure simultaneously.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of illumination efficiency higher, more reliable and stable, the colour of easy operating and black white image acquisition device, retain visualization function simultaneously, with the cell comprehensive analysis device with high efficiency illumination that ensures to recheck under mirror, and as the attachment typed gray scale camera for micrometron of digital middleware.
Technical scheme of the present invention is as follows:
A kind of attachment typed gray scale camera for micrometron, comprise light inlet, light-emitting window, and coaxial 45 ° of band reduction of fractions to a common denominator light color separation films, lens devices, the gray level image sensor arranging, light is injected by light inlet, form two-way light through 45 ° of band reduction of fractions to a common denominator light color separation films, be divided into two-way according to different wavelength, wherein the light of the logical wavelength of specific band is reflected and exposes to gray level image sensor and carry out imaging, the light of the wavelength except the logical wavelength of specific band, keep original direction, penetrated by light-emitting window.
As preferably, the 45 ° of logical wavelength of band and nuclear colouring methods with the reflection of reduction of fractions to a common denominator light color separation film match.
As preferably, be with reduction of fractions to a common denominator light color separation films for 45 °, the light that reflection wavelength is 585-595nm, transmission peak wavelength is the light of 400-585nm, 595-700nm.
As preferably, 45 ° of light wavelengths with reduction of fractions to a common denominator light color separation film foveal reflex are 590nm, and bandwidth is 10nm.
There is a cell comprehensive analysis device for high efficiency illumination, comprise micrometron, colour imagery shot, above-mentioned gray scale camera, image analysis system; Gray scale camera, as annex, is arranged between the eyepiece of micrometron and the light path of object lens, and colour imagery shot is arranged on eyepiece top; Micrometron, colour imagery shot, gray scale camera are connected with image analysis system respectively, carry out data collection and analysis.
As preferably, colour imagery shot, gray scale camera adopt not identical communication interface with the connection of image analysis system.
As preferably, the object space sampling resolution that colour imagery shot is corresponding with gray scale camera is identical.
As preferably, the field of view center that colour imagery shot is corresponding with gray scale camera overlaps.
As preferably, image analysis system gathers the coloured image of colour imagery shot and the gray level image of gray scale camera simultaneously, carry out after spatial registration, with coloured image and gray level image, same cell is carried out to qualitative analysis and quantitative test respectively, draw analysis result in conjunction with qualitative analysis and quantitative test.
Beneficial effect of the present invention is as follows:
The invention provides a kind of cell comprehensive analysis device with high efficiency illumination that can simultaneously carry out the quantitative detection of cellular morphology Epidemiological Analysis and nucleus DNA, utilize device of the present invention can gather the required coloured image of qualitative analysis and the required gray level image of quantitative test simultaneously, improve the accuracy of cell classification and analysis.
The present invention also provides a kind of attachment typed gray scale camera for micrometron, can on the basis of micrometron, realize the collection of gray level image.Gray scale camera of the present invention and cell comprehensive analysis device, retained the original visual function of microscope, meets the needs of checking under mirror; Gray scale camera, as digital intermediate, adopts attachment version design, is easy to install, and does not need to adjust after installation; Maintain original microstructural stability.
Brief description of the drawings
Fig. 1 is the structural representation of cell comprehensive analysis device;
Fig. 2 is the structural representation of micrometron;
Fig. 3 is the cut-open view of gray scale camera;
Fig. 4 is in the present embodiment, the family curve of 45 ° of light with the reflection of reduction of fractions to a common denominator light color separation film;
In figure: the 100th, observe head, the 101st, eyepiece, the 102nd, object lens, the 103rd, objective table, the 104th, main body, 105 is the 3rd, the 200th, gray scale camera, the 201st, light inlet, the 202nd, light-emitting window, 203 is 45 ° of band reduction of fractions to a common denominator light color separation films, the 204th, lens devices, the 205th, gray level image sensor, the 300th, colour imagery shot, the 400th, image analysis system.
Embodiment
Below in conjunction with drawings and Examples, the present invention is described in further detail.
As shown in Figure 1, a kind of cell comprehensive analysis device, comprises micrometron, colour imagery shot 300, gray scale camera 200, image analysis system 400 as shown in Figure 2; Gray scale camera 200, as annex, is arranged between the eyepiece 101 of micrometron and the light path of object lens 102, and in the present embodiment, gray scale camera 200 is arranged between the observation head 100 and microscope body 104 of automatic digit microscope.Colour imagery shot 300 is arranged on eyepiece 101 tops, and in the present embodiment, colour imagery shot 300 is arranged on micrometron and observes on the 3rd 105 of head 100.Micrometron, colour imagery shot 300, gray scale camera 200 are connected with image analysis system 400 respectively, carry out data collection and analysis.Described micrometron, optical system is infinity chromatic aberration correction CCIS structure, and observing head 100 is three orders, and objective table 103 is electronic, can select the BA610Mot of Motic company.
For the light that enters 105 the colour imagery shot 300 of observing head 100, because the light that only has very narrow spectrum is reflected, most of wavelength light still enters colour imagery shot 300, so color intensity remains unchanged substantially, approach 201110071790.6 twices that disclose a kind of technical scheme of full-automatic double-ccd photosensitive element digital microscope; And for the light that enters grey camera 200, owing to there is no the light splitting of prism, approach 201110071790.6 twices that disclose a kind of technical scheme of full-automatic double-ccd photosensitive element digital microscope.
In the time that communication interface of the same type transmits data at the same time, influence each other and underspeed, colour imagery shot 300, gray scale camera 200 adopt not identical communication interface with the connection of image analysis system 400.For example colour imagery shot 300 can be selected Moticam Pro285E, is 1394 interfaces, and gray scale camera 200 is selected USB interface, or other communication interfaces.
The object space sampling resolution that colour imagery shot 300 is corresponding with gray scale camera 200 is identical.The field of view center that colour imagery shot 300 is corresponding with gray scale camera 200 overlaps.
Described qualitative analysis is to analyze based on coloured image, comprises the classification of cell, cytoplasmic color and size, chromosome thickness in nucleus, Chromosomal arrangement and nucleus and cytoplasmic ratio.Described quantitative test is, based on gray level image, nucleus is carried out to DNA quantitative test, comprises nucleus gray scale, integral optical density information.
When concrete application, first to the processing of dyeing of cutting into slices, comprise to nucleus is carried out Feulgen dyeing and cytoplasm is carried out to EA50 or Yihong dyeing.
After twice dyeing, section is placed on the objective table 103 of micrometron, control objective table 103 by image analysis system 400 and move to interested position focusing, control colour imagery shot 300 and gray scale camera 200 carries out Image Acquisition simultaneously, obtain respectively the coloured image of same area-of-interest and the gray level image of 590nm wavelength, two width images are carried out to spatial registration.
By image analysis system 400, image is carried out to analyzing and processing: classify according to the morphological feature parameter of cell with coloured image, and color and the size of analysis of cells slurry, chromosome thickness in nucleus, Chromosomal arrangement and nucleus and cytoplasmic ratio etc.; Homocellular nucleus is carried out to DNA quantitative test with gray level image, comprise nucleus gray scale, integral optical density information etc.; Finally, according to spatial registration information, comprehensively the morphologic information of the nuclear DNA content of certain cell, cell provides comprehensive analysis report.
Above-described embodiment is only for the present invention is described, and not as limitation of the invention.As long as according to technical spirit of the present invention, to above-described embodiment change, modification etc. all will drop in the scope of claim of the present invention.
Claims (9)
1. the attachment typed gray scale camera for micrometron, it is characterized in that, comprise light inlet, light-emitting window, and coaxial 45 ° of band reduction of fractions to a common denominator light color separation films, lens devices, the gray level image sensor arranging, light is injected by light inlet, form two-way light through 45 ° of band reduction of fractions to a common denominator light color separation films, be divided into two-way according to different wavelength, wherein the light of the logical wavelength of specific band is reflected and exposes to gray level image sensor and carry out imaging, the light of the wavelength except the logical wavelength of specific band, keep original direction, penetrated by light-emitting window.
2. according to claim 1ly it is characterized in that for microscopical attachment typed gray scale camera, the 45 ° of logical wavelength of band and nuclear colouring methods with the reflection of reduction of fractions to a common denominator light color separation film match.
3. according to claim 1ly it is characterized in that for microscopical attachment typed gray scale camera, 45 ° of band reduction of fractions to a common denominator light color separation films, the light that reflection wavelength is 585-595nm, transmission peak wavelength is the light of 400-585nm, 595-700nm.
4. according to claim 3ly it is characterized in that for microscopical attachment typed gray scale camera, 45 ° of light wavelengths with reduction of fractions to a common denominator light color separation film foveal reflex are 590nm, and bandwidth is 10nm.
5. a cell comprehensive analysis device with high efficiency illumination, is characterized in that, comprises micrometron, colour imagery shot, if claim 1 is to gray scale camera, image analysis system as described in claim 4 any one; Gray scale camera, as annex, is arranged between the eyepiece of micrometron and the light path of object lens, and colour imagery shot is arranged on eyepiece top; Micrometron, colour imagery shot, gray scale camera are connected with image analysis system respectively, carry out data collection and analysis.
6. the cell comprehensive analysis device with high efficiency illumination according to claim 5, is characterized in that, colour imagery shot, gray scale camera adopt not identical communication interface with the connection of image analysis system.
7. the cell comprehensive analysis device with high efficiency illumination according to claim 5, is characterized in that, the object space sampling resolution that colour imagery shot is corresponding with gray scale camera is identical.
8. the cell comprehensive analysis device with high efficiency illumination according to claim 5, is characterized in that, the field of view center that colour imagery shot is corresponding with gray scale camera overlaps.
9. the cell comprehensive analysis device with high efficiency illumination according to claim 5, it is characterized in that, image analysis system gathers the coloured image of colour imagery shot and the gray level image of gray scale camera simultaneously, carry out after spatial registration, with coloured image and gray level image, same cell is carried out to qualitative analysis and quantitative test respectively, draw analysis result in conjunction with qualitative analysis and quantitative test.
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| 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 |
| CN112638529A (en) * | 2018-06-13 | 2021-04-09 | 新克赛特株式会社 | Method and system for cell counting |
| 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 |
| US11542461B2 (en) | 2015-10-28 | 2023-01-03 | The University Of Tokyo | Analysis device |
| US11579075B2 (en) | 2015-02-24 | 2023-02-14 | The University Of Tokyo | Dynamic high-speed high-sensitivity imaging device and imaging method |
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| WO2022170950A1 (en) * | 2021-02-10 | 2022-08-18 | 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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