CN102519909B - Air-space low-interference phase microscope based on liquid crystal tunable filter - Google Patents
Air-space low-interference phase microscope based on liquid crystal tunable filter Download PDFInfo
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- CN102519909B CN102519909B CN2011103586514A CN201110358651A CN102519909B CN 102519909 B CN102519909 B CN 102519909B CN 2011103586514 A CN2011103586514 A CN 2011103586514A CN 201110358651 A CN201110358651 A CN 201110358651A CN 102519909 B CN102519909 B CN 102519909B
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- crystal tunable
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Abstract
The invention discloses an air-space low-interference phase microscope based on a liquid crystal tunable filter. The air-space low-interference phase microscope comprises an object stage, a first object lens, a first reflecting mirror, a ductoscopy and a CCD (Charge Coupled Device) image sensor and is characterized by further comprising a white light source, a spectroscope and a liquid crystal tunable filter, wherein the spectroscope is used for receiving white light signals which are focused into an observed sample through the first object lens, reflected scattered signals of the observed sample are projected into the ductoscopy through the first object lens and the first reflecting mirror, the liquid crystal tunable filter is arranged between the ductoscopy and the CCD image sensor, and the liquid crystal tunable filter is used for projecting microscope images of different wavelengths onto the CCD image sensor. The air-space low-interference phase microscope has the obvious effects that: a light path structure is simple, the control is convenient, a system has high air-space refractive index and fluctuation sensitivity and can be used for processing a clinical sample slide directly so as to realize high-precision imaging for a cell nano-structure, and the full-field rapid measurement is realized by utilizing electro-optic liquid crystal tunning.
Description
Technical field
The present invention relates to a kind of phase microscope, specifically, is the low coherent phase microscope in a kind of spatial domain based on liquid crystal tunable filter, belongs to optical image technology.
Background technology
Because traditional optical microscope is based on absorption or the fluorescent effect of sample to the imaging of sample, thus resolution lower (＜500nm), can't detect the variation of sample nanostructured.Phase microscope and differential interference difference microscope based on optics can be surveyed small cell nano structural change, but lack quantitative information, have limited its range of application.In recent years, the quantitative phase microscope utilizes interference effect can obtain the systematic parameters relevant with the cell nano architectural characteristic such as phase place, amplitude, quantitative information is provided, though such microscope can be surveyed the cell nano structure, but its speed, sensitivity and precision, and the specific (special) requirements of sample all limited its popularization to clinical practice.
Summary of the invention
The purpose of this invention is to provide a kind of fast, high sensitivity and high-precision low coherent phase microscope, can directly observe clinical cell tissue sample slide, and therefrom extract the effective nanostructured characteristic of cell.
For achieving the above object, scheme of the present invention is as follows:
The low coherent phase microscope in a kind of spatial domain based on liquid crystal tunable filter, comprise objective table, first object lens, first catoptron, pipe mirror and ccd image inductor, its key is: also comprise white light source, spectroscope and liquid crystal tunable filter plate, described spectroscope is installed between first object lens and first catoptron, this spectroscope is used for receiving the white light signal that described white light source produces, this white light signal sees through described first object lens and focuses in the observation sample on the described objective table, the reflection of this observation sample and scattered signal see through described first object lens and spectroscope is incident upon on described first catoptron, this first catoptron arrives Guan Jingzhong with the signal reflex that receives, described liquid crystal tunable filter plate is installed between Guan Jingyu ccd image inductor, is projected on the described ccd image inductor by the micro-image of this liquid crystal tunable filter plate with different wave length.
System by the tuning backreflection image projection with different wave length of liquid crystal to the ccd image inductor, can directly handle the clinical sample slide, with cell as observation sample, the micro-image of cell and the spectral signal of each pixel can be obtained simultaneously, phase place and the amplitude information of cell can be got through spectral analysis.System utilizes white light source and low aperture object lens 3 D complex scattering target can be resolved into a plurality of one dimension channels, removed speckle noise, because propagating to produce repeatedly, one dimension light interferes, the reflected signal that one dimension is propagated has very high sensitivity for the refractive index fluctuation of any yardstick, so system has the refractive index fluctuation sensitivity of high spatial domain.The liquid crystal tunable filter plate adopts electrooptical liquid crystal tuning, has substituted standard machinery scanning, has realized that the whole audience measures fast.The ccd image inductor collects the cell image of different wave length, thereby can extract the spectral information of each pixel.Thereby extract the effective nanostructured characteristic of cell by spectrum being carried out Fourier transform, realize the high precision imaging.
Adjust collimation for the white light that white light source is sent, be provided with collimated light path between described white light source and the spectroscope, this collimated light path is arranged in order by condenser, first lens, first diaphragm, second lens and second diaphragm to be formed.
In order to survey the transmission signal of observation sample, system also is provided with second object lens, second catoptron, the 3rd catoptron and removable catoptron, described second object lens are used for receiving the transmitted light information of described observation sample, and this transmitted light information projects described Guan Jingzhong through behind described second catoptron, the 3rd catoptron and the removable catoptron successively.
Described white light source is xenon source, and the white light intensity of sending is big, and degree of coherence is little.
The surface of described liquid crystal tunable filter plate is coated with the anti-reflection film of 400nm～700nm wave band, is used for improving the signal to noise ratio (S/N ratio) of spectral signal.
Remarkable result of the present invention is: light channel structure is simple, control is convenient, system has the refractive index fluctuation sensitivity of high spatial domain, can directly handle the clinical sample slide, realize the high precision imaging of cell nano structure, and utilize the tuning realization whole audience of electrooptical liquid crystal to measure fast, utilize the spectral information of each pixel that simple Fourier transform obtains the ccd image inductor to carry out phase place and the amplitude information that spectral analysis can be obtained observation sample, thereby determine the nanostructured of cell.
Description of drawings
Fig. 1 is system of the present invention design drawing;
Reference numeral:
1 white light source, 2 condensers, 3 first lens, 4 first diaphragms, 5 second lens, 6 second diaphragms, 7 first catoptrons, 8 spectroscopes, 9 first object lens, 10 objective tables, 11 second object lens, 12 second catoptrons, 13 the 3rd catoptrons, 14 removable catoptrons, 15 pipe mirrors, 16 liquid crystal tunable filter plates, 17 ccd image inductors.
Embodiment
Below in conjunction with the drawings and specific embodiments the present invention is described in further detail.
As shown in Figure 1, the low coherent phase microscope in a kind of spatial domain based on liquid crystal tunable filter, comprise white light source 1, spectroscope 8, first object lens 9, objective table 10, first catoptron 7, pipe mirror 15, liquid crystal tunable filter plate 16 and ccd image inductor 17, for the light to white light source is adjusted collimation, also be provided with collimated light path between described white light source 1 and the spectroscope 8, this collimated light path is arranged in order by condenser 2, first lens 3, first diaphragm 4, second lens 5 and second diaphragm 6 to be formed.
White light source 1 adopts xenon source, and the white light intensity of sending is big, and degree of coherence is little.The white light that white light source 1 sends is behind collimated light path, project on the spectroscope 8, this spectroscope 8 projects the white light that receives on first object lens 9, seeing through first object lens 9 at last focuses in the observation sample, first object lens 9 adopt low aperture object lens, utilize white light source and low aperture object lens 3 D complex scattering target can be resolved into a plurality of one dimension channels, removed speckle noise, because propagating to produce repeatedly, one dimension light interferes, the reflected signal that one dimension is propagated has very high sensitivity for the refractive index fluctuation of any yardstick, so system has the refractive index fluctuation sensitivity of high spatial domain.
Described spectroscope 8 is installed between first object lens 9 and first catoptron 7, not only can receive the white light signal that described white light source 1 produces, and the observation sample signal reflex signal that first object lens 9 can be received is transmitted on first catoptron 7, propagate simultaneously in the light path between spectroscope 8 and first object lens 9 incident light and reflected light are arranged, system adopts coaxial reflective-mode, has reduced the influence that absorbs light and external disturbance.Described first catoptron 7 with the signal reflex that receives in pipe mirror 15, described liquid crystal tunable filter plate 16 is installed between pipe mirror 15 and ccd image inductor 17, project on the described ccd image inductor 17 by the micro-image of this liquid crystal tunable filter plate 16 with different wave length, in order to improve the signal to noise ratio (S/N ratio) of spectral signal, the surface of described liquid crystal tunable filter plate 16 is coated with the anti-reflection film of 400nm～700nm wave band.
Owing to adopt xenon source, the diameter of illumination beam is much larger than the size (the cell size is micron order) of observation sample, size and surface curvature for individual cells, incident light can be approximated to be plane wave, wideband low-coherence light source irradiation observation sample by almost plane can obtain the image that the photon by backreflection forms.By the tuber function of described liquid crystal tunable filter plate 16, can collect the cell image of specific wavelength and project in the ccd image inductor 17, handle the data volume that the image information of ccd image inductor 17 outputs can be set up a three-dimensional by data
, wherein
Represent a pixel of observation sample,
Represent the spectral information of this pixel, directly right at last
Carry out spectral analysis.
System adopts Labview software programming control liquid crystal tunable filter plate 16 and ccd image inductor 17 synchronous, because liquid crystal tunable filter plate 16 adopts electrooptical liquid crystals tuning, has substituted standard machinery scanning, can realize that the whole audience measures fast.Liquid crystal is adjustable filter plate 16 with the backreflection image projection of different wave length to ccd image inductor 17, system scans nano particle, the spectral signal of gathering is analyzed, utilize the system spectrum of microslide as a reference, spectral information is carried out normalized, determine the wavelength coverage that needs are handled according to the intensity of spectrum.The spectral information of gathering is removed the high-frequency electronic noise by Butterworth LPF, and filtered spectral signal is carried out Fourier transform, obtains PHASE DISTRIBUTION figure accurately by optimizing algorithm, realizes the calibration of system.Native system can directly be handled the clinical sample slide, with cell as observation sample, can obtain the micro-image of cell and the spectral signal of each pixel simultaneously, can get phase place and the amplitude information of cell through spectral analysis, thereby extract the effective nanostructured characteristic of cell, realize the high precision imaging.
In order to survey the transmission signal of observation sample, obtaining traditional micro-image analyzes, system also is provided with second object lens 11, second catoptron 12, the 3rd catoptron 13 and removable catoptron (14), described second object lens 11 are used for receiving the transmitted light information of described observation sample, this transmitted light information projects in the described pipe mirror 15 through behind described second catoptron 12, the 3rd catoptron 13 and the removable catoptron 14 successively, finally puts in the described ccd image inductor 17 by pipe mirror 15.
In order to reduce aberration to the influence of system, in specific implementation process, described first lens 3, second lens 5 and pipe mirror 15 adopt achromatic doublet, described spectroscope 8 adopts wide spectrum spectroscope, described first object lens 9 and second object lens 11 adopt achromatic objective, and the catoptron in the system also all adopts broadband mirrors, thereby reduces aberration to the influence of system, the degree of accuracy of raising system finally reaches purpose of the present invention.
Claims (4)
1. the coherent phase microscope is hanged down in the spatial domain based on liquid crystal tunable filter, comprise objective table (10), first object lens (9), first catoptron (7), Guan Jing (15) and ccd image inductor (17), it is characterized in that: also comprise white light source (1), spectroscope (8), liquid crystal tunable filter plate (16), second object lens (11), second catoptron (12), the 3rd catoptron (13) and removable catoptron (14), described spectroscope (8) is installed between first object lens (9) and first catoptron (7), this spectroscope (8) is used for receiving the white light signal that described white light source (1) produces, this white light signal sees through described first object lens (9) and focuses in the observation sample on the described objective table (10), the reflection of this observation sample and scattered signal see through described first object lens (9) and spectroscope (8) is incident upon on described first catoptron (7), this first catoptron (7) arrives the signal reflex that receives among the Guan Jing (15), described liquid crystal tunable filter plate (16) is installed between Guan Jing (15) and ccd image inductor (17), is projected on the described ccd image inductor (17) by the micro-image of this liquid crystal tunable filter plate (16) with different wave length;
Described second object lens (11) are used for receiving the transmitted light information of described observation sample, and this transmitted light information projects among the described Guan Jing (15) after passing through described second catoptron (12), the 3rd catoptron (13) and removable catoptron (14) successively.
2. the coherent phase microscope is hanged down in the spatial domain based on liquid crystal tunable filter according to claim 1, it is characterized in that: be provided with collimated light path between described white light source (1) and the spectroscope (8), this collimated light path is arranged in order by condenser (2), first lens (3), first diaphragm (4), second lens (5) and second diaphragm (6) to be formed.
3. the coherent phase microscope is hanged down in the spatial domain based on liquid crystal tunable filter according to claim 1, and it is characterized in that: described white light source (1) is xenon source.
4. the coherent phase microscope is hanged down in the spatial domain based on liquid crystal tunable filter according to claim 1, and it is characterized in that: the surface of described liquid crystal tunable filter plate (16) is coated with the anti-reflection film of 400nm～700nm wave band.
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| CN103808337A (en) * | 2012-11-12 | 2014-05-21 | 中国航空工业第六一八研究所 | White light interference optical sensor noise signal processing method |
| CN103235408A (en) * | 2013-04-24 | 2013-08-07 | 中国科学院上海光学精密机械研究所 | Sb2Te3 nonlinear super-resolution cover glass |
| CN104330363B (en) * | 2014-10-30 | 2016-12-07 | 厦门市美亚柏科信息股份有限公司 | Spectral imaging analysis system, the method using this system imaging and analysis |
| CN105158888B (en) * | 2015-09-29 | 2020-09-11 | 南京理工大学 | Programmable microscope condenser device based on LCD (liquid crystal display) panel and imaging method thereof |
| KR20200074316A (en) * | 2018-12-14 | 2020-06-25 | 삼성전자주식회사 | Spectral system, Optical inspection apparatus and Method for manufacturing the semiconductor device |
| CN109470685A (en) * | 2019-01-10 | 2019-03-15 | 华东师范大学 | A kind of Raman lines scanning Hyperspectral imager of integrated reflection and transmission |
| CN109557074A (en) * | 2019-01-10 | 2019-04-02 | 华东师范大学 | A kind of acquisition method of Raman high spectrum image |
| CN109799222A (en) * | 2019-01-17 | 2019-05-24 | 华东师范大学 | A kind of image-pickup method of Raman face battle array EO-1 hyperion |
| CN113984631A (en) * | 2021-10-12 | 2022-01-28 | 桂林电子科技大学 | Wide-field multispectral fluorescence microscopic imaging method and system based on dynamic speckle illumination |
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| DE4231439A1 (en) * | 1992-09-19 | 1994-03-24 | Leica Mikroskopie & Syst | Illumination device for microscopes |
| US6403947B1 (en) * | 1999-03-18 | 2002-06-11 | Cambridge Research & Instrumentation Inc. | High-efficiency multiple probe imaging system |
| CN1289918A (en) * | 2000-10-08 | 2001-04-04 | 暨南大学 | Method and device for multi-channel spectrophotometric detection of rapid microscope |
| US7408636B2 (en) * | 2002-10-31 | 2008-08-05 | Chemimage Corporation | Method and apparatus for dark field chemical imaging |
| TWI232955B (en) * | 2002-12-30 | 2005-05-21 | Ind Tech Res Inst | Microscopic image apparatus for flat-top intensity distribution |
| CN100470190C (en) * | 2006-03-14 | 2009-03-18 | 清华大学 | Array vertical cavity-surface transmission laser confocal microscopic system |
| CN1837890A (en) * | 2006-03-29 | 2006-09-27 | 哈尔滨工业大学 | Image recovery and pupil filtering type transverse super-resolution confocal microscopic imaging method and apparatus |
| JP2010038800A (en) * | 2008-08-07 | 2010-02-18 | Nagase & Co Ltd | Device and method for observing viable cell |
| CN101726844B (en) * | 2008-10-21 | 2011-11-23 | 财团法人工业技术研究院 | Interference phase difference microscope |
| CN101795339B (en) * | 2010-01-25 | 2014-04-16 | 赖博 | Confocal optical scanner |
| WO2011105618A1 (en) * | 2010-02-26 | 2011-09-01 | 独立行政法人科学技術振興機構 | Microscope device, optical pickup device, and light irradiation device |
| CN101819069B (en) * | 2010-04-23 | 2012-05-23 | 浙江大学 | White light interferometer with fast zero-setting system |
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