CN105424189A - Scanning type multifunctional microscopic-spectral imaging method - Google Patents
Scanning type multifunctional microscopic-spectral imaging method Download PDFInfo
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- CN105424189A CN105424189A CN201510885021.0A CN201510885021A CN105424189A CN 105424189 A CN105424189 A CN 105424189A CN 201510885021 A CN201510885021 A CN 201510885021A CN 105424189 A CN105424189 A CN 105424189A
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- light
- sample
- beam splitter
- fluorescence
- electric
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- 238000000034 method Methods 0.000 title claims abstract 4
- 238000000701 chemical imaging Methods 0.000 title abstract description 4
- 230000003287 optical effect Effects 0.000 claims abstract description 31
- 238000001069 Raman spectroscopy Methods 0.000 claims abstract description 28
- 238000001228 spectrum Methods 0.000 claims abstract description 24
- 230000003595 spectral effect Effects 0.000 claims abstract 4
- 238000003384 imaging method Methods 0.000 claims description 28
- 238000002381 microspectrum Methods 0.000 claims description 21
- 238000010276 construction Methods 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000005272 metallurgy Methods 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/2823—Imaging spectrometer
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/44—Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/44—Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
- G01J3/4406—Fluorescence spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
- G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
- G01J3/28—Investigating the spectrum
- G01J3/44—Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
- G01J3/4412—Scattering spectrometry
Abstract
The invention provides a scanning type multifunctional microscopic-spectral imaging method, which adopts equipment comprising a pulse laser light source or continuous spectrum light source, a computer, a three-dimensional electric precise translation object stage, a photoelectric detector, a heat mirror-beam splitter switching device, an optical scanning device, objective lenses, a spectrograph, beam splitters, mechanical optical switches, reflectors and a data acquisition system. The scanning type multifunctional microscopic-spectral imaging method comprises the steps that: the computer controls the optical scanning device or the three-dimensional electric precise translation object stage to perform point-by-point scanning, a CCD camera in the spectrograph acquires spectral information of backward or forward fluorescent light (Raman light) and scattered light of a sample synchronously, the data acquisition system can record the spectral information of the spectrograph at each scanning point, the computer generates a microscopic image of the fluorescent light (Raman light) or the scattered light of the sample after finishing data acquisition in a previous region to be tested on a sample plane (xy plane), and the computer generates microscopic images at different spectral band and a three-dimensional microscopic-spectral image of the sample.
Description
Technical field
The present invention relates to micro-imaging field, be specifically related to a kind of scan-type Multifunctional display low-light spectrum imaging method.
Background technology
Micro-imaging technique is long-standing, and it is the most often applied to the sample that observation naked eyes cannot be observed, and application comprises the fields such as biomedicine, chemistry, physics, metallurgy, measurement, has unique position in the development in a lot of field.Meanwhile, micro-imaging technique is also in constantly development is improved, and the field formula microscope from, to confocal microscope afterwards, it has had important breakthrough in enlargement factor, noise resistance etc.But, no matter be traditional field formula microscope, or confocal microscope, be all utilize the light intensity of sample scattering light to imaging of samples, traditional micro-imaging can only obtain the structural images of sample.Even if fluorescence imaging also just obtains the micro-image of a certain wave band fluorescent intensity of sample, the complete spectrum characteristic information of sample can not be obtained.And traditional micro imaging system can not realize multiple imaging function simultaneously.
Summary of the invention
For the deficiency of traditional microscopic system, in order to obtain the more characteristic information of sample, the present invention proposes a kind of scan-type Multifunctional display low-light spectrum imaging method combined with microtechnic by spectral imaging technology, by recording fluorescence (Raman light) spectrum that excites in the sample to which or scattered light spectrum, realize that fluorescence (Raman light) to sample is micro-, fluorescence (Raman light) microspectrum or scattered light is micro-and microspectrum imaging.
Technical scheme of the present invention is as follows:
The invention provides a kind of scan-type Multifunctional display low-light spectrum imaging method, include computing machine, three-D electric accurate translation stage, the pulsed laser light source of collimation, photodetector, the continuous spectrum light source of collimation, heat mirror-beam splitter switching device shifter, light scanning apparatus, object lens, spectrometer, beam splitter, mechanical optical switch, catoptron, data acquisition system (DAS).
Computing machine connects three-D electric accurate translation stage, pulsed laser light source, photodetector, light scanning apparatus and data acquisition system (DAS) simultaneously, and data acquisition system (DAS) connects the CCD camera of spectrometer.The pulse laser that pulsed laser light source sends is divided into two bundles after beam splitter, a branch ofly by photoelectric detector, electric signal passed to computing machine; Another bundle is divided into two bundles again by beam splitter, a branch of sample on mechanical optical switch, heat mirror-beam splitter switching device shifter, light scanning apparatus, the accurate translation stage of object lens arrival three-D electric; The sample of another bundle on mechanical optical switch, catoptron, the accurate translation stage of object lens arrival three-D electric.The light that continuous spectrum light source sends reflects through catoptron, is divided into two bundles by through beam splitter, a branch of sample on mechanical optical switch, heat mirror-beam splitter switching device shifter, light scanning apparatus, the accurate translation stage of object lens arrival three-D electric.The sample of another bundle on mechanical optical switch, catoptron, the accurate translation stage of object lens arrival three-D electric.
Advantageous Effects of the present invention is:
The present invention is when unbalanced pulse LASER Light Source, trigger pip is provided to data acquisition system (DAS) by photodetector, open in two mechanical optical switches, by the accurate translation stage scanning of light scanning apparatus or three-D electric, sample can be obtained dorsad or the fluorescence of forward direction (Raman light) is micro-and fluorescence (Raman light) microspectrum image.
The present invention, when opening continuous spectrum light source, by opening in two mechanical optical switches, by the accurate translation stage scanning of light scanning apparatus or three-D electric, can obtain sample dorsad or the spectrum picture that the scattered light of forward direction is micro-and scattered light is micro-.
Accompanying drawing explanation
Fig. 1 is system construction drawing of the present invention.
Fig. 2 be gather dorsad fluorescence (Raman light) time, the structural drawing of system.
Fig. 3 is when gathering forward direction fluorescence (Raman light), the structural drawing of system.
Fig. 4 is when gathering backscattering light, the structural drawing of system.
Fig. 5 is when gathering forward scattering light, the structural drawing of system.
[reference numeral explanation] 1. computing machine; 2. the accurate translation stage of three-D electric; 3. pulsed laser light source; 4. photodetector; 5. spectrometer; 6. beam splitter; 7. beam splitter; 8. heat mirror-beam splitter switching device shifter; 9. light scanning apparatus; 10. object lens; 11. mechanical optical switches; 12. catoptrons; 13. catoptrons; 14. object lens; 15. continuous spectrum light sources; 16. object lens; 17. mechanical optical switches; 18. data acquisition system (DAS)s.
Embodiment
Below in conjunction with accompanying drawing, the invention will be further described.
As shown in Figure 1, scan-type Multifunctional display low-light spectrum imaging system of the present invention comprises computing machine 1, three-D electric accurate translation stage 2, pulsed laser light source 3, photodetector 4, spectrometer 5, beam splitter 6,7, heat mirror-beam splitter switching device shifter 8, light scanning apparatus 9, object lens 10,14, mechanical optical switch 11,17, catoptron 12,13,16, continuous spectrum light source 15, data acquisition system (DAS) 18.
As shown in Figure 1, computing machine 1 connects three-D electric accurate translation stage 2, pulsed light laser light source 3, photodetector 4, light scanning apparatus 9 and data acquisition system (DAS) 18 simultaneously, and data acquisition system (DAS) 18 connects the CCD camera of spectrometer 5.The pulse laser that pulsed laser light source 3 sends is divided into two bundles after beam splitter 6, is a branch ofly received by photodetector 4 and electric signal is passed to computing machine 1; Another bundle is divided into two bundles again by beam splitter 7, a branch of sample arrived through mechanical optical switch 17, heat mirror-beam splitter switching device shifter 8, light scanning apparatus 9, object lens 10 on the accurate translation stage 2 of three-D electric; Another bundle arrives the sample on the accurate translation stage 2 of three-D electric through mechanical optical switch 11, catoptron 12,13, object lens 14.The light that continuous spectrum light source 15 sends reflects through scintilloscope 16, is divided into two bundles via beam splitter 7, a branch of sample arrived through mechanical optical switch 17, heat mirror-beam splitter switching device shifter 8, light scanning apparatus 9, object lens 10 on the accurate translation stage 2 of three-D electric.Another bundle arrives the sample on the accurate translation stage 2 of three-D electric through mechanical optical switch 11, catoptron 12,13, object lens 14.
Scan-type Multifunctional display low-light spectrum imaging method of the present invention can realize two kinds of functions, and often kind of function has two kinds of mode of operations respectively.
1, micro-, fluorescence (Raman light) the microspectrum imaging function of fluorescence (Raman light)
In the system construction drawing shown in Fig. 1, open pulsed laser light source 3, open photodetector 4, close mechanical optical switch 11, open mechanical optical switch 17, heat mirror-beam splitter switching device shifter 8 is arranged on heat mirror position.To be scanned by light scanning apparatus or the scanning of electric precision translation stage realizes that fluorescence (Raman light) is micro-, fluorescence (Raman light) microspectrum imaging function, system is reduced to as shown in Figure 2, now, the fluorescence of fluorescence (Raman light) (Raman light) is micro-, fluorescence (Raman light) microspectrum imaging pattern gathering dorsad for system works.
In the system construction drawing shown in Fig. 1, open pulsed laser light source 3, open photodetector 4, open mechanical optical switch 11, close mechanical optical switch 17, heat mirror-beam splitter switching device shifter 8 is arranged on heat mirror position.Realize that fluorescence (Raman light) is micro-, fluorescence (Raman light) microspectrum imaging function by electric precision translation stage 2 scanning, system simplifies as shown in Figure 3, now, micro-, fluorescence (Raman light) the microspectrum imaging pattern of the system works fluorescence (Raman light) that gathers fluorescence (Raman light) at forward direction.
2, scattered light micro-imaging, scattered light microspectrum imaging function
In the system construction drawing shown in Fig. 1, open continuous spectrum light source 15, close mechanical optical switch 11, open mechanical optical switch 17, heat mirror-beam splitter switching device shifter 8 is arranged on beam splitter position.Scanned by light scanning apparatus or electric precision translation stage scanning realize scattered light micro-imaging, scattered light microspectrum imaging function, system is reduced to as shown in Figure 3, now, system works, scattered light microspectrum imaging pattern micro-at the scattered light of collection of scattered light dorsad.
In the system construction drawing shown in Fig. 1, open continuous spectrum light source 15, open mechanical optical switch 11, close mechanical optical switch 17, heat mirror-beam splitter switching device shifter 8 is arranged on beam splitter position.Realize that scattered light is micro-, scattered light microspectrum imaging function by electric precision translation stage 2 scanning, system simplifies as shown in Figure 4, and now, system works is micro-at the scattered light of forward direction collection of scattered light, scattered light microspectrum imaging pattern.
Above-described is only the preferred embodiment of the present invention, the invention is not restricted to above embodiment.Be appreciated that the oher improvements and changes that those skilled in the art directly derive without departing from the basic idea of the present invention or associate, all should think and be included within protection scope of the present invention.
Claims (2)
1. a scan-type Multifunctional display low-light spectrum imaging method, it is characterized in that: the accurate translation stage (2) of computing machine (1), three-D electric, pulsed laser light source (3), photodetector (4), continuous spectrum light source (15), heat mirror-beam splitter switching device shifter (8), light scanning apparatus (9), object lens (10,14), spectrometer (5), beam splitter (6,7), mechanical optical switch (11,17), catoptron (12,13,16), data acquisition system (DAS) (18).
Computing machine (1) connects the accurate translation stage (2) of three-D electric, pulsed laser light source (3), photodetector (4), light scanning apparatus (9) and data acquisition system (DAS) (18) simultaneously, and data acquisition system (DAS) (18) connects the CCD camera of spectrometer (5).The pulse laser that pulsed laser light source (3) sends is divided into two bundles after beam splitter (6), a branch of by photodetector (4) receive and electric signal is passed to computing machine (1); Another bundle is divided into two bundles again by beam splitter (7), a branch of sample on mechanical optical switch (17), heat mirror-beam splitter switching device shifter (8), light scanning apparatus (9), the accurate translation stage (2) of object lens (10) arrival three-D electric; The sample of another bundle on mechanical optical switch (11), catoptron (12,13), the accurate translation stage (2) of object lens (14) arrival three-D electric.The light that continuous spectrum light source (15) sends reflects through catoptron (16), two bundles are divided into, a branch of sample on mechanical optical switch (17), heat mirror-beam splitter switching device shifter (8), light scanning apparatus (9), the accurate translation stage (2) of object lens (10) arrival three-D electric via beam splitter (7).The sample of another bundle on mechanical optical switch (11), catoptron (12,13), the accurate translation stage (2) of object lens (14) arrival three-D electric.
2. a scan-type Multifunctional display low-light spectrum imaging method, is characterized in that:
1) by adopting pulsed laser light source or continuous spectrum light source to obtain respectively, the fluorescence (Raman light) of sample is micro-, fluorescence (Raman light) microspectrum, scattered light are micro-and microspectrum image.
2) combinationally using by mechanical optical switch (11,17), forward direction, the dorsad fluorescence (Raman light) that can obtain sample as required are respectively micro-, fluorescence (Raman light), scattered light microspectrum image.
3) by adopting light scanning apparatus (9) and the accurate translation stage (2) of three-D electric, photoscanning or Sample Scan can be realized according to actual needs.
4) gathered by spectrometer (5), data acquisition system (DAS) (18) record, computing machine (1) to the fluorescence (Raman light) of sample of record or the spectral information process of scattered light, can realize that fluorescence (Raman light) is micro-, fluorescence (Raman light) microspectrum, scattered light are micro-and microspectrum image.
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| CN201510885021.0A CN105424189B (en) | 2015-12-04 | 2015-12-04 | A kind of multi-functional microspectrum imaging system of scan-type |
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| CN201510885021.0A CN105424189B (en) | 2015-12-04 | 2015-12-04 | A kind of multi-functional microspectrum imaging system of scan-type |
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| CN105424189B CN105424189B (en) | 2017-11-07 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106248539A (en) * | 2016-09-22 | 2016-12-21 | 中国矿业大学 | A kind of rotary-ferrograph spectral slice photo densitometry analyzes system and method |
| CN106769881A (en) * | 2016-12-16 | 2017-05-31 | 中国科学院工程热物理研究所 | A kind of thermal conductivity scanning system that heat reflection technology is detected based on pumping |
| CN108627494A (en) * | 2018-05-09 | 2018-10-09 | 吉林大学 | A kind of system for the imaging of fast two-dimensional Raman spectroscopy scans |
| CN109724962A (en) * | 2017-10-27 | 2019-05-07 | 中国科学院大连化学物理研究所 | A kind of more acquisition devices and its acquisition method for chiral Raman spectrometer |
| CN110441235A (en) * | 2019-09-13 | 2019-11-12 | 中国科学院新疆理化技术研究所 | A kind of Multiple modes coupling original position microspectrum imaging system |
| CN110715732A (en) * | 2019-10-17 | 2020-01-21 | 北京理工大学 | Multifunctional Stokes-Mueller imaging and spectrum detection system and detection method |
| CN111220615A (en) * | 2019-10-29 | 2020-06-02 | 怀光智能科技（武汉）有限公司 | Inclined three-dimensional scanning microscopic imaging system and method |
| CN114879002A (en) * | 2022-05-07 | 2022-08-09 | 北京科技大学 | Single-pixel image recognition system based on Van der Waals photoelectric detector |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106248539A (en) * | 2016-09-22 | 2016-12-21 | 中国矿业大学 | A kind of rotary-ferrograph spectral slice photo densitometry analyzes system and method |
| CN106769881A (en) * | 2016-12-16 | 2017-05-31 | 中国科学院工程热物理研究所 | A kind of thermal conductivity scanning system that heat reflection technology is detected based on pumping |
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| CN111220615A (en) * | 2019-10-29 | 2020-06-02 | 怀光智能科技（武汉）有限公司 | Inclined three-dimensional scanning microscopic imaging system and method |
| CN114879002A (en) * | 2022-05-07 | 2022-08-09 | 北京科技大学 | Single-pixel image recognition system based on Van der Waals photoelectric detector |
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