CN1158555C - Confocal laser scanning microscope - Google Patents
Confocal laser scanning microscope Download PDFInfo
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- CN1158555C CN1158555C CNB011081376A CN01108137A CN1158555C CN 1158555 C CN1158555 C CN 1158555C CN B011081376 A CNB011081376 A CN B011081376A CN 01108137 A CN01108137 A CN 01108137A CN 1158555 C CN1158555 C CN 1158555C
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- light path
- angle mirror
- mirror combination
- acquisition
- laser scanning
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Abstract
The present invention relates to a laser confocal scanning microscope which comprises a laser excitation light path, a signal detecting light path, a micro positioner, a microscopic optical unit, a signal processing and data converting unit and a computer. The light axis of the laser excitation light path and the light axis of the signal detecting light path partially coincide. The laser excitation light path and the signal detecting light path share an X-Y scanner, a collimating lens and a combined angle mirror composed of two reflecting mirrors which are arranged according to a certain angle. The present invention eliminates factors possibly resulting in the maladjustment of the laser excitation light path and the signal detecting light path and can be stably operated for long time.
Description
Technical field
The present invention relates to a kind of microscope, particularly a kind of confocal laser scanning microscope.
Background technology
Existing confocal laser scanning microscope all has three basic light paths, that is: by laser instrument through reflecting surface to the laser excitation light path that is detected object, by acquisition of signal light path and the micro optical system light path of reflecting surface to detector.The above two constitute the core-scanner head of confocal laser scanning microscope.Because micro optical system generally all is standard commodity of choosing each producer, so the improvement of all confocal laser scanning microscopes and innovation all concentrate on the scanner head.
Chinese patent CN1229927A discloses a kind of confocal laser scanning microscope, and it excites aperture, laser excitation optical system lens in the laser excitation light path, surveys aperture, and the object lens of acquisition of signal optical system are in the acquisition of signal light path.Two system's components and parts in use any loosening and error separately all can cause the disappearance that excites aperture and survey the confocal effect of aperture, thereby instrument can not steady operation.Especially in the time need testing with different optical maser wavelength, credit union's element of two light paths is often wanted several groups and is replaced use, and in frequent replacement operation, must there be error in the simple reflector position; Thereby its emergent ray optical axis can cause the angle error of twice.This is the ubiquitous problem of prior art.
Summary of the invention
The purpose of this invention is to provide and a kind ofly can eliminate laser excitation light path and acquisition of signal light path imbalance factor, guarantee the confocal scan microscope of system's long-term stable operation.
The present invention includes the laser excitation light path, the acquisition of signal light path, micro positioner, the microoptic unit, signal Processing and Date Conversion Unit, computing machine, micro positioner is arranged on the worktable of microoptic unit, its action is by computer control, reflected light that the laser excitation light path excites or fluorescence convert electric signal to through acquisition of signal light path and signal Processing and Date Conversion Unit, the input computer storage, the part optical axis coincidence that it is characterized in that laser excitation light path and acquisition of signal light path, both also have a shared collimation lens and an angle mirror combination that is made of by fixed angle two catoptrons except that having one to shared being subjected to the computer-controlled X-Y scanner.
The present invention has eliminated the factor that may cause laser excitation light path and the imbalance of acquisition of signal light path, but long-term stable operation.Simultaneously, because aforementioned two light path part optical axis coincidences, thereby simplify the structure, reduced cost, make also that accordingly manufacturing process of the present invention and assembling are all simple than prior art.
Description of drawings
Fig. 1 is a structural representation of the present invention.
Fig. 2 is the acquisition of signal light path synoptic diagram of the present invention with hyperchannel detecting function.
Embodiment
As shown in Figure 1, the present invention is made of laser excitation light path, acquisition of signal light path, micro positioner, microoptic unit, signal Processing and Date Conversion Unit and computing machine.Micro positioner is arranged on the worktable of microoptic unit, and it moves by computer control, and reflected light that the laser excitation light path excites or fluorescence convert electric signal to through acquisition of signal light path and signal Processing and Date Conversion Unit, the input computer storage.
Laser excitation optical routing laser instrument 1 and condenser lens 2. smallcolumn diaphragms 3, neutral colour filter combination 4, exciter filter 7, angle mirror combination, collimation lens 9 and the X-Y scanner that are successively set on the same optical axis constitute.The X-Y scanner that the acquisition of signal optical routing sets gradually on same optical axis, collimation lens 9, angle mirror combination, radiation filter 8, smallcolumn diaphragm 17 and detector 18 constitute.Because shared X-Y scanner of above-mentioned two light paths and collimation lens 9 and angle mirror combination, therefore the part optical axis coincidence of two light paths.Simultaneously, press the fixed angle setting owing to constitute two catoptrons 6,7 of angle mirror combination, therefore, be combined to whole components and parts between the sample object point 0 from angle mirror, its any error is equal to the effect of two light paths, can not cause the inconsistent imbalance that brings of two systems.So just eliminated the factor that may cause the imbalance of two light paths.
The shared collimation lens 9 of two light paths can be single or single group lens, also can be a plurality of or many groups.
As shown in Figure 2, the purposes of surveying simultaneously for the wide wavestrip fluorescent image that is fit to medical science, the acquisition of signal light path can add one or more angle mirror combination again and match with it between angle mirror combination and smallcolumn diaphragm 17 detector 18, the transmission output terminal and the reflection output terminal of the angle mirror combination of each increase all are furnished with corresponding radiation filter, its reflection output terminal is furnished with a detector equally, so just can constitute second channel or hyperchannel and survey.
In addition.In order to adapt to biomedicine and multiple use, can settle the coupling of a plurality of different wavelength of laser devices and laser excitation light path in the position of lasing light emitter 1, different wavelength of laser is provided.
Below principle of work of the present invention is described.
Power is that the Argon ion laser 1 of tens milliwatts sends laser, and line focus lens 2 converge to the about tens microns smallcolumn diaphragm in aperture 3, passes neutral colour filter or optical filter combination 4 then, and it is suitable that laser intensity is adjusted to.Laser through overregulating is through exciter filter 7, after picking out the laser (for example 488nm or 514.5nm laser) of specified wavelength, (angle of two catoptrons can be the fixed angle of handled easilies such as 30 °, 45 °, 60 ° to constitute the angle mirrors combination through two catoptrons 5,6 again, present embodiment is 45 °), through collimation lens 9, collimation enters micro optical system 12 for directional light through X-Y scanner two reflectings surface 10,11.Through the object lens of micro optical system 12, laser convergence is to the object point 0 of sample, and this microcobjective can be 40 times, numerical aperture 0.65 or 100 multiple value apertures 1.25 (oil immersion) etc.
From 0 laser that is reflected, or 0 fluorescence that is excited is as flashlight, return through collimation lens 9 along original optical path, pass catoptron 6 and radiation filter 8 in the angle mirror combination, converge to smallcolumn diaphragm 17 places, thereby being detected device 18---photomultiplier is surveyed, and after signal Processing and Date Conversion Unit 19 convert electric signal to, sends into computing machine 16 and stores.
Under computing machine 16 controls, the X-Y scanner is done the scanning vibration, and laser focusing point is scanning vibration fast in this test surface, and then computing machine stores the light signal of all differences on this test surface in turn.Simultaneously, under computing machine 16 control, the micro positioner 15 on the worktable 14 drives samples and makes the depth micro-displacement, and computing machine then stores on the sample light signal on the different depth test surface in turn, thereby computing machine 16 just can recover the three-dimensional image of sample.
Claims (7)
1, a kind of confocal laser scanning microscope, comprise the laser excitation light path, the acquisition of signal light path, micro positioner, the microoptic unit, signal Processing and Date Conversion Unit, computing machine, micro positioner is arranged on the worktable of microoptic unit, the action of micro positioner is by computer control, reflected light that the laser excitation light path excites or fluorescence convert electric signal to through acquisition of signal light path and signal Processing and Date Conversion Unit, the input computer storage, the part optical axis coincidence that it is characterized in that laser excitation light path and acquisition of signal light path, both also have a shared collimation lens and an angle mirror combination that is made of by fixed angle two catoptrons except that shared being subjected to the computer-controlled X-Y scanner.
2, confocal laser scanning microscope as claimed in claim 1 is characterized in that laser excitation optical routing laser instrument (1) and is successively set on condenser lens (2) on the same optical axis, smallcolumn diaphragm (3), neutral colour filter combination (4), exciter filter (7), angle mirror combination, collimation lens (9) and X-Y scanner (10,11) constituting.
3, confocal laser scanning microscope as claimed in claim 1 is characterized in that X-Y scanner (10,11), collimation lens (9), angle mirror combination, radiation filter (8), smallcolumn diaphragm (17) and detector (18) that the acquisition of signal optical routing sets gradually constitute on same optical axis.
4, confocal laser scanning microscope as claimed in claim 3, it is characterized in that the acquisition of signal light path is after the angle mirror combination, can increase an angle mirror combination again, the transmission output terminal and the reflection output terminal of the angle mirror combination that increases all are furnished with corresponding radiation filter, and its reflection output terminal is furnished with a detector (18) equally.
5, confocal laser scanning microscope as claimed in claim 3, it is characterized in that the acquisition of signal light path is after the angle mirror combination, can increase more than one angle mirror combination again, the transmission output terminal and the reflection output terminal of the angle mirror combination of each increase all are furnished with corresponding radiation filter, and its reflection output terminal is furnished with a detector (18) equally.
6,, it is characterized in that collimation lens (9) can be a plurality of lens as claim 2 or 3 described confocal laser scanning microscopes.
7, as claim 1 or 2 or 3 or 4 or 5 described confocal laser scanning microscopes, the angle that it is characterized in that constituting between two catoptrons of angle mirror combination should be the fixed angle of handled easily.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011081376A CN1158555C (en) | 2001-03-16 | 2001-03-16 | Confocal laser scanning microscope |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011081376A CN1158555C (en) | 2001-03-16 | 2001-03-16 | Confocal laser scanning microscope |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1312480A CN1312480A (en) | 2001-09-12 |
| CN1158555C true CN1158555C (en) | 2004-07-21 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011081376A Expired - Fee Related CN1158555C (en) | 2001-03-16 | 2001-03-16 | Confocal laser scanning microscope |
Country Status (1)
| Country | Link |
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| CN (1) | CN1158555C (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175143B (en) * | 2011-02-21 | 2012-11-28 | 哈尔滨工业大学 | Line scanning differential confocal measuring device based on light path of pillar lens |
| CN102566023B (en) * | 2012-01-11 | 2015-09-30 | 麦克奥迪实业集团有限公司 | A kind of digital slide real time scanning automatic focusing system and method thereof |
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2001
- 2001-03-16 CN CNB011081376A patent/CN1158555C/en not_active Expired - Fee Related
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| Publication number | Publication date |
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| CN1312480A (en) | 2001-09-12 |
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| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20040721 |