CN205620304U - Two -photon fluorescence microscope's multimode scanning device - Google Patents
Two -photon fluorescence microscope's multimode scanning device Download PDFInfo
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- CN205620304U CN205620304U CN201521128944.3U CN201521128944U CN205620304U CN 205620304 U CN205620304 U CN 205620304U CN 201521128944 U CN201521128944 U CN 201521128944U CN 205620304 U CN205620304 U CN 205620304U
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- input path
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- galvanometer
- photon fluorescence
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Abstract
The utility model discloses a two -photon fluorescence microscope's multimode scanning device, this scanning device includes: first laser instrument for launch first laser source, an input path includes: the first galvanometer type mirror that shakes, the 2nd input path includes: the resonance type mirror that shakes, the emitting light path includes: second galvanometer type shake mirror and microcobjective, first rotation speculum for feed through a first laser instrument and an input path or the 2nd input path, the second rotates the speculum for intercommunication emitting light path and an input path or the 2nd input path, first laser source switches over the selection as the incident light through first rotation speculum and second rotation speculum, and the selection incident light gets into the emitting light path or gets into the emitting light path through the 2nd input path through an input path. The utility model relates to a two -photon fluorescence microscope's multimode scanning device is with low costs, efficient, the precision is good, and scanning of the real -time switching overall situation and partial sweep are switched over to the user of being convenient for.
Description
Technical field
This utility model belongs to microscopy field, is specifically related to a kind of two-photon fluorescence micro-
The multi-modal scanning means of mirror.
Background technology
Two-photon fluorescence micro-imaging technique utilize near infrared femtosecond laser as light source, permissible
Thicker biological tissue is carried out biology microscope imaging, have that imaging depth is big, photic damage is little,
Spatial resolution and contrast high, the photostimulation simultaneously coordinating another wavelength laser again is real
Test, may be used for studying kinetic characteristic and the molecule dynamic of protein in living cells.
Scanning system can be generally divided into two kinds: a kind of mode is that displacement based on nanoscale is put down
Platform, is scanned sample by the most mobile sample being fixed on sample stage, another kind side
Formula is to use vibration mirror scanning, with the use of scanning lens and tube lens, by two axle galvanometers
Deflection realizes the scanning of the two dimensional surface to sample.
Two ways is respectively provided with different features, and nanometer displacement platform precision is high, but speed is slow,
Range of observation is little, it is adaptable to material science, if being applied to biomedical sector, this
Method has its shortcoming, is mainly reflected in scanning speed the slowest, it is impossible to observation is complete for a long time
Motor process.And in some samples that cannot fix, such as the sample in the solution of suspending,
Displacement platform moving process can produce relative displacement, the particularly XY in level with sample put down
In face, imaging can produce the most serious conditions of streaking.These all have a strong impact on its image quality,
Limit the application at biomedical sector.
Use vibration mirror scanning the most with the obvious advantage, the shortcoming which obviating above-mentioned nanometer displacement platform,
And range of observation is big, sweep limits can be entered by galvanometer is loaded different voltage
Row accurately controls, and image taking speed is substantially better than nanometer displacement platform simultaneously.Scanning galvanometer is divided into again two
Kind, one is galvanometer type galvanometer, and it drives coil therein according to the size of input voltage,
In magnetic field, produce moment, thus drive galvanometer to rotate a certain angle so that incident illumination is formed
The line scanning in one-dimensional direction.Another kind is resonance type galvanometer, under the driving of voltage, carries out non-
Linear simple harmonic motion, adjusts sweep limits according to different magnitudes of voltage, and its speed is considerably beyond inspection
Flowmeter galvanometer, rate of scanning typically can reach 8KHz.
But, traditional galvanometer scanning device structure unification, use single galvanometer type to shake
Mirror or resonance type galvanometer, be not easy to user and all scan the switching with partial sweep.
Therefore one low cost, efficiency are high, precision is good, it is simple to user is all scanned and local
The microscopical multi-modal scanning means of two-photon fluorescence of the switching of scanning urgently proposes.
Utility model content
In order to solve above-mentioned technical problem, the utility model proposes a kind of two-photon fluorescence and show
The multi-modal scanning means of micro mirror, this scanning means low cost, efficiency are high, precision is good, it is simple to
User switches real-time switching whole scan and partial sweep.
In order to achieve the above object, the technical solution of the utility model is as follows:
A kind of microscopical multi-modal scanning means of two-photon fluorescence includes: the first laser instrument, uses
In launching the first lasing light emitter；First input path, including: the first galvanometer type galvanometer；Second
Input path, including: resonance type galvanometer；Emitting light path, including: the second galvanometer type galvanometer
And microcobjective；First rotates reflecting mirror, for connection the first laser instrument and the first incident illumination
Road or the second input path；Second rotates reflecting mirror, is used for connecting emitting light path and the first incidence
Light path or the second input path；First lasing light emitter as incident illumination through first rotate reflecting mirror and
Second rotates reflecting mirror switches over selection, selects incident illumination to enter out through the first input path
Penetrate light path or enter emitting light path through the second input path.
The microscopical multi-modal scanning means simple in construction of this utility model one two-photon fluorescence,
The first galvanometer type galvanometer, resonance type galvanometer and the second galvanometer type galvanometer is used to realize sweeping
Retouching, resonance type vibration mirror scanning frequency is high, and galvanometer type vibration mirror scanning is more accurate.And use first
Rotate reflecting mirror and light path can effectively be switched over by the second rotation reflecting mirror, use resonance type
Galvanometer carries out whole scan observation to whole sample, uses the first galvanometer type galvanometer to be used for playing a game
Region, portion carries out finer scanning, meets the polymorphic demand of user, operation of being more convenient for
Personnel carry out photostimulation experiment to sample.And this utility model one two-photon fluorescence is microscopical
In multi-modal scanning means, the first input path and emitting light path form a complete optical path, and second enters
Penetrating light path and form a complete optical path with emitting light path, its two light paths share emitting light path, more save
Cost-saving.
On the basis of technique scheme, also can do and improve as follows:
As preferred scheme, the first input path also includes: the first scanning lens and first
Tube lens, incident illumination sequentially passes through the first galvanometer type galvanometer, the first scanning lens and
One tube lens enters emitting light path.
Above-mentioned preferred scheme, the first scanning lens and the first tube lens is used to be swept by one dimensional line
The laser beam retouched is imaged on same burnt product face, and laser beam is carried out certain collimation and expansion
Bundle.
As preferred scheme, the second input path also includes: the second scanning lens and second
Tube lens, incident illumination sequentially passes through resonance type galvanometer, the second scanning lens and the second lens barrel
Lens enter emitting light path.
Above-mentioned preferred scheme, the second scanning lens and the second tube lens is used to be swept by one dimensional line
The laser beam retouched is imaged on same burnt product face, and laser beam is carried out certain collimation and expansion
Bundle.
As preferred scheme, emitting light path also includes: the 3rd scanning lens and trinocular tube
Lens, the incident illumination entering emitting light path sequentially passes through the second galvanometer type galvanometer, the 3rd sweeps
Retouch lens and trinocular tube lens, be incident upon microcobjective.
Using above-mentioned preferred scheme, one dimensional line is swept by the 3rd scanning lens and trinocular tube lens
The laser beam retouched is imaged on same burnt product face, and laser beam is carried out certain collimation and expansion
Bundle.
As preferred scheme, first rotates reflecting mirror and second rotates reflecting mirror respectively with the
One driving means and the second driving means are in transmission connection, and the first driving means drives the first rotation
Reflecting mirror rotates, and the second driving means drives the second rotation reflecting mirror to rotate.
Use above-mentioned preferred scheme, it is simple to the first input path is carried out with the second input path
Switching automatically.
As preferred scheme, the microscopical multi-modal scanning means of two-photon fluorescence also wraps
Including: dichroic mirror, the first lasing light emitter enters the first input path after dichroic mirror transmission
Or second input path.
Use above-mentioned preferred scheme, the light beam of different wave length is carried out the sieve of transmission and refraction
Choosing.
As preferred scheme, the microscopical multi-modal scanning means of two-photon fluorescence also wraps
Including: second laser, for launching the second lasing light emitter, the second lasing light emitter is through dichroic mirror
The first input path or the second input path is entered after reflection, and through dichroic mirror transmission
First lasing light emitter is consistent with the second lasing light emitter light path reflected through dichroic mirror.
Use above-mentioned preferred scheme, it is ensured that the stability of its light path.
As preferred scheme, the microscopical multi-modal scanning means of two-photon fluorescence also wraps
Include: second laser, for launching the second lasing light emitter；Dichroic mirror, the first lasing light emitter warp
Crossing dichroic mirror transmission, the second lasing light emitter reflects through dichroic mirror, saturating through dichroic mirror
The first lasing light emitter penetrated carries out closing bundle with the second lasing light emitter through dichroic mirror reflection.
Use above-mentioned preferred scheme, can effectively realize sample is carried out photostimulation experiment.
As preferred scheme, the first laser instrument is femtosecond pulse laser.
Use above-mentioned preferred scheme, effective.
As preferred scheme, second laser is optical fiber laser.
Use above-mentioned preferred scheme, effective.
Accompanying drawing explanation
A kind of two-photon fluorescence that Fig. 1 provides for this utility model embodiment is microscopical multi-modal
One of structural representation of scanning means.
The structural representation of the first rotation reflecting mirror that Fig. 2 provides for this utility model embodiment.
The structural representation of the second rotation reflecting mirror that Fig. 3 provides for this utility model embodiment.
A kind of two-photon fluorescence that Fig. 4 provides for this utility model embodiment is microscopical multi-modal
The index path of the first mode scanning of scanning means.
A kind of two-photon fluorescence that Fig. 5 provides for this utility model embodiment is microscopical multi-modal
The index path of the second mode scanning of scanning means.
A kind of two-photon fluorescence that Fig. 6 provides for this utility model embodiment is microscopical multi-modal
The two of the structural representation of scanning means.
A kind of two-photon fluorescence that Fig. 7 provides for this utility model embodiment is microscopical multi-modal
The index path of the 3rd modality scans of scanning means.
Wherein: 1. the first laser instrument, 2. dichroic mirror, 3. first rotate reflecting mirror, 4. second
Laser instrument, 5. resonance type galvanometer, 6. the second tube lens, 7. the second scanning lens, 8. second turn
Dynamic reflecting mirror, 9. the first tube lens, 10. the first scanning lens, 11. first galvanometer type galvanometers,
12. second galvanometer type galvanometers, 13. the 3rd scanning lenses, 14. trinocular tube lens, 15. show
Speck mirror, 16. sample stages, 17. first driving means, 18. second driving means.
Detailed description of the invention
Describe preferred implementation of the present utility model below in conjunction with the accompanying drawings in detail.
In order to reach the purpose of this utility model, a kind of two-photon fluorescence is microscopical multi-modal to be swept
In the some of them embodiment of imaging apparatus,
As Figure 1-3, a kind of microscopical multi-modal scanning means of two-photon fluorescence includes:
First laser instrument 1, for launching the first lasing light emitter, in an embodiment, the first laser instrument
For sending out the femtosecond pulse laser setting 850nm wavelength；
Dichroic mirror 2, the first lasing light emitter carries out transmission through dichroic mirror 2；
First input path, including: first galvanometer type galvanometer the 11, first scanning lens 10
With the first tube lens 9；
Second input path, including: resonance type galvanometer the 5, second scanning lens 6 and the second mirror
Cylinder lens 7；
Emitting light path, including: the second galvanometer type galvanometer 12, the 3rd scanning lens 13 and
Trinocular tube lens 14 and microcobjective 15, the position phase of microcobjective 15 and sample stage 16
Corresponding；
First rotates reflecting mirror 3, for connection the first laser instrument 1 and the first input path or the
Two input paths, first rotates reflecting mirror 3 and the first driving means 17, the first driving means
17 drive the first rotation reflecting mirror 3 to rotate 90 degree, first rotate reflecting mirror in the present embodiment
3 is silvered mirror；
Second rotates reflecting mirror 8, is used for connecting emitting light path and the first input path or second and enters
Penetrating light path, second rotates reflecting mirror 8 is in transmission connection with the second driving means 18, and second drives
Device 18 drives the second rotation reflecting mirror 8 to rotate 90 degree, second rotates instead in the present embodiment
Penetrate mirror 8 for silvered mirror.
As shown in Figure 4, in first mode scanning, the first driving means 17 drives the first rotation anti-
Penetrate mirror 3 to rotate, the first rotation reflecting mirror 3 is placed in scanning optical path, the second driving means 18
Drive the second rotation reflecting mirror 8 to rotate, the second rotation reflecting mirror 8 is not interposing in scanning optical path.
The laser beam of 850nm wavelength that first laser instrument 1 is launched warp again after dichroic mirror 2 transmission
Cross the first rotation reflecting mirror 3 to reflect, incide on resonance type galvanometer 5, by galvanometer axle
The upset of reflecting mirror, forms one-dimensional linear scanning in X-direction.Anti-through resonance type galvanometer 5
Laser beam after penetrating sequentially passes through the second scanning lens 6 and the second tube lens 7, by one-dimensional
The laser beam of line scanning is imaged on same burnt product face.Through the second scanning lens 6 and second
After tube lens 7, laser beam incides on the second galvanometer type galvanometer 12, passes through galvanometer
The upset of the reflecting mirror on axle, forms one-dimensional linear scanning in Y direction, thus and resonance type
Galvanometer 5 coordinates, and forms two dimensional surface scanning, and plane reflection light enters the 3rd scanning lens therewith
13, trinocular tube lens 14 and microcobjective 15, by laser beam focusing at sample stage 16
On, the two-photon excitation of complete paired samples.
As it is shown in figure 5, in second mode scanning, the first driving means 17 drives the first rotation anti-
Penetrate mirror 3 to rotate, the first rotation reflecting mirror 3 is not interposing in scanning optical path, the second driving means
18 drive the second rotation reflecting mirror 8 to rotate, and are placed in scanning optical path by the second rotation reflecting mirror 8.
The laser beam of 850nm wavelength that first laser instrument 1 is launched after dichroic mirror 2 transmission through
The X-direction that is modulated at of the first galvanometer type galvanometer 11 forms one-dimensional linear scanning, then passes through
Second rotates reflecting mirror 8 reflects, and incides on the second galvanometer type galvanometer 12, passes through
The cooperation of two galvanometer type galvanometers, forms two dimensional surface scanning.In this scanning mode, due to
The controlled range of galvanometer type galvanometer, can facilitate user that sample large area scanning is being observed it
After part interested carry out the most careful observation.
The microscopical multi-modal scanning means simple in construction of this utility model one two-photon fluorescence,
Use the first galvanometer type galvanometer 11, resonance type galvanometer 5 and the second galvanometer type galvanometer 12
Realizing scanning, resonance type galvanometer 5 rate of scanning is high, and galvanometer type vibration mirror scanning is more accurate.
And use the first rotation reflecting mirror 3 and the second rotation reflecting mirror 8 effectively light path to be cut
Change, use resonance type galvanometer 5 that whole sample carries out whole scan observation, use the first inspection stream
Meter type galvanometer 11 carries out finer scanning for localized region, meets user's multiform
The demand of state, the operator that are more convenient for carry out photostimulation experiment to sample.And this utility model
Plant the first input path and emitting light path in the microscopical multi-modal scanning means of two-photon fluorescence
Forming a complete optical path, the second input path and emitting light path form a complete optical path, its two
Light path shares emitting light path, more cost-effective.All it is taken up in order of priority after each galvanometer and is furnished with
Surface sweeping lens and tube lens combination, the laser beam that one dimensional line scans is imaged on same Jiao by it
On product face, and laser beam is carried out certain collimation and expands.
As shown in Figure 6, in order to optimize implementation result of the present utility model further, additionally
In some embodiments, remaining feature technology is identical, and difference is, two-photon fluorescence shows
The multi-modal scanning means of micro mirror also includes: second laser 4, for launching the second lasing light emitter,
Second lasing light emitter enters the first input path or the second incident illumination after dichroic mirror 2 reflects
Road, and through dichroic mirror 2 transmission the first lasing light emitter with through the of dichroic mirror 2 reflection
Two laser sources light path is consistent.In an embodiment, second laser 4 sets 1064nm wavelength for sending out
Optical fiber laser.
As it is shown in fig. 7, in the 3rd modality scans, the first driving means 17 drives the first rotation anti-
Penetrate mirror 3 to rotate, the first rotation reflecting mirror 3 is not interposing in scanning optical path, the second driving means
18 drive the second rotation reflecting mirror 8 to rotate, and are placed in scanning optical path by the second rotation reflecting mirror 8.
The laser beam of 1064nm wavelength is reflected in scanning optical path through dichroic mirror 2.Logical
Cross the cooperatively forming given zone of the first galvanometer type galvanometer 11 and the second galvanometer type galvanometer 12
The two dimensional surface scanning in territory so that it is real that user can carry out photostimulation to the specific region of sample
Test.
Merit attention and be, due to through the first lasing light emitter of dichroic mirror 2 transmission and process
Second lasing light emitter light path of dichroic mirror 2 reflection is consistent, and light beam conduct does not occur deviation.Therefore
In the present embodiment, second mode scanning can be carried out with the 3rd modality scans simultaneously, utilizes the 3rd
Modality scans realizes photostimulation experiment, and the carrying out utilizing second mode to scan is observed, therefore permissible
Effectively observing the photostimulation impact on the biological nature of sample, experimental data is more accurate, halfway without
Organism need to be carried out the change of position.
Novel preferred implementation, it is noted that for those of ordinary skill in the art
For, on the premise of creating design without departing from this utility model, it is also possible to make some changes
Shape and improvement, these broadly fall into protection domain of the present utility model.
Claims (9)
1. the microscopical multi-modal scanning means of two-photon fluorescence, it is characterised in that including:
First laser instrument, for launching the first lasing light emitter；
First input path, including: the first galvanometer type galvanometer；
Second input path, including: resonance type galvanometer；
Emitting light path, including: the second galvanometer type galvanometer and microcobjective；
First rotates reflecting mirror, is used for connecting described first laser instrument and described first input path or described second input path；
Second rotates reflecting mirror, is used for connecting described emitting light path and described first input path or described second input path；
Described first lasing light emitter rotates reflecting mirror as incident illumination through the first rotation reflecting mirror and described second and switches over selection, selects described incident illumination enter described emitting light path through described first input path or enter described emitting light path through described second input path.
The microscopical multi-modal scanning means of two-photon fluorescence the most according to claim 1, it is characterized in that, described first input path also includes: the first scanning lens and the first tube lens, and incident illumination sequentially passes through described first galvanometer type galvanometer, described first scanning lens and described first tube lens and enters described emitting light path.
The microscopical multi-modal scanning means of two-photon fluorescence the most according to claim 1, it is characterized in that, described second input path also includes: the second scanning lens and the second tube lens, and incident illumination sequentially passes through described resonance type galvanometer, described second scanning lens and described second tube lens and enters described emitting light path.
The microscopical multi-modal scanning means of two-photon fluorescence the most according to claim 1, it is characterized in that, described emitting light path also includes: the 3rd scanning lens and trinocular tube lens, the incident illumination entering described emitting light path sequentially passes through described second galvanometer type galvanometer, described 3rd scanning lens and described trinocular tube lens, is incident upon described microcobjective.
5. according to the microscopical multi-modal scanning means of the two-photon fluorescence described in any one of claim 1-4, it is characterized in that, described first rotation reflecting mirror and described second rotates reflecting mirror and is in transmission connection with the first driving means and the second driving means respectively, described first driving means drives described first to rotate reflecting mirror and rotates, and described second driving means drives described second to rotate reflecting mirror and rotates.
The microscopical multi-modal scanning means of two-photon fluorescence the most according to claim 5, it is characterised in that the microscopical multi-modal scanning means of described two-photon fluorescence also includes:
Dichroic mirror, described first lasing light emitter enters described first input path or described second input path after described dichroic mirror transmission.
The microscopical multi-modal scanning means of two-photon fluorescence the most according to claim 6, it is characterised in that the microscopical multi-modal scanning means of described two-photon fluorescence also includes:
Second laser, for launching the second lasing light emitter, described second lasing light emitter enters described first input path or described second input path after described dichroic mirror reflects, and consistent with the described second lasing light emitter light path reflected through described dichroic mirror through described first lasing light emitter of described dichroic mirror transmission.
The microscopical multi-modal scanning means of two-photon fluorescence the most according to claim 6, it is characterised in that described first laser instrument is femtosecond pulse laser.
The microscopical multi-modal scanning means of two-photon fluorescence the most according to claim 7, it is characterised in that described second laser is optical fiber laser.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201521128944.3U CN205620304U (en) | 2015-12-30 | 2015-12-30 | Two -photon fluorescence microscope's multimode scanning device |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201521128944.3U CN205620304U (en) | 2015-12-30 | 2015-12-30 | Two -photon fluorescence microscope's multimode scanning device |
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| CN205620304U true CN205620304U (en) | 2016-10-05 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527261A (en) * | 2015-12-30 | 2016-04-27 | 中国科学院苏州生物医学工程技术研究所 | A multi-mode scanning device of a two-photon fluorescence microscope |
| CN109211855A (en) * | 2018-08-10 | 2019-01-15 | 国家纳米科学中心 | Multiple beam multi-photon microscopic imaging device |
-
2015
- 2015-12-30 CN CN201521128944.3U patent/CN205620304U/en not_active Withdrawn - After Issue
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105527261A (en) * | 2015-12-30 | 2016-04-27 | 中国科学院苏州生物医学工程技术研究所 | A multi-mode scanning device of a two-photon fluorescence microscope |
| CN105527261B (en) * | 2015-12-30 | 2018-07-17 | 中国科学院苏州生物医学工程技术研究所 | A kind of microscopical multi-modal scanning means of two-photon fluorescence |
| CN109211855A (en) * | 2018-08-10 | 2019-01-15 | 国家纳米科学中心 | Multiple beam multi-photon microscopic imaging device |
| CN109211855B (en) * | 2018-08-10 | 2021-11-16 | 国家纳米科学中心 | Multi-beam multi-photon microscopic imaging device |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| AV01 | Patent right actively abandoned |
Granted publication date: 20161005 Effective date of abandoning: 20180717 |
|
| AV01 | Patent right actively abandoned |
Granted publication date: 20161005 Effective date of abandoning: 20180717 |