CN108845411A - Based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method and apparatus - Google Patents
Based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method and apparatus Download PDFInfo
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- CN108845411A CN108845411A CN201810712234.7A CN201810712234A CN108845411A CN 108845411 A CN108845411 A CN 108845411A CN 201810712234 A CN201810712234 A CN 201810712234A CN 108845411 A CN108845411 A CN 108845411A
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- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0036—Scanning details, e.g. scanning stages
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- G—PHYSICS
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- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
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- G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
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Abstract
It is according to the present invention to be based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method and apparatus, device includes beam splitter, frequency modulator, the first reflecting mirror, the second reflecting mirror, first lens unit, the second lens unit, half wave plate, polarization splitting prism, dichroscope, 2-D vibration mirror, focusing objective len, detector array.Beam splitter, first reflecting mirror is set gradually along primary optic axis line, beam splitter, frequency modulator, second reflecting mirror is set gradually along the second optical axis, first reflecting mirror, frequency modulator, half wave plate and polarization splitting prism are set gradually along third optical axis, second reflecting mirror, polarization splitting prism, polyhedral prism, dichroscope and 2-D vibration mirror are set gradually along the 4th optical axis, 2-D vibration mirror, first lens unit, focusing objective len is set gradually along the 5th optical axis, dichroscope, second lens unit, detector array is set gradually along the 6th optical axis.
Description
Technical field
The invention belongs to optical technical fields, and in particular to one kind is high based on polyhedral prism and light beam frequency modulation high-resolution
Fast imaging method and device.
Background technique
Optical imagery is widely used in life science, material science etc. and grinds due to having the characteristics that non-contact, non-destructive
Study carefully field.And optical resolution is then an important indicator of optical imaging system, the higher the better for usual resolution ratio.Optical resolution
Rate includes lateral resolution and axial resolution, can be limited each other between the two.I.e. lateral resolution is higher, axial resolution
It is lower；Axial resolution is higher, and lateral resolution is lower.Therefore, how to improve lateral resolution and axial resolution is research
The target of personnel's ongoing effort.There is important application valence if it can be achieved at the same time high lateral resolution and high axial resolution
Value.
There is important research significance in the fields such as optical microscopy imaging using the vectorial property of linearly polarized light.For example, altogether
Focusing microscope system is the point-to-point imaging of image conjugation, and the laser beam of focusing is scanned in sample surfaces, while photoelectric detector
Part receives the fluorescence (or fluorescence of transmission) of sample reflection, and the variation of sample structure changes the fluorescence intensity of excitation, thus makes
The output electric current of photoelectric detector changes, and by signal processing, simultaneous display is on the computer screen.Due to the linear polarization of irradiation
Light by the lens focus of high-NA, generation be area very little ellipse light spot.If along ellipse light spot short-axis direction
To Sample Scan, according to Rayleigh criterion, confocal microscope scanning step be twice of ellipse short shaft apart from when, photodetector
The change for responding intensity of reflected light, that is, tell the difference of two o'clock, systemic resolution is very high.If along ellipse light spot long axis direction
To Sample Scan, and less than twice transverse of scanning step apart from when, according to Rayleigh criterion, photodetector be will not respond to
The change of intensity of reflected light can not tell the difference of two o'clock.Therefore the resolution ratio of system is decided by focal beam spot long axis size.
In first technology, referring to " K.A.Serrels, E.Ramsay, R.J.Warburton and D.T.Reid, Nanoscale
Optical microscopy in the vectorial focusing regime, nature photonics, vol.2,
May2008,311-314 ", in order to improve resolution ratio, when scanning long axis direction mechanical insertion half wave plate change into
The polarization direction of ray polarised light, but this can reduce the sweep speed and system resolution precision when system changes scanning direction,
And since wherein a branch of incident light have passed through a half wave plate more, the incident power of this two beams crossed polarized light
Difference will increase systematic error so that focus on light beam power be made to change, and system stability is not high.
Summary of the invention
In view of the deficiencies of the prior art, one of the objects of the present invention is to provide a kind of polyhedral prism and light beam frequency modulation are high
High resolution speed imaging method and device, by a kind of effective optical texture, while it is orthogonal to construct two beam polarization directions
Linearly polarized light illumination, and the elliptical light formed after two beam orhtogonal linear polarizaiton light focus is distinguished using the frequency modulation(PFM) of dual-beam
The overlapping region of spot and other Non-overlapping Domains, single-row, equidistant multifocal is generated using polyhedral prism, utilizes array detection
It is corresponding to extract the overlapping region of the ellipse light spot formed after two beam orhtogonal linear polarizaiton light focus that device receives the frequency demodulation of signal
Useful signal, realize improve dimensional resolution purpose.
The present invention provides one kind to be based on polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device, has in this way
Feature, including beam splitter, frequency modulator, the first reflecting mirror, the second reflecting mirror, the first lens unit, the second lens unit,
Half wave plate, polarization splitting prism, dichroscope, 2-D vibration mirror, focusing objective len, detector array, wherein beam splitter,
First reflecting mirror is set gradually along primary optic axis line, and beam splitter, frequency modulator, the second reflecting mirror are along vertical with primary optic axis line
The second optical axis set gradually, the first reflecting mirror, frequency modulator, half wave plate and polarization splitting prism are along with the
The vertical third optical axis of one optical axis is set gradually, and third optical axis is parallel with the second optical axis, the second reflecting mirror, polarization point
Light prism, polyhedral prism, dichroscope and 2-D vibration mirror are set gradually along the 4th optical axis vertical with third optical axis,
2-D vibration mirror, the first lens unit, focusing objective len are set gradually along the 5th optical axis vertical with the 4th optical axis, dichroic
Mirror, the second lens unit, detector array are set gradually along the 6th optical axis vertical with the 4th optical axis, the first lens list
Member, the second lens unit include the combination of a lens or multiple lens.
Provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device, feature exists
In, it further include the third lens unit being arranged on the second optical axis and between frequency modulator and the second reflecting mirror, the
Three lens units include the combination of a lens or multiple lens.
The present invention provides one kind to be existed based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, feature
In including the following steps：
Step 1, beam splitter, the first reflecting mirror are set gradually along primary optic axis line；
Step 2, by beam splitter, frequency modulator, the second reflecting mirror along second optical axis vertical with primary optic axis line according to
Secondary setting；
Step 3, by the first reflecting mirror, frequency modulator, half wave plate and polarization splitting prism edge and the first light
The vertical third optical axis of axis is set gradually, and third optical axis is parallel with the second optical axis；
Step 4, by dichroscope described in the second reflecting mirror, polarization splitting prism, polyhedral prism and 2-D vibration mirror edge
Fourth optical axis vertical with third optical axis is set gradually,
Step 5, by 2-D vibration mirror, the first lens unit, focusing objective len and sample along vertical with the 4th optical axis the
Five optical axis are set gradually,
Step 6, dichroscope, the second lens unit, detector array are along sixth optical axis vertical with the 4th optical axis
It sets gradually, wherein the second lens unit faces the reflecting surface of dichroscope；
Step 7, a branch of to carry out incident first direction incident ray polarized light after beam splitter along primary optic axis line direction
Export the first light beam and the second light beam that two beams have first direction linearly polarized light；
Step 8, the first light beam exports the first carrier frequency after frequency modulator applies carrier frequency f1 along the second optical axis direction
Light beam, the first carrier frequency light beam are reflected along the 4th optical axis direction through the second reflecting mirror by polarization splitting prism transmission output first
Carrier frequency transmitted light beam, the second light beam after the reflection of the first reflecting mirror, are passed through along primary optic axis line direction along third optical axis direction
Overfrequency modulator exports the second carrier frequency light beam after applying carrier frequency f2, and the second carrier frequency light beam exports the after half wave plate
Two direction linearly polarized lights, second direction linearly polarized light export along the 4th optical axis direction after polarization splitting prism reflects
Two carrier frequency the reflected beams；
Step 9, the first carrier frequency transmitted light beam and the superposition synthesis output of the second carrier frequency the reflected beams have mixing first direction
With the linear polarization carrier frequency mixed light beam of second direction；
Step 10, carrier frequency mixed light beam obtains multi beam refraction carrier frequency mixed light beam after polyhedral prism reflects；
Step 11, multi beam refraction carrier frequency mixed light beam passes through dichroscope, 2-D vibration mirror, the first lens unit, conglomeration
Multiple focal beam spots are generated after mirror on sample；
Step 12, multiple focal beam spot excitation samples generate fluorescence, form multiple fluorescence hot spots corresponding with focal beam spot；
Step 13, fluorescence hot spot passes through focusing objective len, the first lens unit, 2-D vibration mirror, dichroscope, the second lens list
After member, multiple fluorescent foci hot spots are formed on detector array；
Step 14, using frequency demodulation algorithm, extract the received overlapping of detector array with carrier frequency f1 and f2
Fluorescent foci spot signal, and pass through the intensity of fluorescent foci hot spot on analysis detector array and sweeping with 2-D vibration mirror
Variation is retouched, the fluorescence signal of each self-excitation of focus with different modulating frequency can be distinguished, to reconstruct reflection sample
The two dimensional image of information.
Provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, can also have
There is such feature：Wherein, first direction linearly polarized light is orthogonal with second direction linearly polarized light.
In addition, being gone back provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high speed imaging method
It can have such feature：Wherein, polyhedral prism is the cylinder with bottom surface and multiple faceted pebbles, and cross section is polygon
Shape, the direction of carrier frequency mixed light beam and plane perpendicular and carrier frequency mixed light beam are introduced into bottom surface, and focal beam spot is one-dimensional array point
Cloth, the quantity of focal beam spot and the quantity of faceted pebble are identical.
In addition, being gone back provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high speed imaging method
It can have such feature：Wherein, the first lens unit is used for optical beam transformation, and the multi beam being emitted from polyhedral prism is reflected
Carrier frequency mixed light beam is full of the entrance pupil of focusing objective len always, realizes the optimal imaging performance of focusing objective len, the first lens unit packet
Include the combination of two optical beam transformation lens or multiple optical beam transformation lens, the angle of dichroscope and the 4th optical axis is 45 degree.
In addition, being gone back provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high speed imaging method
It can have such feature：Wherein, when polyhedral prism be the cone prism with bottom surface and multiple faceted pebbles, the bottom surface of prism with
Carrier frequency mixed light beam direction is vertical, and focal beam spot is two-dimensional array distribution, and the quantity of focal beam spot and the quantity of faceted pebble are identical.
In addition, being gone back provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high speed imaging method
It can have such feature：It wherein, further include being arranged on the second optical axis and being located at frequency modulator and the in step 2
The third lens unit for being used to improve axial resolution of two-mirror.
In addition, being gone back provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high speed imaging method
It can have such feature：Wherein, the third lens unit includes the combination of a lens or multiple lens, for carrying first
The focussing plane of frequency light beam and the focussing plane of the second carrier frequency light beam are separated by a distance.
In addition, being gone back provided by the invention based in polyhedral prism and light beam frequency modulation high-resolution high speed imaging method
It can have such feature：Wherein, in step 14, using frequency demodulation algorithm, the received overlapping of detector array is extracted
The fluorescent foci spot signal with carrier frequency f1 and f2, and by analysis detector array on fluorescent foci hot spot intensity
And with the scan variations of 2-D vibration mirror, the fluorescence signal of each self-excitation of focus with different modulating frequency can be distinguished,
To reconstruct the 3-D image of reflection sample message.
The action and effect of invention
It is based on polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device involved according to the present invention, including divides
Shu Jing, frequency modulator, the first reflecting mirror, the second reflecting mirror, the first lens unit, the second lens unit, half wave plate,
Polarization splitting prism, dichroscope, 2-D vibration mirror, focusing objective len, detector array, the present invention utilize the frequency modulation(PFM) of dual-beam
It distinguishes the overlapping region of ellipse light spot formed after two beam orhtogonal linear polarizaiton light focus and other Non-overlapping Domains, utilizes photoelectricity
Detector receives the frequency demodulation of signal to extract the overlapping region of the ellipse light spot formed after two beam orhtogonal linear polarizaiton light focus
Corresponding useful signal, to realize the purpose for improving two-dimensional resolution.And simultaneously using polyhedral prism in focusing objective len
Array focal beam spot is generated on focusing surface, is realized multiple spot scanning imagery simultaneously, is significantly improved laser confocal scanning imaging
Frame speed, achievees the purpose that high speed imaging.
Alternatively, it is also possible to improve the pixel number of single-frame images in the case where frame speed is constant.Theoretically, compared to tradition
Simple scan imaging, for a frame image of same pixel, the image taking speed of the multi-point scanning imaging of N number of focal beam spot can be with
Improve N times；Identical for frame speed, the pixel number of a frame image can be improved N times.For example, single-point in current industry can be swashed
The canonical parameter 512*32 pixel/frame of light confocal scanning imaging, frame fast 400 frames/second rise to 512*32 pixel/frame, frame speed
400N frame/second, or be 512*32N pixel/frame, frame fast 400 frames/second.
Detailed description of the invention
Fig. 1 is to improve transverse direction using the vectorial property of dual-beam linearly polarized light and frequency modulation(PFM) in the embodiment of the present invention
The schematic illustration of resolution ratio；
Fig. 2 is the principle signal for improving axial resolution in the embodiment of the present invention using the frequency modulation(PFM) of dual-beam
Figure；
Fig. 3 be in the embodiment of the present invention based on the warbled three-dimension high-resolution laser confocal scanning of dual-beam at
As schematic diagram；And
Fig. 4 is to improve resolution ratio when scanning sample in the embodiment of the present invention while improving the schematic diagram of image taking speed；
Fig. 5 is the light beam schematic diagram of prism in the embodiment of the present invention two；
Fig. 6 is the focal beam spot distribution schematic diagram in the embodiment of the present invention two on sample；
Fig. 7 is prism section and focal beam spot distribution schematic diagram in the embodiment of the present invention three；
Fig. 8 is prism section and focal beam spot distribution schematic diagram in the embodiment of the present invention four；
Fig. 9 is prism section and focal beam spot distribution schematic diagram in the embodiment of the present invention five；And
Figure 10 is six neutral body prism of the embodiment of the present invention and focal beam spot distribution schematic diagram.
Specific embodiment
It is real below in order to be easy to understand the technical means, the creative features, the aims and the efficiencies achieved by the present invention
Example combination attached drawing is applied to have to of the invention based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method and apparatus work
Body illustrates.
Embodiment one
If optical system uses linear polarization coherent light illumination, for high-NA optical system, on focussing plane
Focal beam spot distribution significantly influenced by the polarization characteristic of illumination light.As shown in Figure 1, if using the direction y linear polarization coherent light
It illuminates, the focal beam spot on focussing plane is ellipse distribution, and the direction of elliptical short axle and linearly polarized light (is at this time y
Direction) vertically, i.e., elliptical short-axis direction is in the x-direction at this time.If focused flat using the direction x linear polarization coherent light illumination
The short-axis direction of oval focal beam spot on face is in the y-direction.Optical system lateral resolution is decided by the size of focal beam spot.It is aobvious
So, if using the direction y linear polarization coherent light illumination, the direction optical system x has higher lateral resolution；If using
The direction x linear polarization coherent light illumination, then the direction optical system y has higher lateral resolution.Obviously, if it is possible to while benefit
With the direction y linearly polarized light and x direction line polarized illumination, according to principle of stacking, then the focal beam spot on focussing plane is distributed as
In Fig. 1 shown in right figure, it is clear that its shadow region has smaller sized fraction in the direction x and the direction y simultaneously, if using shadow region
Domain then can significantly improve lateral resolution in the direction x and the direction y as effective focal beam spot.But, right figure in Fig. 1 at this time
Shown non-hatched area can also deteriorate lateral resolution.In order to solve this problem, we are by the direction y linear polarization coherent light illumination
Apply frequency be f1 carrier frequency (referred to as illumination light 1z), the direction x linear polarization coherent light illumination apply frequency be f2 carrier frequency (referred to as
Illumination light 2z), then the carrier frequency of shadow region hot spot shown in right figure is f1+f2 in Fig. 1, the carrier frequency of other non-hatched areas is distinguished
For f1 and f2.In this way carrier frequency be f1+f2 the fluorescence signal that is excited on sample of shadow region hot spot there is also carrier frequency f1+f2,
By frequency demodulation algorithm, then the fluorescence signal of carrier frequency f1+f2 can be extracted.To reach the mesh for improving lateral resolution
's.
As in Fig. 1, the direction y linear polarization coherent light illumination is applied carrier frequency (the referred to as illumination light that frequency is f1 by we
1z), linear polarization coherent light illumination in the direction x applies frequency as the carrier frequency (referred to as illumination light 2z) of f2, while increasing in illumination light 1z
Add a lens, finely tune its focussing plane, the focussing plane of the focussing plane of illumination light 1z and illumination light 2z is made to separate a spacing
From then as shown in Fig. 2, the focal beam spot of the focal beam spot (dashed region) of illumination light 1z and illumination light 2z are in axial direction (z-axis) side
To being separated by a distance, the carrier frequency of shadow region hot spot is f1+f2 in figure, the carrier frequency of other non-hatched areas be respectively f1 and
f2.There is also carrier frequency f1+f2 for the fluorescence signal that the shadow region hot spot that carrier frequency is f1+f2 in this way excites on sample, pass through frequency
Rate demodulating algorithm can then extract the fluorescence signal of carrier frequency f1+f2.To achieve the purpose that improve axial resolution.
As shown in figure 3, the plane where Fig. 3 is the face yz, x-axis is perpendicular to the face yz.Based on the warbled three-dimensional of dual-beam
High resolution imaging apparatus includes beam splitter 1, frequency modulator 2, the third lens unit 3, the second reflecting mirror 4, the first reflecting mirror
5, half wave plate 6, polarization splitting prism 7, polyhedral prism 8, dichroscope 9,2-D vibration mirror 10, the first lens unit
(lens 11, lens 12), focusing objective len 13, the second lens unit 15, detector array 16.
Wherein, the third lens unit 3, the second lens unit 15 are the combination of a lens or multiple lens.
Beam splitter 1, the first reflecting mirror 5 are set gradually along primary optic axis line, beam splitter 1, frequency modulator 2, the third lens
Unit 3, the second reflecting mirror 4 are set gradually along the second optical axis vertical with primary optic axis line, the first reflecting mirror 5, frequency modulation(PFM)
Device 2, half wave plate 6 and polarization splitting prism 7 are set gradually along the third optical axis vertical with primary optic axis line, third
Optical axis is parallel with the second optical axis, the second reflecting mirror 4, polarization splitting prism 7, polyhedral prism 8, dichroscope 9 and two
Dimension galvanometer 10 is set gradually along the 4th optical axis vertical with third optical axis, and 2-D vibration mirror 10, lens 12, focuses lens 11
Object lens 13, sample 14 are set gradually along the 5th optical axis vertical with the 4th optical axis, dichroscope 9, the second lens unit 15,
Detector array 16 is set gradually along the 6th optical axis vertical with the 4th optical axis.
The linearly polarized light in a branch of direction y is divided into II liang of beam polarised light of light beam I and light beam after beam splitter 1.Y direction line is inclined
Vibration light light beam I has been applied carrier frequency f1 after frequency modulator 2, reflects by the second lens 3, the second reflecting mirror 4, by inclined
Vibration Amici prism 7 exports the first carrier frequency transmitted light beam, the quilt after frequency modulator 2 of the direction y linearly polarized light light beam II after transmiting
It is applied with carrier frequency f2, light beam II becomes the direction x linearly polarized light after half wave plate 6, and the effect of half wave plate 6 is
It is rotated by 90 ° the linear polarization of light beam II relative to the linear polarization direction of light beam I, therefore light beam I and light beam II are by polarizing
It is equivalent to synthesize light beam after prism 7, there are two orthogonal linear polarizations for light beam tool.It is anti-by polarization splitting prism 7
Penetrating to be superimposed synthesis with the first carrier frequency transmitted light beam afterwards and export has the linear polarization carrier frequency of mixing first direction and second direction mixed
Light combination beam, the carrier frequency mixed light beam obtain multi beam refraction carrier frequency mixed light beam after the polyhedral prism 8 refraction；Multi beam
Refraction carrier frequency mixed light beam focuses on sample after dichroscope 9,2-D vibration mirror 10, lens 11, lens 12, focusing objective len 13
On 14.Light beam I and light beam II are linearly polarized light, and in embodiment, the linear polarization of the linear polarization of light beam I and light beam II is just
It hands over, and light beam I and light beam II generate simultaneously, on the sample that the focal beam spot of the two focuses simultaneously.
According to the reversible principle of optic path, multi beam refraction carrier frequency mixed light beam is after focusing objective len 13 on sample 14
The fluorescence on multiple focal beam spot excitation samples 14 with different modulating frequency generated, forms corresponding with multiple focal beam spots
Fluorescence hot spot, the fluorescence hot spot of sending is by focusing objective len 13, lens 12, lens 11,2-D vibration mirror 10, dichroscope 9, thoroughly
Mirror 15 forms multiple fluorescent foci hot spots in detector array 16.Pass through multiple fluorescent foci light on analysis detector array 16
The intensity of spot with 2-D vibration mirror 10 scan variations, so that it may reconstruct reflection sample message Two-Dimensional Reconstruction image.Implement
In example, fluorescent foci hot spot is two,
Frequency modulator 2 for applying different carrier frequency to light beam I and light beam II respectively.Frequency modulator 2 can be liquid
Brilliant chopper, mechanical chopper or other schemes that can be modulated to incident beam real-time frequency.
The third lens unit 3 can be simple lens, or lens group, effect is the focusing surface and light for making light beam I
The focusing surface of beam II is in axial separation a certain distance, in two focal beam spots for being axially formed similar Fig. 2.Illumination light 1z's is poly-
Burnt hot spot (dashed region) and the focal beam spot of illumination light 2z are separated by a distance in axial direction (z-axis) direction, shadow region in figure
The carrier frequency of hot spot is f1+f2, and the carrier frequency of other non-hatched areas is respectively f1 and f2.In this way carrier frequency be f1+f2 shadow region
There is also carrier frequency f1+f2 for the fluorescence signal that hot spot excites on sample, by frequency demodulation algorithm, then can extract carrier frequency f1
The fluorescence signal of+f2.To achieve the purpose that improve axial resolution.
The effect of half wave plate 6 is to be rotated by 90 ° the linear polarization direction of light beam II, to make half wave plate
The linear polarization of 6 outgoing beam II and the linear polarization direction of light beam I are orthogonal.
Dichroscope 9 is placed between polyhedral prism 8 and 2-D vibration mirror 10, for reflexing to the fluorescence signal of return
It is formed on detector array 16 and the equal number of fluorescent foci hot spot of focal beam spot on sample 14 after mirror 15；In embodiment,
The angle of dichroscope 9 and the 4th optical axis is 45 degree.
2-D vibration mirror 10 is mechanically fixed as quadrature arrangement mode by two one-dimensional galvanometers, and one-dimensional galvanometer can be
Galvanometer galvanometer or resonance galvanometer, such as Cambridge Technology company Galvanometer Optical Scanner
6230H, with CRS 8kHz Resonant Scanner.
First lens unit includes the combination of two lens (lens 11, lens 12) or multiple lens.Lens 11, lens 12
An optical beam transformation lens group is formed, effect has been optical beam transformation effect, and the multi beam oblique incidence for being emitted polyhedral prism 8 is flat
Row light is full of the entrance pupil of focusing objective len 13 always, realizes the optimal imaging performance of focusing objective len 13.In practical applications, lens
11, the first lens unit two the first lens units, two lens as shown in Fig. 3 schematic diagram that lens 12 form
Composition can form for more lens, to realize the effect of optical beam transformation.
Detector array 16 is photodetector, including point type photodetector and array optical electric explorer, such as photoelectricity
Multiplier tube, avalanche diode, electron multiplication CCD etc..
In embodiment, the first lens unit includes two lens (lens 11, lens 12), and the third lens unit 3, second is saturating
Mirror unit 15 is simple lens, and frequency modulator 2 uses Liquid Crystal Chopper, and polyhedral prism 8 uses cylinder prism, two dimension vibration
Two one-dimensional galvanometers in mirror 10 are all made of resonance galvanometer, and detector array 16 uses photomultiplier tube.
By properly selecting the optical parameter of lens 3, lens 3 can make the focussing plane of light beam I and the focusing of light beam II
Plane is separated by a distance, and forms focal beam spot shown in Fig. 2 in axial direction (z-axis direction).Since the light beam I with carrier frequency f1 is
The direction y linear polarization, the light beam II with carrier frequency f2 is the direction x linear polarization, so light beam I is focused on sample 14 with light beam II
Transverse focusing hot spot as shown in Fig. 1 right figure.The fluorescence letter that the shadow region hot spot that carrier frequency is f1+f2 in this way excites on sample
Number there is also carrier frequency f1+f2, fluorescence signal is anti-by focusing objective len 13, lens 12, lens 11,2-D vibration mirror 10, dichroscope 9
It penetrates, then is focused on detector array 16 by lens 15.By frequency demodulation algorithm, then detector array 16 can be extracted
The fluorescence signal of received carrier frequency f1+f2.Therefore, the direction the y linearly polarized light beam I with carrier frequency f1 and the side x for having carrier frequency f2
Effective focal beam spot (the carrier frequency f1+f2 formed to three-dimensional (laterally and axially) of the linearly polarized light beam II on sample 14
Shadow region hot spot) significantly less than do not use the invention patent when focal beam spot, so as to significantly improve three-dimensional resolution
Rate (laterally and axially).
By analyze detector array 16 spot intensity with 2-D vibration mirror 10 scan variations, while utilize frequency solution
Adjust algorithm, so that it may distinguish and locally put in the fluorescence signal such as Fig. 4 of multiple each self-excitations of focus with different modulating frequency
Big figure D, to reconstruct the 3-D image (assuming that scanning n row n arranges a point) of reflection sample message as shown in Figure 4, while benefit
Array focal beam spot is generated on the focusing surface of focusing objective len with polyhedral prism, is realized multiple spot scanning imagery simultaneously, is significantly mentioned
The high frame speed of laser confocal scanning imaging, achievees the purpose that high speed imaging.
One kind being based on polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device method, includes the following steps：
Step 1, beam splitter, the first reflecting mirror are set gradually along primary optic axis line；
Step 2, by beam splitter, frequency modulator, the third lens unit, the second reflecting mirror along vertical with primary optic axis line
Second optical axis is set gradually；
Step 3, by the first reflecting mirror, frequency modulator, half wave plate and polarization splitting prism edge and the first light
The vertical third optical axis of axis is set gradually, and third optical axis is parallel with the second optical axis；
Step 4, by the second reflecting mirror, polarization splitting prism, polyhedral prism, dichroscope and 2-D vibration mirror along with the
The 4th vertical optical axis of three optical axis is set gradually；
Step 5, by 2-D vibration mirror, the first lens unit, focusing objective len and sample along vertical with the 4th optical axis the
Five optical axis are set gradually；
Step 6, dichroscope, the second lens unit, detector array are along sixth optical axis vertical with the 4th optical axis
It sets gradually, wherein the second lens unit faces the reflecting surface of dichroscope；
Step 7, a branch of to carry out incident first direction incident ray polarized light after beam splitter along primary optic axis line direction
Export the first light beam and the second light beam that two beams have first direction linearly polarized light；
Step 8, the first light beam exports the first carrier frequency after frequency modulator applies carrier frequency f1 along the second optical axis direction
Light beam, the first carrier frequency light beam are reflected along the 4th optical axis direction through the second reflecting mirror by polarization splitting prism transmission output first
Carrier frequency transmitted light beam；Second light beam after the reflection of the first reflecting mirror, is passed through along primary optic axis line direction along third optical axis direction
Overfrequency modulator exports the second carrier frequency light beam after applying carrier frequency f2, and the second carrier frequency light beam exports the after half wave plate
Two direction linearly polarized lights, second direction linearly polarized light export along the 4th optical axis direction after polarization splitting prism reflects
Two carrier frequency the reflected beams；
Step 9, the first carrier frequency transmitted light beam and the superposition synthesis output of the second carrier frequency the reflected beams have mixing first direction
With the linear polarization carrier frequency mixed light beam of second direction；
Step 10, carrier frequency mixed light beam obtains multi beam refraction carrier frequency mixed light beam after polyhedral prism reflects；
Step 11, multi beam refraction carrier frequency mixed light beam passes through dichroscope, 2-D vibration mirror, the first lens unit, conglomeration
Multiple focal beam spots are generated after mirror on sample；
Step 12, multiple focal beam spot excitation samples generate fluorescence, form multiple fluorescence hot spots corresponding with focal beam spot；
Step 13, fluorescence hot spot passes through focusing objective len, the first lens unit, 2-D vibration mirror, dichroscope, the second lens list
After member, multiple fluorescent foci hot spots are formed on detector array；
Step 14, using frequency demodulation algorithm, extract the received overlapping of detector array with carrier frequency f1 and f2
Fluorescent foci spot signal, and pass through the intensity of fluorescent foci hot spot on analysis detector array and sweeping with 2-D vibration mirror
Variation is retouched, the fluorescence signal of each self-excitation of focus with different modulating frequency can be distinguished, to reconstruct reflection sample
The 3-D image of information.
Embodiment two
As shown in figure 5, incident light g, refraction light is g1, g2, if the folder of two faceted pebble m1 and m2 and bottom surface of prism 8
Angle is respectively θ1And θ2, the refractive index of prism 9 is n, then the refraction light g1 of available faceted pebble m1 and primary optic axis line (z-axis)
Angle is θ1'=asin (nsin θ1-θ1), as shown in fig. 6, distance h of the focal beam spot A to the 5th optical axis (x-axis)1=fsin
[asin(nsinθ1-θ1)], wherein f is the focal length of focusing objective len 2.Similarly, distance h of the focal beam spot B to the 5th optical axis2=
fsin[asin(nsinθ2-θ2)].It therefore, can be the angle of n, faceted pebble m1, m2 and z-axis by the refractive index of prism, and
The focal length f of condenser lens 13 can accurately control the position of focal beam spot.If plane m1 and m2 be not right about primary optic axis line
Claim, then focal beam spot A is different from the intensity of focal beam spot B.The intensity of focal beam spot A and focal beam spot B is decided by faceted pebble m1, m2
Area and the ratio between entire incident beam sectional area.
Embodiment three
The present embodiment is identical as other structures in embodiment one and setting, and only polyhedral prism changes into the present embodiment
As shown in the left side in Fig. 7 polyhedral prism.The polyhedral prism is four sides cylinder prism, has bottom surface and three faceted pebbles,
Three faceted pebbles are symmetrical arranged along primary optic axis line.
Focal beam spot is distributed as in the present embodiment：Three edges on x/y plane such as the right side in Fig. 7 are formed on sample 14
Y-axis arrangement focal beam spot.If faceted pebble is vertical with light beam g and is symmetrical arranged along primary optic axis line, obtained by the faceted pebble
Focal beam spot on the origin of reference axis.
Further, it is assumed that cofocus scanning imaging will finally obtain the image of spoke n row n column, utilize the more of the present embodiment
Face body prism generates the column distribution focus of 3 points, then need to only scan n/3 row, image taking speed can be imaged than existing single focus
Speed improves 3 times.
Example IV
The present embodiment is identical as other structures in embodiment one and setting, and only polyhedral prism changes into the present embodiment
As shown in the left side in Fig. 8 polyhedral prism.The polyhedral prism is five face cylinder prisms, has bottom surface and four faceted pebbles,
Four faceted pebbles are symmetrical arranged along primary optic axis line.
Focal beam spot is distributed as in the present embodiment：Four edges on x/y plane such as the right side in Fig. 8 are formed on sample 14
Y-axis arrangement focal beam spot.
Further, it is assumed that cofocus scanning imaging will finally obtain the image of spoke n row n column, utilize the more of the present embodiment
Face body prism generates the column distribution focus of four points, then need to only scan n/4 row, image taking speed can be imaged than existing single focus
Speed improves 4 times.
Embodiment five
The present embodiment is identical as other structures in embodiment one and setting, and only polyhedral prism changes into the present embodiment
As shown in the left side in Fig. 9 polyhedral prism.The polyhedral prism is six face cylinder prisms, has bottom surface and five faceted pebbles,
Five faceted pebbles are symmetrical arranged along primary optic axis line.
Focal beam spot is distributed as in the present embodiment：Five edges on x/y plane such as the right side in Fig. 9 are formed on sample 14
Y-axis arrangement focal beam spot.
Further, it is assumed that cofocus scanning imaging will finally obtain the image of spoke n row n column, utilize the more of the present embodiment
Face body prism generates the column distribution focus of five points, then need to only scan n/5 row, image taking speed can be imaged than existing single focus
Speed improves 5 times.
Embodiment six
The present embodiment is identical as other structures in embodiment one and setting, and only polyhedral prism changes into the present embodiment
As shown in the left side in Figure 10 polyhedral prism.The polyhedral prism is cone prism, has bottom surface and four faceted pebbles, four ribs
Face then generates the focal beam spot of the two-dimensional array about z-axis rotation distribution along axis y rotary setting.
Focal beam spot is distributed as in the present embodiment：Four edges on x/y plane such as the right side in Figure 10 are formed on sample 14
The focal beam spot that uniformly arranges of x, y-axis.
If the face of polyhedral prism is changed along two dimensions, the focal beam spot of two-dimensional array is obtained.
The action and effect of embodiment
Polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device are based on according to involved in the present embodiment, including
Beam splitter, frequency modulator, the first reflecting mirror, the second reflecting mirror, the first lens unit, the second lens unit, half wave
Piece, polarization splitting prism, dichroscope, 2-D vibration mirror, focusing objective len, detector array, the present invention utilize the frequency of dual-beam
It modulates to distinguish the overlapping region of the ellipse light spot formed after two beam orhtogonal linear polarizaiton light focus and other Non-overlapping Domains, utilizes
Photodetector receives the frequency demodulation of signal to extract the overlapping of the ellipse light spot formed after two beam orhtogonal linear polarizaiton light focus
The corresponding useful signal in region, to realize the purpose for improving dimensional resolution.And simultaneously using polyhedral prism in conglomeration
On the focusing surface of mirror generate array focal beam spot, realize multiple spot simultaneously scanning imagery, significantly improve laser confocal scanning at
The frame speed of picture, achievees the purpose that high speed imaging.
Alternatively, it is also possible to improve the pixel number of single-frame images in the case where frame speed is constant.Theoretically, compared to tradition
Simple scan imaging, for a frame image of same pixel, the image taking speed of the multi-point scanning imaging of N number of focal beam spot can be with
Improve N times；Identical for frame speed, the pixel number of a frame image can be improved N times.For example, single-point in current industry can be swashed
The canonical parameter 512*32 pixel/frame of light confocal scanning imaging, frame fast 400 frames/second rise to 512*32 pixel/frame, frame speed
400N frame/second, or be 512*32N pixel/frame, frame fast 400 frames/second.
It further, further include being arranged on the second optical axis and being located at frequency modulator and the second reflecting mirror in embodiment
For improving the third lens unit of axial resolution.It is that the focusing surface of light beam I and the focusing surface of light beam II is made to exist that it, which is acted on,
Axial separation a certain distance is being axially formed two focal beam spots, is improving axial resolution.It is differentiated to realize that raising is three-dimensional
The purpose of rate.
Further, when polyhedral prism is prism, if generating the column distribution focus of 2 points using prism,
N/2 row then need to be only scanned, image taking speed can improve 1 times than existing image taking speed.
Further, when polyhedral prism is four prism, if generating the column distribution focus of 3 points using four prisms,
N/3 row then need to be only scanned, image taking speed can improve 3 times than existing image taking speed.
It is further possible to improve the pixel number of single-frame images in the case where frame speed is constant.Theoretically, compared to
Traditional simple scan imaging, for a frame image of same pixel, the image taking speed of the multi-point scanning imaging of N number of focal beam spot
It can be improved N times；Identical for frame speed, the pixel number of a frame image can be improved N times.For example, can will be single in current industry
The canonical parameter 512*32 pixel/frame of dot laser confocal scanning imaging, frame fast 400 frames/second, rise to 512*32 pixel/frame,
Frame speed 400N frame/second, or be 512*32N pixel/frame, frame fast 400 frames/second.
Above embodiment is preferred case of the invention, the protection scope being not intended to limit the invention.
Claims (10)
1. one kind is based on polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device, which is characterized in that including：Beam splitting
Mirror, frequency modulator, the first reflecting mirror, the second reflecting mirror, the first lens unit, the second lens unit, half wave plate, partially
Shake Amici prism, dichroscope, 2-D vibration mirror, focusing objective len, detector array,
Wherein, the beam splitter, first reflecting mirror are set gradually along primary optic axis line,
The beam splitter, the frequency modulator, second reflecting mirror are along second optical axis vertical with the primary optic axis line
Line is set gradually,
First reflecting mirror, the frequency modulator, the half wave plate and polarization splitting prism edge and institute
The vertical third optical axis of primary optic axis line to be stated to set gradually, the third optical axis is parallel with second optical axis,
Second reflecting mirror, the polarization splitting prism, the polyhedral prism, the dichroscope and the two dimension vibration
Mirror is set gradually along the 4th optical axis vertical with the third optical axis,
The 2-D vibration mirror, first lens unit, the focusing objective len are along fiveth light vertical with the 4th optical axis
Axis is set gradually,
The dichroscope, second lens unit, the detector array are along the vertical with the 4th optical axis the 6th
Optical axis is set gradually,
First lens unit, second lens unit include the combination of a lens or multiple lens.
2. according to claim 1 be based on polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device device,
It is characterized in that, further includes：
The third lens on second optical axis and between the frequency modulator and second reflecting mirror are set
Unit,
The third lens unit includes the combination of a lens or multiple lens.
3. one kind is based on polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device method, using described in claim 1
Based on polyhedral prism and light beam frequency modulation high-resolution high-speed imaging device, which is characterized in that include the following steps：
Step 1, the beam splitter, first reflecting mirror are set gradually along primary optic axis line；
Step 2, by the beam splitter, the frequency modulator, second reflecting mirror along vertical with the primary optic axis line
Second optical axis is set gradually；
Step 3, by first reflecting mirror, the frequency modulator, half wave plate and polarization splitting prism edge and institute
It states the vertical third optical axis of primary optic axis line to set gradually, the third optical axis is parallel with second optical axis；
Step 4, by dichroscope described in second reflecting mirror, the polarization splitting prism, polyhedral prism and described two
Dimension galvanometer is set gradually along the 4th optical axis vertical with the third optical axis；
Step 5, by the 2-D vibration mirror, first lens unit, the focusing objective len and sample edge and the 4th light
The 5th vertical optical axis of axis is set gradually；
Step 6, the dichroscope, second lens unit, the detector array are along vertical with the 4th optical axis
The 6th optical axis set gradually, wherein second lens unit face the dichroscope reflecting surface；
Step 7, a branch of to carry out incident first direction incident ray polarized light by the beam splitting along primary optic axis line direction
The first light beam and the second light beam that two beams have first direction linearly polarized light are exported after mirror；
Step 8, first light beam exports after the frequency modulator applies carrier frequency f1 along second optical axis direction
First carrier frequency light beam, the first carrier frequency light beam are reflected through second reflecting mirror along described in the process of the 4th optical axis direction
Polarization splitting prism transmission the first carrier frequency transmitted light beam of output,
Second light beam along primary optic axis line direction by first reflecting mirror reflection after, along the third optical axis
Direction exports the second carrier frequency light beam after the frequency modulator applies carrier frequency f2, and the second carrier frequency light beam passes through described two
Second direction linearly polarized light is exported after/mono- wave plate, the second direction linearly polarized light is reflected by the polarization splitting prism
The second carrier frequency the reflected beams along the 4th optical axis direction are exported afterwards；
Step 9, the first carrier frequency transmitted light beam and the second carrier frequency the reflected beams superposition synthesis output have mixing first
The linear polarization carrier frequency mixed light beam in direction and second direction；
Step 10, the carrier frequency mixed light beam obtains multi beam refraction carrier frequency mixed light beam after polyhedral prism refraction；
Step 11, refraction carrier frequency mixed light beam described in multi beam passes through the dichroscope, the 2-D vibration mirror, first lens
Multiple focal beam spots are generated on to the sample after unit, the focusing objective len；
Step 12, multiple focal beam spots excite the sample to generate fluorescence, are formed multiple corresponding with the focal beam spot
Fluorescence hot spot；
Step 13, the fluorescence hot spot passes through the focusing objective len, first lens unit, the 2-D vibration mirror, described two
To after Look mirror, second lens unit, multiple fluorescent foci hot spots are formed on the detector array；
Step 14, using frequency demodulation algorithm, extract the received overlapping of the detector array with carrier frequency f1 and f2
The fluorescent foci spot signal, and the intensity by analyzing the fluorescent foci hot spot on the detector array and with
The scan variations of the 2-D vibration mirror can distinguish the fluorescence signal of each self-excitation of focus with different modulating frequency, from
And reconstruct the two dimensional image of reflection sample message.
4. according to claim 3 be based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, feature
It is：
Wherein, the first direction linearly polarized light is orthogonal with the second direction linearly polarized light.
5. according to claim 3 be based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, feature
It is：
Wherein, the polyhedral prism is the cylinder with bottom surface and multiple faceted pebbles, and cross section is polygon, the load
The direction of frequency mixed light beam is with the plane perpendicular and the carrier frequency mixed light beam is introduced into the bottom surface, the focusing light
Spot is one-dimensional array distribution, and the quantity of the focal beam spot is identical as the quantity of the faceted pebble.
6. according to claim 3 be based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, feature
It is：
Wherein, first lens unit is used for optical beam transformation, carries reflecting described in the multi beam being emitted from the polyhedral prism
Frequency mixed light beam is full of the entrance pupil of the focusing objective len always, realizes the optimal imaging performance of the focusing objective len,
First lens unit includes the combination of two optical beam transformation lens or multiple optical beam transformation lens,
The angle of the dichroscope and the 4th optical axis is 45 degree.
7. according to claim 3 be based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, feature
It is：
Wherein, when the polyhedral prism is the cone prism with bottom surface and multiple faceted pebbles, the bottom surface of the prism and the load
Frequency mixed light beam direction is vertical, and the focal beam spot is two-dimensional array distribution, the quantity of the focal beam spot and the faceted pebble
Quantity is identical.
8. according to claim 3 be based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, feature
It is：
It wherein, further include being arranged on second optical axis and being located at the frequency modulator and described the in the step 2
The third lens unit for being used to improve axial resolution of two-mirror.
9. according to claim 8 be based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method, feature
It is：
Wherein, the third lens unit includes the combination of a lens or multiple lens, is used for the first carrier frequency light beam
The focussing plane of focussing plane and the second carrier frequency light beam be separated by a distance.
10. according to claim 8 or claim 9 based on polyhedral prism and light beam frequency modulation high-resolution high speed imaging method,
It is characterized in that：
Wherein, in step 14, using frequency demodulation algorithm, extract the received overlapping of the detector array has carrier frequency f1
With the fluorescent foci spot signal of f2, and the intensity by analyzing the fluorescent foci hot spot on the detector array
And with the scan variations of the 2-D vibration mirror, the fluorescence letter of each self-excitation of focus with different modulating frequency can be distinguished
Number, to reconstruct the 3-D image of reflection sample message.
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