CN108982433A - A layer device is cut using the optics of optical interference microscopy and fluorescence microscopy - Google Patents
A layer device is cut using the optics of optical interference microscopy and fluorescence microscopy Download PDFInfo
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- CN108982433A CN108982433A CN201710416404.2A CN201710416404A CN108982433A CN 108982433 A CN108982433 A CN 108982433A CN 201710416404 A CN201710416404 A CN 201710416404A CN 108982433 A CN108982433 A CN 108982433A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/41—Refractivity; Phase-affecting properties, e.g. optical path length
- G01N21/45—Refractivity; Phase-affecting properties, e.g. optical path length using interferometric methods; using Schlieren methods
Abstract
A kind of optics using optical interference microscopy and fluorescence microscopy cuts a layer device, comprising: an optical splitter can make an incident beam be divided into a reflected beams and a transmitted light beam；One broad band light source device, is used to provide the described incident beam；One refers to end unit, for making the transmitted light beam return to the optical splitter through an adjustable light path；One shortwave light supply apparatus；One the 1st color optical splitter, first side face the shortwave light supply apparatus, and third edge surface is to the optical splitter, and its light beam that wavelength can be made to be shorter than a preset wavelength can not penetrate；One object lens face the second side of the one or the two color optical splitter with a directional light side；One sample load bearing unit, in face of an optically focused side of the object lens；And projecting lens and a sensing unit, for receiving an output beam of the optical splitter.
Description
Technical field
Layer device is cut the present invention relates to a kind of optics more particularly to a kind of using optical interference microscopy and fluorescence microscopy
Optics cut a layer device.
Background technique
When carrying out tumor resection, must often pathologists be waited to be examined with frozen section (frozen section)
To determine whether tumour cuts off completely, the time is not only expended, it is also possible to fully validated can not arrive all orientation due to it is pressed for time
Tumor tissues cut off.
During carrying out frozen section, for the sample of juicy, generated ice crystal (crystal after freezing
Ice institutional framework can) be destroyed；For the sample of fattiness (fat), in general tissue freezing solidification temperature (about -20 DEG C),
Its adipose tissue is easy to fall off from slice, causes biopsy tissues imperfect because of not yet freezing solidification.
Optical coherence Tomography (optical coherence tomography, OCT), is in recent years emerging one kind
Optical image technology, working principle is similar to ultrasonic, but resolution ratio ultrasonic is higher, mainly using each tissue to light
Reflection, absorption and scattering power difference and sample is imaged and is differentiated through principle of optical interference.Because can be directly
Tissue under room temperature is scanned, is not required to using the programs such as freezing and slice, so being avoided that juicy or fattiness group
The structure distortion (morphological artifacts) generated when being woven in frozen section, to maintain the integrality of tissue samples,
Therefore the accuracy that pathologic finding can be improved also is contributed to operating time is shortened with operation effect is promoted.However due to the light
The depth of field (depth of focus) for learning imaging technique is too big, and tissue must be made to entity slice (real when so checking
Sectioning), thickness is usually 4~5gm, is coincided together with to avoid the tissue image of different depth, and then is obtained clear
Clear image.
Known technology, if Authorization Notice No. is US9185357 B2, entitled " Optical tissue
The United States Patent (USP) of sectioning using full field optical coherence tomography ", discloses one kind
For observing the whole audience formula OCT device of histotomy, it is characterised in that: include a whole audience imaging interferometer and an optical segmentation
Imaging system.By optical interdferometer the problem of improving the depth of field, the structure image in tissue can be directly obtained, the image is made
Resolution ratio laterally and longitudinally is attained by within 1 μm, so that saving tissue fixes (fixation) program.Such as: freezing or stone
The program of wax (paraffin) cladding and entity slice (sectioning).
In addition, traditional H&E stained slice (H&E section) is using hematoxylin (hematoxylin) and Yihong
(eosin) two kinds of stains respectively to nucleus (nucleus) and cytoplasm (cytoplasm) bluish violet and pink, so
And the signal of royal purple is presented in nucleus in the image obtained, is not easy to show endonuclear details.Such as: kernel (nucleolus)
Or heterochromatin (heterochromotin), the patent are to supplement nucleus thin portion image by fluorescent staining mode to meet
Needed for pathologic finding.
However, in the patent framework: (1), shortwave light beam (such as: ultraviolet light) is in the power meeting after optical splitter spectro-film
It differs be left original luminous intensity 10~40%；(2), the antireflection plated film of optical splitter is less than the ultraviolet light wave of 400nm in wavelength
Section penetrance is lower.Under the influence of this two kinds of factors, the fluorescence signal that will lead to sample becomes very weak, for enhancing fluorescence letter
Number intensity shortens the time for exposure, and then accelerates capture speed, and this field needs a novel optics and cuts a layer device.
Summary of the invention
A purpose of the present invention is that disclosing a kind of optics cuts a layer device, wherein the one or two color optical splitter be placed in optical splitter with
Between first object lens, the shortwave light beam for making to be irradiated on sample will not be split device light splitting, and enhance fluorescence signal intensity,
Shorten the time for exposure, and then accelerates capture speed.
Another object of the present invention is to disclose a kind of optics to cut a layer device, wherein fluorescent light beam emitted by the sample is passing through
When the one or two color optical splitter, the one or two color optical splitter can filter shortwave light beam in advance to obtain the preferable fluorescence letter of contrast
Number, shorten the time for exposure, and then accelerate capture speed.
Another object of the present invention is to disclose a kind of optics to cut a layer device, wherein there is sensing unit long wave to lead to optical filter
It filters the shortwave light beam further to promote fluorescence signal intensity, shortens the time for exposure, and then accelerate capture speed.
In order to achieve the above objectives, propose that a kind of optics using optical interference microscopy and fluorescence microscopy cuts a layer device,
It is included
One optical splitter has a first side, a second side, a third side and a four side, and it can make
The reflected beams being pierced by by the second side are divided into and by the third by an incident beam of the first side incidence
The transmitted light beam that side is pierced by；
One broad band light source device irradiates the first side of the optical splitter for generating a broad band light beam；
One refers to end unit, for making the transmitted light beam return to the optical splitter through an adjustable light path；
One shortwave light supply apparatus, for generating a shortwave light beam；
One the 1st color optical splitter has a first side, a second side and a third side, the first side face
To the shortwave light supply apparatus, the third edge surface is to the second side of the optical splitter, and the one or two color optical splitter is used
It can not be penetrated in the light beam for making wavelength be shorter than a preset wavelength, and the wavelength of the shortwave light beam is less than the preset wavelength；
One first object lens have a directional light side and an optically focused side, and the directional light side is to the one or two color optical splitter
The second side；
One sample load bearing unit, in face of first object lens optically focused side and for carrying a sample for being infected with fluorescer；
One projecting lens has an incident side and a light emission side, the four side of the light incident sides to the optical splitter；
And one sensing unit face the projecting lens the light emission side.
In one embodiment, this includes: an optical path delay device with reference to end unit, has a first side and a second side
Side, the first side face the third side of the optical splitter；One second object lens have a directional light side and an optically focused
Side, the second side of the directional light side to the optical path delay device；And a reflecting mirror, described in second object lens
Optically focused side, for reflecting the transmitted light beam, wherein the optical path delay device is for adjusting the adjustable light path so that the adjustable light path pair
Claim in being formed by a sample light path by the sample load bearing unit, first object lens and the one or two color optical splitter.
In one embodiment, the broad band light source device, the shortwave light supply apparatus include a light source；An or light source and one
Grating；Or a light source, a grating and one can modulation tilt angle turning mirror；An or LED strip shape distribution light source.
In one embodiment, which it is appropriate to provide first object lens one further to have a white light source
Penetrate brightness, the white light source include a white light LEDs, a white light halogen lamp or a tengsten lamp.
In one embodiment, which includes one the 2nd 2 color optical splitter, a color 2 D photosensory assembly, a long wave
Logical optical filter and a single color two-dimension photosensory assembly, the two or two color optical splitter have a first side, a second side and one the
Three sides, the first side faces the projecting lens, by the fluorescent light beam and the white light beam via the third side
It reflects and images in the color 2 D photosensory assembly, and the broad band light beam is transmitted via the second side and images in the list
Color two dimension photosensory assembly；Or the sensing unit includes a turnable turning mirror, a color 2 D photosensory assembly, the logical optical filtering of a long wave
Device and a single color two-dimension photosensory assembly, when which rights, which images in photosensitive group of the color 2 D
Part, then the broad band light beam and the fluorescent light beam successively image in single color two-dimension photosensory assembly when opening, which leads to optical filter and set
Be placed between the projecting lens and the reversible turning mirror, between the projecting lens and the two or two color optical splitter, this may be reversed
Between formula turning mirror and the single color two-dimension photosensory assembly or between the two or two color optical splitter and the single color two-dimension photosensory assembly.
In one embodiment, the wave-length coverage of the broad band light beam is in 470nm to 800nm, the wavelength model of the shortwave light beam
365nm is trapped among between 460nm, in 400nm between 800nm, the one or two color is divided the operating wavelength range of the optical splitter
Device and the long wave lead to the cutoff wavelength range of optical filter in 400nm between 470nm.
In one embodiment, the wave-length coverage of the broad band light beam is in 650nm to 1000nm, the wavelength model of the shortwave light beam
365nm is trapped among between 630nm, the operating wavelength range of the optical splitter is divided in 400nm to the one or two color between 1000nm
Device, the two or two color optical splitter and the long wave lead to the cutoff wavelength range of optical filter in 400nm between 650nm.
In one embodiment, further there is one first polarizer, the list between the broad band light source device and the optical splitter
Further there is one second polarizer, between first object lens and the one or two color optical splitter further in front of color two dimension photosensory assembly
With one first quarter-wave plate, further there is one second quarter-wave between the optical path delay device and second object lens
Piece, first polarizer have one first polarization direction, which has one second polarization direction, first polarizer
Direction and second polarization direction are orthogonal, first quarter-wave plate have a primary optic axis direction, the two or four point
One of wave plate there is one second optical axis direction, the primary optic axis direction and second optical axis direction are between first polarization direction
Between second polarization direction, for providing the effect of an enhancing interference efficiency and the quality of image.
It in one embodiment, further include an information processing unit, for executing an image processing program.
In one embodiment, this further has an axial platform with reference to end unit, which further has
There is a three-dimensional mobile platform, with this refers to second object lens of end unit and the reflecting mirror, by being somebody's turn to do by axial platform movement
Mobile this of three-dimensional mobile platform is infected with sample and the modulation optical path delay device of fluorescer, and the information processing unit is enable to count
Calculate a 3-dimensional image of the sample.
Detailed description of the invention
Fig. 1 is that the optics using optical interference microscopy and fluorescence microscopy that one embodiment of the invention proposes cuts a layer device
Structural schematic diagram；
Fig. 2 is the light splitting of Fig. 1 and the schematic diagram of focusing；
Fig. 3 is the structural schematic diagram of an embodiment of the reference end unit of Fig. 1；
Fig. 4 is the receipts light of Fig. 1 and the schematic diagram for converging rear projection process；
Fig. 5 a is the structural schematic diagram of an embodiment of the sensing unit of Fig. 1；
Fig. 5 b is the structural schematic diagram of another embodiment of the sensing unit of Fig. 1；
Fig. 5 c is the structural schematic diagram of the another embodiment of the sensing unit of Fig. 1；
Fig. 5 d is the structural schematic diagram of the another embodiment of the sensing unit of Fig. 1；
Fig. 6 is that the optics using optical interference microscopy and fluorescence microscopy that another embodiment of the present invention proposes cuts layer dress
The structural schematic diagram set；
Fig. 7 is that the optics using optical interference microscopy and fluorescence microscopy of yet another embodiment of the invention cuts a layer device
Structural schematic diagram.
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with specific embodiment, and reference
Attached drawing, the present invention is described in further detail.
Please with reference to Fig. 1 to Fig. 2, wherein Fig. 1 is one embodiment of the invention using optical interference microscopy and fluorescence
The optics of microscopy cuts the structural schematic diagram of layer device, and Fig. 2 is the light splitting of Fig. 1 and the schematic diagram of focusing.
As shown, the optics using optical interference microscopy and fluorescence microscopy of the present embodiment cuts a layer device, packet
It includes: a broad band light source device 100；One shortwave light supply apparatus 200；One optical splitter 300；One the 1st color optical splitter 400；One first
Object lens 500；One sample load bearing unit 600；One projecting lens 700；One refers to end unit 800；An and sensing unit 900.
Wherein, the broad band light source device 100 is for generating a broad band light beam 10, and the direction of travel of the broad band light beam is in one
Nominal arrow indicates；For the shortwave light supply apparatus 200 for generating a shortwave light beam 20, the direction of travel of the shortwave light beam is real with one
Line arrow indicates；For the sample load bearing unit 600 for carrying a sample 610 for being infected with fluorescer, this is infected with the sample of fluorescer
Fluorescer in 610 can be released a fluorescent light beam 30, the direction of travel of the fluorescent light beam 30 by the shortwave light beam 20 irradiation
It is indicated with dotted arrow；The sample load bearing unit 600 further has a white light source 620 to provide first object lens 500 1
Brightness suitably is penetrated, the direction of travel of a white light beam 40 of the white light source 620 release is indicated with a nominal arrow.
The optical splitter 300 has a first side S301, a second side S302, a third side S303 and one the 4th side
Side S304；The broad band light source device 100 is for generating a broad band light beam 10 to irradiate the first side of the optical splitter 300
S301, the optical splitter 300 can make to be divided by the broad band light beam 10 of the first side S301 incidence by the second side
S302 is reflected and is transmitted by the third side S303.
The shortwave light supply apparatus 200 has one first for generating the 20, the 1st color optical splitter 400 of a shortwave light beam
Side S401, a second side S402 and a third side S403, the first side S401 face the shortwave light supply apparatus 200
And the shortwave light beam 20 is reflected, the third side S403 faces the second side S302 of the optical splitter 300 and for saturating
Penetrate the broad band light beam 10.
First object lens 500, have a directional light side S501 and an optically focused side S502, the directional light side S501 in face of this
The second side S402 of one or two color optical splitters 400；The sample load bearing unit 600, in face of the optically focused of first object lens 500
Side S502 and for carrying a sample 610 for being infected with fluorescer, the sample load bearing unit 600, first object lens 500 and this first
Two color optical splitters 400 form a sample light path.
There is the projecting lens 700 an incident side S701 and a light emission side S702, incident side S701 to face the optical splitter
The 300 four side S304；The sensing unit 900 faces the light emission side S702 of the projecting lens 700.
Wherein, which for example but is not limited to include a white light LEDs, a white light halogen lamp or a tengsten lamp,
The operating wavelength range of first object lens 500 for example but is not limited to 350~1000nm, the broad band light source device 100, the short wavelength light
Source device 200 includes a light source；Or light source and a grating；Or a light source, a grating and one can modulation tilt angle turn
Fold mirror；Or a LED strip shape distribution light source (being not shown in figure), it is known technology, therefore is not repeated to describe herein.
Referring to figure 3., be Fig. 1 one embodiment of reference end unit structural schematic diagram.
As shown, this includes: an optical path delay device 810 with reference to end unit 800；One second object lens 820 and a reflection
Mirror 830.
The optical path delay device 810, second object lens 820 and the reflecting mirror 830 form an adjustable light path, and the reference end is single
Member 800 is for making the broad band light beam 10 return to 300 (not shown) of optical splitter via the adjustable light path.
Wherein, the optical path delay device 810 has an a first side S811 and second side S812, the first side
S811 faces the third side S303 (being not shown in figure) of the optical splitter 300；Second object lens 820, it is parallel with one
Light side S821 and an optically focused side S822, the directional light side S821 face the second side S812 of the optical path delay device 810；It should
Reflecting mirror 830 faces the optically focused side S822 of second object lens 820, for reflecting the broad band light beam 10.Second object lens 820
Operating wavelength range for example but be not limited to 350~1000nm.
The optical path delay device 810 for example, by but be not limited to change a turning mirror displacement (displacement), at this
Adjustable light path is symmetrical under the sample light path state, for generating a continuous interference carrier wave (continuous interference
Carrier wave), carrier number can be one or more, then be recorded in each pixel by single color two-dimension photosensory assembly 940, do
Strength Changes caused by relating to capture cross section (en-face) interference strength shadow after r.m.s. and intensity are averagely waited and calculated
Picture, wherein the interference carrier wave can also be generated by three-dimensional mobile platform 630.It is known technology, and details are not described herein.
It referring to figure 4., is the receipts light of Fig. 1 and the schematic diagram for converging rear projection process.
As shown, the sample load bearing unit 600 can reflect the shortwave light beam 20 and the broad band light beam 10, this is infected with fluorescence
After fluorescer in 610 (not shown) of sample of agent is irradiated by the shortwave light beam 20, a fluorescent light beam 30 can be released,
620 (not shown) of white light source that the sample load bearing unit 600 has can also release a white light beam 40.
The light beam that one or two color optical splitter 400 is used to that wavelength to be made to be shorter than a preset wavelength can not penetrate, and the short wavelength light
The wavelength of beam 20, which is less than the preset wavelength namely the shortwave light beam 20, to be penetrated, only the broad band light beam 10, the fluorescent light beam
30 and the white light beam 40 can penetrate the one or two color optical splitter 400 respectively.
The third side S303 of the optical splitter 300 is used to reflect the broad band light beam 10 with reference to end unit 800 in face of this；
The second side S302 is in face of the one or two color optical splitter 400 for transmiting the broad band light beam 10, the fluorescent light beam 30 respectively
And the white light beam 40.
Wherein, since the adjustable light path and the sample light path are symmetrical (between the coherent
Length), so that the broad band light beam reflected via the adjustable light path reflects broad band light beam conjunction with via the sample light path
And and generate an optical interference phenomena, be known technology, details are not described herein.
There is the projecting lens 700 an incident side S701 and a light emission side S702, incident side S701 to face the optical splitter
The 300 four side S304, light emission side S702 are used to project the broad band light beam 10, the fluorescence to the sensing unit 900
Light beam 30 and the white light beam 40.
Please with reference to Fig. 5 a~Fig. 5 b, wherein Fig. 5 a is the structural schematic diagram of an embodiment of the sensing unit of Fig. 1；Figure
5b is the structural schematic diagram of another embodiment of the sensing unit of Fig. 1.
The sensing unit 900 includes: a turnable turning mirror 915；One color 2 D photosensory assembly 920；The logical filter of one long wave
Light device 930 and a single color two-dimension photosensory assembly 940.
Wherein the long wave leads to optical filter 930 for further filtering 20 (not shown) of shortwave light beam, this can be turned over
Formula turning mirror 915 have a turnable bracket (flip mount, be not shown in the figure) to be adjusted to one right (flip on) or
One opens the state of (flip off), is known technology, details are not described herein.
As shown in Figure 5 a, the logical optical filter 930 of the long wave is set to the projecting lens 700 and the reversible turning mirror 915
Between, when the turnable turning mirror 915 rights (flip on), which images in the color 2 D photosensory assembly
920, when the turnable turning mirror 915 opens (flip off), then the broad band light beam 10 and the fluorescent light beam 30 are successively imaged
In single color two-dimension photosensory assembly 940.
As shown in Figure 5 b, which leads to the setting reversible turning mirror 915 of optical filter 930 and photosensitive group of the single color two-dimension
Between part 940, when the turnable turning mirror 915 rights (flip on)) when, which images in the color 2 D sense
Optical assembly 920, when the turnable turning mirror 915 opens (flip off), then the broad band light beam 10 and the fluorescent light beam 30 are first
After image in single color two-dimension photosensory assembly 940.
Please with reference to Fig. 5 c~Fig. 5 d, wherein Fig. 5 c is the structural schematic diagram of the another embodiment of the sensing unit of Fig. 1；
Fig. 5 d is the structural schematic diagram of the another embodiment of the sensing unit of Fig. 1.
The sensing unit 900 includes: one second dichroic beam splitter 910；One color 2 D photosensory assembly 920；One long wave is logical
Optical filter 930 and a single color two-dimension photosensory assembly 940, wherein the long wave leads to optical filter 930 and is used for the shortwave light beam 20 (in figure
It is not shown) further filtering.
As shown in Figure 5 c, the logical optical filter 930 of the long wave is set to the projecting lens 700 and second dichroic beam splitter 910
Between, by the two or two color optical splitter 910 by the fluorescent light beam 30 and the white light beam 40 via the third side S903
It reflects and images in the color 2 D photosensory assembly 920；And the broad band light beam 10 is transmitted via the second side S902
And image in the single color two-dimension photosensory assembly 940.
As fig 5d, which leads to the two or two color optical splitter 910 of the setting of optical filter 930 and photosensitive group of the single color two-dimension
Between part 940, by the two or two color optical splitter 910 by the fluorescent light beam 30 and the white light beam 40 via the third side
S903 reflects and images in the color 2 D photosensory assembly 920；And by the broad band light beam 10 via the second side S902
It transmits and images in the single color two-dimension photosensory assembly 940.
Wherein, when the wave-length coverage of the broad band light beam 10 is in 470nm to 800nm, the turnable turning mirror need to be used
915, in 365nm between 460nm, which (does not show the wave-length coverage of 20 (not shown) of shortwave light beam in figure
In 400nm between 800nm, the cutoff wavelength range of the one or two color optical splitter 400 exists operating wavelength range out)
400nm is between 470nm.
When the wave-length coverage of the broad band light beam 10 is in 650nm to 1000nm, the wave-length coverage of the shortwave light beam 20 exists
365nm is between 630nm, and the operating wavelength range of the optical splitter 300 is in 400nm to the turnable between 1000nm, can be used
Turning mirror 915 or the two or two color optical splitter 910, the one or two color optical splitter 400, the two or two color optical splitter 910 and the length
Wave leads to the cutoff wavelength range of optical filter 930 in 400nm between 650nm.
Fig. 6 is please referred to, is the optics using optical interference microscopy and fluorescence microscopy of another embodiment of the present invention
Cut the structural schematic diagram of layer device.
As shown, further having one first polarizer 950 between the broad band light source device 100 and the optical splitter 300；
940 front of single color two-dimension photosensory assembly further has one second polarizer 960；One or two color optical splitter 400 with this
Further there is one first quarter-wave plate 970 between one object lens 500；Between the optical path delay device 810 and the second object lens 820 into
One step has one second quarter-wave plate 980.
Wherein, which has one first polarization direction, which has one second polarization
Direction, first quarter-wave plate 970 have a primary optic axis direction, which has one second
Optical axis direction, the first polarizer direction and second polarization direction are orthogonal, the primary optic axis direction and the second optical axis side
To between first polarization direction and second polarization direction.
The first polarizer of this in figure direction is vertical；Second polarization direction is level；The primary optic axis direction and this
Two optical axis directions are 45 degree, and but not limited to this.When the broad band light beam 10 that the broad band light source device 100 generates by this
One polarizer 950 becomes vertical polarization directions, refers to end unit by this respectively after being divided via the optical splitter 300
800 the second quarter-wave plate 980 and by between first object lens 500 and the one or two color optical splitter 400 this first
It after quarter-wave plate 970, can become forward circular polarisation direction, become reverse circular polarisation direction after reflection, respectively once again
Become horizontal polarization directions, and the single color two-dimension after the second quarter-wave plate 980 and first quarter-wave plate 970
Second polarizer, 960 tolerable injury level polarization direction broad band light beams 10 in 940 front of photosensory assembly pass through, and further isolate out
The reflected light of 300 anti-reflective film of spectroscope, for providing the effect of an enhancing interference efficiency and the quality of image.
Fig. 7 is please referred to, is the optics using optical interference microscopy and fluorescence microscopy of yet another embodiment of the invention
Cut the structural schematic diagram of layer device.
As shown, this further has an axial platform 840 with reference to end unit 800；The sample load bearing unit 600 into one
Step has a three-dimensional mobile platform 630；This cuts a layer device using the optics of optical interference microscopy and fluorescence microscopy, more into
One step includes an information processing unit (not shown), to execute an image processing program.
By mobile second object lens 820 of axial platform 840 and the reflecting mirror 830, modulation optical path delay device 500 and with three
Mobile this of dimension mobile platform 630 is infected with the sample 610 of fluorescer, and the information processing unit (not shown) is enable to calculate
A 3-dimensional image (not shown) of the sample out, is known technology, details are not described herein.
By above-mentioned disclosed design, the present invention is had the advantage that
1, optics of the invention cuts a layer device, wherein the one or two color optical splitter is placed between optical splitter and the first object lens, uses
Shorten the time for exposure to enhance fluorescence signal intensity in making to be irradiated to the device light splitting that will not be split of the shortwave light beam on sample,
Accelerate capture speed.
2, optics of the invention cuts a layer device, wherein fluorescent light beam emitted by the sample pass through the one or two color optical splitter when,
One or two color optical splitter can filter shortwave light beam in advance to obtain the preferable fluorescence signal of contrast, shorten the time for exposure, add
Fast capture speed.
3, optics of the invention cuts a layer device, and wherein there is sensing unit long wave to lead to optical filter, further filters the shortwave
Light beam shortens the time for exposure to enhance fluorescence signal intensity, accelerates capture speed.
Particular embodiments described above has carried out further in detail the purpose of the present invention, technical scheme and beneficial effects
Describe in detail bright, it should be understood that the above is only a specific embodiment of the present invention, is not intended to restrict the invention, it is all
Within the spirit and principles in the present invention, any modification, equivalent substitution, improvement and etc. done should be included in protection of the invention
Within the scope of.
Claims (10)
1. a kind of optics using optical interference microscopy and fluorescence microscopy cuts a layer device, comprising:
One optical splitter has a first side, a second side, a third side and a four side, and it can make by institute
The incident beam for stating first side incidence is divided into the reflected beams being pierced by by the second side and by the third side
The transmitted light beam being pierced by；
One broad band light source device irradiates the first side of the optical splitter for generating a broad band light beam；
One refers to end unit, for making the transmitted light beam return to the optical splitter through an adjustable light path；
One shortwave light supply apparatus, for generating a shortwave light beam；
One the 1st color optical splitter, has a first side, a second side and a third side, and the first side faces institute
State shortwave light supply apparatus, the second side of the third edge surface to the optical splitter, the one or the two color optical splitter use
It can not be penetrated in the light beam for making wavelength be shorter than a preset wavelength, and the wavelength of the shortwave light beam is less than the preset wavelength；
One first object lens have a directional light side and an optically focused side, and the directional light side is to the one or the two color optical splitter
The second side；
One sample load bearing unit, in face of first object lens optically focused side and for carrying a sample for being infected with fluorescer；
One projecting lens has an incident side and a light emission side, the four side of the light incident sides to the optical splitter；
And
One sensing unit, in face of the light emission side of the projecting lens.
2. cutting a layer device using the optics of optical interference microscopy and fluorescence microscopy as described in claim 1, wherein described
Include: with reference to end unit
One optical path delay device, has a first side and a second side, and the first side is described in face of the optical splitter
Third side；
One second object lens have a directional light side and an optically focused side, and the directional light side is to described in the optical path delay device
Second side；And
One reflecting mirror, in face of the optically focused side of second object lens, for reflecting the transmitted light beam, wherein the light path prolongs
Slow device is for adjusting the adjustable light path so that the adjustable light path is symmetrical with by the sample load bearing unit, first object lens
And the one or the two color optical splitter is formed by a sample light path.
3. cutting a layer device using the optics of optical interference microscopy and fluorescence microscopy as described in claim 1, wherein described
Broad band light source device, the shortwave light supply apparatus include a light source；Or light source and a grating；Or a light source, a grating and
One can modulation tilt angle turning mirror；An or LED strip shape distribution light source.
4. cutting a layer device using the optics of optical interference microscopy and fluorescence microscopy as described in claim 1, wherein described
Sample load bearing unit further has a white light source, and to provide, first object lens one are appropriate to penetrate brightness, the white light light
Source includes a white light LEDs, a white light halogen lamp or a tengsten lamp.
5. cutting a layer device using the optics of optical interference microscopy and fluorescence microscopy as described in claim 1, wherein described
Sensing unit includes one the 2nd 2 color optical splitter, a color 2 D photosensory assembly, the logical optical filter of a long wave and a single color two-dimension sense
Optical assembly, the two or the two color optical splitter have a first side, a second side and a third side, the first side face
To the projecting lens, the fluorescent light beam and the white light beam are reflected via the third side and imaged in described
Color 2 D photosensory assembly, and the broad band light beam is transmitted via the second side and to image in the single color two-dimension photosensitive
Component；Or the sensing unit includes that a turnable turning mirror, a color 2 D photosensory assembly, the logical optical filter of a long wave and one are single
Color two dimension photosensory assembly, when the turnable turning mirror rights, the white light beam images in the color 2 D photosensory assembly,
Then the broad band light beam and the fluorescent light beam successively image in single color two-dimension photosensory assembly when opening, and the long wave leads to optical filter
Be set between the projecting lens and the reversible turning mirror, between the projecting lens and the two or two color optical splitter,
Between the reversible turning mirror and the single color two-dimension photosensory assembly or the two or the two color optical splitter and described monochromatic two
It ties up between photosensory assembly.
6. cutting a layer device using the optics of optical interference microscopy and fluorescence microscopy as claimed in claim 5, wherein described
For the wave-length coverage of broad band light beam in 470nm to 800nm, the wave-length coverage of the shortwave light beam is in 365nm to the described 1st
Color optical splitter and the long wave lead to the cutoff wavelength range of optical filter in 400nm between 470nm.
7. cutting a layer device using the optics of optical interference microscopy and fluorescence microscopy as claimed in claim 5, wherein described
The wave-length coverage of broad band light beam in 650nm to 1000nm, the wave-length coverage of the shortwave light beam in 365nm between 630nm,
The operating wavelength range of the optical splitter in 400nm between 1000nm, the one or the two color optical splitter, the two or two color optical splitter
And long wave leads to the cutoff wavelength range of optical filter in 400nm between 650nm.
8. the optics using optical interference microscopy and fluorescence microscopy as described in claim 2 or 5 cuts a layer device, wherein
Further there is one first polarizer, before the single color two-dimension photosensory assembly between the broad band light source device and the optical splitter
Side further has one second polarizer, further has one the 1st between first object lens and the one or the two color optical splitter
/ mono- wave plate, further has one second quarter-wave plate between the optical path delay device and second object lens, and described the
One polarizer has one first polarization direction, and second polarizer has one second polarization direction, the first polarizer side
To orthogonal with second polarization direction, first quarter-wave plate has a primary optic axis direction, and described second
Quarter-wave plate has one second optical axis direction, and the primary optic axis direction and second optical axis direction are between described the
Between one polarization direction and second polarization direction, for providing the effect of an enhancing interference efficiency and the quality of image.
9. a layer device is cut using the optics of optical interference microscopy and fluorescence microscopy as claimed in claim 2, more into one
Step includes an information processing unit, for executing an image processing program.
10. cutting a layer device using the optics of optical interference microscopy and fluorescence microscopy as claimed in claim 9, wherein institute
Stating further has an axial platform with reference to end unit, and the sample load bearing unit further has a three-dimensional mobile platform, leads to
Cross optical path delay device described in mobile second object lens of the axial platform and reflecting mirror, modulation and by described three-dimensional mobile flat
It is infected with the sample of fluorescer described in platform is mobile, and the information processing unit is enable to calculate a three-dimensional shadow of the sample
Picture.
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