CN102540439B - Confocal axial scanning device and confocal axial scanning method based on reflection type liquid crystal spatial light modulator - Google Patents
Confocal axial scanning device and confocal axial scanning method based on reflection type liquid crystal spatial light modulator Download PDFInfo
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Abstract
A confocal axial scanning device and a confocal axial scanning method based on a reflection type liquid crystal spatial light modulator relate to a confocal microscope and a scanning method of the confocal microscope, which solve the problems that in an existing axial scanning device and a method, the device is complex in structure and a sample is poor in repeatability of measurement due to the fact that defocus of a measured object is achieved by means of a lens or movement of the measured object along an axial direction. By means of the confocal axial scanning device, the reflection type liquid crystal spatial light modulator is added between a polarization beam splitter and the lens of the existing device, an axial scanning mechanism in a traditional confocal axial scanning device is eliminated simultaneously, and the reflection type liquid crystal spatial light modulator is utilized to replace the axial scanning mechanism. The confocal axial scanning method adopts a mode of utilizing a computer to directly modulate a phase gray-scale figure to adjust wavefront of a light wave in front of the lens so that zoom is achieved, and axial scanning to the measured object can be achieved without axially moving the measurement lens and the measured object. The confocal axial scanning device and the confocal axial scanning method based on the reflection type liquid crystal spatial light modulator are applied to the confocal microscope and the scanning method of the confocal microscope.
Description
Technical field
The present invention relates to a kind of confocal microscope and scan method thereof.
Background technology
Since the 1980s, microelectric technique, bioengineering, micro-optic and Micro-Opto-Electro-Mechanical Systems technology have entered the stage of a develop rapidly, people pay attention to the quantitative test to three-dimensional state more in the microscale field, and three-dimensional Fast Ultra-Precision Measurement has become the important topic in modern test technology and the instrument research.And confocal measurement method is because of its high precision, high resolution, noncontact be easy to realize that the digitized unique advantage of three-dimensional imaging becomes the important research direction that three-dimensional microstructures is measured.A kind of method that adopts confocal measurement method to realize that three-dimensional microstructures is measured is that testee is moved axially with respect to the focal position, form defocus signal at detector surface, the light intensity that detects according to detector realizes the measurement to the testee three-dimensional microstructures.Typical measurement mechanism is as follows:
Harbin Institute of Technology's automatic test has proposed a kind of confocal device that can realize axial scan with the Wang Fusheng doctor of control system in its PhD dissertation " based on the three-dimension measuring system research of the micro-Detection Techniques of differential confocal ", the formation of this device is to comprise: laser instrument, convergent lens, pin hole 3, beam expander, polarization spectroscope, λ/4 wave plates, object lens, testee also comprise: condenser, spectroscope, pin hole 1, photodetector 1, pin hole 2, photodetector 2.The parallel beam of laser instrument emission is imaged onto pin hole 3 positions through plus lens and forms pointolite, the light beam of pointolite emission incides on the polarization spectroscope through beam expander is parallel, transmission is crossed the light beam of polarization spectroscope through being imaged onto the testee surface by object lens behind λ/4 wave plates and reflecting along former road, again reflex to condenser formation converging beam through being polarized spectroscope behind λ/4 wave plates, converging beam is divided into the two-way differential wave by spectroscope, pass respectively burnt before and surveyed by photodetector behind defocused two pin holes and form the differential confocal system.This device moves forward and backward the realization axial scan by Z-direction driver element carrying object lens along optical axis direction.
Doctor Huang Xiangdong of Harbin Institute of Technology's automatic test and control system has introduced the principle of work of reflective laser scanning confocal microscope in its PhD dissertation " array space super-resolution confocal microscan Detection Techniques and theoretical research ", the formation of this device is to comprise: laser instrument, pin hole 1, spectroscope, object lens, testee also comprise: pin hole 2, detector.The light beam of laser instrument emission forms pointolite through needle passing hole 1, and the light beam of pointolite emission is imaged onto the testee surface and along the reflection of former road, reflection ray is reflexed to pin hole 3 positions by spectroscope again through behind the object lens again through object lens.This device comes the realization system to the axial scan of testee by the axial location of adjusting testee.
These two pieces of PhD dissertation have all proposed a kind of confocal microscopy and scan method that can realize axial scan, the characteristics that they have are that the testee out of focus is that this method just requires measurement mechanism to comprise axial scan mechanism by object lens or in axial direction mobile realization of testee.First shortcoming with axial scan mechanism arrangement is that apparatus structure is complicated, realizes that precision measurement needs routing motion mechanism, control gear and feedback element; Second shortcoming be if axially movable be sample, and deformation can constantly occur in this sample in moving process, such as cell, not only reduce the measuring accuracy of this sample, and bad measurement repeatability.
Summary of the invention
The objective of the invention is to realize because the testee out of focus in axial direction moves by object lens or testee in order to solve existing axial scan device and method, thereby cause the problem of bad measurement repeatability of complex structure, the sample of device, a kind of confocal axial scan device and confocal axial scanning method based on the reflection type liquid crystal spatial light modulator is provided.
Confocal axial scan device based on the reflection type liquid crystal spatial light modulator, it comprises laser instrument, convergent lens, the first pin hole, illumination objective lens, diaphragm, polarization spectroscope, λ/4 wave plates, the reflection type liquid crystal spatial light modulator, measured object, measure object lens, spectroscope, collect object lens, the second pin hole and light intensity detector, the parallel beam of laser instrument emission is imaged onto the first pin hole place by convergent lens, light beam by the first pin hole collimates the rear parallel polarization spectroscope that incides by illumination objective lens, polarization spectroscope is divided into reflected light and transmitted light to incident light, described reflected light is S light, described transmitted light is P light, transmitted light is by arriving the reflection type liquid crystal spatial light modulator behind λ/4 wave plates, the reflection type liquid crystal spatial light modulator is arriving light total reflection, the total reflection light that obtains is back into by original optical path and is incident upon polarization spectroscope, reflected light through this polarization spectroscope reflection is incident to spectroscope, reflected light through this spectroscope reflection is incident to the measurement object lens, be imaged onto on the measured object by these measurement object lens, be back into along original optical path through the reflected light of measured object reflection and be incident upon spectroscope, transmitted light through this spectroscope transmission is imaged onto the second pin hole position by collecting object lens, and incides the photosurface of light intensity detector by the second pin hole.
Realize confocal axial scanning method based on above-mentioned confocal axial scan device based on the reflection type liquid crystal spatial light modulator, its concrete steps are as follows:
The relative position of step 2, the maintenance described object to be scanned of step 1 and measurement object lens is motionless, by the phase place gray-scale map variation of computer control reflection type liquid crystal spatial light modulator, so that theoretical focussing distance x corresponding to described phase place gray-scale map
FIn axial direction from 0 μ m to the stepping of 105 μ m with 5 μ m, described phase place gray-scale map and theoretical focussing distance x
FBetween the pass be:
Wherein, f
0Be the focal length of measuring object lens, d is that the reflection type liquid crystal spatial light modulator arrives the light path of measuring object lens, and λ is the wavelength of light wave, and ξ, η represent the space two-dimensional coordinate;
After the phase place gray-scale map of each adjustment reflection type liquid crystal spatial light modulator, adopt the light intensity detector imaging, obtain corresponding intensity signal;
Step 3, each the theoretical focussing distance x that obtains according to step 2
FObtain a corresponding confocal axial scan curve with intensity signal.
Confocal axial scan device of the present invention, between the polarization spectroscope on the existing apparatus basis and object lens, increased the reflection type liquid crystal spatial light modulator, remove simultaneously axial scanning mechanism in traditional confocal axial scan device, replace axial scan mechanism with the reflection type liquid crystal spatial light modulator.The structure of apparatus of the present invention is more simple in structure than the prior art, confocal axial scanning method of the present invention adopts the mode of the direct phase modulation gray-scale map of computing machine to adjust the before wavefront of light wave of object lens, thereby realization zoom, so that measurement object lens and measured object all do not need to move axially the axial scan that can realize measured object, not only avoided the accurate and complicated control of three-dimensional micro-displacement platform, and make confocal axial scanning method operation of the present invention simpler, improve simultaneously measuring accuracy and the repeatability of easy deformation sample, thereby enlarged the sample measurement kind.
Description of drawings
Fig. 1 is based on the confocal axial scan apparatus structure synoptic diagram of reflection type liquid crystal spatial light modulator, and Fig. 2 is focussing distance x
FThe phase place gray-scale map of reflection type liquid crystal spatial light modulator during=10 μ m, Fig. 3 is focussing distance x
FThe phase place gray-scale map of reflection type liquid crystal spatial light modulator during=50 μ m, Fig. 4 is based on the confocal axial response experimental provision structural representation of reflection type liquid crystal spatial light modulator, Fig. 5 is that focussing distance is 40 μ m and the confocal axial scan curve comparison figure when not focusing, wherein solid line is that focussing distance is the curve behind the 40 μ m, and dotted line is the curve of not focusing.
Embodiment
Embodiment one: present embodiment is described in conjunction with Fig. 1, the described confocal axial scan device based on the reflection type liquid crystal spatial light modulator of present embodiment comprises laser instrument 1, convergent lens 2, the first pin hole 3, illumination objective lens 4, diaphragm 5, polarization spectroscope 6, λ/4 wave plates 7, reflection type liquid crystal spatial light modulator 8, measured object 10, measure object lens 11, spectroscope 12, collect object lens 13, the second pin hole 14 and light intensity detector 16, the parallel beam of laser instrument 1 emission is imaged onto the first pin hole 3 places by convergent lens 2, light beam by the first pin hole 3 collimates the rear parallel polarization spectroscope 6 that incides by illumination objective lens 4, polarization spectroscope 6 is divided into reflected light and transmitted light to incident light, described reflected light is S light, described transmitted light is P light, transmitted light is by λ/4 wave plates, 7 rear arrival reflection type liquid crystal spatial light modulators 8, reflection type liquid crystal spatial light modulator 8 is arriving light total reflection, the total reflection light that obtains is back into by original optical path and is incident upon polarization spectroscope 6, reflected light through these polarization spectroscope 6 reflections is incident to spectroscope 12, reflected light through these spectroscope 12 reflections is incident to measurement object lens 11, be imaged onto on the measured object 10 by these measurement object lens 11, be back into along original optical path through the reflected light of measured object 10 reflection and be incident upon spectroscope 12, transmitted light through these spectroscope 12 transmissions is imaged onto the second pin hole 14 positions by collecting object lens 13, and incides the photosurface of light intensity detector 16 by the second pin hole 14.
Embodiment two: present embodiment is described in conjunction with Fig. 1, present embodiment and embodiment one described based on the confocal axial scan device of reflection type liquid crystal spatial light modulator different be that it also comprises displacement platform 9, measured object 10 places on the displacement platform 9, and displacement platform 9 can be made one-dimensional movement along the optical axis direction of measuring object lens 11.Other composition is identical with embodiment one with annexation.
Embodiment three: present embodiment is described in conjunction with Fig. 1, present embodiment and embodiment one described based on the confocal axial scan device of reflection type liquid crystal spatial light modulator different be that it also comprises three-dimensional precision displacement table 15, light intensity detector 16 places on the three-dimensional precision displacement table 15, and three-dimensional precision displacement table 15 can be done three-dimensional motion.Other composition is identical with embodiment one with annexation.
Embodiment four: the effective pixel area of embodiment one described reflection type liquid crystal spatial light modulator 8 is 7.68mm * 7.68mm, and each pixel size is 15 μ m * 15 μ m.
Embodiment five: adopt embodiment one described confocal axial scan device based on the reflection type liquid crystal spatial light modulator to realize confocal axial scanning method, it comprises that concrete steps are as follows:
The relative position of step 2, the maintenance described object to be scanned of step 1 and measurement object lens 11 is motionless, by the phase place gray-scale map variation of computer control reflection type liquid crystal spatial light modulator 8, so that theoretical focussing distance x corresponding to described phase place gray-scale map
FIn axial direction from 0 μ m to the stepping of 105 μ m with 5 μ m, described phase place gray-scale map and theoretical focussing distance x
FBetween the pass be:
Wherein, f
0Be the focal length of measuring object lens, d is that the reflection type liquid crystal spatial light modulator arrives the light path of measuring object lens, and λ is the wavelength of light wave, and ξ, η represent the space two-dimensional coordinate;
After the phase place gray-scale map of each adjustment reflection type liquid crystal spatial light modulator 8, adopt light intensity detector 16 imagings, obtain corresponding intensity signal;
Step 3, each the theoretical focussing distance x that obtains according to step 2
FObtain a corresponding confocal axial scan curve with intensity signal.
Embodiment six: in conjunction with Fig. 4 explanation, present embodiment is a specific embodiment of embodiment five, in the present embodiment, based on adopting stepper motor to realize the driving of displacement platform 9 in the confocal axial scan device of reflection type liquid crystal spatial light modulator, described stepper motor adopts SC300-2A type stepper motor driver to drive;
Described method is:
Step 2, the selected theoretical focussing distance x of basis
FBe respectively 5 μ m, 10 μ m, 15 μ m, 20 μ m, 25 μ m, 30 μ m, 35 μ m, 40 μ m, 45 μ m, 50 μ m, 75 μ m, 100 μ m, 175 μ m and 250 μ m, by computer control reflection type liquid crystal spatial light modulator 8, the phase place gray-scale map of reflection type liquid crystal spatial light modulator 8 is satisfied:
Wherein, f
0Be the focal length of measuring object lens 11, d is that reflection type liquid crystal spatial light modulator 8 arrives the light path of measuring object lens 11, and λ is the wavelength of light wave;
Fig. 2 is theoretical focussing distance x
FThe phase place gray-scale map of reflection type liquid crystal spatial light modulator during=10 μ m; Fig. 3 is theoretical focussing distance x
FThe phase place gray-scale map of reflection type liquid crystal spatial light modulator during=50 μ m.
Step 3, according to the selected series of theories focussing distance x of step 2
F, with the theoretical focussing distance x of imageing sensor at range observation object lens 11
FNear in axial direction move with the step-length of 1 μ m, until hot spot brightness reaches maximum, imageing sensor present position range observation object lens 11 focuses apart from x ' when hot spot brightness is maximum
FBe actual focussing distance;
X '
FBe respectively 5 μ m, 10 μ m, 17.5 μ m, 25 μ m, 27.5 μ m, 35 μ m, 37.5 μ m, 42.5 μ m, 47.5 μ m, 55 μ m, 80 μ m, 105 μ m, 205 μ m and 340 μ m;
Theoretical focussing distance x
FWith actual focussing distance x '
FAnd the error size between them is as shown in the table.
Theoretical focussing distance x
FWith actual focussing distance x '
FThe Data Comparison table
Can illustrate by the data in the form, focussing distance in 5～105 mu m ranges, theoretical focussing distance x
FWith actual focussing distance x '
FThe error maximum be no more than 5 μ m, and focussing distance is when 100 μ m are above, theoretical focussing distance x
FWith actual focussing distance x '
FError increase suddenly; The focussing distance error allowed band of proof present embodiment institute corresponding intrument is when 5 μ m are following, and focussing distance can reach 100 μ m.
Step 4, the theoretical focussing distance x of setting
F=0, measured object 10 is configured to object to be scanned, adjust object to be scanned and be in the place, focal position of measuring object lens 11; Add successively and collect object lens 13, the second pin hole 14, three-dimensional precision displacement table 15, light intensity detector 16, so that the light beam that returns along former road from the scanning object body position, respectively through after measuring object lens 11, transmission and crossing spectroscope 12, be imaged onto the second pin hole 14 positions by collecting object lens 13, and pass behind the second pin hole 14 by light intensity detector 16 imagings; Formation is based on the confocal axial scan device of reflection type liquid crystal spatial light modulator, as shown in Figure 1;
The position of step 5, maintenance step 4 object to be scanned is motionless, according to required actual focussing distance x '
F, x '
FBe respectively 5 μ m, 10 μ m, 17.5 μ m, 25 μ m, 27.5 μ m, 35 μ m, 37.5 μ m, 42.5 μ m, 47.5 μ m, 55 μ m, 80 μ m and 105 μ m, by actual focussing distance x '
FCorresponding theoretical focussing distance x
F, corresponding x
FBe respectively 5 μ m, 10 μ m, 15 μ m, 20 μ m, 25 μ m, 30 μ m, 35 μ m, 40 μ m, 45 μ m, 50 μ m, 75 μ m, 100 μ m, the phase place gray-scale map that obtains the reflection type liquid crystal spatial light modulator is:
Wherein, f
0Be the focal length of measuring object lens 11, d is that the reflection type liquid crystal spatial light modulator arrives the light path of measuring object lens 11, and λ is the wavelength of light wave;
Step 6, according to the axial scan degree of depth of object appearance to be scanned, change the phase place gray-scale map of reflection type liquid crystal spatial light modulator, light intensity detector 16 imagings obtain confocal axial scan curve simultaneously.When wherein, focussing distance is 40 μ m and the confocal axial scan curve comparison figure in when focusing as shown in Figure 5.
Adopt the confocal axial scan device based on the reflection type liquid crystal spatial light modulator of the present invention to realize scanning object pattern method, it comprises that concrete steps are as follows:
The relative position of step 2, the maintenance described object to be scanned of step 1 and measurement object lens 11 is motionless, by the phase place gray-scale map variation of computer control reflection type liquid crystal spatial light modulator 8, so that theoretical focussing distance x corresponding to described phase place gray-scale map
FIn axial direction from 0 μ m to the stepping of 105 μ m with 5 μ m, described phase place gray-scale map and theoretical focussing distance x
FBetween the pass be:
After the phase place gray-scale map of each adjustment reflection type liquid crystal spatial light modulator 8, adopt light intensity detector 16 imagings, obtain corresponding intensity signal;
Step 3, each the theoretical focussing distance x that obtains according to step 2
FObtain a corresponding confocal axial scan curve with intensity signal;
The scanning curve of step 4, one group of different axial depth that step 3 is obtained, the ordinate maximal value is integrated and is obtained object appearance in the junction curve.
After the focusing in the confocal axial scan curve light intensity maximal value peaked horizontal spacing of light intensity when not focusing be focussing distance, if focussing distance and theoretical focussing distance x
FMaximum error in 5 mu m ranges, then think this moment x
FValue is effective value;
It is 2f that measured object is placed into the range observation object lens
0The position.At first, at effective x
FIn the scope, every corresponding gray-scale map measured object is carried out a raster pattern transversal scanning, thereby light intensity detector is exported one group of two-dimensional matrix value, loads continuously different x
FBe worth corresponding gray-scale map, then obtain the two-dimentional light intensity matrix of one group of different axial depth, then with x
FValue is horizontal ordinate, and the corresponding element value of identical row and column is that ordinate is depicted as curve in the matrix, seeks horizontal ordinate x corresponding to ordinate maximal value in the curve
FValue is object at the pattern place at this element place, and the value that each element is corresponding is integrated and obtained object appearance at last.
Claims (4)
1. based on the confocal axial scanning method of the confocal axial scan device of reflection type liquid crystal spatial light modulator, the confocal axial scan device of described reflection type liquid crystal spatial light modulator comprises laser instrument (1), convergent lens (2), the first pin hole (3), illumination objective lens (4), diaphragm (5), polarization spectroscope (6), λ/4 wave plates (7), reflection type liquid crystal spatial light modulator (8), measured object (10), measure object lens (11), spectroscope (12), collect object lens (13), the second pin hole (14) and light intensity detector (16), the parallel beam of laser instrument (1) emission is imaged onto the first pin hole (3) by convergent lens (2) and locates, light beam by the first pin hole (3) collimates the rear parallel polarization spectroscope (6) that incides by illumination objective lens (4), polarization spectroscope (6) is divided into reflected light and transmitted light to incident light, described reflected light is S light, described transmitted light is P light, transmitted light is by arriving reflection type liquid crystal spatial light modulator (8) behind λ/4 wave plates (7), reflection type liquid crystal spatial light modulator (8) is arriving light total reflection, the total reflection light that obtains is back into by original optical path and is incident upon polarization spectroscope (6), reflected light through this polarization spectroscope (6) reflection is incident to spectroscope (12), reflected light through this spectroscope (12) reflection is incident to measurement object lens (11), be imaged onto on the measured object (10) by these measurement object lens (11), be back into along original optical path through the reflected light of measured object (10) reflection and be incident upon spectroscope (12), transmitted light through this spectroscope (12) transmission is imaged onto the second pin hole (14) position by collecting object lens (13), and incides the photosurface of light intensity detector (16) by the second pin hole (14);
It is characterized in that described method comprises that concrete steps are as follows:
Step 1, the theoretical focussing distance x of setting
F=0, measured object is configured to object to be scanned, it is 2f that object to be scanned is placed into range observation object lens (11)
0The position;
The relative position of step 2, the maintenance described object to be scanned of step 1 and measurement object lens (11) is motionless, phase place gray-scale map by computer control reflection type liquid crystal spatial light modulator (8) changes, so that theoretical focussing distance x corresponding to described phase place gray-scale map
FIn in axial direction from 0 μ m to 105 mu m ranges with the step-length stepping of 5 μ m, described phase place gray-scale map and theoretical focussing distance x
FBetween the pass be:
Wherein, f
0Be the focal length of measuring object lens, d is that the reflection type liquid crystal spatial light modulator arrives the light path of measuring object lens, and λ is the wavelength of light wave, and ξ, η represent the space two-dimensional coordinate;
After the phase place gray-scale map of each adjustment reflection type liquid crystal spatial light modulator (8), adopt light intensity detector (16) imaging, obtain corresponding intensity signal;
Step 3, each the theoretical focussing distance x that obtains according to step 2
FObtain a corresponding confocal axial scan curve with intensity signal.
2. the confocal axial scanning method of described confocal axial scan device based on the reflection type liquid crystal spatial light modulator according to claim 1, the confocal axial scan device that it is characterized in that described reflection type liquid crystal spatial light modulator also comprises displacement platform (9), measured object (10) places on the displacement platform (9), and displacement platform (9) can be made one-dimensional movement along the optical axis direction of measuring object lens (11).
3. the confocal axial scanning method of described confocal axial scan device based on the reflection type liquid crystal spatial light modulator according to claim 1, the confocal axial scan device that it is characterized in that described reflection type liquid crystal spatial light modulator also comprises three-dimensional precision displacement table (15), and light intensity detector (16) places on the three-dimensional precision displacement table (15).
4. the confocal axial scanning method of described confocal axial scan device based on the reflection type liquid crystal spatial light modulator according to claim 1, the effective pixel area that it is characterized in that described reflection type liquid crystal spatial light modulator (8) is 7.68mm * 7.68mm, and each pixel size is 15 μ m * 15 μ m.
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