CN113189102A - Dual-wavelength dual-confocal laser microscopic measurement device and measurement method - Google Patents
Dual-wavelength dual-confocal laser microscopic measurement device and measurement method Download PDFInfo
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- CN113189102A CN113189102A CN202110476165.6A CN202110476165A CN113189102A CN 113189102 A CN113189102 A CN 113189102A CN 202110476165 A CN202110476165 A CN 202110476165A CN 113189102 A CN113189102 A CN 113189102A
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Abstract
The invention discloses a dual-wavelength double-confocal laser microscopic measurement device and a measurement method, relates to the technical field of optical precision detection, and aims to simultaneously improve the axial resolution capability and the transverse resolution capability of a laser confocal system. The technical scheme adopted by the invention is as follows: the double-wavelength double-confocal laser microscopic measuring device comprises a laser, an incident light path, a reflection light path and a detection device, wherein a first laser I and a second laser respectively emit laser beams with different wavelengths, the laser beams are converged to a sample through a beam splitter, a collimator, a polarization spectroscope, a quarter glass slide and an objective lens, the reflected light is reflected by the polarization spectroscope after passing through the objective lens and the quarter glass slide, then is divided into two paths by a half-reflecting and half-transmitting lens, and then respectively enters a detector I and a detector II for detection. The detector I and the detector II are respectively arranged at the positions conjugated with the focus of the objective lens, and the output obtained by multiplying the normalized axial response curves of the double detectors is taken as the output of the confocal system, so that the method is suitable for optical high-resolution measurement.
Description
Technical Field
The invention relates to the technical field of optical precision detection, in particular to a laser micro-measurement device and a laser micro-measurement method based on multiplication and adopting a dual-wavelength double-confocal detection structure.
Background
Compared with the common optical microscope, the laser confocal microscope adopts a point illumination-point detection overall optical structure, thereby effectively inhibiting the influence of stray light on system imaging and obviously improving the transverse resolution and the axial resolution.
The traditional laser confocal measurement technology utilizes an axial response curve extreme point of a confocal system to perform focusing and height measurement on the surface displacement of a sample, and because the light intensity change rate of the axial response curve extreme point of the confocal system is zero, system noise can also cause certain fluctuation of the extreme point position, the axial resolution capability of the confocal system based on extreme point detection is limited. In order to improve the axial resolution and imaging efficiency of confocal microscopy systems, Chau-Hwang Lee proposed a differential confocal microscopy method in 1997, which utilizes the hypotenuse region with good linearity of the confocal system axial response curve for sensing measurements, improving the axial resolution of confocal systems (Chau-Hwang Lee, J. nonlinear microscopy with 2-nm depth resolution [ J ]. Optics Communications,1997, V135(4-6): 233-. In 2000, Wangfengsheng et al proposed a differential confocal detection method, which adopts a double confocal detection optical path structure to perform differential detection on the axial response curves before and after the confocal system is focused, so as to effectively suppress system noise and further improve the axial resolution of the system (Wang Fusheng, Tan Jiubin, Zhao Weiqiian. the optical protocol using capacitive technical for surface profile measurement [ C ] SPIE,2000,4222: 194-197). In 2004, Zhaowei et al proposed a laser confocal detection method combining pupil filtering and differential confocal detection, which applied pupil filtering to improve lateral resolution and differential confocal detection to improve axial resolution, and also performed a precise measurement of surface displacement of samples with reflectance differences by normalizing the differential signals (Zhaowei, Changein, Qioli. differential confocal scanning detection method with high spatial resolution [ P ] Chinese, patent No. ZL200410006359.6,2004-02-27).
Among the various confocal technologies, the differential confocal technology mainly improves the axial resolution of the system, and the transverse resolution loss of the system is large; compared with the differential confocal technology, the differential confocal detection method has the advantages of absolute zero point, large measuring range, high signal-to-noise ratio and the like, but the transverse resolution of the system is relatively low because the double detectors are in a defocused state; based on the confocal technology combining the super-resolution pupil filter and the differential confocal technology, the transverse resolution capability and the axial resolution capability of the system are improved, but the imaging system has the advantages of complex optical path, high assembly and adjustment difficulty and relatively high system cost.
Disclosure of Invention
The invention firstly provides a dual-wavelength double-confocal laser microscopic measurement device and a measurement method, and aims to simultaneously improve the axial resolution capability and the transverse resolution capability of a laser confocal system.
The technical scheme adopted by the invention is as follows: double-wavelength double-confocal laser microscopic measuring device, including laser instrument, incident light path, reflection light path and detection device, the laser instrument includes laser instrument I and laser instrument II, and the incident light path is: the laser I and the laser II are respectively arranged in the two incident beam directions of the beam splitter, and the collimator, the polarization beam splitter, the quarter glass, the objective lens and the sample stage are sequentially arranged in the emergent beam direction of the beam splitter; the reflection light path comprises an objective lens, a quarter glass slide, a polarization beam splitter and a semi-reflection and semi-transmission lens arranged along the reflection direction of the polarization beam splitter, wherein a light filter I, a lens I, a pinhole I and a detector I are sequentially arranged in the transmission direction of the semi-reflection and semi-transmission lens, and a light filter II, a lens II, a pinhole II and a detector II are sequentially arranged in the reflection direction of the semi-reflection and semi-transmission lens; the optical filter I corresponds to the wavelength of a laser beam emitted by the laser I, the optical filter II corresponds to the wavelength of a laser beam emitted by the laser II, and the detector I and the detector II are respectively arranged at the positions conjugated with the focus of the objective lens.
Further, the method comprises the following steps: the double-wavelength double-confocal laser micro-measuring device also comprises an objective lens driver, and the objective lens is matched with the objective lens driver.
Further, the method comprises the following steps: the incident directions of the laser I and the laser II are mutually vertical.
Specifically, the method comprises the following steps: the optical filter I and the optical filter II are respectively a laser line purification optical filter or a super narrow band pass optical filter.
Specifically, the method comprises the following steps: the detector I and the detector II are photomultiplier detectors or CCD detectors.
The invention also provides a dual-wavelength double-confocal laser microscopic measurement method, which aims to simultaneously improve the axial resolution capability and the transverse resolution capability of a laser confocal system, and adopts the technical scheme that: the double-wavelength double-confocal laser microscopic measurement method for performing laser microscopic measurement on a sample by using the double-wavelength double-confocal laser microscopic measurement device comprises the following steps:
s1, placing the sample on a sample stage, enabling the first laser I and the second laser II to respectively emit laser beams with different wavelengths, converging the laser beams to the sample through an incident light path, and then respectively entering the detector I and the detector II along a reflection light path.
And S2, adjusting the objective lens, and axially scanning the sample to obtain a sample surface light intensity-axial displacement response signal. The objective lens is adjusted, for example, by an objective lens actuator.
And S3, dividing the light intensity-axial displacement response signals measured by the detector I and the detector II by the maximum light intensity value of each signal respectively to obtain two normalized light intensity-axial displacement response signals.
And S4, multiplying the light intensities of the same axial displacement points in the two normalized light intensity-axial displacement response signals, and taking the multiplied light intensity-axial displacement response signals as the output of the confocal system.
The invention has the beneficial effects that: firstly, the invention can simultaneously improve the axial resolution capability and the transverse resolution capability of the system. The dual-wavelength dual-confocal detection structure is adopted, dual detectors are respectively arranged at the conjugate positions of the dual detectors and the focus of an objective lens, the output of the dual detectors after the normalization axial response curve or the transverse response curve is multiplied is taken as the output of a confocal system, the value of the normalized axial response signal is 1 because the light intensity of the focus center position of the normalized axial response signal is the maximum, and the value of the signal focus center position is 1 because the intensity of the focus center position of the normalized axial response signal is still the maximum after multiplication; with the increase of the defocusing amount, the signal intensity of the confocal system after multiplication is rapidly reduced relative to the signal intensity before multiplication, which causes the half-height widths of the normalized axial response curve and the transverse response curve output by the confocal system to be equal to or lower than the full-width half-maximum, so that the axial resolution capability and the transverse resolution capability are improved.
Secondly, the invention can effectively inhibit the sidelobe noise and improve the signal-to-noise ratio of the signal. The invention adopts dual wavelengths for measurement, and because the normalized axial response curves of different wavelengths are distributed differently, the maximum values of the side-lobe signals are staggered, and after the two are subjected to multiplication, the side-lobe noise is effectively inhibited, thereby being beneficial to improving the signal-to-noise ratio.
Thirdly, the invention improves the adaptability to samples with different reflectivities. The invention adopts a dual-wavelength structure, is beneficial to reducing the influence of the reflectivity difference of the sample on the measurement signal, and improves the signal-to-noise ratio of the measurement signal of the sample with different reflectivity, thereby improving the axial resolution capability and the transverse resolution capability of the sample.
Drawings
Fig. 1 is a schematic diagram of a dual-wavelength double-confocal laser micro-measuring device of the invention.
FIG. 2 is an axial response curve of an embodiment of the present invention.
Fig. 3 is a lateral response curve of an embodiment of the present invention.
Reference numerals: the device comprises a laser I1, a laser II 2, a beam splitter 3, a collimator 4, a polarizing beam splitter 5, a quarter glass 6, an objective lens 7, an objective lens driver 8, a sample stage 9, a half-reflecting and half-transmitting mirror 10, a filter I11, a lens I12, a pinhole I13, a detector I14, a filter II 15, a lens II 16, a pinhole II 17 and a detector II 18.
Detailed Description
The invention will be further explained with reference to the drawings.
As shown in fig. 1, the dual-wavelength double-confocal laser microscopic measuring device of the present invention comprises a laser, an incident light path, a reflection light path and a detecting device, wherein the laser comprises a laser I1 and a laser ii 2, and the incident light path is as follows: the laser I1 and the laser II 2 are respectively arranged in the two incident beam directions of the beam splitter 3, the collimator 4, the polarization beam splitter 5, the quarter glass 6, the objective lens 7 and the sample stage 9 are sequentially arranged in the emergent beam direction of the beam splitter 3, the laser I1 and the laser II 2 respectively emit two laser beams with different wavelengths, for example, the incident directions of the laser I1 and the laser II 2 are mutually vertical, and the laser beams irradiate the surface of a sample on the sample stage 9 after passing through the beam splitter 3, the collimator 4, the polarization beam splitter 5, the quarter glass 6 and the objective lens 7. In order to adjust the position of the objective lens 7, the dual-wavelength double-confocal laser micro-measurement device further comprises an objective lens driver 8, and the objective lens driver 8 is used for adjusting the position of the objective lens 7 and realizing axial scanning of the sample.
The reflection light path comprises an objective lens 7, a quarter glass 6, a polarization beam splitter 5 and a half-reflecting and half-transmitting mirror 10 arranged along the reflection direction of the polarization beam splitter 5, wherein a light filter I11, a lens I12, a pinhole I13 and a detector I14 are sequentially arranged in the transmission direction of the half-reflecting and half-transmitting mirror 10, and a light filter II 15, a lens II 16, a pinhole II 17 and a detector II 18 are sequentially arranged in the reflection direction of the half-reflecting and half-transmitting mirror 10. The objective lens 7, the quarter-slide 6 and the polarizing beam splitter 5 are both components in the incident light path and in the reflected light path. The filter I11 corresponds to the wavelength of the laser beam emitted by the laser I1, the filter II 15 corresponds to the wavelength of the laser beam emitted by the laser II 2, and the detector I14 and the detector II 18 are respectively arranged at the positions conjugated with the focus of the objective lens 7. Filter I11 and filter II 15 are conventional, such as laser line cleaning filters or ultra narrow bandpass filters. The detector I14 and the detector II 18 are photomultiplier detectors or CCD detectors.
The double-wavelength double-confocal laser microscopic measurement method for performing laser microscopic measurement on a sample by using the double-wavelength double-confocal laser microscopic measurement device comprises the following steps:
s1, placing the sample on the sample stage 9, wherein the first laser I1 and the laser II 2 respectively emit laser beams with different wavelengths, the laser beams are converged to the sample through an incident light path, and the reflected light on the surface of the sample respectively enters the detector I14 and the detector II 18 along a reflected light path. Specifically, reflected light on the surface of a sample is reflected by the polarizing beam splitter 5 after passing through the objective lens 7 and the quarter-glass 6, the reflected light is divided into two paths by the half-reflecting and half-transmitting mirror 10, one path is detected by the detector I14 after passing through the optical filter I11, the lens I12 and the pinhole I13, and the other path is detected by the detector II 18 after passing through the optical filter II 15, the lens II 16 and the pinhole II 17.
And S2, adjusting the objective lens 7, for example, adjusting the objective lens 7 through the objective lens driver 8 or adjusting the objective lens 7 in other ways, and performing axial scanning on the sample to obtain a sample surface light intensity-axial displacement response signal.
And S3, dividing the light intensity-axial displacement response signals measured by the detector I14 and the detector II 18 by the maximum light intensity value of the respective signals respectively to obtain two normalized light intensity-axial displacement response signals. Dividing the light intensity-axial displacement response signal of the detector I14 by the maximum value of the light intensity of the signal to obtain a normalized light intensity-axial displacement response signal A; the light intensity-axial displacement response signal measured by the confocal detector 18 is divided by the maximum value of the light intensity of the signal to obtain a normalized light intensity-axial displacement response signal B.
And S4, multiplying the light intensities of the same axial displacement points in the two normalized light intensity-axial displacement response signals, and taking the multiplied light intensity-axial displacement response signals as the output of the confocal system. And multiplying the light intensity of the same axial displacement point in the normalized light intensity-axial response signal A and the normalized light intensity-axial response signal B, and taking the multiplied light intensity-axial response signal as the output of the confocal system.
The axial resolving power of the present invention is demonstrated by the following examples. The double-wavelength double-confocal laser microscopic measurement method comprises the following specific parameters according to the steps: (1) calculating an axial response curve 1 under the parameters of numerical aperture of 0.9 and laser wavelength of 532nm by using an axial response function of a confocal system; (2) calculating an axial response curve 2 under the parameters of the numerical aperture of 0.9 and the laser wavelength of 633nm by using an axial response function of a confocal system; (3) the light intensities with the same axial displacement in the axial response curves 1 and 2 are multiplied to obtain axial response curves as shown in fig. 2.
Correspondingly, the double-wavelength double-confocal laser microscopic measurement method comprises the following specific parameters according to the steps: (1) calculating a transverse response curve 1 under the parameters of numerical aperture 0.9 and laser wavelength 532nm by a transverse response function of a confocal system; (2) calculating a transverse response curve 2 under the parameters of the numerical aperture of 0.9 and the laser wavelength of 633nm by a transverse response function of a confocal system; (3) the transverse response curves obtained by multiplying the light intensities having the same transverse displacement in the transverse response curves 1 and 2 are shown in fig. 3.
As can be seen from the figures 2 and 3, the invention can effectively inhibit the sidelobe noise, improve the signal-to-noise ratio, simultaneously improve the axial resolution capability and the transverse resolution capability of the system, and has important application value in the field of optical high-resolution measurement.
Claims (7)
1. The double-wavelength double-confocal laser microscopic measuring device comprises a laser, an incident light path, a reflection light path and a detecting device, and is characterized in that: the laser comprises a laser I (1) and a laser II (2), and an incident light path is as follows: the laser I (1) and the laser II (2) are respectively arranged in the two incident beam directions of the beam splitter (3), the collimator (4), the polarizing beam splitter (5), the quarter glass (6), the objective lens (7) and the sample stage (9) are sequentially arranged in the emergent beam direction of the beam splitter (3), and the objective lens (7) is further provided with an objective lens driver (8); the reflection light path comprises an objective lens (7), a quarter glass (6), a polarization beam splitter (5) and a half-reflecting and half-transmitting mirror (10) arranged along the reflection direction of the polarization beam splitter (5), wherein a light filter I (11), a lens I (12), a pinhole I (13) and a detector I (14) are sequentially arranged in the transmission direction of the half-reflecting and half-transmitting mirror (10), and a light filter II (15), a lens II (16), a pinhole II (17) and a detector II (18) are sequentially arranged in the reflection direction of the half-reflecting and half-transmitting mirror (10); the filter I (11) corresponds to the wavelength of a laser beam emitted by the laser I (1), the filter II (16) corresponds to the wavelength of a laser beam emitted by the laser II (2), and the detector I (14) and the detector II (18) are respectively arranged at the positions conjugated with the focus of the objective lens (7).
2. The dual wavelength dual confocal laser microscopy measurement device of claim 1, wherein: the device also comprises an objective lens driver (8), and the objective lens (7) is matched with the objective lens driver (8).
3. The dual wavelength dual confocal laser microscopy measurement device of claim 1, wherein: the incidence directions of the laser I (1) and the laser II (2) are mutually vertical.
4. The dual wavelength dual confocal laser microscopy measurement device of claim 1, 2 or 3, wherein: the filter I (11) and the laser II (2) are respectively a laser line purification filter or an ultra-narrow band-pass filter.
5. The dual wavelength dual confocal laser microscopy measurement device of claim 1, 2 or 3, wherein: the detector I (14) and the detector II (18) are photomultiplier detectors or CCD detectors.
6. The double-wavelength double-confocal laser microscopic measurement method is characterized by comprising the following steps of: the dual wavelength dual confocal laser micro-measuring device of any one of the preceding claims 1 to 5 for performing laser micro-measurement on a sample, comprising the steps of:
s1, placing the sample on a sample stage (9), wherein a first laser I (1) and a laser II (2) respectively emit laser beams with different wavelengths, converge the laser beams to the sample through an incident light path, and then respectively enter a detector I (14) and a detector II (18) along a reflection light path;
s2, adjusting an objective lens (7), and axially scanning the sample to obtain a sample surface light intensity-axial displacement response signal;
s3, dividing the light intensity-axial displacement response signals measured by the detector I (14) and the detector II (18) by the maximum light intensity value of the respective signals respectively to obtain two normalized light intensity-axial displacement response signals;
and S4, multiplying the light intensities of the same axial displacement points in the two normalized light intensity-axial displacement response signals, and taking the multiplied light intensity-axial displacement response signals as the output of the confocal system.
7. The dual wavelength dual confocal laser microscopy measurement method of claim 6, wherein: the dual-wavelength bi-confocal laser micro-measurement device further comprises an objective lens driver (8), and in S2, the objective lens (7) is adjusted by the objective lens driver (8).
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| CN114894224A (en) * | 2022-07-12 | 2022-08-12 | 之江实验室 | Sensitivity-adjustable long working distance differential confocal system |
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