WO2017070997A1 - Système et procédé d'imagerie microscopique tridimensionnelle à haute vitesse - Google Patents

Système et procédé d'imagerie microscopique tridimensionnelle à haute vitesse Download PDF

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WO2017070997A1
WO2017070997A1 PCT/CN2015/095429 CN2015095429W WO2017070997A1 WO 2017070997 A1 WO2017070997 A1 WO 2017070997A1 CN 2015095429 W CN2015095429 W CN 2015095429W WO 2017070997 A1 WO2017070997 A1 WO 2017070997A1
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module
pulse
signal
optical
time domain
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王卫
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南京巨鲨显示科技有限公司
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
    • G01B11/00—Measuring arrangements characterised by the use of optical techniques
    • G01B11/24—Measuring arrangements characterised by the use of optical techniques for measuring contours or curvatures
    • G01B11/2441—Measuring arrangements characterised by the use of optical techniques for measuring contours or curvatures using interferometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
    • G01B11/00—Measuring arrangements characterised by the use of optical techniques
    • G01B11/24—Measuring arrangements characterised by the use of optical techniques for measuring contours or curvatures
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements

Definitions

  • the present invention relates to a high speed three dimensional microscopic imaging system and method.
  • the detection rate is about 1000 cells per second, and it takes about 2 months to detect 1 mL of blood cells. This is not advisable.
  • real-time continuous line scan imaging technology is required for the detection of product size measurement and classification, bar code, optical character recognition, non-contact size measurement and spectroscopy.
  • the imaging analysis of the appearance structure of the measured object is limited by the traditional imaging sensing technology, and the detection rate is generally in the KHz magnitude category.
  • the imaging rate can generally be achieved on the order of 100 KHz and below.
  • the highest reported frame rate CMOS imaging sensor is currently up to 1 MHz.
  • the main factors limiting the CCD/CMOS imaging rate include the following two points: 1.
  • the limitation of the mechanical scanning rate the value of which is generally limited to the 10KHz category; 2.
  • the limitation of the carrier download rate which is generally limited to the KHz category.
  • the imaging rate is continuously increased, the exposure time of each frame is shortened, and the number of detectable photons is correspondingly reduced, which greatly reduces the sensor detection sensitivity.
  • the signal-to-noise ratio of the resulting image will drop poorly so that the image cannot be resolved. It can be seen that there is a contradiction between the imaging frame rate and the detection sensitivity.
  • the object of the present invention is to provide a high-speed three-dimensional microscopic imaging system and method for performing high-frame-rate continuous line scan imaging detection on a high-speed moving target object, and obtaining an object to be measured by using an image restoration algorithm.
  • the target three-dimensional structure while achieving an imaging frame rate of the order of magnitude of MHz to tens of MHz.
  • the fast three-dimensional microscopic imaging technology realized by the ultra-short pulse light source effectively improves the detection sensitivity of the imaging system by directly amplifying the signal light in the optical domain, and obtains the space-time mapping method of each optical pulse spectrum.
  • the higher speed imaging rate is superior to the traditional CCD/CMOS imaging technology, and the three-dimensional structure distribution of the surface of the measured object can be effectively recovered by introducing the reference beam interference technique and the back-end image restoration processing algorithm.
  • a high-speed three-dimensional microscopic imaging system comprising: a broadband ultrashort pulse laser source, a time domain dispersion module connected to the broadband ultrashort pulse laser source, and connected to the output of the time domain dispersion module
  • an image recovery module connected to the signal acquisition module, wherein the broadband ultrashort pulse laser source outputs a time domain ultrashort optical pulse train having a certain spectral bandwidth, and then the spatial domain dispersion module realizes spatial dispersion effect on the ultrashort optical pulse train in the time domain.
  • the time-domain dispersive component module realizes the time dispersion effect on the ultra-short optical pulse train in the time domain
  • the reference arm module realizes the three-dimensional information interference recording of the image
  • the signal acquisition module completes the acquisition process of mapping the three-dimensional image information
  • the image restoration module is based on The algorithm recovers the recorded three-dimensional image information.
  • the broadband ultrashort pulse laser source has a certain spectral width
  • the repetition frequency of the pulse is in the range of MHz to 10 s MHz
  • the pulse time domain width under the conversion limit is on the order of ten femtoseconds to hundred femtoseconds, space
  • the imaging process of the imaging module utilizes a spectrally encoded mapping imaging method of the light source.
  • the broadband ultrashort pulse laser source utilizes the principles of fiber amplification and mode locking techniques for stable output.
  • the reference arm module records three-dimensional information of the object to be measured by completing a coherent process of the imaging beam and the reference beam.
  • the laser in the broadband ultrashort pulse laser source is composed of a fiber ring cavity, and the optical amplification effect is realized by using an optical fiber doped with a rare earth element, and the output of the ultrashort pulse in the time domain is realized by using a mode locking medium.
  • the gain output of the laser requires the use of an injection of a pumping source and a stable optical pulse output by adjusting the polarization state of the optical signal in the fiber.
  • the broadband ultrashort pulse laser source is split before imaging, wherein one path is used as a reference beam, and no operation is performed. After interference with the imaging beam, the two beams realize the beat frequency and the beat signal. In the middle, the three-dimensional information of the measured object is recorded.
  • the time-frequency analysis processing of the signal is performed, and the real-time spectrum of the signal is analyzed by using the short-time Fourier transform operation, thereby analyzing the three-dimensional image information of the object.
  • the second technical solution of the present invention is specifically as follows: a high-speed three-dimensional microscopic imaging method, comprising the steps of: spatially dispersing a short pulse spectrum by an ultrashort pulse laser source, so that spectral information is mapped to a spatial domain, and Spectral coding imaging of the target object; at the same time, using the operation of interference with the reference light, the beat frequency optical signal records the three-dimensional information of the object; then, by performing the time dispersion operation of the signal light pulse, the spectral information is mapped to the time domain, that is, The information recorded by the imaging is mapped to the time domain signal, and then the signal acquisition module is used to collect and store the measured image information, and combined with the image restoration algorithm, finally recovering the three-dimensional structure contour of the measured object.
  • a high-speed three-dimensional microscopic imaging method which comprises the following steps:
  • the laser used here is ultrashort pulse light based on passive mode locking of the optical fiber, and the gain fiber uses an erbium doped fiber;
  • the pulse output from the pulse source is sent to the time domain dispersion module, that is, into the dispersion compensation fiber for time domain dispersion transmission;
  • the optical pulse of the time domain dispersion is optically amplified by an erbium doped fiber amplifier
  • the amplified pulse source enters the spatial optical link, first passes through a half-wave plate and a quarter-wave plate, and rotates the arbitrary input linearly polarized light at any angle.
  • the use of a spatial grating, the role of the grating is to achieve a frequency-to-air mapping of the light pulse, and a convex lens to converge the divergent beam on the space object;
  • the illumination light pulse is reflected back from the target path back to the link and interferes with the second path beam reflected from the reference surface.
  • the two beams are different in frequency shift under different delay conditions, based on different frequency shift information. Recording longitudinal concave-convex displacement information of the target;
  • the combined optical pulse signal is coupled into the optical fiber from the original path of the spatial optical link, and a high-speed photodetector realizes electrical conversion of the optical signal;
  • the detected time domain pulse signal can be calculated by the short time Fourier transform to obtain the longitudinal structure distribution of the measured object;
  • the time domain signal strength marks the lateral structure information of the object to be measured, and the three-dimensional structure image of the object to be measured is obtained by moving in the horizontal direction of the scanning direction.
  • the imaging rate is basically limited to the hundred KHz range, and the imaging rate and imaging sensitivity also exist.
  • the relationship of mutual constraints that is, the higher the imaging rate, the worse the imaging signal-to-noise ratio.
  • a technique of encoding imaging using laser pulse spectroscopy can be employed. This technique uses a pulse spectrum to map the target image, and its imaging frame rate is equivalent to the repetition frequency of the pulse. By amplifying the pulse spectrum in the optical domain, the constraint relationship between the imaging frame rate and the detection sensitivity is broken. Under the condition that the frame rate is not reduced, the signal-to-noise ratio of imaging is effectively improved;
  • the pulse spectrum encodes and images the target object, and the intensity information of the spectrum records the horizontal gray structure distribution of the target object, and by introducing a reference beam to interfere with the imaging signal light, the combined beat frequency is
  • the longitudinal structure distribution of the target is recorded on the frequency, so the intensity and frequency information of the combined light records the three-dimensional structure of the target;
  • the longitudinal distribution of the measured object is recorded in the beat frequency information of the beat frequency beam.
  • the frequency component obtained from the analysis can be Calculate the displacement difference between the surface of the target and the reference surface.
  • Figure 1 is a schematic diagram of the system of the present invention.
  • FIG. 2 is a schematic diagram showing the principle of spatial mapping of a light source according to the present invention.
  • FIG. 3 is a block diagram of the system of the present invention.
  • Figure 4 is a graph showing the results of imaging a planar object of the present invention.
  • Fig. 5 is a view showing the result of imaging an uneven surface of the present invention.
  • the present invention provides a high-speed three-dimensional microscopic imaging system comprising: a broadband ultrashort pulse laser source, a time domain dispersion module connected to a broadband ultrashort pulse laser source, and time
  • the optical dispersion module connected to the output of the domain dispersion module, the spatial domain dispersion module connected to the output of the optical amplification module, the spatial imaging module connected to the output of the spatial domain dispersion module, and the output of the spatial imaging module a reference arm module, a signal acquisition module connected to the reference arm module, and an image recovery module connected to the signal acquisition module, wherein the broadband ultrashort pulse laser source outputs a time domain ultrashort optical pulse train having a certain spectral bandwidth, followed by a spatial domain
  • the dispersion module realizes the spatial dispersion effect on the ultra-short optical pulse train in the time domain
  • the time domain dispersion component module realizes the time dispersion effect on the time domain ultrashort optical pulse train
  • the reference arm module realizes
  • the ultrashort pulse laser source is used for emitting ultrashort pulse optical signals;
  • the time domain dispersion module is used for realizing the frequency-time mapping process of the pulse spectrum;
  • the optical amplification module realizes the amplification operation of the signal spectrum;
  • the spatial dispersion module is used for Realizing the frequency-to-air mapping operation process of the pulse spectrum;
  • the spatial imaging module is used for realizing focusing the optical signal to the target sample for spectral imaging;
  • the reference arm module is used for realizing the interference of the imaging signal light and the reference light, The three-dimensional information of the target object is recorded;
  • the signal acquisition module is configured to realize high-speed acquisition and storage of the imaging signal; and
  • the image recovery module is configured to implement analysis and image processing of the imaging signal.
  • a space-frequency-time mapping operation on the target sample is realized, that is, mapping the spatial information of the object to the pulse spectrum, and then mapping the pulse spectrum to the time.
  • the domain signal is then recorded by the reference beam and the combined beat frequency of the signal light, and finally the three-dimensional structure information of the measured object is recorded.
  • the ultrashort pulse laser source of the present invention is a femtosecond ultrashort pulse laser source with a fiber passive lock mode, and the structure thereof is composed of the following parts: ordinary single mode fiber, erbium doped fiber, fiber coupler, fiber Polarizers, fiber optic circulators, saturated absorbers, pump sources, etc.
  • the output characteristics of the ultrashort pulse source based on the fiber passive lock mode satisfy the following characteristics: the time domain pulse width does not exceed the magnitude of the hundred femtoseconds under the transformation limit conditions, the spectral bandwidth ranges from several nm to several tens of nm, and the pulse repetition frequency is generally In the range of several MHz to tens of MHz.
  • the pulse center spectrum of the output using the erbium-doped fiber is around 1550 nm
  • the pulse time domain width is about 300 fs
  • the pulse spectrum 10 dB bandwidth is about 10 nm
  • the pulse repetition frequency is 48.8 MHz
  • the average optical power of the output pulse is about 0dBm.
  • the ultrashort pulse laser output by the light source first performs time domain dispersion operation, enters the time dispersion module, and delays the spectrum of a certain width of the pulse in the time domain, thereby reducing the peak power of the pulse.
  • both the time domain dispersion operation and the spatial domain dispersion operation are linear reversible operations, so the order of the two does not affect the imaging result.
  • the peak optical power of the ultrashort pulse is very large, which may cause damage to the subsequent optical device.
  • the time domain dispersion operation of the pulse is placed at the initial end of the system process.
  • the dispersion used in the time domain dispersion module is the dispersion compensation.
  • this type of fiber has a large time dispersion value for different frequency optical signals near 1550 nm.
  • the fiber undergoes a large dispersion value and considers only the second-order dispersion effect of the fiber.
  • the variation process satisfies the following:
  • f( ⁇ ) represents the exit spectrum of the laser source
  • C 1 is the coefficient of variation, related to fiber dispersion and loss
  • f(t) represents the time domain signal of the output pulse.
  • the ultra-short optical pulse passes through the time domain dispersion module to complete the frequency-time mapping operation of the signal. Since the process loses a certain amount of optical power, the spectral energy compensation can be achieved by the optical amplification method.
  • the optical signal amplification module utilizes an erbium-doped fiber amplifier to achieve efficient amplification of the spectrum around 1550 nm.
  • the gain spectrum of the optical amplifier can be extended from 1530 nm to 1600 nm.
  • the erbium-doped fiber amplifier in this embodiment can amplify an optical signal having an average power of about -10 dBm to 10 dBm.
  • the optical pulse After the time domain optical pulse amplification operation, the optical pulse enters the spatial optical link module.
  • a one-half wavelength polarizing plate and a quarter-wavelength polarizing plate are used successively, and the line of the optical signal can be adjusted by adjusting two polarizing plates.
  • the polarization direction is arbitrarily output within a range of 360°.
  • the spatial dispersion module in this embodiment is a spatial diffraction grating whose first-order diffracted light linearly disperses the incident pulse spectrum in a one-dimensional spatial domain.
  • the diffraction efficiency of this element is affected by the polarization of the incident light.
  • the spatial diffraction grating used in this embodiment has a blaze wavelength of 1060 nm and a scribe line density of 1200 lines/mm.
  • the optical signal with a certain spectral bandwidth realizes the dispersion mapping of the spectral information into the spatial domain by the diffraction grating, and the spatially dispersed spectrum is then concentrated by a circular convex lens and focused on the surface of the sample to be tested.
  • the light bar size focused on the target sample in this embodiment was 2000 ⁇ m ⁇ 30 ⁇ m.
  • the optical pulse carrying the sample space information reflected back from the target sample returns from the optical path system and interferes with the reflected light of the reference arm. During this space-frequency mapping operation, the mapping process satisfies the following:
  • f(x) represents the spatial distribution of the object under test
  • C 2 is the conversion coefficient of the spatial domain to the spectral domain
  • f( ⁇ ) represents the imaging spectrum
  • the target returns to the spectrally encoded signal light path and interferes with the reference light. This gives a beat light signal.
  • the coherent optical signal records the horizontal two-dimensional gray-scale information of the object on its intensity (visible by equation (2)), and the beat information records the relative delay of the two beams. That is, the displacement information of the longitudinal surface of the target object relative to the reference surface, that is, the third-dimensional information component of the object.
  • the above method of acquiring the longitudinal information of the target object by using two laser beams to interfere is obtained by time-frequency analysis of the acquired signal.
  • the scheme uses a linear mapping of the pulse spectrum to the spatial domain, that is, each point of the spatial position corresponds to a spectral frequency, as shown in FIG. 2, and the position x n corresponds to the spectral frequency ⁇ n .
  • the ultrashort pulse used in the scheme passes through the time domain dispersion, its time domain component characterizes the spectral information of the pulse, that is to say, each time point of the signal corresponds to a spectral frequency, which is visible by equation (1).
  • the temporal dispersion and spatial dispersion of the optical pulse are identical to the imaging optical pulse, that is, the mapping of its spectrum in the time domain is consistent with the imaging optical signal.
  • the difference between the two optical signals is that they have different displacements in the spatial domain, thus causing a delay of a certain size for the two signals, and the following expression is satisfied between the time domain delay amount and the displacement difference:
  • c is the transmission rate of light in vacuum
  • n is the refractive index of the transmission medium
  • ⁇ x represents the spatial displacement difference of the two beams
  • ⁇ t represents the delay amount of the two beams.
  • ⁇ d represents the displacement difference between the object plane and the reference mirror
  • C3 represents the conversion coefficient of the beat frequency difference and the displacement difference.
  • the frequency difference here can be obtained by processing the combined time domain signal, and the specific processing method is the time-frequency analysis method using the signal, and the short-time Fourier transform method is adopted in the scheme.
  • the short-time Fourier transform is a method of time-frequency analysis of time domain signals.
  • the basics of the method The operation process is to discretize a continuous time domain signal, that is, the long time segment signal is divided into many short time signals in different time periods, and each short time signal is subjected to Fourier transform operation to calculate The instantaneous spectrum information of the short-term signal of the segment is obtained.
  • the frequency distribution of the entire long-term signal at each time position can be obtained by the Fourier transform operation of all the short-time signals, that is, the time-frequency distribution of the time domain signal is obtained. From the time-frequency distribution obtained by the analysis, the instantaneous frequency of the time domain signal at all times can be obtained more obviously.
  • the frequency of the time domain signal calculated from the short-time Fourier transform can be expressed as:
  • f(t) represents the time domain signal
  • ⁇ represents the short-time Fourier transform operation
  • F is the time-frequency distribution of the signal.
  • the continuous beat frequency information of the pulse signal can be obtained, and the displacement difference between the signal light and the reference light can be calculated by the equation (6).
  • the different frequency components of the optical pulse map information in different locations in the space, and this information is finally mapped to the time domain. It can be seen that the horizontal information of the target object is recorded at different time positions of the time domain signal, and the instantaneous frequency information of the signal records the longitudinal information of the target object, that is, the displacement difference between the surface and the reference mirror.
  • the instantaneous frequency of the time domain signal obtained by the interference changes after the short-time Fourier transform, and the longitudinal concave-convex change of the surface of the measured object can be obtained by calculating the frequency of the change, that is, the object The third dimension of information.
  • the invention also provides a high-speed three-dimensional microscopic imaging method, which first uses an ultrashort pulse laser source to spatially disperse a short pulse spectrum, so that spectral information is mapped to a spatial domain, and the target object is spectrally coded and imaged, and simultaneously utilized.
  • the operation of interference with the reference light causes the beat frequency optical signal to record three-dimensional information of the object.
  • the spectral information is mapped to the time domain, that is, the information recorded by the imaging is mapped to the time domain signal, and then the signal acquisition module is used to collect and store the measured image information, and combine the images.
  • the algorithm is restored, and finally the three-dimensional structure contour of the measured object can be recovered.
  • the signal light can be directly amplified in the optical domain, which will effectively ensure the detection sensitivity of the system, which overcomes the CCD. / CMOS imaging between high frame rate and high sensitivity Mutual constraints.
  • the invention also provides a high speed three-dimensional microscopic imaging method comprising the following steps:
  • the laser used here is ultrashort pulse light based on passive mode locking of the optical fiber, which is used by the gain fiber. It is an erbium-doped fiber, and its output pulse spectrum is around 1550 nm.
  • the 10 dB pulse spectrum obtained in this example is at 1560-1570 nm, the time domain width of the pulse at the transformation limit is 300 fs, and the time interval of adjacent pulses is about 20 ns.
  • the average power of the light pulse is about 0 dBm;
  • the pulse output from the pulse source is sent to the time dispersion module, that is, into the dispersion compensation fiber for time domain dispersion transmission.
  • the dispersion fiber used here has a large dispersion effect on the spectrum near 1550 nm, and the dispersion value is about 1000 ps/ Nm, the dispersion fiber used has a certain optical loss of about 10 dB.
  • the function of the dispersive fiber not only realizes the linear mapping of the ultrashort pulse spectrum to the time domain, but also reduces the peak power of the pulse and avoids damage to the optical components at the back end;
  • the optical pulse of the time domain dispersion is optically amplified by an erbium doped fiber amplifier
  • the amplified pulse source enters the spatial optical link. First, it passes through a half-wave plate and a quarter-wave plate. Their function can rotate any input linearly polarized light at any angle. The output, followed by a spatial grating, the grating is used to achieve frequency-to-air mapping of the optical pulses, and a convex lens is used to converge the diverging beams onto the spatial target;
  • the illumination light pulse is reflected back from the target path back to the link and interferes with the second path beam reflected from the reference surface.
  • the two beams are different in frequency shift under different delay conditions, based on different frequency shift information. Recording longitudinal concave-convex displacement information of the target;
  • the combined optical pulse signal is coupled into the optical fiber from the original path of the spatial optical link, and a high-speed photodetector realizes electrical conversion of the optical signal.
  • the bandwidth of the photodetector used here is 25 GHz, and after being converted into an electrical signal, , enter the signal acquisition module, this example uses a real-time oscilloscope, the sampling rate is 20GS / s, the sampling bandwidth is 7GHz;
  • the detected time domain pulse signal can be calculated by the short time Fourier transform to obtain the longitudinal structure distribution of the object under test.
  • the spectral intensity of the signal, and the signal intensity of the real-time domain mark the lateral structure information of the object to be measured.
  • the scanning imaging adopted in this scheme can obtain the three-dimensional structure image of the measured object by moving in the horizontal and vertical directions of the scanning direction. .

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Abstract

L'invention concerne un système et un procédé d'imagerie microscopique tridimensionnelle à haute vitesse. Le système comprend une source laser à impulsions ultracourtes à large bande, un module de dispersion de domaine temporel, un module d'amplification de lumière, un module de dispersion de domaine spatial, un module d'imagerie aérienne, un module de bras de référence, un module de collecte de signal et un module d'extraction d'image. L'invention rompt la relation restrictive entre une fréquence de trames d'imagerie et une sensibilité de détection. Sans réduire une fréquence de trames, un rapport signal sur bruit d'imagerie est accru de manière efficace, et est bien supérieur à celui de la technologie d'imagerie CCD/CMOS classique. Par l'introduction d'une technologie d'interférence de faisceau lumineux de référence et d'un algorithme de traitement de récupération d'image d'arrière-plan, la distribution structurelle tridimensionnelle d'une surface d'un objet détecté est efficacement extraite, et la technologie améliore considérablement le rendement de détection d'objet dans la production industrielle et, en même temps, permet la capture dynamique tridimensionnelle d'un objet dynamique à haute vitesse.
PCT/CN2015/095429 2015-10-30 2015-11-24 Système et procédé d'imagerie microscopique tridimensionnelle à haute vitesse WO2017070997A1 (fr)

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