CN110231245B - High-flux detection system for obtaining trace mass and molecular structure information - Google Patents
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Abstract
The invention discloses a high-flux detection system for obtaining trace mass and molecular structure information, wherein a micro-cantilever array fixed on a piezoelectric ceramic driver is driven by a voice coil motor to translate along the width direction of the micro-cantilever, so that laser beams cast the micro-cantilever array one by one, laser reflected by a tip is detected by a photoelectric position sensitive detector to obtain bending signals of the free end of each micro-cantilever, and the piezoelectric ceramic driver is used for adjusting the vibration frequency of the micro-cantilever; setting a micro-Raman spectrum unit which consists of an observation optical path subunit and a Raman laser optical path subunit working in a time-sharing way; obtaining the micro-morphology of the surface sample of the free end of the micro-cantilever by using the observation optical path subunit; and obtaining a Raman spectrum by utilizing the Raman laser optical path subunit so as to further obtain the molecular structure information of the sample. The invention combines micro-cantilever beam sensing and micro-Raman technology to realize trace mass change detection, and is used for observing the appearance of a sample and detecting the change of a molecular structure in the reaction process in real time.
Description
Technical Field
The invention relates to the field of micro-cantilever beam detection and the field of Raman spectrum analysis, in particular to a high-flux detection system for obtaining trace mass and molecular structure information.
Background
The micro-cantilever sensing technology is a new sensing method which is rapidly developed after an atomic force microscope and a micro system appear, and is always a hot spot for research of the micro-nano sensing technology as the simplest micro-mechanical element. The micro-beam sensor can measure the biochemical reaction process with specificity in real time, and when the biochemical reaction occurs on the surface of the micro-beam, the micro-beam generates bending deformation due to the stress difference of the upper surface and the lower surface. The sensing technology is widely researched in the fields of biology, chemistry and the like as a real-time, high-sensitivity and non-calibration sensing method.
There are generally two modes of operation for micro-cantilevers: static mode and dynamic mode. The dynamic mode working principle is that the resonance frequency and the amplitude of the micro-cantilever beam are changed by changing the mass, the damping coefficient, the environment and other factors of the micro-cantilever beam, and the change process of relevant parameters is recorded, so that the change condition of an external signal can be obtained; the micro-cantilever in the dynamic mode has higher sensitivity than that in the static mode and is less influenced by environmental factors.
On the basis of a single micro-cantilever detection system, in order to further eliminate background noise influences such as environmental temperature drift, solution refractive index change and the like and realize rapid parallel detection of various target molecules, micro-cantilever sensing technology is gradually developing to multi-arrays. The micro-cantilever array can reduce experimental errors and realize high-flux detection.
Raman is a light scattering technique. When the incident light of the laser light source is scattered by molecules, most of the scattered light has the same wavelength as the incident laser light, and the scattering is called rayleigh scattering; however, there is a very small portion of scattered light that has a wavelength different from the incident light, and the change in wavelength is determined by the structure of the sample, which is called raman scattering. By analyzing the spectra, specific structural features or characteristic groups can be identified.
In the prior art, from the equipment perspective, the continuous updating and improving system aims at detecting the trace mass change of the micro-cantilever; from the perspective of detection mechanism, no matter how good the repeatability of the micro-cantilever bending signal is, only a single micro-cantilever free end bending response signal can be obtained, and other information of the material to be detected, such as structural information, cannot be detected in real time, so that the mechanism cannot be analyzed from the perspective of structure and mechanical properties.
Disclosure of Invention
The invention aims to avoid the defects of the prior art and provides a high-flux detection system for obtaining trace mass and molecular structure information, which makes full use of the advantages of non-labeling, real-time and trace detection of stress and mass change of a micro-cantilever beam, combines the advantages of chemical identification, form and phase research and analysis of Raman spectrum, realizes the correspondence of the change of stress, mass or heat and the like of a material with the molecular structure information in the detection process, realizes the real-time and synchronous detection of the trace mass change performance and structure, and more comprehensively analyzes the material performance.
The invention adopts the following technical scheme for realizing the purpose of the invention:
the high-flux detection system for obtaining the trace mass and molecular structure information has the characteristics that: the micro-Raman spectrometer consists of a micro-cantilever beam sensing unit and a micro-Raman spectrum unit;
in the micro-cantilever sensing unit, a plurality of micro-cantilevers are arranged in parallel to form a micro-cantilever array, and laser emitted by a laser is projected to the micro-cantilever array according to a set angle through a laser guide device; the micro-cantilever array is fixedly arranged on the piezoelectric ceramic driver, the piezoelectric ceramic driver is fixedly arranged on the voice coil motor, the voice coil motor drives the piezoelectric ceramic driver to translate, so that the micro-cantilever array is driven to translate along the width direction of the micro-cantilever, laser beams emitted by the laser are projected to the tip part of the free end of each micro-cantilever in the micro-cantilever array one by one along with the translation of the micro-cantilever array, and laser reflected beams reflected by the tip part are received by a receiving target surface of the photoelectric position sensitive detector; exciting the micro-cantilever array to generate oscillation by a piezoelectric ceramic driver, and performing data processing on output information of the photoelectric position sensitive detector by using a first computer to obtain the vibration frequency of the free end of each micro-cantilever; when the object to be measured is adsorbed on the surface of the free end of the micro-cantilever beam, the surface stress of the micro-cantilever beam changes, so that the vibration frequency of the free end of the micro-cantilever beam changes, and the variable quantity of the trace mass of the object to be measured is obtained by analyzing the variation quantity of the vibration frequency;
the micro-Raman spectrum unit consists of an observation optical path subunit and a Raman laser optical path subunit which work in a time-sharing manner;
the observation optical path subunit has the structural form that: white light output by a white light source sequentially passes through the semi-transparent semi-reflective mirror and the objective lens, so that the white light is focused on the micro-cantilever array, white light reflected light on the micro-cantilever array sequentially passes through the objective lens, the semi-transparent semi-reflective mirror, the reflector and the lens sleeve and is transmitted to the CCD sensor, a second computer connected with the CCD sensor is used for observing the micro-cantilevers, and the micro-morphology of the sample on the surface of the free end of each micro-cantilever is obtained one by one;
the structural form of the Raman laser optical path subunit is as follows: laser emitted by a Raman laser is guided into a collimator through an incident optical fiber, stray light is filtered by a laser filter after beam expansion is carried out on the laser through the collimator, the stray light is focused on the surface of the free end of the micro-cantilever array through an objective lens after being reflected by a dichroic mirror, scattered light reflected by the micro-cantilever array penetrates through the dichroic mirror after being collected by the objective lens, rayleigh stray light is filtered by a trap filter to obtain Raman scattered light, the Raman scattered light is coupled to the emergent optical fiber through a coupler and is guided into a spectrometer, a Raman spectrum is obtained by a third computer after passing through a CCD (charge coupled device) image processor in the spectrometer, and molecular structure information of samples on the surface of the free end of each micro-cantilever can be obtained according to Raman spectrum analysis.
The high-flux detection system for obtaining the trace mass and molecular structure information is also characterized in that: the micro-cantilever array and the piezoelectric ceramic driver form a dynamic micro-cantilever array, a driving signal of the piezoelectric ceramic driver comes from the function signal generator, and the driving signal is changed by adjusting related parameters of the function signal generator so as to excite the micro-cantilever array to generate oscillation; the method is characterized in that sensitive molecules are modified on the surface of the free end of the micro-cantilever beam, when a molecule to be detected is in contact with the sensitive molecules, an adsorption effect is generated, so that the effective mass of the micro-cantilever beam is changed, the increase of the mass of the micro-cantilever beam causes the reduction of the vibration frequency, the magnitude of frequency offset is obtained by measuring the bending signal of the micro-cantilever beam and processing data, the mass of the molecule to be detected adsorbed by the micro-cantilever beam is reflected, and the nano-scale resolution and sensitivity can be obtained.
The high-flux detection system for obtaining the trace mass and molecular structure information is also characterized in that: the observation optical path subunit and the Raman laser optical path subunit form a coaxial optical path system by using an objective lens, and the semi-transparent semi-reflecting mirror and the reflecting mirror are plug-in detachable devices.
The high-flux detection system for obtaining the trace mass and molecular structure information is also characterized in that: the white light source is a cold light source.
The high-flux detection system for obtaining the trace mass and molecular structure information is also characterized in that: the Raman laser is a single longitudinal mode laser, the wavelength is 785nm, and the Raman laser is suitable for inorganic material detection.
The high-flux detection system for obtaining the trace mass and molecular structure information is also characterized in that: the laser filter is a laser narrow-band filter and is used for filtering plasma rays and other stray signal light in a laser source, and a MaxLine laser filter of Semrock company is adopted, wherein the center wavelength of the MaxLine laser filter is 785nm.
The high-flux detection system for obtaining the trace mass and molecular structure information is also characterized in that: the dichromatic directional mirror is a 785nm BrightLine single-edge laser dichromatic directional mirror from Semrock.
The high-flux detection system for obtaining the trace mass and molecular structure information is also characterized in that: the notch filter is used for attenuating or blocking scattered energy from reaching an exit area, and a 785nm StopLine single notch filter of Semrock company is adopted.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention combines the micro-cantilever beam sensing technology and the micro-Raman technology, can detect the trace mass change, can also observe the appearance of a sample, can detect the change of a molecular structure in the reaction process in real time, realizes the real-time, in-situ and non-labeled detection, and can more comprehensively analyze the material performance.
2. Compared with a static mode, the micro-cantilever dynamic working mode has the advantages of higher sensitivity, better detection effect, strong anti-interference capability to the outside and the like;
3. the array scanning device realizes array scanning by utilizing the voice coil motor to drive the clamping device to translate, and has high efficiency, simple system construction and easy control;
4. the invention can ensure that the energy projected by the laser beam to the free end of each micro-cantilever beam is the same, and ensures the conditions to be consistent, thereby improving the precision;
5. the micro-Raman system has the advantages of high flexibility and high signal-to-noise ratio.
Drawings
Fig. 1 is a schematic view of the overall structure of the present invention.
FIG. 2 is a schematic diagram of a micro-cantilever sensing unit according to the present invention.
FIG. 3 is a schematic diagram of a micro-Raman spectroscopy unit according to the present invention.
FIG. 4 is a schematic view of an observation light path in the micro-Raman spectroscopy unit according to the present invention.
FIG. 5 is a schematic diagram of a laser path in the micro-Raman spectroscopy unit of the present invention.
Reference numbers in the figures: the system comprises a power supply 1, a laser 2, a laser 3, a laser guiding device 4, a micro-cantilever array 5, a piezoceramic driver 6, a photoelectric position sensitive detector 6, a first computer 7, a white light source 8, a semi-transparent and semi-reflective mirror 9, an objective lens 10, a lens sleeve 11, a CCD sensor 12, a second computer 13, a Raman laser 14, an incident optical fiber 15, a collimator 16, a laser optical filter 17, a dichroic mirror 18, a notch optical filter 19, a coupler 20, an emergent optical fiber 21, a spectrometer 22, a CCD image processor 23, a third computer 24, a reflector 25, a voice coil motor 26 and a function signal generator 27.
Detailed Description
Referring to fig. 1, the high-throughput detection system for obtaining trace mass and molecular structure information in this example is: the micro-Raman spectrum micro-sensor consists of a micro-cantilever beam sensing unit and a micro-Raman spectrum unit.
Referring to fig. 1 and fig. 2, in the micro-cantilever sensing unit of the present embodiment, a plurality of micro-cantilevers are arranged in parallel to form a micro-cantilever array 4, a power supply 1 is turned on, and laser light emitted by a laser 2 is projected to the micro-cantilever array 4 through a laser guide device 3 at a set angle; the micro-cantilever array 4 is fixedly arranged on the piezoelectric ceramic driver 5, the piezoelectric ceramic driver 5 is fixedly arranged on the voice coil motor 26, the voice coil motor 26 drives the piezoelectric ceramic driver 5 to translate, so that the micro-cantilever array 4 is driven to translate along the width direction of the micro-cantilever, laser beams emitted by the laser 2 are projected on the tip parts of the free ends of the micro-cantilevers in the micro-cantilever array 4 one by one along with the translation of the micro-cantilever array 4, and laser reflected beams reflected by the tips are received by a receiving target surface of the photoelectric position sensitive detector 6; the micro-cantilever array 4 is excited by the piezoelectric ceramic driver 5 to generate oscillation, and the computer 7 is used for processing data aiming at the output information of the photoelectric position sensitive detector 6 to obtain the vibration frequency of the free end of each micro-cantilever; when the object to be measured is adsorbed on the surface of the free end of the micro-cantilever, the surface stress of the micro-cantilever changes, so that the vibration frequency of the free end of the micro-cantilever changes, and the variable quantity of the trace mass of the object to be measured is obtained by analyzing the variation quantity of the vibration frequency.
The micro-Raman spectrum unit consists of an observation optical path subunit and a Raman laser optical path subunit which work in a time-sharing manner;
referring to fig. 1, 3 and 4, the structural form of the observation optical path subunit in the present embodiment is: white light output by the white light source 8 sequentially passes through the semi-transparent semi-reflecting mirror 9 and the objective lens 10, so that the white light is focused on the micro-cantilever array 4, white light reflected light on the micro-cantilever array 4 is sequentially transmitted to the first CCD sensor 12 through the objective lens 10, the semi-transparent semi-reflecting mirror 9, the reflecting mirror 25 and the lens sleeve 11, the micro-cantilevers are observed by the second computer 13 connected with the CCD sensor 12 in an online mode, and the micro-morphology of samples on the surfaces of the free ends of the micro-cantilevers is obtained one by one.
Referring to fig. 1, 3 and 5, the raman laser optical path subunit in this embodiment is in the form of: laser emitted by a Raman laser 14 is guided into a collimator 16 through an incident optical fiber 15, after being expanded by the collimator 16, stray light is filtered by a laser filter 17, reflected by a dichroic mirror 18 and then focused on the surface of the free end of a micro-cantilever array 4 through an objective lens 10, scattered light reflected by the micro-cantilever array 4 is collected by the objective lens 10 and then passes through the dichroic mirror 18, rayleigh stray light is filtered by a notch filter 19 to obtain Raman scattered light, the Raman scattered light is coupled to an exit optical fiber 21 through a coupler 20 and guided into a spectrometer 22, a Raman spectrum is obtained by a third computer 24 after passing through a CCD image processor 23 in the spectrometer 22, and molecular structure information of a sample on the surface of the free end of each micro-cantilever is obtained according to Raman spectrum analysis.
In specific implementation, the micro-cantilever array 4 and the piezoelectric ceramic driver 5 form a dynamic micro-cantilever array, a driving signal of the piezoelectric ceramic driver 5 comes from the function signal generator 27, and the driving signal is changed by adjusting related parameters of the function signal generator 27, so that the micro-cantilever array 4 is excited to oscillate; the method is characterized in that sensitive molecules are modified on the surface of the free end of the micro-cantilever beam, when a molecule to be detected is in contact with the sensitive molecules, an adsorption effect is generated, so that the effective mass of the micro-cantilever beam is changed, the increase of the mass of the micro-cantilever beam causes the reduction of the vibration frequency, the magnitude of frequency offset is obtained by measuring the bending signal of the micro-cantilever beam and processing data, the mass of the molecule to be detected adsorbed by the micro-cantilever beam is reflected, and the nano-scale resolution and sensitivity can be obtained.
The corresponding settings also include: the observation optical path subunit and the Raman laser optical path subunit form a coaxial optical path system by using an objective lens 10, and the semi-transparent semi-reflecting mirror 9 and the reflecting mirror 25 are plug-in detachable devices; the white light source 8 is a cold light source; the Raman laser 14 is a single longitudinal mode laser, adopts the wavelength of 785nm, and is suitable for inorganic material detection; the laser filter 17 is a laser narrow-band filter used for filtering plasma rays and other stray signal light in a laser source, and adopts a MaxLine laser filter of Semrock company, wherein the central wavelength of the MaxLine laser filter is 785nm; the dichroic mirror 18 is a 785nm BrightLine single-edge laser dichroic mirror from Semrock; the notch filter 19 is used for attenuating or blocking scattered energy from reaching the exit area, and a 785nm StopLine single notch filter of Semrock is adopted.
The micro-Raman spectrum system in the embodiment consists of an observation light path and a laser light path, wherein the observation light path is used for realizing white light illumination and microscopic observation of the overall appearance of a sample or a measurement area of the sample, the function of the micro-Raman spectrum system is equal to that of a complete optical microscope, the structure is simple and compact, and the spatial position is convenient to adjust; the whole external optical path system combines the observation optical path and the laser optical path together through the pluggable semi-transparent semi-reflecting mirror and the pluggable reflecting mirror, the observation optical path and the laser optical path share one microscope objective, the sample position determined by the observation optical path is directly analyzed by Raman, the sample or the instrument position does not need to be moved, the operation is simplified, and the accuracy is improved. In the micro-cantilever sensor system, the laser light source must irradiate the tip of the micro-cantilever, but because the micro-cantilever has small volume, the observation of human eyes is not obvious, and the adjustment before the test is very difficult.
In the invention, the whole external optical path system shares the same microscope objective, and the observation optical path and the laser optical path are combined together through the pluggable semi-transparent semi-reflecting mirror 9 and the pluggable reflecting mirror 25, so that the semi-transparent semi-reflecting mirror 9 and the pluggable reflecting mirror 25 need to be dismantled before the detection of the Raman signals.
In the invention, laser emitted by a Raman laser 14 enters a system after being expanded by a collimator 16, stray light is filtered by a laser filter 17, the stray light is reflected to an objective lens 10 by a dichroic directional mirror 18 and is focused on the surface of a micro-cantilever array 4, scattered light returned from the micro-cantilever array 4 is collected by the objective lens 10, rayleigh stray light is filtered by a notch filter 19 after penetrating through the dichroic directional mirror 18, the remained Raman scattered light is coupled to an emergent optical fiber 21 by a coupler 20 and is led into a spectrometer 22, and a Raman spectrum is obtained after processing; the system effectively utilizes the energy of the light source, eliminates stray light and Rayleigh scattered light, and collects Raman scattered light to the maximum extent.
Raman spectroscopy is very sensitive to molecular bonding and the structure of the sample, so each molecule or sample has its own spectral "fingerprint". These "fingerprints" can be used for chemical identification, morphological and phase studies and composition studies.
Claims (1)
1. A high throughput detection system for obtaining trace mass and molecular structure information, characterized by: the micro-Raman spectrum spectrometer consists of a micro-cantilever beam sensing unit and a micro-Raman spectrum unit;
in the micro-cantilever sensing unit, a plurality of micro-cantilevers are arranged in parallel to form a micro-cantilever array (4), and laser emitted by a laser (2) is projected to the micro-cantilever array (4) through a laser guide device (3) according to a set angle; the micro-cantilever array (4) is fixedly arranged on the piezoelectric ceramic driver (5), the piezoelectric ceramic driver (5) is fixedly arranged on the voice coil motor (26), the voice coil motor (26) drives the piezoelectric ceramic driver (5) to translate, so that the micro-cantilever array (4) is driven to translate along the width direction of the micro-cantilever, laser beams emitted by the laser (2) are projected on the tip parts of the free ends of the micro-cantilevers in the micro-cantilever array (4) one by one along with the translation of the micro-cantilever array (4), and laser reflection beams reflected by the tips are received by a receiving target surface of the photoelectric position sensitive detector (6); the micro-cantilever array (4) is excited by the piezoelectric ceramic driver (5) to generate oscillation, and data processing is carried out on output information of the photoelectric position sensitive detector (6) by using a first computer (7) to obtain the vibration frequency of the free end of each micro-cantilever; when the object to be measured is adsorbed on the surface of the free end of the micro-cantilever beam, the surface stress of the micro-cantilever beam changes, so that the vibration frequency of the free end of the micro-cantilever beam changes, and the variable quantity of the trace mass of the object to be measured is obtained by analyzing the variation quantity of the vibration frequency;
the micro-Raman spectrum unit consists of an observation optical path subunit and a Raman laser optical path subunit which work in a time-sharing manner;
the observation optical path subunit has the structural form that: white light output by a white light source (8) sequentially passes through a half-transmitting half-reflecting mirror (9) and an objective lens (10) to focus the white light on a micro-cantilever array (4), white light reflected light on the micro-cantilever array (4) sequentially passes through the objective lens (10), the half-transmitting half-reflecting mirror (9), a reflecting mirror (25) and a lens sleeve (11) to be transmitted to a CCD (charge coupled device) sensor (12), and a second computer (13) which is connected with the CCD sensor (12) is used for observing the micro-cantilevers to obtain the micro-morphology of a sample on the surface of the free end of each micro-cantilever one by one;
the structural form of the Raman laser optical path subunit is as follows: laser emitted by a Raman laser (14) is guided into a collimator (16) through an incident optical fiber (15), after being expanded by the collimator (16), stray light is filtered by a laser filter (17), reflected by a dichroic mirror (18) and focused on the surface of the free end of a micro-cantilever array (4) through an objective lens (10), scattered light reflected by the micro-cantilever array (4) is collected by the objective lens (10) and then penetrates through the dichroic mirror (18), rayleigh stray light is filtered by a notch filter (19) to obtain Raman scattered light, the Raman scattered light is coupled to an emergent optical fiber (21) through a coupler (20) and is guided into a spectrometer (22), a Raman spectrum is obtained by a third computer (24) after passing through a CCD image processor (23) in the spectrometer (22), and molecular structure information of a sample on the surface of the free end of each micro-cantilever is obtained according to Raman spectrum analysis;
the micro-cantilever array (4) and the piezoelectric ceramic driver (5) form a dynamic micro-cantilever array, a driving signal of the piezoelectric ceramic driver (5) comes from the function signal generator (27), and the driving signal is changed by adjusting related parameters of the function signal generator (27), so that the micro-cantilever array (4) is excited to oscillate; modifying sensitive molecules on the surface of the free end of the micro-cantilever beam, wherein when a molecule to be detected is in contact with the sensitive molecules, an adsorption effect is generated, so that the effective mass of the micro-cantilever beam is changed, the increase of the mass of the micro-cantilever beam causes the reduction of the vibration frequency, and the magnitude of frequency offset is obtained by measuring a bending signal of the micro-cantilever beam and processing data, so that the mass of the molecule to be detected adsorbed by the micro-cantilever beam is reflected, and the nano-scale resolution and sensitivity can be obtained;
the observation optical path subunit and the Raman laser optical path subunit form a coaxial optical path system by using an objective lens (10), and the semi-transparent semi-reflecting mirror (9) and the reflecting mirror (25) are plug-in detachable devices;
the white light source (8) is a cold light source; the Raman laser (14) is a single longitudinal mode laser, the wavelength is 785nm, and the Raman laser is suitable for inorganic material detection; the laser filter (17) is a laser narrow-band filter, the center wavelength of the laser narrow-band filter is 785nm, and the laser narrow-band filter is used for filtering plasma rays and other stray signal light in the laser source; the two-color directional mirror (18) is a single-edge laser two-color directional mirror; the notch filter (19) is a single notch filter;
the laser filter (17) adopts a MaxLine laser filter of Semrock company; the dichromatic directional mirror (18) adopts a 785nm BrightLine single-edge laser dichromatic directional mirror of Semrock company; the notch filter (19) is used for attenuating or blocking scattered energy from reaching an exit area, and a 785nm StopLine single notch filter of Semrock company is adopted.
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