CN107991233B - Extinction spectrum measuring device for noble metal nano array and sensing detection method thereof - Google Patents
Extinction spectrum measuring device for noble metal nano array and sensing detection method thereof Download PDFInfo
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Abstract
The invention discloses a noble metal nano array extinction spectrum measuring device and a sensing detection method thereof, wherein the device comprises a white light source, a light beam lifter, a microscopic component, a sample clamp, a light signal collector, a spectrometer and a computer terminal, wherein light beams emitted by the white light source are incident on the microscopic component after changing a propagation path through the light beam lifter, focused light beams emitted by the microscopic component are incident on a measured sample, the light signal collector receives the focused light beams transmitted by the measured sample and transmits the focused light beams to the spectrometer, the spectrometer outputs spectrum signals to the computer terminal, and the computer terminal calculates and obtains the concentration of an organic solution according to the change of the extinction spectrum of the measured sample relative to the extinction peak position of the extinction spectrum of a noble metal nano array; the device can realize accurate positioning measurement on the extinction spectrum of the noble metal nano array with a complex structure and micron-sized size; the device is simple in process of detecting the concentration of the organic solution, low in cost and high in sensitivity.
Description
Technical Field
The invention relates to a sensing detection technology of organic solution concentration, in particular to a noble metal nano-array extinction spectrum measuring device and a method for realizing sensing detection of organic solution concentration by using the same.
Background
It is well known that the extinction properties of noble metal nanoparticles are strongly dependent on their own size, morphology and environmental medium. The extinction spectrum measurement of the noble metal nano-particles can characterize and analyze the extinction characteristics of the noble metal nano-particles. Generally, the extinction spectrum of the noble metal nanoparticles is obtained by measuring the absorption spectrum of a colloidal solution of the noble metal nanoparticles, and for the extinction spectrum of a noble metal nano array formed by self-assembly of the noble metal nanoparticles or a noble metal nano array prepared by a micro-nano etching technology, the scattering spectrum and the absorption spectrum of the noble metal nano array need to be measured at the same time. At present, the scattering spectrum of a noble metal nano array needs to be measured by an integrating sphere, the absorption spectrum of the noble metal nano array needs to be measured by an ultraviolet-visible near-infrared spectrophotometer, wherein the measuring method by the integrating sphere needs to perform multistep luminous flux and spectrum calibration, and the measuring method by the ultraviolet-visible near-infrared spectrophotometer needs to perform a series of steps such as self-checking, dark current and baseline calibration, so that the two methods have the defects of long measuring time consumption and complex operation, and the integrating sphere and the ultraviolet-visible near-infrared spectrophotometer can only measure the integral spectral characteristics of a sample, namely cannot perform local accurate positioning measurement on the sample, so that the method is not suitable for extinction spectrum measurement of a noble metal nano array sample with a complex structure.
On the other hand, for the sensing of the concentration of organic solutions, there are currently many common methods, such as: an absorption spectrum method based on Lambert-beer law, a gas/liquid chromatography based on a substance separation and adsorption principle, a molecular mass spectrometry based on a molecular ionization technology, a Raman spectrum detection method based on light and substance inelastic scattering and the like. Although the common methods can effectively realize the sensing detection of the concentration of the organic solution, the common methods also have respective defects, such as: the absorbance spectroscopy method based on the lambert-beer law is only suitable for the detection of low concentration (<0.01mol/L) solutions; the detection cost of gas/liquid chromatography based on the principle of substance separation and adsorption and the molecular mass spectrometry based on the molecular ionization technology is high; the Raman spectrum detection method based on the inelastic scattering of light and substances is easily influenced by fluorescence and optical system parameters, and the detection precision is not ideal enough.
Therefore, the device capable of accurately measuring the extinction spectrum of the detected sample of the noble metal nano array with micron-sized size and a complex structure is designed, the characteristic that the extinction spectrum of the noble metal nano array strongly depends on the size, the shape and the environment medium of the noble metal nano array is utilized, accurate sensing detection of the concentration of the organic solution is carried out, and the device has important application value.
Disclosure of Invention
The first technical problem to be solved by the invention is to provide a simple-structured and convenient-to-operate extinction spectrum measurement device for a precious metal nano array, which can realize accurate positioning measurement on a measured sample of the precious metal nano array with micron-sized dimensions and a complex structure.
The second technical problem to be solved by the invention is to provide a method for realizing the sensing detection of the concentration of the organic solution by using a noble metal nano-array extinction spectrum measuring device, which has the advantages of simple detection process, low detection cost and high detection sensitivity.
The technical scheme adopted by the invention for solving the first technical problem is as follows: a noble metal nano array extinction spectrum measuring device is characterized in that: the device comprises a white light source, a light beam lifter for changing a light beam propagation path, a microscopic component, a sample clamp for clamping a measured sample to enable the measured sample to be horizontally placed, an optical signal collector, a spectrometer and a computer terminal, wherein light beams emitted by the white light source are incident on the microscopic component after the propagation path of the light beams is changed by the light beam lifter, focused light beams emitted by the microscopic component are incident on the measured sample, the optical signal collector receives the focused light beams transmitted through the measured sample and transmits the focused light beams to the spectrometer, and the spectrometer outputs spectral signals to the computer terminal.
The light beam lifter comprises a first support, a first reflecting mirror and a second reflecting mirror, wherein the first support is vertically arranged, the first reflecting mirror is arranged at the lower part of the first support, the second reflecting mirror is arranged at the upper part of the first support, light beams emitted by the white light source are incident on the first reflecting mirror, the light beams reflected by the first reflecting mirror are incident on the second reflecting mirror, and the light beams reflected by the second reflecting mirror are incident on the microscopic component. The beam lifter is used for lifting the beam from the low position to the high position, and can be realized by utilizing the two reflectors.
The first reflector with the second reflector all personally submit 45 degrees angles with the level, first reflector with the second reflector parallel arrangement, the plane of reflection of first reflector with the plane of reflection of second reflector just right. Here, the mounting positions of the first mirror and the second mirror are defined to ensure that the light beam reflected by the first mirror is received entirely by the second mirror and that the light beam reflected by the second mirror is received entirely by the microscopic assembly.
The microscope assembly comprises a second support, a half-mirror and an objective lens which are vertically arranged, and a light absorption plate which is connected to the second support from top to bottom and is sequentially connected to the second support, wherein the half-mirror and the objective lens are arranged on the second support and are positioned on a transmission path of a light beam transmitted by the half-mirror, the half-mirror and the objective lens are positioned right above the sample clamp, the light beam emitted by the light beam lifter is incident on the half-mirror, the light beam reflected by the half-mirror is incident on the objective lens, a focused light beam emitted by the objective lens is incident on the sample to be detected, the light beam transmitted by the half-mirror is incident on the light absorption plate, and the light absorption plate absorbs the light beam transmitted by the half-mirror. The beam reflected by the half mirror is vertically incident on the objective lens, and the beam transmitted by the half mirror is incident on the light absorption plate and absorbed by the light absorption plate; then the objective lens focuses the light beam reflected by the semi-transparent and semi-reflective mirror, and the micro-assembly has a simple structure and is convenient to realize functions.
The semi-transparent semi-reflecting mirror and the horizontal plane form an angle of 45 degrees, and the reflecting surface of the semi-transparent semi-reflecting mirror is opposite to the reflecting surface of the second reflecting mirror; the vertical setting of extinction board with semi-transparent half mirror be 45 degrees angles. Here, by limiting the relative positions of the second mirror and the half mirror, it is possible to ensure that all the light beam reflected by the second mirror is received by the half mirror; by limiting the relative position of the half mirror and the light absorbing plate, it is possible to ensure that all the light beams transmitted by the half mirror are absorbed by the light absorbing plate.
The second bracket is also connected with a CCD (Charge-coupled Device) camera, the CCD camera is positioned right above the half-transmitting and half-reflecting mirror, the output end of the CCD camera is connected with the computer terminal, and the CCD camera acquires the appearance of the noble metal nano array in the detected sample observed through the objective lens. The CCD camera is arranged right above the objective lens, and is used for acquiring the shape of the noble metal nano array serving as a detected sample, so that the function of the device is expanded.
The device for measuring the extinction spectrum of the precious metal nano array further comprises a first three-dimensional position adjusting device for adjusting the position of the sample clamp, wherein the first three-dimensional position adjusting device consists of a first positioning plate, a first three-dimensional displacement platform and a first connecting rod, the first three-dimensional displacement platform is installed on the first positioning plate, the first connecting rod is connected with the first three-dimensional displacement platform and the sample clamp, and the position of the sample clamp is adjusted through the first three-dimensional displacement platform so that the sample to be measured is located right below the objective lens. The position of the sample clamp is adjusted by utilizing the first three-dimensional displacement platform, so that the sample to be measured is positioned right below the objective lens, and the focused light beam emitted by the objective lens is completely incident on the sample to be measured.
The optical signal collector consists of an optical fiber probe clamp and an optical fiber probe clamped by the optical fiber probe clamp, the optical fiber probe is vertically placed, a receiving end of the optical fiber probe is positioned right below a measured sample, the optical fiber probe receives a focused light beam transmitted through the measured sample, an output end of the optical fiber probe is connected with an input end of the spectrometer through a conducting optical fiber, and the optical fiber probe transmits the focused light beam transmitted through the measured sample to the spectrometer;
the device for measuring the extinction spectrum of the precious metal nano array further comprises a second three-dimensional position adjusting device for adjusting the position of the optical signal collector, wherein the second three-dimensional position adjusting device consists of a second positioning plate, a second three-dimensional displacement platform installed on the second positioning plate and a second connecting rod for connecting the second three-dimensional displacement platform with the optical fiber probe clamp, and the position of the optical fiber probe clamp is adjusted through the second three-dimensional displacement platform, so that the optical fiber probe is positioned under the detected sample and close to the lower surface of the detected sample. The position of the optical fiber probe clamp is adjusted by using the second three-dimensional displacement platform, so that the optical fiber probe is positioned under the detected sample and close to the lower surface of the detected sample, and the information acquired by the optical fiber probe is more accurate.
The device for measuring the extinction spectrum of the noble metal nano array further comprises a base, the white light source and the spectrometer are placed on the base, the bottom of the first support and the bottom of the second support are fixedly connected with the base respectively, and the first positioning plate and the second positioning plate are fixedly connected with the base respectively. All the components are integrated by the base to form a whole.
The technical scheme adopted by the invention for solving the second technical problem is as follows: the sensing detection method of the noble metal nano array extinction spectrum measuring device is characterized by comprising the following steps of: the method for sensing and detecting the concentration of the organic solution by using the noble metal nano array extinction spectrum measuring device specifically comprises the following steps:
① preparing a noble metal nano array with a specific pattern on the conductive glass deposited with the conductive film by using an electron beam etching technology, and then taking the conductive glass with the noble metal nano array with the specific pattern as a tested sample;
② turning on white light source, adjusting the position of the optical fiber probe clamp by the second three-dimensional displacement platform to make the receiving port of the optical fiber probe be at the focal plane of the objective lens, so that the receiving port of the optical fiber probe can be clearly displayed in the display of the computer terminal by the objective lens and the CCD camera, turning on the spectrometer and setting signal acquisition parameters, wherein the spectrum signal output by the spectrometer is the spectrum of the white light source in the state of turning on the white light source, marked as I (lambda), and transmitted to the computer terminal for storage by the computer terminal, wherein lambda represents wavelength;
③ the position of the optical fiber probe clamp is adjusted by the second three-dimensional displacement platform to make the optical fiber probe move vertically downwards for the measured sample to be displayed on the focal plane, the optical fiber probe generally moves 2cm downwards because the conductive glass is very thin, then the sample clamp is used to clamp the measured sample and make the measured sample horizontally placed, then the position of the sample clamp is adjusted by the first three-dimensional displacement platform to make the noble metal nano array in the measured sample to be located on the focal plane of the objective lens and ensure the noble metal nano array image in the measured sample to be clearly displayed in the display of the computer terminal, then the position of the optical fiber probe clamp is adjusted by the second three-dimensional displacement platform to make the optical fiber probe be located right below the measured sample and make the receiving port of the optical fiber probe approach the lower surface of the measured sample as much as possible, at this time, the spectrum signal output by the spectrometer in the white light source on state is the transmission spectrum of the noble metal nano array in the measured sample and is marked as T1(lambda), and transmitting to the computer terminal for storage by the computer terminal;
④ turning off the white light source, and taking the spectrum signal output by the spectrometer in the off state of the white light source as the first background spectrum marked as B1(lambda), and transmitting to the computer terminal for storage by the computer terminal;
⑤ computer terminal utilizes formula E according to the definition of extinction spectrum1(λ)＝I(λ)-T1(λ)+B1(lambda), calculating to obtain an extinction spectrum E of the noble metal nano array in the tested sample1(λ)；
⑥ dropping the organic solution to be detected onto the noble metal nano array in the sample to be detected, and taking the spectral signal output by the spectrometer in the off state of the white light source as a second background spectrum marked as B2(lambda), and transmitting to the computer terminal for storage by the computer terminal;
⑦ turning on the white light source, wherein the spectrum signal output by the spectrometer is precious gold dropwise added with the organic solution to be detected when the white light source is turned onTransmission spectrum of nano array, marked as T2(lambda), and transmitting to the computer terminal for storage by the computer terminal;
⑧ computer terminal utilizes formula E according to the definition of extinction spectrum2(λ)＝I(λ)-T2(λ)+B2(lambda), calculating to obtain extinction spectrum E of the noble metal nano array dripped with the organic solution to be detected2(λ)；
⑨ mixing E1(lambda) as a reference spectrum, according to E2Extinction peak in (λ) relative to E1And (lambda) changing the position or the intensity of the extinction peak to obtain the concentration of the organic solution to be detected.
Compared with the prior art, the invention has the advantages that:
1) the device of the invention adopts few mechanical parts and optical elements, so that the structure of the device is simpler and is easy to realize.
2) When the device is used, the measurement can be realized only by turning on and off the white light source and the spectrometer, the operation is simple, and the measurement time is short.
3) The device of the invention directly uses the optical signal collector to receive the focused light beam transmitted through the tested sample to obtain the extinction spectrum of the noble metal nano array, and the detection process is rapid and convenient.
4) The device utilizes the objective lens to focus light beams on the surface of a sample to be measured and utilizes the CCD camera to clearly image, thereby realizing the accurate positioning measurement of extinction spectra of various noble metal nano array structures under the micron scale.
5) After the device disclosed by the invention is used for realizing the sensing detection of the concentration of the organic solution, the used noble metal nano array can be repeatedly used after being cleaned, and the detection cost is reduced.
6) The method for realizing the sensing detection of the concentration of the organic solution by utilizing the noble metal nano array extinction spectrum measuring device has the advantages of simple process, low detection cost and high detection sensitivity.
Drawings
FIG. 1 is a schematic diagram of the structure of a noble metal nano-array extinction spectrum measurement device according to the present invention;
FIG. 2 is a schematic diagram of a first three-dimensional position adjusting device in the extinction spectrum measuring device of the precious metal nano-array according to the present invention;
FIG. 3 is a schematic diagram of a second three-dimensional position adjusting device in the extinction spectrum measuring device of the precious metal nano-array according to the present invention;
FIG. 4 is a scanning electron microscope photograph of a gold nano-array having a periodic structure in which gold nano-particles have a diameter of 200 nm, a height of 75 nm, and an interval between the gold nano-particles of 50 nm;
FIG. 5 is extinction spectra of gold nano-arrays to which 20. mu.l of ethanol solutions having concentrations of 20%, 40%, 60% and 80% and absolute ethanol having a concentration of 99.7% were added dropwise, respectively, on gold nano-arrays in example three, obtained after addition of 20. mu.l of ethanol solutions having concentrations of 20%, 40%, 60% and 80% and absolute ethanol having a concentration of 99.7%;
FIG. 6 is a graph showing the response of ethanol solution and absolute ethanol concentration in example III;
FIG. 7 is extinction spectra of gold nano-arrays to which enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L were added dropwise after dropping 20. mu.l of the enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L, respectively, on the gold nano-arrays in example four;
FIG. 8 is a graph of the concentration response of different concentrations of enrofloxacin solutions from example four;
FIG. 9 is a scanning electron microscope photograph of a gold nano-array having a periodic structure in which gold nanoparticles have a diameter of 200 nm, a height of 75 nm, and a spacing of 100 nm therebetween;
fig. 10 is extinction spectra of gold nano-arrays to which ethanol solutions having concentrations of 20%, 40%, 60% and 80% and absolute ethanol having a concentration of 99.7% were added dropwise, respectively, after 20 μ l of ethanol solutions having concentrations of 20%, 40%, 60% and 80% and absolute ethanol having a concentration of 99.7% were added dropwise on the gold nano-arrays in example five;
FIG. 11 is a graph showing the response of ethanol solution and absolute ethanol of different concentrations in the fifth example;
FIG. 12 is extinction spectra of gold nano-arrays to which enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L were added dropwise, respectively, after 20 microliters of enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L were added dropwise onto the gold nano-arrays in example six;
FIG. 13 is a graph showing the concentration response of various enrofloxacin solutions of example six.
Detailed Description
The invention is described in further detail below with reference to the accompanying examples.
The first embodiment is as follows:
a precious metal nano array extinction spectrum measuring device provided in this embodiment, as shown in fig. 1, includes a white light source 2, a light beam lifter 3 for changing a light beam propagation path, a microscopic component 4, a sample holder 5 for holding a measured sample and horizontally placing the measured sample, an optical signal collector 6, a spectrometer 7, and a computer terminal 8, where a light beam emitted by the white light source 2 is incident on the microscopic component 4 after changing the propagation path by the light beam lifter 3, a focused light beam emitted by the microscopic component 4 is incident on the measured sample 9, the optical signal collector 6 receives the focused light beam transmitted through the measured sample 9 and transmits the focused light beam to the spectrometer 7, and the spectrometer 7 outputs a spectrum signal to the computer terminal 8.
In this embodiment, the beam lifter 3 is composed of a first support 31 vertically disposed, a first reflector 32 disposed at a lower portion of the first support 31, and a second reflector 33 disposed at an upper portion of the first support 31, wherein the light beam emitted from the white light source 2 is incident on the first reflector 32, the light beam reflected by the first reflector 32 is incident on the second reflector 33, and the light beam reflected by the second reflector 33 is incident on the micro-assembly 4; the light beam lifter 3 is used for lifting the light beam from a low position to a high position, and can be realized by utilizing two reflectors.
In this embodiment, the first reflector 32 and the second reflector 33 both form an angle of 45 degrees with the horizontal plane, the first reflector 32 and the second reflector 33 are arranged in parallel, and the reflecting surface of the first reflector 32 is opposite to the reflecting surface of the second reflector 33; the positioning of the first mirror 32 and the second mirror 33 is defined to ensure that the light beam reflected by the first mirror 32 is received by the second mirror 33 and that the light beam reflected by the second mirror 33 is received by the microscopic assembly 4.
In this embodiment, the microscope assembly 4 comprises a second support 41 vertically disposed, a half mirror 42 and an objective 43 sequentially connected to the second support 41 from top to bottom, and an absorption plate 44 connected to the second support 41 and located on a propagation path of a light beam transmitted by the half mirror 42, wherein the half mirror 42 and the objective 43 are located right above the sample holder 5, the light beam emitted from the light beam lifter 3 is incident on the half mirror 42, the light beam reflected by the half mirror 42 is incident on the objective 43, a focused light beam emitted by the objective 43 is incident on the sample 9 to be tested, the light beam transmitted by the half mirror 42 is incident on the absorption plate 44, and the absorption plate 44 absorbs the light beam transmitted by the half mirror 42; the half mirror 42 is used to change the propagation path of the light beam reflected from the second reflector 33, so that the light beam reflected by the half mirror 42 is vertically incident on the objective lens 43, and the light beam transmitted by the half mirror 42 is incident on the light-absorbing plate 44 and absorbed by the light-absorbing plate 44; the beam reflected by the half mirror 42 is then focused by the objective lens 43, such a microscopic assembly 4 being simple in construction and convenient in functional implementation.
In this embodiment, the half mirror 42 forms an angle of 45 degrees with the horizontal plane, and the reflective surface of the half mirror 42 is opposite to the reflective surface of the second reflector 33; the light absorption plate 44 is vertically arranged to form an angle of 45 degrees with the semi-transparent semi-reflecting mirror 42; by limiting the relative positions of the second mirror 33 and the half mirror 42, it is possible to ensure that the entire light beam reflected by the second mirror 33 is received by the half mirror 42; by limiting the relative positions of the half mirror 42 and the light absorbing plate 44, it is possible to ensure that all the light beams transmitted by the half mirror 42 are absorbed by the light absorbing plate 44.
In this embodiment, the second support 41 is further connected to a CCD (Charge-coupled Device) camera 45, the CCD camera 45 is located right above the half mirror 42, an output end of the CCD camera 45 is connected to the computer terminal 8, and the CCD camera 45 acquires the feature of the precious metal nano array in the measured sample 9 observed through the objective lens 43; the CCD camera 45 is arranged right above the objective lens 43, and the appearance of the noble metal nano array serving as the detected sample 9 is obtained by the CCD camera 45, so that the function of the device is expanded.
In this embodiment, the optical signal collector 6 is shown in fig. 3 and is composed of an optical fiber probe clamp 64 and an optical fiber probe 65 clamped by the optical fiber probe clamp 64, the optical fiber probe 65 is vertically disposed, a receiving end of the optical fiber probe 65 is located right below the sample 9 to be measured, the optical fiber probe 65 receives the focused light beam transmitted through the sample 9 to be measured, an output end of the optical fiber probe 65 is connected with an input end of the spectrometer 7 through a conducting optical fiber 66, and the optical fiber probe 65 transmits the focused light beam transmitted through the sample 9 to the spectrometer 7.
Example two:
the extinction spectrum measurement device for the precious metal nano array provided by the embodiment is a further improvement of the device of the first embodiment, as shown in fig. 2 and fig. 3, that is, it further includes a first three-dimensional position adjustment device for adjusting the position of the sample holder 5, a second three-dimensional position adjustment device for adjusting the position of the optical signal collector 6, and a base 1 (see fig. 1), the first three-dimensional position adjustment device is composed of a first positioning plate 51, a first three-dimensional displacement platform 52 installed on the first positioning plate 51, and a first connecting rod 53 connecting the first three-dimensional displacement platform 52 and the sample holder 5, and the position of the sample holder 5 is adjusted by a first micrometer screw 521, a second micrometer screw 522, and a third micrometer screw 523 in the first three-dimensional displacement platform 52, so that the sample 9 to be measured is located right below the objective 43; the second three-dimensional position adjusting device consists of a second positioning plate 61, a second three-dimensional displacement platform 62 arranged on the second positioning plate 61, and a second connecting rod 63 connecting the second three-dimensional displacement platform 62 and the optical fiber probe clamp 64, and the position of the optical fiber probe clamp 64 is adjusted by a first micro-wire rod 621, a second micro-wire rod 622 and a third micro-wire rod 623 in the second three-dimensional displacement platform 62, so that the optical fiber probe 65 is positioned right below the sample 9 to be detected and is close to the lower surface of the sample 9 to be detected; the white light source 2 and the spectrometer 7 are placed on the base 1, the bottom of the first support 31 and the bottom of the second support 41 are respectively and fixedly connected with the base 1, and the first positioning plate 51 and the second positioning plate 61 are respectively and fixedly connected with the base 1.
The position of the sample clamp 5 is adjusted by using the first three-dimensional displacement platform 52, so that the measured sample 9 is positioned right below the objective lens 43, and the focused light beam emitted by the objective lens 43 is completely incident on the measured sample 9; the position of the optical fiber probe clamp 64 is adjusted by using the second three-dimensional displacement platform 62, so that the optical fiber probe 65 is positioned right below the tested sample 9 and close to the lower surface of the tested sample 9, and the information acquired by the optical fiber probe 65 is more accurate; all the components are gathered together by the base 1 to form a whole.
In the first and second embodiments, the white light source 2 adopts the existing xenon lamp light source or the bromine tungsten lamp light source or other white light sources in the prior art; the sample clamp 5 adopts the existing clamping equipment which can stably and reliably clamp the object; the spectrometer 7 adopts the prior art; the computer terminal 8 adopts the prior art and is provided with a display, the computer terminal 8 is internally provided with the existing spectrometer control software, the existing image processing software and the existing noble metal nano-array extinction spectrum calculation software, the computer terminal 8 controls the spectrometer 7 to collect signals through the spectrometer 7 control software and processes the spectrum signals output by the spectrometer 7, the computer terminal 8 calculates through the noble metal nano-array extinction spectrum calculation software to obtain the extinction spectrum of the noble metal nano-array, and the computer terminal 8 processes the noble metal nano-array obtained by the CCD camera 45 through the image processing software; the first three-dimensional displacement platform 52 and the second three-dimensional displacement platform 62 both adopt the existing three-dimensional displacement platform and are provided with three micrometer screws; the optical fiber probe clamp 64 adopts the existing clamping equipment which can stably and reliably clamp the object; the fiber optic probe 65 employs multimode plastic optical fiber or other dielectric optical fiber as is known in the art.
Example three:
the embodiment provides a method for realizing the sensing detection of the concentration of an ethanol solution by using the noble metal nano array extinction spectrum measuring device of the second embodiment, which comprises the following steps:
① gold nano-arrays with periodic structures are prepared on conductive glass deposited with conductive films by using the existing electron beam etching technology, the gold nano-particles are nano-cylinders with the diameter of 200 nm and the height of 75 nm, the distance between the gold nano-particles is 50 nm, the Scanning Electron Microscope (SEM) picture is shown in figure 4, and then the conductive glass with the gold nano-arrays is used as the tested sample.
The conductive glass can be a glass substrate deposited with a conductive film, or an organic glass substrate deposited with a conductive film, wherein the conductive film is an ITO (Indium-Tin Oxide) film, or an FTO (SnO2: F, namely fluorine-doped Tin dioxide) film; the noble metal material is gold or silver.
② turning on a white light source, adjusting the position of a fiber probe clamp by a second three-dimensional displacement platform to make the receiving port of the fiber probe be at the focal plane of the objective lens, so that the receiving port of the fiber probe is clearly displayed in a display of the computer terminal by the objective lens and the CCD camera, turning on the spectrometer, and setting signal acquisition parameters, such as setting the integration time to be 1 second, wherein the spectrum signal output by the spectrometer in the white light source on state is the spectrum of the white light source and is marked as I (lambda), and is transmitted to the computer terminal and stored by the computer terminal, wherein lambda represents the wavelength.
③ the position of the optical fiber probe clamp is adjusted by the second three-dimensional displacement platform to make the optical fiber probe move vertically downwards, the optical fiber probe moves downwards for the tested sample to be displayed on the focal plane, the conductive glass is very thin, so the optical fiber probe generally moves downwards by 2cm, then the tested sample is clamped by the sample clamp and is horizontally placed, then the position of the sample clamp is adjusted by the first three-dimensional displacement platform to make the gold nano array in the tested sample to be located on the focal plane of the objective lens and ensure that the gold nano array image in the tested sample is clearly displayed in the display of the computer terminal by the objective lens and the CCD camera, the position of the optical fiber probe clamp is adjusted by the second three-dimensional displacement platform to make the optical fiber probe be located under the tested sample and make the receiving port of the optical fiber probe approach the receiving port of the tested sample as much as possibleA lower surface; at the moment, the spectrum signal output by the spectrometer in the state of opening the white light source is the transmission spectrum of the gold nano array in the tested sample and is marked as T1And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
④ turning off the white light source, and taking the spectrum signal output by the spectrometer in the off state of the white light source as the first background spectrum marked as B1And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
⑤ computer terminal utilizes formula E according to the definition of extinction spectrum1(λ)＝I(λ)-T1(λ)+B1(lambda), calculating to obtain an extinction spectrum E of the gold nano array in the tested sample1(λ)。
⑥ transferring 20 μ l ethanol solution as organic solution to be detected with a liquid transfer gun, dripping onto gold nano array in the sample to be detected, and taking the spectrum signal outputted by the spectrometer in the off state of the white light source as a second background spectrum, and recording as B2And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
⑦ turning on the white light source, wherein the spectrum signal output by the spectrometer in the state of turning on the white light source is the transmission spectrum of the gold nano array dripped with the organic solution to be detected and marked as T2And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
⑧ computer terminal utilizes formula E according to the definition of extinction spectrum2(λ)＝I(λ)-T2(λ)+B2(lambda), calculating to obtain extinction spectrum E of the gold nano array dripped with the organic solution to be detected2(λ)。
⑨ mixing E1(lambda) as a reference spectrum, according to E2Extinction peak in (λ) relative to E1And (lambda) changing the position or the intensity of the extinction peak to obtain the concentration of the organic solution to be detected.
Fig. 5 shows extinction spectra of gold nano-arrays to which 20. mu.l of ethanol solutions having concentrations of 20%, 40%, 60% and 80% and absolute ethanol having a concentration of 99.7% were added dropwise, respectively, on the gold nano-arrays in example three, obtained after addition of ethanol solutions having concentrations of 20%, 40%, 60% and 80% and absolute ethanol having a concentration of 99.7% was added dropwise. Ethanol solutions of varying concentrations were prepared by formulating ethanol with a purity greater than 99.99% with deionized water. As can be seen from FIG. 5, the position of the extinction peak of the extinction spectrum is red-shifted with the increase in the concentration of the ethanol solution.
FIG. 6 is a graph showing the response of the concentration of ethanol solution and absolute ethanol in different concentrations in example three. After obtaining the extinction spectrum of the gold nano array dropwise added with the ethanol solution, the concentration of the dropwise added ethanol solution can be obtained according to fig. 6.
Example four:
the embodiment provides a method for realizing concentration sensing detection of an enrofloxacin solution by using the noble metal nano array extinction spectrum measuring device of the second embodiment, which comprises the following steps:
① gold nano-arrays with periodic structures are prepared on conductive glass deposited with conductive films by using the existing electron beam etching technology, the gold nano-particles are nano-cylinders with the diameter of 200 nm and the height of 75 nm, the distance between the gold nano-particles is 50 nm, the Scanning Electron Microscope (SEM) picture is shown in figure 4, and then the conductive glass with the gold nano-arrays is used as the tested sample.
The conductive glass can be a glass substrate deposited with a conductive film, or an organic glass substrate deposited with a conductive film, wherein the conductive film is an ITO (Indium-Tin Oxide) film, or an FTO (SnO2: F, namely fluorine-doped Tin dioxide) film; the noble metal material is gold or silver.
② turning on a white light source, adjusting the position of a fiber probe clamp by a second three-dimensional displacement platform to make the receiving port of the fiber probe be at the focal plane of the objective lens, so that the receiving port of the fiber probe is clearly displayed in a display of the computer terminal by the objective lens and the CCD camera, turning on the spectrometer, and setting signal acquisition parameters, such as setting the integration time to be 1 second, wherein the spectrum signal output by the spectrometer in the white light source on state is the spectrum of the white light source and is marked as I (lambda), and is transmitted to the computer terminal and stored by the computer terminal, wherein lambda represents the wavelength.
③ the position of the optical fiber probe clamp is adjusted by the second three-dimensional displacement platform to make the optical fiber probe move vertically downwards for the tested sample to be displayed on the focal plane, the optical fiber probe can move downwards for 2cm under normal condition because the conductive glass is very thin, then the sample clamp is used to clamp the tested sample and make the tested sample horizontally placed, then the position of the sample clamp is adjusted by the first three-dimensional displacement platform to make the gold nano array in the tested sample to be located on the focal plane of the objective lens and ensure that the gold nano array image in the tested sample is clearly displayed in the self-carried display of the computer terminal by the objective lens and the CCD camera, the position of the optical fiber probe clamp is adjusted by the second three-dimensional displacement platform to make the optical fiber probe to be located right below the tested sample and make the receiving port of the optical fiber probe to be as close to the lower surface of the tested sample as possible, at this time, the spectrum signal output by the spectrometer in the state of turning on the white light source is the transmission spectrum of the gold nano array in the tested sample and is marked as T1And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
④ turning off the white light source, and taking the spectrum signal output by the spectrometer in the off state of the white light source as the first background spectrum marked as B1And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
⑤ computer terminal utilizes formula E according to the definition of extinction spectrum1(λ)＝I(λ)-T1(λ)+B1(lambda), calculating to obtain an extinction spectrum E of the gold nano array in the tested sample1(λ)。
⑥ transferring 20 μ l of enrofloxacin solution as organic solution to be measured with a liquid-transferring gun, dripping onto gold nano array in the sample to be measured, and taking the spectral signal outputted by the spectrometer in the off state of the white light source as a second background spectrum marked as B2And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
⑦ turning on the white light source, the spectrum signal output by the spectrometer is dropped with the to-be-detected spectrum signal under the state of turning on the white light sourceTransmission spectrum of gold nano array of organic solution, marked as T2And (lambda) and transmitting the data to the computer terminal for storage by the computer terminal.
⑧ computer terminal utilizes formula E according to the definition of extinction spectrum2(λ)＝I(λ)-T2(λ)+B2(lambda), calculating to obtain extinction spectrum E of the gold nano array dripped with the organic solution to be detected2(λ)。
⑨ mixing E1(lambda) as a reference spectrum, according to E2Extinction peak in (λ) relative to E1And (lambda) changing the position or the intensity of the extinction peak to obtain the concentration of the organic solution to be detected.
FIG. 7 shows extinction spectra of gold nanoarrays to which enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L were added dropwise, respectively, after 20. mu.l of the enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L was added dropwise in example four. Solutions of enrofloxacin at various concentrations were prepared by formulating enrofloxacin powder having a purity greater than or equal to 98% (HPLC) with an analytically pure ethanol solution. As can be seen from fig. 7, as the concentration of the enrofloxacin solution increased, the position of the extinction peak in the extinction spectrum red-shifted and the peak intensity increased.
FIG. 8 is a graph showing the concentration response of different concentrations of enrofloxacin solution from example four. After obtaining the extinction spectrum of the gold nano array with the enrofloxacin solution added dropwise, the concentration of the dropwise added enrofloxacin solution can be obtained according to the figure 8.
Example five:
the embodiment provides a method for sensing and detecting the concentration of an ethanol solution by using the noble metal nano-array extinction spectrum measuring device in the second embodiment, which has the same specific steps as those of the method for sensing and detecting the concentration of the ethanol solution in the third embodiment, and the difference is that: different from the gold nano-arrays with periodic structures, in this embodiment, the gold nanoparticles are nanocylinders with a diameter of 200 nm and a height of 75 nm, the spacing between the gold nanoparticles is 100 nm, and a Scanning Electron Microscope (SEM) photograph thereof is shown in fig. 9.
Fig. 10 shows extinction spectra of gold nano-arrays to which 20. mu.l of ethanol solutions having concentrations of 20%, 40%, 60% and 80% and absolute ethanol having a concentration of 99.7% were added dropwise, respectively, on the gold nano-arrays in example five, obtained after addition of the ethanol solutions having concentrations of 20%, 40%, 60% and 80% and the absolute ethanol having a concentration of 99.7% was added dropwise. Ethanol solutions of varying concentrations were prepared by formulating ethanol with a purity greater than 99.99% with deionized water. As can be seen from fig. 10, as the concentration of the ethanol solution increases, the position of the extinction peak of the extinction spectrum is red-shifted therewith; and it was found that the amount of change in the position of the extinction peak of the extinction spectrum in example five was much smaller than that in example three, which is associated with the decrease in Localized Surface Plasmon Resonance (LSPR) due to the increase in the inter-particle distance of the gold nano-array in example five.
FIG. 11 is a graph showing the response of the concentration of ethanol solution and absolute ethanol in different concentrations in example five. After obtaining the extinction spectrum of the gold nano array dropwise added with the ethanol solution, the concentration of the dropwise added ethanol solution can be obtained according to fig. 11.
Example six:
the embodiment provides a method for realizing the sensing detection of the concentration of the enrofloxacin solution by using the device for measuring the extinction spectrum of the noble metal nano array in the second embodiment, and the specific steps of the method for realizing the sensing detection of the concentration of the enrofloxacin solution in the fourth embodiment are the same as those of the method for realizing the sensing detection of the concentration of the enrofloxacin solution in the fourth embodiment, except that: different from the gold nano-arrays with periodic structures, in this embodiment, the gold nanoparticles are nanocylinders with a diameter of 200 nm and a height of 75 nm, the spacing between the gold nanoparticles is 100 nm, and a Scanning Electron Microscope (SEM) photograph thereof is shown in fig. 9.
FIG. 12 shows extinction spectra of gold nanoarrays to which enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L were added dropwise, respectively, after 20. mu.l of the enrofloxacin solutions having concentrations of 5mg/L, 10mg/L, 15mg/L, 20mg/L, and 25mg/L were added dropwise in example six. Solutions of enrofloxacin at various concentrations were prepared by formulating enrofloxacin powder having a purity greater than or equal to 98% (HPLC) with an analytically pure ethanol solution. As can be seen from fig. 12, as the solution concentration of enrofloxacin increased, the position of extinction peak in the extinction spectrum was red-shifted and the peak intensity increased; and it was found that the amount of change in the position and peak intensity of the extinction peak in the extinction spectrum in example six was much smaller than that in the extinction spectrum in example four, which is related to the decrease in Localized Surface Plasmon Resonance (LSPR) due to the increase in the inter-particle distance of the gold nano-array in example six.
FIG. 13 shows the concentration response curves of the enrofloxacin solutions of different concentrations in example six. After obtaining the extinction spectrum of the gold nano array with the enrofloxacin solution added dropwise, the concentration of the dropwise added enrofloxacin solution can be obtained according to the graph 13.
Claims (1)
1. A sensing detection method of a precious metal nano array extinction spectrum measuring device comprises a white light source, a light beam lifter for changing a light beam propagation path, a microscopic component, a sample clamp for clamping a measured sample to enable the measured sample to be horizontally placed, an optical signal collector, a spectrometer and a computer terminal, wherein light beams emitted by the white light source are incident on the microscopic component after the propagation path of the light beams is changed by the light beam lifter, focused light beams emitted by the microscopic component are incident on the measured sample, the optical signal collector receives the focused light beams transmitted by the measured sample and transmits the focused light beams to the spectrometer, and the spectrometer outputs spectrum signals to the computer terminal; the light beam lifter consists of a first support, a first reflecting mirror and a second reflecting mirror, wherein the first support is vertically arranged, the first reflecting mirror is arranged at the lower part of the first support, the second reflecting mirror is arranged at the upper part of the first support, light beams emitted by the white light source are incident on the first reflecting mirror, the light beams reflected by the first reflecting mirror are incident on the second reflecting mirror, and the light beams reflected by the second reflecting mirror are incident on the microscopic component; the first reflector and the second reflector form an angle of 45 degrees with the horizontal plane, the first reflector and the second reflector are arranged in parallel, and the reflecting surface of the first reflector is opposite to the reflecting surface of the second reflector; the microscope assembly comprises a second support, a half-mirror and an objective lens which are vertically arranged, and a light absorption plate which is connected to the second support and is positioned on a transmission path of a light beam transmitted by the half-mirror from top to bottom, wherein the half-mirror and the objective lens are positioned right above the sample clamp; the semi-transparent semi-reflecting mirror and the horizontal plane form an angle of 45 degrees, and the reflecting surface of the semi-transparent semi-reflecting mirror is opposite to the reflecting surface of the second reflecting mirror; the light absorption plate is vertically arranged to form an angle of 45 degrees with the semi-transparent semi-reflecting mirror; the second bracket is also connected with a CCD camera, the CCD camera is positioned right above the semi-transparent semi-reflective mirror, the output end of the CCD camera is connected with the computer terminal, and the CCD camera acquires the appearance of the noble metal nano array in the detected sample observed through the objective lens; the extinction spectrum measurement device for the precious metal nano array further comprises a first three-dimensional position adjusting device for adjusting the position of the sample clamp, wherein the first three-dimensional position adjusting device consists of a first positioning plate, a first three-dimensional displacement platform installed on the first positioning plate and a first connecting rod for connecting the first three-dimensional displacement platform with the sample clamp, and the position of the sample clamp is adjusted through the first three-dimensional displacement platform so that the sample to be measured is located right below the objective lens; the optical signal collector consists of an optical fiber probe clamp and an optical fiber probe clamped by the optical fiber probe clamp, the optical fiber probe is vertically placed, a receiving end of the optical fiber probe is positioned right below a measured sample, the optical fiber probe receives a focused light beam transmitted through the measured sample, an output end of the optical fiber probe is connected with an input end of the spectrometer through a conducting optical fiber, and the optical fiber probe transmits the focused light beam transmitted through the measured sample to the spectrometer; the device for measuring the extinction spectrum of the precious metal nano array further comprises a second three-dimensional position adjusting device for adjusting the position of the optical signal collector, wherein the second three-dimensional position adjusting device consists of a second positioning plate, a second three-dimensional displacement platform installed on the second positioning plate and a second connecting rod for connecting the second three-dimensional displacement platform with the optical fiber probe clamp; the device for measuring the extinction spectrum of the noble metal nano array further comprises a base, the white light source and the spectrometer are placed on the base, the bottom of the first support and the bottom of the second support are respectively and fixedly connected with the base, and the first positioning plate and the second positioning plate are respectively and fixedly connected with the base; the method is characterized in that: the method for sensing and detecting the concentration of the organic solution by using the noble metal nano array extinction spectrum measuring device specifically comprises the following steps:
① preparing a noble metal nano array with a specific pattern on the conductive glass deposited with the conductive film by using an electron beam etching technology, and then taking the conductive glass with the noble metal nano array with the specific pattern as a tested sample;
② turning on white light source, adjusting the position of the optical fiber probe clamp by the second three-dimensional displacement platform to make the receiving port of the optical fiber probe be at the focal plane of the objective lens, so that the receiving port of the optical fiber probe can be clearly displayed in the display of the computer terminal by the objective lens and the CCD camera, turning on the spectrometer and setting signal acquisition parameters, wherein the spectrum signal output by the spectrometer is the spectrum of the white light source in the state of turning on the white light source, marked as I (lambda), and transmitted to the computer terminal for storage by the computer terminal, wherein lambda represents wavelength;
③ adjusting the position of the fiber probe clamp by the second three-dimensional displacement platform to move the fiber probe vertically downwards, clamping the sample to be tested by the sample clamp and horizontally placing the sample to be tested, adjusting the position of the sample clamp by the first three-dimensional displacement platform to make the noble metal nano array in the sample to be tested positioned at the focal plane of the objective lens and ensure that the noble metal nano array image in the sample to be tested is clearly displayed in the display of the computer terminal by the objective lens and the CCD camera, adjusting the position of the fiber probe clamp by the second three-dimensional displacement platform to make the fiber probe positioned under the sample to be tested and make the receiving port of the fiber probe as close as possible to the lower surface of the sample to be tested, and the spectrum signal output by the spectrometer in the white light source on state is the transmission spectrum of the noble metal nano array in the sample to be tested and is marked as T1(lambda), and transmitting to the computer terminal for storage by the computer terminal;
④ turning off the white light source, and taking the spectrum signal output by the spectrometer in the off state of the white light source as the first background spectrum marked as B1(lambda), and transmitting to the computer terminal for storage by the computer terminal;
⑤ computer terminal utilizes formula E according to the definition of extinction spectrum1(λ)＝I(λ)-T1(λ)+B1(lambda), calculating to obtain an extinction spectrum E of the noble metal nano array in the tested sample1(λ)；
⑥ dropping the organic solution to be detected onto the noble metal nano array in the sample to be detected, and taking the spectral signal output by the spectrometer in the off state of the white light source as a second background spectrum marked as B2(lambda), and transmitting to the computer terminal for storage by the computer terminal;
⑦ turning on the white light source, wherein the spectrum signal output by the spectrometer in the state of turning on the white light source is the transmission spectrum of the noble metal nano array dripped with the organic solution to be detected and is marked as T2(lambda), and transmitting to the computer terminal for storage by the computer terminal;
⑧ computer terminal utilizes formula E according to the definition of extinction spectrum2(λ)＝I(λ)-T2(λ)+B2(lambda), calculating to obtain extinction spectrum E of the noble metal nano array dripped with the organic solution to be detected2(λ)；
⑨ mixing E1(lambda) as a reference spectrum, according to E2Extinction peak in (λ) relative to E1And (lambda) changing the position or the intensity of the extinction peak to obtain the concentration of the organic solution to be detected.
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