CN115824976A - Photoacoustic micro-fluidic detection system based on planar waveguide - Google Patents
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Abstract
The invention is suitable for the field of photoacoustic microfluidic detection, and provides a photoacoustic microfluidic detection system based on a planar waveguide, which comprises: the device comprises a micro-fluidic chip module, a laser excitation module, a laser detection module block and a signal processing module; the micro-fluidic chip module comprises a micro-channel and a planar waveguide arranged above the micro-channel; the laser excitation module is used for exciting a target sample in the micro-channel below the planar waveguide to emit ultrasonic waves; the laser detection module is used for receiving a photoacoustic signal generated after the target sample is excited and converting the photoacoustic signal into a voltage signal; and the signal processing module is used for receiving and processing the voltage signal of the laser detection module to obtain an optical signal of the target sample. The invention has simple structure, low cost, high integration level and wide detection range, can realize large-area rapid scanning detection of biological or chemical samples, obtains photoacoustic signals of multiple points of the samples and further performs rapid imaging on the samples.
Description
Technical Field
The invention belongs to the technical field of photoacoustic microfluidic detection, and particularly relates to a photoacoustic microfluidic detection system based on a planar waveguide.
Background
Microfluidics, as a system on a chip, enables biological or chemical experiments to be performed in micron-scale spaces. Therefore, the method has great significance for detecting the sample in the microfluidic system in real time. However, the current microfluidic detection technologies, such as laser-induced fluorescence method, electrochemical detection method, etc., have the defects of sample pollution and insufficient detection sensitivity. Therefore, a real-time detection method with high sensitivity, fast response speed and no mark is needed for detecting the sample in the microfluidic chip. Photoacoustic detection is a detection mode based on photoacoustic effect, and people begin to apply photoacoustic detection to the field of microfluidic detection due to the characteristics of high sensitivity, good contrast, high response speed, simple structure and no mark.
Currently, the existing photoacoustic microfluidic system uses an ultrasonic transducer as a probe for detecting photoacoustic waves, if we have studied to detect tumor cells in Real time ("Capture of circulating tumor cells using photoacoustic flow and two phase flow", biological Optics 17 (6), 061221 (June 2012)), the technology of flowing droplet imaging has also been developed ("optical-fluidic for a fluidic label-free detection of droplets and cells in microorganisms", lab Chip,2018,18, 1292), further studies have been carried out to count red cells in Real time ("read-time bonded and fluidic analysis using a microfluidic-based probe", lab Chip, 251, 2586), but these are not influenced by the complex process of making the piezoelectric transducer and the microfluidic Chip is inevitably affected by the defects of the piezoelectric transducer. Aiming at the defects of limited detection range, difficulty in miniaturization, poor compatibility effect, high cost and the like of the existing photoacoustic microfluidic system, researchers develop a pure optical detection method based on polarized light and combined with a microfluidic chip to realize the detection of photoacoustic signals of a sample, but the existing microfluidic chip is formed by combining a microchannel and a prism, only single reflection of the prism is utilized, the detection light and excitation light are coincided at one point, the single-point photoacoustic signal measurement of the sample is realized, the detection area is small, and the range is narrow. In order to better research a biological or chemical sample and image the sample, the sample needs to be scanned and detected to obtain more photoacoustic signals, so that the research and development of a photoacoustic microfluidic system which is simple, feasible, economical and practical, large in detection area, rapid and high in performance has important significance for the application and popularization of photoacoustic detection in microfluidic detection.
Disclosure of Invention
The invention aims to provide a photoacoustic microfluidic detection system based on a planar waveguide, which is simple, feasible, economical, practical, large in detection area, rapid and high in performance.
The invention is realized in such a way that a photoacoustic microfluidic detection system based on a planar waveguide is characterized in that the system comprises: the device comprises a micro-fluidic chip module, a laser excitation module, a laser detection module block and a signal processing module; wherein,
the micro-fluidic chip module comprises a micro-channel and a planar waveguide arranged above the micro-channel;
the laser excitation module is used for exciting a target sample in the micro-channel below the planar waveguide to emit ultrasonic waves;
the laser detection module is used for receiving a photoacoustic signal generated after the target sample is excited and converting the photoacoustic signal into a voltage signal;
the signal processing module is used for receiving and processing the voltage signal of the laser detection module to obtain an optical signal of the target sample.
The further technical scheme of the invention is as follows: the laser excitation module comprises a two-dimensional galvanometer, and the two-dimensional galvanometer is used for controlling excitation light to rapidly scan the target sample in x and y vertical directions.
The further technical scheme of the invention is as follows: the planar waveguide is a one-dimensional planar waveguide or a two-dimensional planar waveguide.
The further technical scheme of the invention is as follows: the reflecting mirror structures on two sides in the two-dimensional planar waveguide are provided with metal films on the surfaces on two sides of the waveguide, and the mirror surfaces are vertically arranged.
The further technical scheme of the invention is as follows: the upper cladding of the one-dimensional planar waveguide and the two-dimensional planar waveguide is the same as the liquid medium in the micro-channel in the micro-fluidic chip.
The further technical scheme of the invention is as follows: the laser detection module introduces a phase type photoacoustic sensor and adopts a phase type detection mode to perform photoacoustic imaging.
The photoacoustic microfluidic detection system based on the planar waveguide has the advantages that:
the invention has simple structure, low cost, high integration level and wide detection range, can realize large-area rapid scanning detection of biological or chemical samples, obtains photoacoustic signals of multiple points of the samples and further performs rapid imaging on the samples.
Drawings
FIG. 1 is a schematic structural diagram of an embodiment 1 and an embodiment 2 of the photoacoustic microfluidic detection system based on a planar waveguide according to the present invention;
FIG. 2 is a schematic structural diagram of a microfluidic chip in embodiment 1 of the photoacoustic microfluidic detection system based on planar waveguide according to the present invention;
FIG. 3 is a schematic diagram of a three-dimensional structure of a microfluidic chip in embodiment 2 of the photoacoustic microfluidic detection system based on planar waveguides according to the present invention;
FIG. 4 is a three-dimensional front view of a microfluidic chip in embodiment 2 of the photoacoustic and microfluidic detection system based on planar waveguide according to the present invention;
FIG. 5 is a top view of a three-dimensional structure of a microfluidic chip in embodiment 2 of the photoacoustic microfluidic detection system based on planar waveguides according to the present invention;
FIG. 6 is a schematic diagram of a simulation of an embodiment of the photoacoustic microfluidic detection system based on the planar waveguide according to the present invention;
FIG. 7 is a schematic diagram of theoretical calculation and simulation results of a photoacoustic microfluidic detection system according to an embodiment of the present invention;
fig. 8 is a schematic structural diagram of embodiment 3 of the photoacoustic microfluidic detection system based on the planar waveguide according to the present invention.
Detailed Description
The invention provides a photoacoustic microfluidic detection system based on a planar waveguide, which comprises: the device comprises a microfluidic chip die, a laser excitation module, a laser detection module block and a signal processing module.
The micro-fluidic chip module comprises a micro-channel and a planar waveguide arranged above the micro-channel; the laser excitation module is used for exciting a target sample in the micro-channel below the planar waveguide to emit ultrasonic waves; the laser detection module is used for receiving a photoacoustic signal generated after the target sample is excited and converting the photoacoustic signal into a voltage signal; and the signal processing module is used for receiving and processing the voltage signal of the laser detection module to obtain and display the optical signal of the target sample.
The planar waveguide with simple structure is composed of three layers of uniform media, the medium layer in the middle is called a waveguide layer or a core layer, and the medium layers on the upper side and the lower side of the core layer are called cladding layers. By utilizing the principle of multiple total internal reflection in the waveguide, each reflection point is used as a detection point, the detection area can be enlarged, and the advantages of simple manufacture and low cost are achieved.
The photoacoustic microfluidic detection system based on the planar waveguide has the advantages of simple structure, low cost, high integration level and wide detection range, can realize large-area rapid scanning detection on biological or chemical samples, obtains photoacoustic signals of multiple points of the samples and further performs rapid imaging on the samples.
Furthermore, in the photoacoustic microfluidic detection system based on the planar waveguide, the laser excitation module comprises a two-dimensional galvanometer, and the two-dimensional galvanometer is used for controlling the excitation light to rapidly scan the target sample in x and y vertical directions.
Further, the planar waveguide is a one-dimensional planar waveguide or a two-dimensional planar waveguide.
Furthermore, the two sides of the reflecting mirror structure in the two-dimensional planar waveguide are plated with metal films on the surfaces of the two sides of the waveguide, and the mirror surface is vertically arranged.
Further, the upper cladding of the one-dimensional planar waveguide and the two-dimensional planar waveguide is the same as the liquid medium in the micro-channel in the micro-fluidic chip.
Furthermore, a phase type photoacoustic sensor is introduced into the laser detection module, and photoacoustic imaging is carried out in a phase type detection mode.
The structure and the operation principle of the photoacoustic microfluidic detection system based on planar waveguide according to the present invention will be further explained with reference to fig. 1 to 8.
Example 1
As shown in fig. 1 and 2, the laser excitation module includes a pulse excitation light source 5, a beam expander 6, an adjustable attenuator 7, a beam splitter 8, a two-dimensional galvanometer G, an objective lens 10, and a photoelectric probe. A pulse laser beam is emitted from a pulse excitation light source 5, is collimated and amplified by a beam expander 6, is filtered by an adjustable attenuator 7, is split by a beam splitter 8, and one beam of light enters a photoelectric probe 9 to serve as a trigger signal of a subsequent signal processing module. The other beam of light 532 is reflected by the two-dimensional galvanometer G and focused on the sample 300 in the micro-fluidic chip micro-channel 200 through the objective lens 10 to excite the target sample to emit ultrasonic waves.
The laser detection module comprises a continuous laser light source 1, a polarizer 2, a 1/4 wave plate 3, a reflecting mirror M, a polarization beam splitter PBS and an optical probe 11. The continuous laser source emits a continuous laser beam, which is converted into linearly polarized light by the polarizer 2 and converted into circularly polarized light by the 1/4 wave plate 3. After passing through the reflector M, the circularly polarized light is incident into the planar waveguide from the side surface of the planar waveguide at a certain angle and undergoes total internal reflection for multiple times. As shown in fig. 2, the one-dimensional planar waveguide includes a core layer 100 and an upper cladding layer 110, and the microfluidic chip is formed by packaging a lower micro channel 200 and a one-dimensional planar waveguide above the micro channel. When ultrasonic waves exist in the micro-channel on the lower surface of the planar waveguide, the refractive index of liquid around the corresponding reflection point is changed, the energy of P polarized light and S polarized light in the reflected light of the corresponding point generates nonlinear change with different responses, and the reflectivity of the P polarized light and the S polarized light of other points is the same. In the range where the incident angle is close to the total reflection angle, the P-polarized light and the S-polarized light are most sensitive to the change of the refractive index of water, and the signal difference between the two is also the largest. The angle of incidence is then reduced to just no total reflection by fine tuning the mirrors of the input planar waveguide, with this angle close to the angle of total reflection as the angle of incidence. The reflected light 633 reflected by the planar waveguide for multiple times is reflected by the mirror M, and the S-polarized light and the P-polarized light in the reflected light are separated into two beams by the polarization beam splitter 9, and received by the two probes of the high-bandwidth balanced photodetector 11.
Each reflection point on the lower surface of the core layer in the planar waveguide is a detection point, so that the detection area is enlarged, the two-dimensional galvanometer G can control two coordinate systems for exciting the vertical coordinate of laser, the galvanometer G controls the exciting light 532 to carry out rapid scanning in the x direction (horizontal direction), the photoacoustic signals of a plurality of points in the x direction on a sample are excited, one photoacoustic signal can be generated by each excitation, and the optical probe can receive each photoacoustic signal to obtain the photoacoustic signals of the plurality of points in the x direction of the sample.
The signal processing module 12 includes a filter, an amplifier, and an oscilloscope. The detected change of the intensity difference of the P polarized light and the S polarized light is converted into a voltage signal by a balance light detector in the laser detection module, an interference signal is filtered by a filter, the signal is amplified by an amplifier, and finally the signal is acquired by a high-bandwidth oscilloscope. The oscilloscope takes the photoelectric signals received by the photoelectric probe in the excitation module as triggers, the photoacoustic signals are displayed finally, each photoacoustic signal can be rapidly acquired by the oscilloscope, and then rapid imaging of the sample is realized.
Example two:
as shown in fig. 1 and 3, the laser excitation module includes a pulse excitation light source 5, a beam expander 6, an adjustable attenuator 7, a beam splitter 8, a two-dimensional galvanometer G, an objective lens 10, and a photoelectric probe. A pulse laser beam is emitted from a pulse excitation light source 5, is collimated and amplified by a beam expander 6, is filtered by an adjustable attenuator 7, is split by a beam splitter 8, and one beam of light enters a photoelectric probe 9 to serve as a trigger signal of a subsequent signal processing module. The other beam of light 532 is reflected by the two-dimensional galvanometer G and focused on the sample 300 in the micro-fluidic chip micro-channel 400 through the objective lens 10 to excite the sample to emit ultrasonic waves.
The laser detection module comprises a continuous laser light source 1, a polarizer 2, a 1/4 wave plate 3, a reflecting mirror M, a polarization beam splitter PBS and an optical probe 11. The continuous laser source emits a continuous laser beam, which is converted into linearly polarized light by the polarizer 2 and converted into circularly polarized light by the 1/4 wave plate 3. After passing through the reflector M, the circularly polarized light enters the planar waveguide from the side surface of the planar waveguide at a certain angle and undergoes total internal reflection for multiple times, and a detection point is formed at the reflection point of each lower surface. As shown in fig. 3,4 and 5, the two-dimensional planar waveguide includes four parts of a core layer 500, an upper cladding layer 510 and reflectors M disposed on two sides, and the microfluidic chip is formed by packaging the lower micro-channel 200 and the two-dimensional planar waveguide above the micro-channel. When ultrasonic waves exist in the micro-channel on the lower surface of the planar waveguide, the refractive index of liquid around the corresponding reflection point is changed, the energy of the P polarized light and the S polarized light in the reflected light of the corresponding point generates nonlinear change with different responses, and the reflectivity of the P polarized light and the S polarized light of other reflection points is kept unchanged. In the range where the incident angle is close to the total reflection angle, the P-polarized light and the S-polarized light are most sensitive to the change of the refractive index of water, and the signal difference between the two is also the largest. The angle of incidence is then reduced to just no total reflection by fine tuning the mirrors of the input planar waveguide, with this angle close to the angle of total reflection as the angle of incidence. The reflected light 634 reflected back and forth multiple times in the planar waveguide is reflected by the mirror M, and the S-polarized light and the P-polarized light in the reflected light are separated into two beams by the polarization beam splitter 9, and are received by the two probes of the high-bandwidth balanced light detector 11.
As shown in figures 4 and 5 of the drawings,
representing the reflection point of the upper surface in the two-dimensional planar waveguide core,
the two-dimensional plane waveguide core layer is characterized in that reflection points of the middle lower surface of the two-dimensional plane waveguide core layer are represented, each reflection point of the lower surface of the two-dimensional plane waveguide core layer is a detection point, the detection area is further enlarged, the two-dimensional galvanometer G can control two coordinate systems for exciting the vertical coordinate of laser, the galvanometer G controls the exciting light 532 to carry out x and y fast scanning in two vertical directions, photoacoustic signals of multiple points in two vertical directions of a sample x and y are excited, each excitation can generate one photoacoustic signal, each photoacoustic signal can be received by the optical probe, and the photoacoustic signals of the multiple points in two directions of the sample x and y are obtained.
The signal processing module comprises a filter, an amplifier and an oscilloscope. The detected change of the intensity difference of the P polarized light and the S polarized light is converted into a voltage signal by a balance light detector in the laser detection module, an interference signal is filtered by a filter, the signal is amplified by an amplifier, and finally the signal is acquired by a high-bandwidth oscilloscope. The oscilloscope takes the photoelectric signals received by the photoelectric probe in the excitation module as triggers, the photoacoustic signals are displayed finally, each photoacoustic signal can be rapidly acquired by the oscilloscope, and then rapid imaging of the sample is realized.
Example three:
as shown in fig. 2, 3 and 7, the laser excitation module includes a pulse excitation light source 3, a beam expander 4, an adjustable attenuator 5, a beam splitter 6, a two-dimensional galvanometer G, an objective 8 and a photoelectric probe 7. A pulse laser beam is emitted from a pulse excitation light source 3, is collimated and amplified by a beam expander 4, is filtered by an adjustable attenuator 5, is split by a beam splitter 6, and one beam of light enters a photoelectric probe 7 to serve as a trigger signal of a subsequent signal processing module 10. The other beam of light 532 is reflected by the two-dimensional galvanometer G and focused on the sample 300 in the micro-flow control chip micro-channel through the objective lens 8 to excite the sample to emit ultrasonic waves.
The laser detection module comprises a continuous laser light source 1, a polaroid P, a 1/2 wave plate HWP, a 1/4 wave plate QWP, a reflecting mirror M and a phase type photoelectric detector 9. The continuous laser source emits a beam of continuous laser, the linear polarization ratio of the beam is improved through the linear polarizer P, and the ratio of the P polarization component to the S polarization component is adjusted through the 1/2 wave plate and the 1/4 wave plate. After passing through the reflector M, the probe light enters the planar waveguide from the side surface of the planar waveguide at a certain angle and undergoes total internal reflection for multiple times, and a probe point is formed at the reflection point of each lower surface. When ultrasonic waves exist in a micro-channel on the lower surface of the planar waveguide, the refractive index of liquid around the corresponding reflection point is changed, the phases of the P polarized light and the S polarized light in the reflected light of the corresponding point generate nonlinear changes with different responses, and the phase difference of the P polarized light and the S polarized light of other reflection points is kept unchanged. The reflected light of the detection light after repeatedly reflecting back and forth on the planar waveguide is reflected by the reflecting mirror M and enters the phase type photoelectric detector through the linear polarizer by taking an angle larger than a total reflection angle (arcsin (1.33/1.457)) as an incident angle.
Referring to fig. 2 and 3, the micro-fluidic chip based on the one-dimensional and two-dimensional planar waveguide structures can be used for large-area detection of samples, and the galvanometer can be used for fast scanning the samples, so that the samples can be fast scanned and subjected to photoacoustic detection.
The signal processing module comprises a filter, an amplifier and an oscilloscope. The phase-type photoelectric detector converts the detected change of the phase difference between the P polarized light and the S polarized light into a voltage signal, the interference signal is filtered by a filter, the signal is amplified by an amplifier, and finally the signal is acquired by a high-bandwidth oscilloscope. The oscilloscope takes the photoelectric signal received by the photoelectric probe in the excitation module as trigger, and finally displays the photoelectric signal as a photoacoustic signal, so that the rapid photoacoustic imaging of the sample is realized.
The invention carries out theoretical calculation and simulation on the condition of the one-dimensional waveguide, and concretely comprises the following steps:
taking the refractive index of the planar waveguide core layer as 1.457, the refractive index of the upper cladding water as 1.333, and the refractive index of the lower cladding water as 1.33301, calculating that the total reflection angle under the refractive index of water as arcsin (1.333/1.457), the probe light is incident on the planar waveguide at the angle of (pi/2-arcsin (1.333/1.457)) rad, calculating that the reflectivities of S-polarized light and P-polarized light under the refractive index of water as 1.33301 are Rs =0.9654, rp =0.9589, and calculating that the reflection is performed n times (Rs) after reflection is performed n -Rp n ) And (Rs) n +Rp n ) The ratio of (a) to (b). Then, we use a planar waveguide with a three-layer structure as a model, that is, the refractive index of the waveguide core layer is 1.457, the width of the waveguide core layer is 350um, the upper boundary is an impedance boundary, the refractive index is 1.333, which represents the upper cladding layer, the lower boundary is also an impedance boundary condition, the refractive index is 1.33301, which represents the influence of the photoacoustic effect on the refractive index of water, the width of the waveguide core layer is 350um, the length changes with the increase of the reflection times, and the incident light is a gaussian beam. Comsol software is used for simulating and calculating that the probe light enters the planar waveguide at an angle of (pi/2-arcsin (1.333/1.457)) rad, and after n reflections (Rs) in the waveguide n -Rp n ) And (Rs) n +Rp n ) The ratio of (R), theoretical and simulation results are shown in FIG. 6, which shows (Rs) n -Rp n ) And (Rs) n +Rp n ) The ratio of (A) to (B) being a function of the number of reflectionsThe increase is basically linear, and the theory is highly consistent with the simulation result, so that the method is proved to be feasible.
The working principle of the photoacoustic microfluidic detection system based on the planar waveguide is explained below.
Recording the incident light intensity I of the detection light, wherein the light intensities of the S polarized light and the P polarized light in the incident light are Is, ip and Is = Ip respectively, the reflectivities of the S polarized light and the P polarized light of each reflection point are Rs1, rs2, rs3
Namely, it is
Note book
Then there is
Namely, it is
Taking the logarithm of X, i.e.
When the reflectances Rs, rp of all the dots are the same, (3) becomes the formula
Then
When the reflectivity Rsx and Rpx of only one reflection point are changed and the reflectivity of other points is kept unchanged, the formula (3) is changed into
Then
I.e. a change in the reflectivity of each reflected spot of S-polarized light and P-polarized light causes a change in the reflected light Isn, ipn.
The photoacoustic microfluidic detection system based on the planar waveguide has the advantages that:
the invention has simple structure, low cost, high integration level and wide detection range, can realize large-area rapid scanning detection of biological or chemical samples, obtains photoacoustic signals of multiple points of the samples and further performs rapid imaging on the samples.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (6)
1. A photoacoustic microfluidic detection system based on planar waveguides, the system comprising: the device comprises a micro-fluidic chip module, a laser excitation module, a laser detection module block and a signal processing module; wherein,
the micro-fluidic chip module comprises a micro-channel and a planar waveguide arranged above the micro-channel;
the laser excitation module is used for exciting a target sample in the micro-channel below the planar waveguide to emit ultrasonic waves;
the laser detection module is used for receiving the photoacoustic signal generated after the target sample is excited and converting the photoacoustic signal into a voltage signal;
the signal processing module is used for receiving and processing the voltage signal of the laser detection module to obtain an optical signal of the target sample.
2. The planar waveguide-based photoacoustic microfluidic detection system according to claim 1, wherein the laser excitation module comprises a two-dimensional galvanometer for controlling the excitation light to perform fast scanning of the target sample in two perpendicular directions x and y.
3. The photoacoustic, microfluidic detection system based on planar waveguide of claim 1, wherein the planar waveguide is a one-dimensional planar waveguide or a two-dimensional planar waveguide.
4. The photoacoustic micro-fluidic detection system based on planar waveguide of claim 3, wherein the two-dimensional planar waveguide has mirror structures on both sides, the surfaces on both sides of the waveguide are plated with metal films, and the mirror surfaces are vertically arranged.
5. The photoacoustic micro-fluidic detection system based on planar waveguides of claim 3, wherein the upper cladding of the one-dimensional planar waveguide and the two-dimensional planar waveguide is the same as the liquid medium in the micro-channel of the micro-fluidic chip.
6. The photoacoustic and microfluidic detection system based on planar waveguides of claim 1, wherein the laser detection module incorporates a phase-type photoacoustic sensor for photoacoustic imaging by means of phase-type detection.
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| CN116499975A (en) * | 2023-06-29 | 2023-07-28 | 之江实验室 | Prism device for optical surface wave sensor and design and installation method thereof |
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| CN116499975A (en) * | 2023-06-29 | 2023-07-28 | 之江实验室 | Prism device for optical surface wave sensor and design and installation method thereof |
| CN116499975B (en) * | 2023-06-29 | 2023-09-22 | 之江实验室 | Prism device for optical surface wave sensor and design and installation method thereof |
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