CN206132579U - Measurement device for single -particle is restrainted scattering light intensity and is distributed - Google Patents

Measurement device for single -particle is restrainted scattering light intensity and is distributed Download PDF

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Publication number
CN206132579U
CN206132579U CN201621077977.4U CN201621077977U CN206132579U CN 206132579 U CN206132579 U CN 206132579U CN 201621077977 U CN201621077977 U CN 201621077977U CN 206132579 U CN206132579 U CN 206132579U
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light source
micro
fluidic chip
light
input duct
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丁驰竹
戴杰
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Huazhong Agricultural University
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Huazhong Agricultural University
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Abstract

The utility model discloses a measurement device for single -particle is restrainted scattering light intensity and is distributed, it includes light source, beam split light path, light -receiving and detection subassembly and micro -fluidic chip subassembly, the light source volume of including main survey light source, assistant measuring light source and system adjustment light source, the beam split light path includes spectroscope and PIN pipe, the light -receiving includes 90 off axis parabolic mirror, telescope mirror group, diaphragm, light filter, ICCD detector, signal detection and generating circuit, compound light filter, PMT detector, oscilloscope and computer with the detection subassembly, the micro -fluidic chip subassembly includes micro -fluidic chip, screen, triaxial regulation utensil and miniflow pump. Additionally, the utility model also discloses a methods of measurement that the single -particle is restrainted the scattering light intensity and is distributed.

Description

The measurement apparatus of single-particle beam scattered light intensity distribution
Technical field
The utility model is related to optics and fields of measurement, more particularly to a kind of measurement of single-particle beam scattered light intensity distribution Device.
Background technology
Counting micro particles and grain diameter measurement play an important role in clinical diagnosis, industry and environment measuring in liquid.Wherein, flow Formula cell art is the conventional Multi-parameter Measurement Method of Bio-clinical quick diagnosis and field of cell analysis.Testing sample suspension is in sheath By nozzle under the constraint of liquid, slender cytosol stream is formed, and by incident laser radiation.Photomultiplier receives dissipating for sample particulate Light or fluorescence signal are penetrated, process is analyzed to detection data by computer.Compared with population is integrally measured, flow cytometry More accurate result can be obtained.However, the sample size of flow cytometry needs is larger, instrument is complicated, operation and maintenance is inconvenient.
Traditional counting and measurement side are compared with the method for grain diameter measurement due to carrying out counting micro particles using micro-fluidic chip Method has very low sample consumption, can greatly shorten time of measuring, simplify operation and be easy to make portable equipment and be applied to The advantages of on-the-spot test, existing researcher is proposed based on the flow cytometry measure device and method of microfluidic chip technology. In measurement process, sample particulate is limited in by microchannel center flow by fluid, makes sample form simple grain subflow, so as to Avoid channel block, sample by conduit wall it is viscous glutinous or absorb, sample is overlapped the problems such as.
However, the existing flow cytometry measure device based on micro-fluidic chip is mostly flat in two dimension simply by sheath fluid Sample stream is limited on face, thus sample stream can not be made to become cylindrical fluid, and sample particulate easily deviates sample stream Central axis, have impact on measuring accuracy.Further, since photomultiplier is employed as light receiving element, so as to limit The measurement angle of scattered light, it is impossible to realize the measurement that scattered light intensity is distributed.
The content of the invention
Defect of the utility model for prior art, there is provided a kind of single-particle beam scattering based on micro-fluidic chip is surveyed Amount device, it can in real time measure scattered light intensity distribution of single particulate in the single-particle beam for flow through microchannel, and measuring speed it is fast, High precision.
For achieving the above object, the utility model provides a kind of measurement apparatus of single-particle beam scattered light intensity distribution, it Including light source, light splitting optical path, light-receiving and probe assembly and micro-fluidic chip component, the light source include main measurement light source, Subsidiary light source and system call interception light source；The light splitting optical path includes that spectroscope and PIN are managed；The light-receiving and probe assembly Including 90 ° of off-axis parabolic mirrors, telescope microscope group, diaphragm, optical filter, ICCD detectors, signal detection and occur circuit, Composite filter mating plate, PMT detectors, oscillograph and computer；The micro-fluidic chip component include micro-fluidic chip, optical screen, three Axle adjusts tool and miniflow pump；Wherein, the main measurement light source, the spectroscope, 90 ° of off-axis parabolic mirrors and described Three axles adjust tool and are successively set in same first straight line, and the laser of the main measurement light source transmitting is divided into master by the spectroscope Light path and reference path, the main optical path overlaps with the first straight line, and the reference path is vertical with the main optical path, described PIN pipe be located at the reference path on, the system call interception light source, the telescope microscope group, the diaphragm, the optical filter and The ICCD detectors are successively set in same second straight line, and the system call interception light source is anti-with 90 ° of off axis paraboloid mirrors The parabola for penetrating mirror is relative, and the optical screen is arranged on three axle and adjusts tool above and positioned at 90 ° of off-axis parabolic mirrors Focal point, the micro-fluidic chip is arranged on three axle and adjusts in tool, and the miniflow pump is connected with the micro-fluidic chip, The subsidiary light source is located at the left side of the micro-fluidic chip, and the composite filter mating plate and the PMT detectors set successively In the right side of the micro-fluidic chip, the PIN pipes, the oscillograph, the PMT detectors, the signal detection and generation Circuit, the ICCD detectors and the computer are sequentially connected.
Further, the micro-fluidic chip include annular sheath fluid input duct, linear sample liquid input duct and Linear sprue, the linear sample liquid input duct and the linear sprue are located on same 3rd straight line, institute State annular sheath fluid input duct symmetrical with regard to the 3rd straight line, one end of the annular sheath fluid input duct is provided with sheath fluid Input hole, the other end of the annular sheath fluid input duct is connected with the linear sprue, the sample liquid inlet flow Road is surrounded and connected with the sprue by the sheath fluid input duct, and the sample liquid input duct is provided with sample liquid input Hole, the sprue is provided with delivery outlet.
Further, the diameter of the sample liquid input duct and the sheath fluid input duct is respectively less than the sprue Diameter.
Further, the middle part of the sprue is the area of observation coverage of the micro-fluidic chip, the sight of the micro-fluidic chip Survey face is the face of cylinder, the face of cylinder be located in the area of observation coverage and axis and the sprue on the face of cylinder axis weight Close, the bottom surface of the micro-fluidic chip is plane.
Further, the main measurement light source and the subsidiary light source are laser instrument, the system call interception light source For parallel light tube.
In addition, the utility model additionally provides a kind of measuring method of single-particle beam scattered light intensity distribution, the method includes Following steps：
(1) system call interception light source, 90 ° of off-axis parabolic mirrors, optical screen and three axles are configured and adjusts tool, described 90 ° off-axis Parabolic mirror and three axle adjust tool and are located along the same line, the system call interception light source and described 90 ° off-axis parabolics The optical axis of face speculum is parallel, and the optical screen is arranged on three axle and adjusts tool above and positioned at 90 ° of off axis paraboloid mirrors reflection The focal point of mirror；
(2) PMT detectors and oscillograph are configured, the PMT detectors are located at the right side that three axle adjusts tool, will be described PMT detectors are connected with the oscillograph, and according to the oscillographic reading position and direction of the PMT detectors are adjusted, The PMT detectors are made to be directed at the focus of 90 ° of off-axis parabolic mirrors；
(3) optical screen is removed, is adjusted in three axle and micro-fluidic chip, the observation of the micro-fluidic chip are installed on tool Facing to 90 ° of off-axis parabolic mirrors, the area of observation coverage of the micro-fluidic chip and 90 ° of off-axis parabolic mirrors Optical axis in sustained height, the position that tool adjusts the X-axis, Y-axis and Z axis of the micro-fluidic chip, root are adjusted by three axle According to the oscillographic reading, the output signal for making the PMT detectors reaches maximum, described micro-fluidic after the completion of regulation Chip is located at the focal point of 90 ° of off-axis parabolic mirrors；
(4) remove the system call interception light source and configure main measurement light source and spectroscope, the spectroscope is located at the master Between measurement light source and 90 ° of off-axis parabolic mirrors, the spectroscope divides the laser of the main measurement light source transmitting For main optical path and reference path, the main optical path, the main measurement light source, 90 ° of off-axis parabolic mirrors and described three Axle adjusts tool and is located along the same line, and the reference path is vertical with the main optical path；
(5) position and direction of the main measurement light source are adjusted according to the oscillographic reading, the PMT detectors are made Output signal reach maximum, complete main measurement light source and 90 ° of off-axis parabolic mirrors and described micro-fluidic The alignment of chip is adjusted；
(6) PIN pipes are configured, the PIN pipes are located in spectroscopical reference path, while the PIN is managed It is connected with the oscillograph, with the light-intensity variation of the main laser of main measurement light source transmitting described in real-time monitoring；
(7) subsidiary light source is configured, the subsidiary light source is located at the left side that three axle adjusts tool, adjusts described The position and direction of subsidiary light source so as to which the auxiliary laser of transmitting is irradiated on the area of observation coverage of the micro-fluidic chip, and Point of irradiation is slightly above the point of irradiation of the main measurement light source, so that the PMT detectors are received by the micro-fluidic chip The auxiliary laser launched by the subsidiary light source of scattering；
(8) adjust in the PMT detectors and three axle and configure between tool composite filter mating plate, adjust the composite filter The position of piece and the height of the subsidiary light source, make the PMT detectors be dissipated by the microflow hole chip while receiving The main laser and the auxiliary laser of subsidiary light source transmitting by the main measurement light source transmitting penetrated；
(9) configure miniflow pump, the miniflow pump be connected with the micro-fluidic chip, sheath fluid pass through the miniflow pump via The sheath fluid input hole of the micro-fluidic chip pumps into the micro-fluidic chip, and sample liquid is by the miniflow pump via the miniflow The sample liquid input hole of control chip is pumped in the micro-fluidic chip, and the sheath fluid surrounds the sample liquid, and limits the sample The flowing of product liquid, so that the sample liquid becomes single-particle beam；
(10) when the sample liquid flows through the area of observation coverage of the micro-fluidic chip, according to two shown on the oscillograph The distance of the light hole of the time difference of individual adjacent peak and the composite filter mating plate calculates the flow velocity of the sample liquid；
(11) configure signal detection and circuit and ICCD detectors occur, successively visit the PMT detectors, the signal Circuit and ICCD detectors connection are surveyed and occur, the receiving plane of the ICCD detectors is anti-with 90 ° of off axis paraboloid mirrors The optical axis for penetrating mirror is vertical, and the PMT detectors send light intensity signal to the signal detection and circuit, the signal detection occur And there is circuit transmission detection trigger to the ICCD detectors, to start the ICCD detectors, and the signal Detection and generation circuit are from the reception light intensity signal to the time difference sent the detection trigger by the sample The flow velocity of liquid is determined；
(12) configure telescope microscope group, diaphragm and optical filter, the telescope microscope group, the diaphragm, the optical filter and The ICCD detectors are sequentially located on same straight line, the light of the telescope microscope group and 90 ° of off-axis parabolic mirrors Axle is parallel and towards 90 ° of off-axis parabolic mirrors, and the ICCD detectors are connected with computer, the ICCD detections Device obtains the scattering pattern of sample particulate in the sample liquid, and the scattering pattern of the sample particulate is sent to the calculating Machine；
(13) trigger is manually fed to start the ICCD detectors, so as to obtain background patterns and by the background Pattern is sent to the computer；
(14) computer deducts the intensity of the background patterns with the intensity of the scattering pattern of the sample particulate, obtains Scattered light intensity to single-particle beam is distributed.
Further, the micro-fluidic chip include annular sheath fluid input duct, linear sample liquid input duct and Linear sprue, the linear sample liquid input duct and the linear sprue are located along the same line, the circle Annular sheath fluid input duct is symmetrical with regard to the straight line, and one end of the annular sheath fluid input duct is provided with sheath fluid input hole, The other end of the annular sheath fluid input duct is connected with the linear sprue, and the sample liquid input duct is described Sheath fluid input duct is surrounded and connected with the sprue, and the sample liquid input duct is provided with sample liquid input hole, described Sprue is provided with delivery outlet.
Further, the middle part of the sprue is the area of observation coverage of the micro-fluidic chip, the sight of the micro-fluidic chip Survey face is the face of cylinder, the face of cylinder be located in the area of observation coverage and axis and the sprue on the face of cylinder axis weight Close, the bottom surface of the micro-fluidic chip is plane.
Further, the sheath fluid by silicone oil and paraffin group into the refractive index of the sheath fluid is equal to the micro-fluidic core The refractive index of piece, the sample liquid adds deionized water dilution to form by particle samples solution to be measured, and dilution volume ratio is 1:1000 ～1:10000, the sheath fluid is immiscible with the sample liquid.
Further, the preparation method of the micro-fluidic chip is comprised the following steps：
A () emulates to the structure of the runner of the micro-fluidic chip, to determine the size of the runner；
B () is coated in the first negative optical cement in first substrate with silicon single crystal flake as the first substrate, negative to described first Optical cement and first substrate make the plane template of the observation layer of the micro-fluidic chip by Twi-lithography technique；
C () makes the semi-cylindrical template of the observation layer of acrylic material, with the plane template of the observation layer and Semi-cylindrical template carries out reverse mould to the first dimethyl silicone polymer, and carries out baking and solidification and remove the flat of the observation layer Face die plate and semi-cylindrical template, obtain the observation layer of the micro-fluidic chip；
D () is coated in the second negative optical cement in second substrate with silicon single crystal flake as the second substrate, negative to described second Optical cement and second substrate make the template of the bottom of the micro-fluidic chip by Twi-lithography technique；
E () carries out reverse mould with the template of the bottom to the second dimethyl silicone polymer, and carry out baking and solidification and go Except the template of the bottom, the bottom of the micro-fluidic chip is obtained；
F () carries out ozone process and sealing in the presence of ultraviolet to the bottom and the observation layer, obtain complete The micro-fluidic chip.
The beneficial effects of the utility model are：Micro-fluidic chip of the present utility model is focused on by three dimensional fluid, makes sample Product stream becomes cylindrical fluid, realizes the structure of single-particle beam environment and is accurately positioned；The inspection surface of micro-fluidic chip is in circle Cylindricality, reduces light in chip-impact of the Air Interface refraction to measurement result.Further, since the utility model employ by 90 ° of off-axis parabolic mirrors, telescope microscope group, diaphragm and filter set into light-receiving component, thus include on a large scale Scattered light measurement angle；The utility model employ subsidiary light source, composite filter mating plate, PMT detectors, oscillograph so as to Realize real-time, the accurate measurement of the scattered light intensity distribution of the single particulate for flowing through micro-fluidic chip.
Description of the drawings
Fig. 1 is the schematic top plan view of the measurement apparatus of the utility model single-particle beam scattered light intensity distribution in measuring phases.
Fig. 2 is the schematic top plan view of the measurement apparatus of the utility model particle beams scattered light intensity distribution in the adjusting stage.
Fig. 3 is composite filter mating plate of the present utility model, PMT detectors and oscillographic connection diagram in measuring phases.
Fig. 4 is the structural representation of an angle of micro-fluidic chip of the present utility model.
Fig. 5 is the structural representation of another angle of micro-fluidic chip of the present utility model.
Fig. 6 is the flow chart of the measuring method of single-particle beam scattered light intensity distribution of the present utility model.
Fig. 7 is the flow chart of the preparation method of micro-fluidic chip of the present utility model.
Specific embodiment
In order to preferably explain the utility model, further elucidate below in conjunction with specific embodiment of the present utility model main Content, but content of the present utility model is not limited solely to following examples.
With reference to Fig. 1-3, the measurement apparatus of the single-particle beam scattered light intensity distribution of the present embodiment include：Including light source, light splitting Light path, light-receiving and probe assembly and micro-fluidic chip component.
Specifically, the light source includes main measurement light source 10, subsidiary light source 11 and system call interception light source 12.Described point Light light path includes spectroscope 20 and PIN pipes 21.The light-receiving and probe assembly include 90 ° of off-axis parabolic mirrors 30, hope Remote mirror microscope group 31, diaphragm 32, optical filter 33, ICCD detectors 34, signal detection and generation circuit 35, composite filter mating plate 36, PMT Detector 37, oscillograph 38 and computer 39.The micro-fluidic chip component includes that micro-fluidic chip 40, optical screen 41, three axles are adjusted Section tool 42 and miniflow pump 43.
Wherein, the main measurement light source 10, the spectroscope 20,90 ° of off-axis parabolic mirrors 30 and described three Axle adjusts tool 42 and is successively set in same first straight line, the laser point that the spectroscope 20 launches the main measurement light source 10 For main optical path and reference path, the main optical path overlaps with the first straight line, and the reference path is vertical with the main optical path, The PIN pipes 21 be located at the reference path on, the system call interception light source 12, the telescope microscope group 31, the diaphragm 32, The optical filter 33 and the ICCD detectors 34 are successively set in same second straight line, the system call interception light source 12 and institute The parabola for stating 90 ° of off-axis parabolic mirrors 30 is relative, and the optical screen (not shown) is arranged on three axle and adjusts on tool 42 And positioned at the focal point of 90 ° of off-axis parabolic mirrors 30, the micro-fluidic chip 40 is arranged on three axle and adjusts tool In 42, the miniflow pump 43 is connected with the micro-fluidic chip 40, and the subsidiary light source 11 is located at the micro-fluidic chip 40 left side, the composite filter mating plate 36 and the PMT detectors 37 are sequentially arranged in the right side of the micro-fluidic chip 40, described PIN pipes 21, the oscillograph 38, the PMT detectors 37, the signal detection and generation circuit 35, the ICCD detectors 34 and the computer 39 be sequentially connected.
Further, the main measurement light source 10 and the subsidiary light source 11 are laser instrument, the system call interception Light source 12 is parallel light tube.The main measurement light source 10 is different with the wavelength of the subsidiary light source 11.The main measurement light Source 10 is provided with main measurement light source and adjusts tool 10a, and the system call interception light source 12 is provided with system call interception light source and adjusts tool 12a, institute State PIN pipes 21 and be provided with PIN pipes and adjust and have 21a, the composite filter mating plate 36 is provided with composite filter mating plate and adjusts tool 36a, and the PMT is visited Survey device 37 and be provided with PMT detectors and adjust and have 37a.
In detail, with reference to Fig. 4-5, the micro-fluidic chip 40 includes annular sheath fluid input duct 401, linear sample Liquid input duct 402 and linear sprue 403, the linear sample liquid input duct 402 and the linear sprue 403 are located along the same line, and the annular sheath fluid input duct 401 is symmetrical with regard to the straight line, and the annular sheath fluid is defeated One end of air stream enter runner 401 is provided with sheath fluid input hole (not shown), the other end of the annular sheath fluid input duct 401 with it is described Linear sprue 403 is connected, the sample liquid input duct 402 by the sheath fluid input duct 401 surround and with the master Runner 403 is connected, and the sample liquid input duct 402 is provided with sample liquid input hole (not shown), sets on the sprue 403 There is delivery outlet (not shown).In a preferred embodiment, the diameter of the sample liquid input duct 402 and the sheath fluid inlet flow The diameter of road 401 is respectively less than the diameter of the sprue 403.Wherein, the sheath fluid by silicone oil with paraffin group into the sheath fluid Refractive index is equal to the refractive index of the micro-fluidic chip 40, and the sample liquid adds deionized water to dilute by particle samples solution to be measured Form, dilution volume ratio is 1:1000～1:10000, the sheath fluid is immiscible with the sample liquid.
Further, the middle part of the sprue 403 is the area of observation coverage C of the micro-fluidic chip 40, the micro-fluidic core The inspection surface of piece is the face of cylinder, and the face of cylinder is located in the area of observation coverage C, the axis on the face of cylinder and the sprue 403 dead in line, the bottom surface of the micro-fluidic chip 40 is plane.
With reference to Fig. 6-7, the measuring method of the single-particle beam scattered light intensity distribution of the present embodiment includes：
Step S1：Configuration 12,90 ° of off-axis parabolic mirrors 30, optical screen (not shown) of system call interception light source and three axles are adjusted Section tool 42,90 ° of off-axis parabolic mirrors 30 and three axle adjust tool 42 and are located along the same line, the system call interception Light source 12 is parallel with the optical axis of 90 ° of off-axis parabolic mirrors 30, and the optical screen is arranged on three axle and adjusts on tool 42 And positioned at the focal point of 90 ° of off-axis parabolic mirrors 30；
Step S2：Configuration PMT detectors 37 and oscillograph 38, the PMT detectors 37 are located at three axle and adjust tool 42 Right side, the PMT detectors 37 are connected with the oscillograph 38, the PMT is adjusted according to the reading of the oscillograph 38 The position and direction of detector 37, makes the PMT detectors 37 be directed at the focus of 90 ° of off-axis parabolic mirrors 30；
Step S3：The optical screen is removed, is adjusted in three axle and install on tool 42 micro-fluidic chip 40, the micro-fluidic core The inspection surface of piece 40 towards 90 ° of off-axis parabolic mirrors 30, the area of observation coverage C of the micro-fluidic chip 40 with described 90 ° The optical axis of off-axis parabolic mirror 30 adjusts tool 42 and adjusts the micro-fluidic chip 40 in sustained height by three axle The position of X-axis, Y-axis and Z axis, according to the reading of the oscillograph 38, the output signal for making the PMT detectors 37 reaches greatly Value, after the completion of regulation, the micro-fluidic chip 40 is located at the focal point of 90 ° of off-axis parabolic mirrors 30；
Step S4：Remove the system call interception light source 12 and configure main measurement light source 10 and spectroscope 20, the spectroscope 20 are located between the main measurement light source 10 and 90 ° of off-axis parabolic mirrors 30, and the spectroscope 20 is by the main survey The laser of the amount transmitting of light source 10 is divided into main optical path and reference path, the main optical path, the main measurement light source 10, described 90 ° from Axle parabolic mirror 30 and three axle adjust tool 42 and are located along the same line, and the reference path is hung down with the main optical path Directly；
Step S5：The position and direction of the main measurement light source 10 are adjusted according to the reading of the oscillograph 38, is made described The output signal of PMT detectors 37 reaches maximum, completes the main measurement light source 10 and 90 ° of off-axis parabolic mirrors 30 and the micro-fluidic chip 40 alignment adjust；
Step S6：Configuration PIN pipes 21, the PIN pipes 21 are located in the reference path of the spectroscope 20, while will The PIN pipes 21 are connected with the oscillograph 38, with the light high-amplitude wave of the main laser of main measurement light source 10 transmitting described in real-time monitoring It is dynamic；
Step S7：Configuration subsidiary light source 11, the subsidiary light source 11 is located at the left side that three axle adjusts tool 42 Side, adjusts the position and direction of the subsidiary light source 11 so as to which the auxiliary laser of transmitting is irradiated to the micro-fluidic chip On 40 area of observation coverage C, and point of irradiation is slightly above the point of irradiation of the main measurement light source 10, so that the PMT detectors 37 connect Receive the auxiliary laser launched by the subsidiary light source 11 scattered by the micro-fluidic chip 40；
Step S8：Adjust in the PMT detectors 37 and three axle and configure between tool 42 composite filter mating plate 36, adjust institute State the position of composite filter mating plate 36 and the height of the subsidiary light source 11, make the PMT detectors 37 and meanwhile receive by The main laser launched by the main measurement light source 10 of the scattering of the microflow hole chip 40 and the subsidiary light source 11 The auxiliary laser of transmitting；
Step S9：Configuration miniflow pump 43, the miniflow pump 43 is connected with the micro-fluidic chip 40, and sheath fluid is by described Miniflow pump 43 pumps into the micro-fluidic chip 40 via the sheath fluid input hole of the micro-fluidic chip 40, and sample liquid is by described micro- Stream pump 43 is pumped in the micro-fluidic chip 40 via the sample liquid input hole of the micro-fluidic chip 40, and the sheath fluid surrounds institute State sample liquid, and limit the flowing of the sample liquid, so that the sample liquid becomes single-particle beam；
Step S10：When the sample liquid flows through the area of observation coverage of the micro-fluidic chip 40, according on the oscillograph 38 The distance of the light hole of the time difference and the composite filter mating plate 36 of two adjacent peaks for showing calculates the sample liquid Flow velocity；
Step S11：Configuration signal detection and there is circuit 35 and ICCD detectors 34, successively by the PMT detectors 37, The signal detection and there is circuit 35 and the ICCD detectors 34 and connect, the receiving plane of the ICCD detectors 34 with it is described The optical axis of 90 ° of off-axis parabolic mirrors 30 is vertical, and the PMT detectors 37 send light intensity signal to the signal detection and send out Raw circuit 35, the signal detection and generation circuit 35 send detection trigger to the ICCD detectors 34, to start The ICCD detectors 34, the signal detection and generation circuit 35 are triggered from the light intensity signal is received to the transmission detection Time difference between signal is determined by the flow velocity of the sample liquid；
Step S12：Configuration telescope microscope group 31, diaphragm 32 and optical filter 33, the telescope microscope group 31, the diaphragm 32nd, the optical filter 33 and the ICCD detectors 34 are sequentially located on same straight line, the telescope microscope group 31 with described 90 ° The optical axis of off-axis parabolic mirror 30 is parallel and towards 90 ° of off-axis parabolic mirrors 30, by the ICCD detectors 34 are connected with computer 39, and the ICCD detectors 34 obtain the scattering pattern of sample particulate in the sample liquid, and will be described The scattering pattern of sample particulate is sent to the computer 39；
Step S13：Trigger is manually fed to start the ICCD detectors 34, so as to obtain background patterns and by institute State background patterns to send to the computer 39；
Step S14：The computer 39 deducts the background patterns with the intensity of the scattering pattern of the sample particulate Intensity, obtains the scattered light intensity distribution of single-particle beam.
Specifically, with reference to Fig. 7, the preparation method of the micro-fluidic chip 40 is comprised the following steps：
A () emulates to the structure of the runner of the micro-fluidic chip 40, to determine the size of the runner；
B () is coated in the first negative optical cement 51 in first substrate 51, to described with silicon single crystal flake as the first substrate 50 First negative optical cement 51 and first substrate 50 make the observation layer 53 of the micro-fluidic chip 40 by Twi-lithography technique Plane template；
C () makes the semi-cylindrical template of the observation layer 53 of acrylic material, with the plane mould of the observation layer 53 Plate and semi-cylindrical template carry out reverse mould to the first dimethyl silicone polymer 52, and carry out baking and solidification and remove the observation The plane template and semi-cylindrical template of layer 53, obtains the observation layer 53 of the micro-fluidic chip 40；
D () is coated in the second negative optical cement 61 in second substrate 60, to described with silicon single crystal flake as the second substrate 60 Second negative optical cement 61 and second substrate 60 make the mould of the bottom 63 of the micro-fluidic chip 40 by Twi-lithography technique Plate；
E () carries out reverse mould with the template of the bottom 63 to the second dimethyl silicone polymer 62, and carry out baking and solidification And the template of the removal bottom 63, obtain the bottom 63 of the micro-fluidic chip 40；
F () carries out ozone process and sealing in the presence of ultraviolet to the observation layer 53 and the bottom 63, obtain The complete micro-fluidic chip 40.
Other unspecified parts are prior art.Although above-described embodiment is made that detailed to the utility model Description, but it is only a part of embodiment of the utility model, rather than whole embodiments, and people can be with according to this enforcement Example obtains other embodiment under the premise of without creativeness, and these embodiments belong to the utility model protection domain.

Claims (5)

1. the measurement apparatus that a kind of single-particle beam scattered light intensity is distributed, it includes light source, light splitting optical path, light-receiving and probe assembly And micro-fluidic chip component, it is characterised in that：
The light source includes main measurement light source (10), subsidiary light source (11) and system call interception light source (12)；
The light splitting optical path includes spectroscope (20) and PIN pipes (21)；
The light-receiving and probe assembly include 90 ° of off-axis parabolic mirrors (30), telescope microscope group (31), diaphragm (32), Optical filter (33), ICCD detectors (34), signal detection and generation circuit (35), composite filter mating plate (36), PMT detectors (37), oscillograph (38) and computer (39)；
The micro-fluidic chip component includes that micro-fluidic chip (40), optical screen, three axles adjust tool (42) and miniflow pump (43)；
Wherein, main measurement light source (10), the spectroscope (20), 90 ° of off-axis parabolic mirrors (30) and described Three axles adjust tool (42) and are successively set in same first straight line, and the spectroscope (20) launches main measurement light source (10) Laser be divided into main optical path and reference path, the main optical path overlaps with the first straight line, the reference path and the master Light path is vertical, and the PIN manages (21) in the reference path, the system call interception light source (12), the telescope microscope group (31), the diaphragm (32), the optical filter (33) and the ICCD detectors (34) are successively set in same second straight line, The system call interception light source (12) is relative with the parabola of 90 ° of off-axis parabolic mirrors (30), and the optical screen is arranged on Three axle is adjusted on tool (42) and positioned at the focal point of 90 ° of off-axis parabolic mirrors (30), the micro-fluidic chip (40) it is arranged on three axle to adjust in tool (42), the miniflow pump (43) is connected with the micro-fluidic chip (40), described auxiliary Measurement light source (11) is helped positioned at the left side of the micro-fluidic chip (40), the composite filter mating plate (36) and the PMT detectors (37) right side of the micro-fluidic chip (40), PIN pipes (21), the oscillograph (38), PMT detections are sequentially arranged in Device (37), the signal detection and generation circuit (35), the ICCD detectors (34) and the computer (39) are sequentially connected.
2. the measurement apparatus that single-particle beam scattered light intensity as claimed in claim 1 is distributed, it is characterised in that the micro-fluidic core Piece (40) includes annular sheath fluid input duct (401), linear sample liquid input duct (402) and linear sprue (403), the linear sample liquid input duct (402) and the linear sprue (403) be on same 3rd straight line, The annular sheath fluid input duct (401) is symmetrical with regard to the 3rd straight line, the annular sheath fluid input duct (401) One end is provided with sheath fluid input hole, the other end of the annular sheath fluid input duct (401) and the linear sprue (403) Connection, the sample liquid input duct (402) is surrounded and connected with the sprue (403) by the sheath fluid input duct (401) Logical, the sample liquid input duct (402) is provided with sample liquid input hole, and the sprue (403) is provided with delivery outlet.
3. the measurement apparatus that single-particle beam scattered light intensity as claimed in claim 2 is distributed, it is characterised in that the sample liquid is defeated The diameter of air stream enter runner (402) and the sheath fluid input duct (401) is respectively less than the diameter of the sprue (403).
4. the measurement apparatus that single-particle beam scattered light intensity as claimed in claim 2 is distributed, it is characterised in that the sprue (403) middle part is the area of observation coverage (C) of the micro-fluidic chip (40), and the inspection surface of the micro-fluidic chip (40) is cylinder Face, the face of cylinder be located in the area of observation coverage (C) and axis and the sprue (403) on the face of cylinder axis weight Close, the bottom surface of the micro-fluidic chip (40) is plane.
5. the measurement apparatus that single-particle beam scattered light intensity as claimed in claim 1 is distributed, it is characterised in that the main measurement light Source (10) and the subsidiary light source (11) are laser instrument, and the system call interception light source (12) is parallel light tube.
CN201621077977.4U 2016-09-22 2016-09-22 Measurement device for single -particle is restrainted scattering light intensity and is distributed Active CN206132579U (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106442278A (en) * 2016-09-22 2017-02-22 华中农业大学 Measurement device and measurement method for scattered light intensity distribution of single particle beam
CN108956391A (en) * 2018-06-12 2018-12-07 西安理工大学 The survey meter and detection method of the gentle aerosol particle size Spectral structure of droplet in atmospheric sounding

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106442278A (en) * 2016-09-22 2017-02-22 华中农业大学 Measurement device and measurement method for scattered light intensity distribution of single particle beam
CN108956391A (en) * 2018-06-12 2018-12-07 西安理工大学 The survey meter and detection method of the gentle aerosol particle size Spectral structure of droplet in atmospheric sounding
CN108956391B (en) * 2018-06-12 2021-02-12 西安理工大学 Detector and detection method for detecting particle size spectrum distribution of fog drops and aerosol in atmosphere

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