CN111157730B

CN111157730B - Optical waveguide multi-micro-channel detection system

Info

Publication number: CN111157730B
Application number: CN202010052556.0A
Authority: CN
Inventors: 陈昌; 刘博�; 豆传国
Original assignee: Shanghai Jinguan Technology Co ltd
Current assignee: Shanghai Jinguan Technology Co ltd
Priority date: 2020-01-17
Filing date: 2020-01-17
Publication date: 2024-09-13
Anticipated expiration: 2040-01-17
Also published as: CN111157730A

Abstract

The invention provides an optical waveguide multi-micro-channel detection system, which comprises: a microfluidic chip; the microfluidic chip includes: a microfluidic group comprising microfluidics; the microfluid comprises an optical waveguide group and a micro-channel, wherein the optical waveguide group comprises an optical waveguide so as to guide light into the micro-channel along the horizontal direction; further comprises: the device comprises a substrate, a lower cladding layer, a waveguide layer, an upper cladding layer and a runner cover plate; the micro-channel penetrates through the upper cladding layer, the waveguide layer and the lower cladding layer from top to bottom and extends into the substrate; the lower cladding layer is silicon dioxide with the thickness of 2-3 mu m, the upper cladding layer is high polymer material with the thickness of 15-30 mu m, and the micro flow channel extends into the substrate with the thickness of 10-15 mu m and the width of 10-100 mu m. Has the beneficial effects that: the structure of the integrated matrix of the optical waveguide and the multiple micro-channels is formed, the analysis performance of higher flux than that of the traditional optical system is realized through the multiple micro-fluid channels and the large-scale matrix optical waveguide, the chip-level on-chip optical detection analysis system of the high-flux biological sample is quickly constructed, and the high-flux chip for biological detection under the micro-nano scale is realized.

Description

Optical waveguide multi-micro-channel detection system

Technical Field

The invention relates to an optical waveguide multi-micro-channel detection system, in particular to an optical waveguide multi-micro-channel biological detection system.

Background

In modern biochemical analytical procedures, high throughput detection devices have been widely used. Most of these devices employ a biochip based on microfluidic technology or microwell arrays, loaded in high performance optical systems, to enable analysis of biological samples of different sizes, such as nucleic acids, proteins, viruses, bacteria, cells, etc. The design of these optical systems is generally based on complex geometric optics, which are bulky, costly, require optical collimation, and are costly to maintain.

In the precise medical age, miniaturized, high performance, low cost, and mobile integrated analytical systems are receiving great attention. Especially, the concept of lab on chip has been developed for decades, and the control of biological samples based on microfluidic technology has been a great progress, but the real lab on chip system still lacks an integrated system for on-chip optical detection and analysis of high-throughput biological samples at micro-nano scale.

Disclosure of Invention

In order to solve the new demands of current modern biochemical analysis instrument such as huge volume, high cost, and miniaturization, portability and integration of the instrument required by the accurate medical era.

The invention provides an optical waveguide multi-micro-channel detection system, which comprises: microfluidic chip, microscope, measurement device and analysis device; it is characterized in that the method comprises the steps of,

The microfluidic chip includes a microfluidic group including a first number of microfluidics;

the micro-fluid comprises a light waveguide group and a micro-channel, the light waveguide group comprises a second number of light waveguides, the light waveguides are used for guiding light into the micro-channel along the horizontal direction, the microscope is used for collecting light signals in the micro-channel and transmitting the light signals to the measuring device, the measuring device is used for processing the light signals and generating signals to be analyzed and transmitting the signals to be analyzed to the analyzing device, and the analyzing device analyzes the signals to be analyzed to form a spectrum;

the microfluidic chip further comprises: the optical waveguide comprises a substrate, a lower cladding layer, a waveguide layer, an upper cladding layer and a runner cover plate which are sequentially arranged from bottom to top, wherein the waveguide layer is made of silicon nitride material and is used for forming the optical waveguide;

the micro-channel penetrates through the upper cladding layer, the waveguide layer and the lower cladding layer from top to bottom and extends into the substrate;

the flow channel cover plate covers the upper opening of the micro flow channel and comprises a liquid injection port for injecting a solution containing biomolecules to be detected into the micro flow channel;

The lower cladding layer is silicon dioxide with the thickness of 2-3 mu m, the upper cladding layer is a high polymer material with the thickness of 15-30 mu m, the micro flow channel extends into the substrate for 10-15 mu m, and the width of the micro flow channel is 10-100 mu m.

Preferably, the optical waveguide assembly further comprises a light guiding structure for providing a light source to the optical waveguide assembly.

Preferably, the optical waveguide group includes a second number of the optical waveguides parallel to each other to guide light into the micro flow channel, and the optical waveguides have a width of 300-600nm.

Preferably, the second number is 1, and the whole or most of the waveguide layers corresponding to the microfluid form one sheet-shaped optical waveguide.

Preferably, the waveguide layer has a thickness of 150-1000nm.

Preferably, the light guiding structure comprises a main light guiding and a light guiding group, the light guiding group is led out from the main light guiding, and the light guiding group is in optical connection with the light waveguide group.

Preferably, the light guide group is led out from the main light guide by adopting a light splitting structure.

Preferably, the trunk light guide comprises a first light guide, the light guide group comprises a second light guide, and the first light guide and the second light guide are intersected through an intersecting cross-layer structure.

Preferably, the cross-layer structure comprises a first light guide overlap region and a second light guide overlap region; the first light guide is broken at the intersection, and two acute-angle light guide end faces are formed at the two opposite ends of the broken first light guide; the second light guide forms an acute light guide surface matched with the acute light guide end surface at the intersection; the first light guide overlapping area comprises the acute light guide end face and an acute light guide surface matched with the acute light guide end face, and the second light guide overlapping area comprises the acute light guide end face and an acute light guide surface matched with the acute light guide end face.

Preferably, the optical waveguide is a coupled optical waveguide;

The coupling optical waveguide comprises an incidence grating to guide the light above the upper cladding into the coupling optical waveguide until the micro-channel is guided; the incident grating protrudes from the waveguide layer and extends upwards into the upper cladding layer, the waveguide layer thickness is 150-1000nm, and the width of the coupling optical waveguide is 300-600nm.

The invention provides an optical waveguide multi-micro-channel detection system, which has the beneficial effects that: the chip-level optical detection and analysis system is produced by an integrated circuit mass production process, the functions of the traditional optical system are realized by integrating optical or on-chip optical devices, the traditional desk-top or even large-scale optical system can be reduced to the chip size, the equal or better analysis performance is ensured, the high-flux chip-level optical detection and analysis integrated system of biological samples under the micro-nano scale is realized, and the system cost is greatly reduced. And a structure of an optical waveguide and multi-micro-channel integrated matrix is formed, the analysis performance of higher flux than that of the traditional optical system is realized through multi-micro-fluid channels and large-scale matrixed optical waveguides, a chip-level on-chip optical detection and analysis integrated system of the high-flux biological sample under the micro-nano scale can be quickly constructed, and the high-flux chip for detecting the biological sample under the micro-nano scale is realized.

Drawings

FIG. 1 is a side view of an optical waveguide multi-microchannel detection system of the present invention;

FIG. 2 is a side view of one microfluidic of the present invention;

FIG. 3 is a side view of another microfluidic according to the present invention;

FIG. 4 is a top view of FIG. 2 or FIG. 3;

FIG. 5 is a top view of the sheet-form optical waveguide microfluidics of FIG. 1;

FIG. 6 is a schematic diagram of a light guide structure;

FIG. 7 is an enlarged view of A in FIG. 6;

FIG. 8 is an enlarged view of B in FIG. 6;

Fig. 9 is a cross-sectional view of fig. 8.

Detailed Description

The following describes the embodiments of the present invention in detail with reference to the drawings.

In the drawings, dimensional proportions of layers and regions are not true proportions for convenience of description. When a layer (or film) is referred to as being "on" another layer or substrate, it can be directly on the other layer or substrate, or intervening layers may also be present. Furthermore, when a layer is referred to as being "under" another layer, it can be directly under, and one or more intervening layers may also be present. In addition, when a layer is referred to as being between two layers, it may be the only layer between the two layers, or one or more intervening layers may also be present. Like numbers refer to like elements throughout. In addition, when two components are referred to as being "connected," it is intended to include physical connection, unless the specification expressly defines otherwise, such physical connection includes, but is not limited to, electrical connection, contact connection, wireless signal connection.

The invention provides a horizontal optical waveguide and microfluidic channel integrated module scheme, and simultaneously provides a multi-microfluidic channel system matrixing scheme, so that a chip-level on-chip optical detection and analysis integrated system of a high-flux biological sample under a micro-nano scale is quickly constructed. Wherein the horizontal optical waveguide refers to an optical waveguide for guiding light into the micro flow channel along the horizontal direction

An optical waveguide multi-microchannel detection system, as shown in fig. 1, comprises: a microfluidic chip (not shown), a microscope 3, a measuring device 4 and an analyzing device 5;

the microfluidic chip comprises a microfluidic group comprising a first number of microfluidics (not shown), as shown in fig. 1, the first number being m;

The microfluid includes an optical waveguide group and a micro flow channel, and one microfluid as shown in fig. 2 includes the optical waveguide group 131 and the micro flow channel 201; the optical waveguide group 131, 132 … m includes a second number of optical waveguides, as shown in fig. 2 and 4, where the second number is n, and the optical waveguide group 131 includes n optical waveguides 1311, 1312 … n to form an n×m matrix detection system.

The optical waveguides 1311, 1312 … n are used for guiding light into the micro-flow channel 201 along the horizontal direction, the microscope 3 is used for collecting optical signals in the micro-flow channel 201, 202 and … m and transmitting the optical signals to the measuring device 4, the measuring device 4 is used for processing the optical signals and generating signals to be analyzed and transmitting the signals to be analyzed to the analyzing device 5, and the analyzing device 5 analyzes the signals to be analyzed to form a spectrum;

The microfluidic chip further comprises: the substrate 11, the lower cladding layer 12, the waveguide layer 13, the upper cladding layer 14 and the runner cover plate 15 are sequentially arranged from bottom to top, the waveguide layer 13 is made of silicon nitride material, and the waveguide layer 13 is used for forming the optical waveguide groups 131, 132 … m;

The micro flow channels 201, 202 … m extend from top to bottom through the upper cladding layer 14, the waveguide layer 13 and the lower cladding layer 12 into the substrate 11;

the runner cover plate 15 covers the upper openings of the micro runners 201, 202 … m, and the micro runner cover plate 15 comprises liquid injection ports 151, 152 and … m for injecting the solution containing the biomolecules to be detected into the micro runners 201, 202 … m; it should be noted that the device further includes a liquid outlet (not shown) to form a circulation system in one-to-one correspondence with the liquid injection ports 151, where the liquid outlet may be an opening on the runner cover 15; the liquid outlet may be an opening at two ends of the micro flow channel, which is not limited herein.

The lower cladding layer 12 is silicon dioxide with the thickness of 2-3 mu m, the upper cladding layer 14 is a high polymer material with the thickness of 15-30 mu m, the micro flow channels 201 and 202 … and 20m extend into the substrate 11 for 10-15 mu m, and the widths of the micro flow channels 201 and 202 … and 20m are 10-100 mu m; and a structure of an optical waveguide and multi-micro-channel integrated matrix is formed, and a chip-level on-chip optical detection and analysis integrated system of the high-flux biological sample under the micro-nano scale is rapidly constructed.

It should be noted that, the first number m of microfluid may form a microfluidic group, and a microfluidic matrix formed by a third number of microfluidic groups may be further constructed, where the third number is k, and the total number of optical waveguides may be n×m×k matrix detection systems; and a structure of an optical waveguide and multi-micro-channel integrated matrix is formed, and a chip-level on-chip optical detection and analysis integrated system of the high-flux biological sample under the micro-nano scale is rapidly constructed.

It should be noted that, the optical waveguide group includes a second number n of optical waveguides parallel to each other, as shown in fig. 1 and 4, the optical waveguide group 131 includes a second number n of optical waveguides 1311, 1312 … n parallel to each other to guide light into the micro flow channel 201 in a horizontal direction, and the width of the optical waveguides is 300-600nm.

Wherein the light source direction is different according to the waveguide group 131, such as: in fig. 2, the light guide group 131 is introduced with a light source from the left end, and fig. 3 is introduced with a light source from above the light guide group 131, and in a multi-micro flow channel, particularly a matrixing detection system, the former needs to be structurally added with a light guide structure as shown in fig. 6 when manufacturing a matrixing chip, and the latter does not need to be added with the light guide structure, and the light guide structure is described below with reference to fig. 1 to 2 and 4 to 9:

As shown in fig. 1 and 5, the optical waveguide group 131, 132 … m may include only one optical waveguide, and then the light guide group 601, 602 … m each include one light guide line to be optically connected with the optical waveguide group 131, 132 … m respectively; one of the optical waveguide layers 13 forms a sheet-like optical waveguide 1311, and the excitation light field introduced by the sheet-like optical waveguide 1311 can reduce the background light signal in the detection labeled biomolecules, thereby greatly improving the detection rate of small biomolecules.

As shown in fig. 1 and fig. 4, the optical waveguide group 131 includes a plurality, for example, n, of optical waveguides 1311, 1312 … n parallel to each other, and then the light guiding group 601 optically connected to the optical waveguides needs n corresponding light guiding lines to guide the light into the micro flow channel 201 along the horizontal direction; in actual detection, for biomolecules with different labels in the micro flow channel 201, the optical waveguides 1311, 1312 … n connected with n light guide lines one by one can respectively guide light with different wavelengths of λ1, λ2 … λn into the micro flow channel 201 along the horizontal direction, and the biomolecules can be identified simultaneously by exciting the labeled biomolecules 21 with the light with different wavelengths, while the non-excited biomolecules 20 in the excitation light field guided by the optical waveguides 1311, 1312 … 131n will not be identified, and the non-excited biomolecules 20 are normal biomolecules without labels or biomolecules with labels but outside the light field and without excitation; wherein, as shown in FIG. 3, the widths of the optical waveguides 1311, 1312 … n are 300-600nm.

As shown in fig. 1-2, the waveguide layer 13 has a thickness of 150-1000nm, i.e., the optical waveguides 1311, 1312 … n in fig. 2, 4-5 have a thickness of 150-1000nm.

The light guide group is optically connected to the light guide group 131, and is further optically connected to the light guides 1311, 1312 … n in the light guide group 131.

With the main light guide 60 providing the light guide group 602..60m to the second micro flow channel 202 … having n light guides up to the m-th micro flow channel 20m, the light guide lines cross, so for the multi-flow channel monitoring system matrixing integrated light guide multi-micro flow channel chip in fig. 1, a specific light guide structure (not shown) needs to be designed, as shown in fig. 6-9, for the detection system matrixing the light guides n x m, the light guide structure as shown in fig. 6 is provided, including the main light guide 60, and the light guide groups 601, 602 … m led out from the main light guide 60 are provided to transmit light sources to the micro flow channels 201, 202 … m respectively; the main light guide 60 includes n first light guides 61 that transmit light having wavelengths λ1, λ2, λ3 … λn, that is, each light guide group includes n second light guides 62 that transmit light to the light guides 1311, 1312 … n in the light guide group 131, respectively. Wherein, the leading-out node A of the light guiding groups 601, 602 … m leading out from the main light guiding 60 and the light guiding crossing node B in the leading-out light guiding and main light guiding 60 need to be specially designed; the light guide groups 601, 602 … m are vertically led out from the main light guide 60 by adopting a light splitting structure, as shown in fig. 7 to 8, and are light splitting structures of led-out nodes a, and only need to lead out the second light guide 62 in the light guide groups 601, 602 … m from the first light guide 61 in the main light guide 60; as shown in fig. 8, which is a cross-layer structure B of a cross node, the trunk light guide 60 includes a first light guide 61, the light guide group 601 includes a second light guide 62, and the first light guide 61 and the second light guide 62 cross through the cross-layer structure; the cross-layer structure includes a first light guide overlap region 610 and a second light guide overlap region 620; the first light guide 61 is broken at the intersection and forms two acute light guide end surfaces at two opposite ends of the broken line; the second light guide 62 forms an acute light guide surface matched with the acute light guide end surface at the intersection; the first light guide overlapping region 610 includes the acute light guide end surface and an acute light guide surface matched with the acute light guide end surface, and the distance between the two surfaces is less than 1 μm; the second light guide overlapping region 620 includes the acute light guide end surface and an acute light guide surface matched with the acute light guide end surface, and the distance between the two surfaces is less than 1 μm; that is, the first light guide 61 is broken at the intersection, an acute light guide end face is formed at each of opposite ends of the break, the second light guide 62 led out from the trunk light guide 60 is formed at the intersection with two acute light guide faces which are matched with the two acute light guide end faces and have a distance of less than 1 μm, so that a first light guide overlapping region 610 and a second light guide overlapping region 620 are formed, and light transmitted from one end of the break of the first light guide 61 enters the second light guide 62 through the first light guide overlapping region 610 and then enters the other end of the break of the first light guide 61 through the second light guide overlapping region 620.

It should be noted that, for the detection system in which the total number of optical waveguides is formed into n×m×k matrix, the light source may be continuously transmitted from the light guide group 601, 602 … m to the next microfluidic group up to the kth microfluidic group by using the light splitting structure a and the cross-layer structure B.

The optical waveguide multi-micro-channel detection system with the light source introduced from above the optical waveguide group 131 does not need special light guide structure design:

As shown in fig. 3, an incident grating (not shown) of silicon nitride material is further included to form a coupling optical waveguide with the optical waveguides 1311, 1312 … n, the light above the upper cladding layer 14 is guided into the optical waveguide until it is guided into the micro flow channel 201, and the upper cladding layer 14 is a light-transmissive layer; the incident grating protrudes from the waveguide layer 13 and extends up into the upper cladding layer 14.

As shown in fig. 3 and 4, a microfluidic device includes an optical waveguide group 131 including a plurality of, for example, n, coupling optical waveguides parallel to each other to guide light into the micro flow channel 201 in a horizontal direction, and in actual detection, for biomolecules containing different labels in the micro flow channel 201, the coupling optical waveguides can guide light with wavelengths λ1 and λ … λn into the micro flow channel 201 in a horizontal direction, respectively, and the biomolecules can be identified simultaneously by exciting the labeled biomolecules 21 of different labels with light of different wavelengths, and the non-excited biomolecules 20 which are not in an excitation field guided by the coupling optical waveguides will not be identified, and the non-excited biomolecules 20 are normal biomolecules which are not labeled or biomolecules which are labeled but are not excited outside the light field; wherein, as shown in fig. 4, the width of the coupling optical waveguide is 300-600nm, and wherein, as shown in fig. 3, the thickness of the waveguide layer 13 is 300-600nm.

In the present invention, the substrate 11 is a silicon substrate; preferably, the substrate 11 is a4, 8, 12 inch silicon wafer.

In the invention, the high molecular polymer material is SU-8 resin, polyimide, polydimethylsilane, polyethylene or styrene-acrylic.

In the present invention, the silicon nitride waveguide layer 13 is a silicon nitride thin film layer having a thickness of 150nm to 500nm formed at a low temperature of 25 to 150 ℃ of deposition temperature; the refractive index of the silicon nitride film is 1.75-2.2. The silicon nitride film may be a film having a uniform refractive index, or may be a film having a non-uniform refractive index, such as a silicon nitride film having a layered refractive index.

Circulating tumor cells are a collective term for various types of tumor cells that leave tumor tissue and enter the blood circulation system of the human body. By detecting trace circulating tumor cells in peripheral blood and monitoring the trend of the type and quantity change, the tumor dynamics can be monitored in real time, the treatment effect can be evaluated, and the real-time individual treatment can be realized. Referring to fig. 1, an embodiment of detecting circulating tumor cells using the above optical waveguide multi-microfluidic detection system in which the total number of optical waveguides is formed in an n×m×k matrix is described below, and the main steps are as follows:

The first step: sorting and enriching various tumor cells possibly existing in the collected m x k patient blood samples by adopting an immunomagnetic bead technology (such as immunomagnetic bead positive sorting) or a microfluidic technology to obtain a solution containing circulating tumor cells, and directly adopting the patient blood samples;

and a second step of: adding an antibody group capable of specifically binding to the surface antigens of various tumor cells or an aptamer group capable of binding to the surfaces of various tumor cells to the solution or blood sample containing the circulating tumor cells, wherein the antibody group and the aptamer group are modified with a label, and the modified label on the antibody or aptamer bound to the specific tumor cells is unique, so that the solution or blood sample containing the labeled circulating tumor cells is obtained; the label is n, and the label can be a target probe of a fluorescent molecule;

And a third step of: as shown in fig. 1, m×k portions of the solution or the blood sample obtained in the second step are added to the micro flow channels 201, 202, … m (incompletely listed, m×k portions of the total number of the liquid injection ports) and 202, … m (incompletely listed, m×k portions of the total number of the micro flow channels) from the liquid injection ports 151, 152, … m (incompletely listed, m×k portions of the total number of the liquid injection ports), respectively, the light guide groups 601, 602, … m (incompletely listed, m×k portions of the total number of the light guide groups) guide n light guides (n×35 m) (n×35 m portions of the total number of the light guide groups, m×k portions of the total number of the light guides are shown in fig. 1 and 4) of n light guides 1311, …, 131n of the total number of the light guide groups 131, n×k portions of the total number of the light guides are guided into the micro flow channels 201, 202, … m in the horizontal direction, the above-mentioned labeled biomolecules 21 containing different fluorescent molecular markers are circulating tumor cells excited by the light of the different wavelengths to emit fluorescence of a specific wavelength, the microscope 3 is used for collecting fluorescence of a specific wavelength (optical signal) and transmitting the fluorescence to the measurement device 4, the measurement device 4 processes and collects fluorescence of a specific wavelength (optical signal) and generates a signal to be analyzed and transmits the signal to be analyzed to the analysis device 5, the analysis device 5 analyzes the signal to be analyzed to form a spectrum of fluorescence of a specific wavelength, the types of circulating tumor cells in a solution or a blood sample can be judged by reading the spectrum, various tumor circulating cells of different patients can be detected at one time, a high-throughput chip for detecting various tumor cells under micro-nano scale is realized, thereby monitoring tumor dynamics and evaluating treatment effect in real time, realizing real-time individual treatment.

The invention provides an optical waveguide multi-micro-channel detection system, which has the beneficial effects that: the structure of the integrated matrix of the optical waveguide and the multi-micro-channel is formed, the analysis performance of higher flux than that of the traditional optical system is realized through the multi-micro-fluid channel and the large-scale matrix optical waveguide, a chip-level on-chip optical detection and analysis integrated system of the high flux biological sample under the micro-nano scale can be quickly constructed, and the high flux chip for detecting the biological sample under the micro-nano scale is realized.

The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims

1. An optical waveguide multi-microchannel detection system comprising: microfluidic chip, microscope, measurement device and analysis device; it is characterized in that the method comprises the steps of,

the lower cladding is silicon dioxide with the thickness of 2-3 mu m, the upper cladding is a high polymer material with the thickness of 15-30 mu m, the micro-channels extend into the substrate by 10-15 mu m, and the width of the micro-channels is 10-100 mu m;

The light guide structure is used for providing a light source for the light waveguide group;

The optical waveguide group comprises a second number of optical waveguides which are parallel to each other so as to guide light into the micro-channel, and the width of the optical waveguides is 300-600nm;

the light guide structure comprises a main light guide and a light guide group, wherein the light guide group is led out from the main light guide and is in optical connection with the light guide group;

The light guide group is led out from the main road light guide by adopting a light splitting structure;

the main light guide comprises a first light guide, the light guide group comprises a second light guide, and the first light guide and the second light guide are crossed through a cross-layer structure;

the cross-layer structure comprises a first light guide overlapping region and a second light guide overlapping region; the first light guide is broken at the intersection, and two acute-angle light guide end faces are formed at the two opposite ends of the broken first light guide; the second light guide forms an acute light guide surface matched with the acute light guide end surface at the intersection; the first light guide overlapping area comprises the acute light guide end face and an acute light guide surface matched with the acute light guide end face, and the second light guide overlapping area comprises the acute light guide end face and an acute light guide surface matched with the acute light guide end face.

2. The system of claim 1, wherein the waveguide layer is 150-1000nm thick.