Integrated sensing device, preparation method and application thereof

Abstract

The invention provides an integrated sensing device and a preparation method thereof, two optical elements including a thicker glass-based medium reflecting mirror with a liquid sample inlet and outlet or extremely thin optical glass are integrally fused with a soft polymer interlayer pre-engraved with a flow channel, and the integrated sensing device is formed. Finally, a stable optical microcavity through which biological specimens such as liquid samples and cells can pass is generated, and the optical microcavity can be used as a practical carrier for single-cell laser emission analysis, single-cell fluorescence spectrum analysis and molecular fluorescence lifetime research, and is wide in application.

Description

Technical Field

[0001] The present application relates to the technical field of bio-optical devices, and in particular to an integrated sensing device, a device, a preparation method and applications thereof. Background Art

[0002] In bioanalysis and cell biology research, analysis of minute amounts of biochemical samples is often necessary. Examples include fluorescent probes, free samples of nucleic acids and proteins dissolved in aqueous or oily buffers, and micro- and nanoscale biological samples in a largely solid state, such as cells. These studies often require supporting microfluidic sample processing devices and biosensor devices that amplify and modulate the information carried by the sample, transforming the microscopic world into perceptible signals and enabling sensitive optical analysis. Optical sensors convert the interaction between the analyte and the sensitive element into an optical signal for detection. These sensors are resistant to external electromagnetic interference, enabling long-range detection. Furthermore, because they can transmit multiple signals using a single pathway, they can detect a large amount of information.

[0003] Existing tests of this type often involve directly dropping dye onto a glass slide (with an open top surface), making it difficult to effectively control the depth and thickness of the sample on the slide. Other teams have also used closed PDMS channels to achieve similar goals, but PDMS polymers exhibit strong autofluorescence, which can easily induce background fluorescence noise.

[0004] Existing parallelism control methods for this type of testing often employ a method that sandwiches a small plastic bead (10-30 μL) between two reflective mirrors. However, this method suffers from poor stability between the mirrors. Furthermore, the quality factor of the region near the plastic bead and that outside the cavity within the cavity differ significantly, hindering the precise quantitative analysis of cells required in practical research. Summary of the Invention

[0005] The embodiments of the present application provide an integrated sensing device that can avoid the lack of stability and repeatability of existing devices.

[0006] In a first aspect, an embodiment of the present application provides an integrated sensing device, comprising a first reflector, a microchannel cavity unit, and a second reflector arranged in sequence from top to bottom, wherein the reflective surfaces of the first reflector and the second reflector are coated with a high-reflectivity coating.

[0007] In some embodiments, the first reflector and the second reflector are made of quartz or optical glass.

[0008] In some embodiments, the high reflectivity coating is a dielectric coating, and the thickness of the high reflectivity coating is 10-1000 nm.

[0009] In some embodiments, the thickness of the first reflector and the second reflector is 0.5-3 mm.

[0010] In some embodiments, the microfluidic channel cavity unit is a microfluidic channel film, and the microfluidic channel film includes PDMS silica gel or SU8 photoresist or photocurable resin.

[0011] In some embodiments, the thickness of the microfluidic cavity unit is 10-200 μm.

[0012] In some embodiments, the microfluidic channel film is provided with an inlet area, a detection area and an outlet area. The inlet area is used for adding liquid, the detection area is used for detecting samples, and the outlet area is used for sample outflow.

[0013] In some embodiments, the sample includes a dissolved aqueous phase, a free sample in an oil phase buffer, or a solid phase micro-nanoscale biological sample, the dissolved aqueous phase includes a fluorescent probe or nucleic acid or protein, and the solid phase micro-nanoscale biological sample includes a virus or a cell.

[0014] In a second aspect, an embodiment of the present application provides an integrated sensing device for fluorescence signal testing, comprising a first optical glass, a microchannel cavity unit, and a second optical glass arranged in sequence from top to bottom.

[0015] In some embodiments, the thickness of the first optical glass and the second optical glass is 50-300 μm.

[0016] In some embodiments, the microfluidic channel cavity unit is a microfluidic channel film, and the microfluidic channel film includes PDMS silica gel or SU8 photoresist or photocurable resin.

[0017] In some embodiments, the thickness of the microfluidic cavity unit is 10-200 μm.

[0018] In some embodiments, the microfluidic channel film is provided with an inlet area, a detection area and an outlet area. The inlet area is used for adding liquid, the detection area is used for detecting samples, and the outlet area is used for sample outflow.

[0019] In some embodiments, the sample includes a dissolved aqueous phase, a free sample in an oil phase buffer, or a solid phase micro-nanoscale biological sample, the dissolved aqueous phase includes a fluorescent probe or nucleic acid or protein, and the solid phase micro-nanoscale biological sample includes a virus or a cell.

[0020] In a third aspect, an embodiment of the present application provides a method for preparing an integrated sensor device, comprising the following steps:

[0021] Assembling the first reflector, the second reflector and the microfluidic cavity unit by plasma bonding or double-sided tape; or

[0022] The first optical glass, the microchannel cavity unit and the second optical glass are assembled by plasma bonding or double-sided tape bonding.

[0023] In some embodiments, the microfluidic cavity unit can be processed by secondary machining of a commercial PDMS film, wherein the secondary machining includes photolithography, 3D printing, silicon etching, wet etching, and knife etching.

[0024] In a fourth aspect, embodiments of the present application provide applications of an integrated sensing device in intracavity laser emission biomolecule analysis.

[0025] In a fifth aspect, embodiments of the present application provide applications of an integrated sensing device in fluorescence lifetime analysis of intracavitary biomolecules.

[0026] The technical solution provided in this application integrates two optical elements, including a thicker glass-based dielectric reflector or extremely thin optical glass, with a soft polymer interlayer pre-engraved with flow channels, ultimately producing a stable optical microcavity through which liquids, cells, viruses and other specimens can pass. This can be a practical vehicle for single-cell laser emission analysis and single-cell fluorescence lifetime research, and has a wide range of applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0028] Figure 1 Schematic diagram of the structure of an integrated sensing device for analyzing intracavity laser emission signals provided in an embodiment of the present application;

[0029] Figure 2 Schematic diagram of the structure of the microfluidic cavity unit provided in the embodiment of the present application;

[0030] Figure 3 Schematic diagram of the structure of an integrated sensing device for fluorescence signal testing provided in an embodiment of the present application;

[0031] Figure 4 This is a schematic diagram of an integrated sensing device provided in an embodiment of the present application for analyzing intracavity laser emission signals. DETAILED DESCRIPTION

[0032] In order to help those skilled in the art better understand the technical solutions of this application, the following is a clear and complete description of the technical solutions in the embodiments of this application in conjunction with the drawings in the embodiments of this application. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the description of the embodiments of this application, all other embodiments obtained by those skilled in the art without making creative efforts are within the scope of protection of this application.

[0033] The terms "first," "second," and so on in the specification and claims of this application and the accompanying drawings are used to distinguish between different objects, not to describe a specific order. In addition, the terms "including," "having," and any variations thereof are intended to cover non-exclusive inclusions. For example, a process, method, software, product, or apparatus comprising a series of steps or elements is not limited to the listed steps or elements but may also include steps or elements not listed, or may include other steps or elements inherent to the process, method, product, or apparatus.

[0034] References herein to "embodiments" mean that a particular feature, structure, or characteristic described in connection with the embodiments may be included in at least one embodiment of the present application. The appearance of this phrase in various places in the specification does not necessarily refer to the same embodiment, nor does it constitute an independent or alternative embodiment that is mutually exclusive of other embodiments. It is understood, both explicitly and implicitly, by those skilled in the art that the embodiments described herein may be combined with other embodiments.

[0035] Example 1

[0036] like Figure 1 The figure shows a schematic diagram of the structure of an integrated sensing device for analyzing intracavity laser emission signals provided by the present application, comprising a first reflector, a microfluidic cavity unit, and a second reflector, wherein the reflective surfaces of the first and second reflectors are coated with a high-reflectivity coating. It should be noted that the intracavity laser emission signal includes spectral information and / or spatial imaging information. The specific implementation thereof is described in detail below.

[0037] In this embodiment, the first and second reflectors are made of quartz or an optical glass substrate, and the reflective surfaces of the reflectors are coated with a high-reflectivity coating. The high-reflectivity coating is a dielectric coating with a thickness of 10-1000 nm. The thickness of the first and second reflectors is 1-3 mm.

[0038] It can be understood that the thickness of the first reflector and the second reflector provided in this embodiment is set between 1-3 mm, which better ensures that they are closer to a rigid body and are not easily deformed.

[0039] It should be noted that the integrated sensing device provided in the above embodiment can be used for laser emission testing. From top to bottom, first, the first reflector 110 is made of square (25mm*25mm) optical glass with a high-reflectivity coating (dielectric coating). The reflector thickness is approximately 1-3mm. For example, the reflector used is 3mm thick. A hole is mechanically punched at a specific location in the first reflector 110, with an aperture diameter of 0.5-2mm.

[0040] See also Figure 2 , which is a structural diagram of the microfluidic cavity unit provided in one embodiment, is a microfluidic channel film, and the microfluidic channel film includes PDMS (refer to Figure 1 PDMS film) silica gel or SU8 photoresist or light curing resin, with a thickness of 10-200um.

[0041] It can be understood that the thickness of the microfluidic cavity unit is determined by the thickness of the single layer of cells. The thickness of the microfluidic cavity unit is selected between 10-200 μm to better meet the actual application requirements.

[0042] Furthermore, the micro-channel cavity unit is formed in a cavity between the reflective surfaces of the first reflector and the second reflector coated with a high reflectivity coating.

[0043] See also Figure 2 The thickness of the selected microfluidic channel film is 50μm, and it is cut by a customized die. The microfluidic channel film is mainly composed of an inlet area, a detection area, and an outlet area. The sampling area is mainly used for liquid sampling. The sampling methods used include but are not limited to mechanical pumps (including pipettes), gravity sampling or mechanical valve sampling. The detection area is a relatively wide flat area, which is mainly used for sample detection, and the outlet area is mainly used for sample outflow. The sample automatically fills the microcavity and flows out to the outside through the outlet area. The samples used include fluorescent probes, free samples in aqueous and oil-phase buffer solutions such as nucleic acids and proteins, and micro-nanoscale biological samples that are roughly solid phase, such as viruses and cells.

[0044] Specifically, the integrated sensing device provided in the above-mentioned embodiment of the present application comprises, from top to bottom, a square (25mm*25mm) quartz reflector with a high-reflectivity coating (dielectric coating). The reflector thickness is approximately 1-3mm, for example, the thickness of the reflector used is 3mm. A hole is mechanically punched at a specific location on the upper mirror, with an aperture of 0.5-2mm, for example, 1mm. The first and second reflectors are then assembled with the microfluidic cavity unit via plasma bonding or double-sided tape. The integrated sensing device provided in this embodiment can be used for laser emission testing.

[0045] It can be understood that in the integrated sensing device provided in this embodiment, there are no special requirements for the structures of the reflecting surfaces of the first reflector and the second reflector. In practice, they can be designed according to application needs. For example, the reflecting surface can be a planar structure or a concave mirror structure.

[0046] It should be noted that the PDMS cavity layer formed between the first and second reflectors largely ensures the parallelism of the upper and lower reflectors, achieving highly stable FP resonant cavity performance. The thickness of the microfluidic channel membrane ensures that the sample (cells, microspheres, etc.) in the detection area is a single layer, making it a practical vehicle for single-cell laser emission analysis.

[0047] Example 2

[0048] See also Figure 3 , is a schematic structural diagram of an integrated sensing device provided in another embodiment, comprising a first optical glass, a microfluidic cavity unit, and a second optical glass. The thickness of the first optical glass and the second optical glass is 50-300 μm.

[0049] The specific implementation of the micro-channel cavity unit provided in this embodiment can be referred to in Example 1 and will not be described in detail here.

[0050] It should be noted that the device used for fluorescence lifetime testing in the above embodiments is constructed from low-fluorescence glass on both the top and bottom surfaces, virtually eliminating interference with autofluorescence. Furthermore, the highly stable flow path ensures a consistent concentration of sample molecules within the test area, further facilitating quantitative results. This device for fluorescence lifetime analysis is well-suited for fluorescence lifetime analysis equipment and accompanying imaging systems.

[0051] Example 3

[0052] The present application also provides a method for preparing an integrated sensing device, comprising the following steps: assembling the first reflector, the second reflector, and the microfluidic cavity unit by plasma bonding or double-sided tape; or

[0053] The first optical glass, the microchannel cavity unit and the second optical glass are assembled by plasma bonding or double-sided tape bonding.

[0054] In this embodiment, the microfluidic channel cavity unit is a microfluidic channel film, and the microfluidic channel film includes PDMS silica gel or SU8 photoresist or photocurable resin.

[0055] Furthermore, the microfluidic channel film can be processed by secondary machining of a commercial PDMS film. For example, secondary machining of a commercial PDMS film can be achieved by photolithography, 3D printing, silicon etching, wet etching or knife etching.

[0056] It can be understood that when using materials with poor viscosity such as SU8 and photocurable resin to manufacture sandwich microfluidic channels, the integrated assembly of the device can be achieved by using stainless steel fixtures, plastic fixtures, double-sided adhesive, etc.

[0057] In this embodiment, the microfluidic channel membrane is primarily composed of an inlet area, a detection area, and an outlet area. The inlet area is primarily used for liquid loading, using methods including but not limited to mechanical pumps (including pipettes), gravity loading, or mechanical valve loading. The detection area is a relatively wide, flat area primarily used for sample detection, while the outlet area is primarily used for sample discharge. Samples automatically fill the microcavity and flow out through the outlet area. Samples used include fluorescent probes, free samples dissolved in aqueous or oily buffers such as nucleic acids and proteins, and micro- and nanoscale biological samples that are generally solid-phase, such as viruses and cells.

[0058] It should be noted that the planar shape of the PDMS microchannel in the microchannel cavity unit, the size of the observation area, the size of the sample inlet and outlet, and the thickness of the interlayer can all be changed. In the manufacturing process, the thin film PDMS can also be replaced with materials such as SU8 photoresist and double-sided tape. In devices used for laser emission observation, the material of the dielectric reflective coating, the reflection wavelength and the limiting reflectivity can all be adjusted according to actual needs. The thickness of the reflector base glass (1-5mm) can also be adjusted as needed. In devices used for fluorescence lifetime observation, the thickness and material of the upper and lower glass slides can be adjusted according to actual needs (50-300μm).

[0059] The preparation method of the integrated sensing device provided in the above-mentioned embodiment of the present application integrates two optical elements, including a relatively thick glass-based dielectric reflector or an extremely thin optical glass, and integrates the optical elements with a soft polymer interlayer with pre-engraved flow channels to ultimately produce a stable optical microcavity through which liquids, cells, viruses and other specimens can pass. The device can be a practical vehicle for intracavity laser emission spectroscopy biomolecular analysis, laser emission imaging analysis, fluorescence imaging of intracavity biological samples and molecular samples, fluorescence spectroscopy analysis and fluorescence lifetime analysis. It has a wide range of applications, a simple preparation method and is suitable for industrial production.

[0060] See also Figure 4, which is a schematic diagram of the integrated sensing device provided in the above embodiment of the present application for intracavity laser emission signal analysis, wherein the first reflector is 3mm, the microchannel cavity unit is 50μm, the second reflector is 3mm, and the thickness of the high reflectivity coating is 50nm. In this integrated sensing device, by adding FITC fluorescent dye aqueous solutions of different concentrations into the microchannel cavity unit, it is observed in a microscopic imaging system with a spectrometer using OPO nanosecond pulse laser (473nm) micro-pump light. In the observation area of ​​the device, stable FP resonant cavity lasers (FSR of about 2.7nm) can appear in different areas. In the experiment, the central area and the area near the edge were tested, and the laser thresholds were not much different, showing good stability and consistency between regions.

[0061] Depend on Figure 4 It can be seen that the integrated sensing device provided by this application realizes the integration of FP laser emission devices for biomolecular analysis under relatively simple conditions through the fusion of multiple materials. The optical path of the FP resonant cavity can be stabilized by industrially produced PDMS films, achieving stabilization and homogenization of the quality factor. The thickness of the flow channel can ensure that the sample (cell) in the detection area is a single layer, further improving the analysis accuracy. This invention not only makes the laser emission measurement results easier to reproduce, but also enables the reuse of the device without damaging the resonant cavity.

[0062] It should be understood that the "at least one" involved in the embodiments of the present application refers to one or more, and "plurality" refers to two or more. "And / or" describes the association relationship of associated objects, indicating that three relationships may exist. For example, A and / or B can represent: the existence of A alone, the existence of A and B at the same time, and the existence of B alone, where A and B can be singular or plural. The character " / " generally indicates that the previous and subsequent associated objects are in an "or" relationship. "At least one of the following items" or similar expressions refers to any combination of these items, including any combination of single items or plural items. For example, at least one of a, b, or c can represent: a, b, c, ab, ac, bc, or abc, where a, b, c can be single or multiple.

[0063] Furthermore, unless otherwise specified, ordinal numbers such as "first" and "second" mentioned in the embodiments of the present application are used to distinguish multiple objects and are not used to limit the order, timing, priority or importance of multiple objects.

[0064] It should be noted that for the aforementioned method embodiments, for the sake of simplicity, they are all expressed as a series of action combinations, but those skilled in the art should be aware that this application is not limited by the order of the actions described, because according to this application, certain steps can be performed in other orders or simultaneously. Secondly, those skilled in the art should also be aware that the embodiments described in the specification are all preferred embodiments, and the actions and modules involved are not necessarily required by this application.

[0065] In the above embodiments, the description of each embodiment has its own focus. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0066] In the several embodiments provided in this application, it should be understood that the disclosed devices can be implemented in other ways. For example, the device embodiments described above are only schematic. For example, the division of the above-mentioned units is only a logical function division. In actual implementation, there may be other division methods, such as multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be through some interfaces, and the indirect coupling or communication connection of devices or units can be electrical or other forms.

[0067] The above is a detailed introduction to the embodiments of the present application. Specific examples are used herein to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the method and core idea of ​​the present application. At the same time, for those skilled in the art, according to the idea of ​​the present application, there may be changes in the specific implementation methods and application scope. In summary, the content of this specification should not be understood as a limitation on the present application.

Claims

1. An integrated sensing device for analyzing intracavity laser emission signals, characterized in that: The invention comprises a first reflector, a microchannel cavity unit and a second reflector which are arranged in sequence from top to bottom. The reflective surfaces of the first reflector and the second reflector are coated with a high reflectivity coating.

2. The integrated sensor device for analyzing intracavity laser emission signals according to claim 1, characterized in that: The first reflector and the second reflector are made of quartz or optical glass.

3. The integrated sensor device for analyzing intracavity laser emission signals according to claim 2, characterized in that: The high reflectivity coating is a dielectric coating, and the thickness of the high reflectivity dielectric coating is 10-1000nm.

4. The integrated sensor device for analyzing intracavity laser emission signals according to claim 2, characterized in that: The thickness of the first reflector and the second reflector is 0.5-3 mm.

5. The integrated sensor device for analyzing intracavity laser emission signals according to claim 1, characterized in that: The microfluidic channel cavity unit is a microfluidic channel film, and the microfluidic channel film includes PDMS silica gel or SU8 photoresist or photocurable resin.

6. The integrated sensor device for analyzing intracavity laser emission signals according to claim 1, characterized in that: The thickness of the microfluidic channel cavity unit is 10-200 μm.

7. The integrated sensor device for analyzing intracavity laser emission signals according to claim 1, characterized in that: The microfluidic channel film is provided with a sample inlet area, a detection area and a sample outlet area. The sample adding area is used for adding liquid, the detection area is used for detecting samples, and the sample outlet area is used for the outflow of samples.

8. The integrated sensor device for analyzing intracavity laser emission signals according to claim 7, characterized in that: The sample includes a free sample in a dissolved water phase, an oil phase buffer or a solid phase micro-nano scale biological sample, the dissolved water phase includes a fluorescent probe or nucleic acid or protein, and the solid phase micro-nano scale biological sample includes a virus or a cell.

9. An integrated sensing device for fluorescence signal testing, characterized in that: The invention comprises a first optical glass, a microchannel cavity unit and a second optical glass which are arranged in sequence from top to bottom.

10. The integrated sensor device for fluorescence signal testing according to claim 9, characterized in that: The thickness of the first optical glass and the second optical glass is 50-300 μm.

11. The integrated sensor device for fluorescence signal testing according to claim 9, characterized in that: The microfluidic channel cavity unit is a microfluidic channel film, and the microfluidic channel film includes PDMS silica gel or SU8 photoresist or photocurable resin.

12. The integrated sensor device for fluorescence signal testing according to claim 9, characterized in that: The thickness of the microfluidic channel cavity unit is 10-200 μm.

13. The integrated sensor device for fluorescence signal testing according to claim 9, characterized in that: The microfluidic channel film is provided with a sample inlet area, a detection area and a sample outlet area. The sample adding area is used for adding liquid, the detection area is used for detecting samples, and the sample outlet area is used for the outflow of samples.

14. The integrated sensor device for fluorescence signal testing according to claim 13, characterized in that: The sample includes a free sample in a dissolved water phase, an oil phase buffer or a solid phase micro-nano scale biological sample, the dissolved water phase includes a fluorescent probe or nucleic acid or protein, and the solid phase micro-nano scale biological sample includes a virus or a cell.

15. A method for preparing the integrated sensor device according to claim 1 or 9, characterized in that: The steps include: Assembling the first reflector, the second reflector and the microfluidic cavity unit by plasma bonding or double-sided adhesive bonding; or The first optical glass, the microchannel cavity unit and the second optical glass are assembled by plasma bonding or double-sided adhesive bonding.

16. The method for preparing the integrated sensor device according to claim 13, characterized in that: The microfluidic cavity unit can be processed by secondary machining of a commercial PDMS film, and the secondary machining includes but is not limited to photolithography, 3D printing, silicon etching, wet etching and knife carving.

17. An application of the integrated sensor device according to claim 1 in intracavity laser emission spectroscopy biomolecule analysis and laser emission imaging analysis.

18. Use of the integrated sensor device according to claim 9.

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