Integrated sensor device, method of manufacture and use thereof

Abstract

The application provides an integrated sensing device and a preparation method. Two optical elements, including a thick glass-based medium mirror with liquid sample inlet and outlet or an extremely thin optical glass, are integrally fused with a soft polymer interlayer pre-engraved with a flow channel, so that a stable optical microcavity through which a liquid sample and biological samples such as cells can pass is finally generated. The device can be a practical carrier for single-cell laser emission analysis, single-cell fluorescence spectrum analysis and molecular fluorescence lifetime research, and has wide application.

Description

Technical Field

[0001] This application relates to the field of bio-optical device technology, and in particular to an integrated sensing device and apparatus, its preparation method and its application. Background Technology

[0002] In bioanalysis and cell biology research, it is often necessary to analyze trace amounts of biochemical samples. Examples include fluorescent probes, free samples of nucleic acids and proteins dissolved in aqueous or oil-based buffer solutions, and micro / nanoscale biological samples such as cells, which are largely solid. These studies often require the use of microfluidic sample processing devices and biosensors that amplify and modulate the information carried in the samples, converting the microscopic world into recognizable signals for sensitive optical analysis. Optical sensors convert the interaction between the analyte and a sensitive element into light signals for detection. These sensors are less susceptible to external electromagnetic interference, can perform long-distance detection, and, because multiple signals can be transmitted through a single channel, they can detect a large amount of information.

[0003] Existing tests of this type often involve directly dropping the dye onto a glass slide (with an open upper surface), making it difficult to effectively control the depth and thickness of the sample on the slide. In addition, some teams have used closed PDMS channels to achieve similar requirements, but PDMS polymer materials have strong autofluorescence properties, which can easily induce background fluorescence noise.

[0004] Existing methods for this type of testing often employ a parallelism control scheme using two mirrors with a small plastic bead (10-30 μL) sandwiched between them. However, this results in poor stability between the mirror groups. Furthermore, there is a significant difference in quality factor between the regions near and outside the plastic bead within the resonant cavity, which is not conducive to the precise quantitative analysis of cells required in practical research. Summary of the Invention

[0005] This application provides an integrated sensing device that avoids the inadequacies in stability and repeatability of existing devices.

[0006] In a first aspect, embodiments of this application provide an integrated sensing device, including a first reflector, a microchannel cavity unit, and a second reflector arranged sequentially 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 and second reflectors are made of quartz or optical glass substrates.

[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 and second reflectors is 0.5-3 mm.

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

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

[0012] In some embodiments, the microfluidic channel membrane has 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 the sample, and the outlet area is used for the sample to flow out.

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

[0014] Secondly, embodiments of this application provide an integrated sensing device for fluorescence signal testing, comprising a first optical glass, a microfluidic cavity unit, and a second optical glass arranged sequentially 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 cavity unit is a microfluidic channel film, which includes PDMS silicone, SU8 photoresist, or photocurable resin.

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

[0018] In some embodiments, the microfluidic channel membrane has 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 the sample, and the outlet area is used for the sample to flow out.

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

[0020] Thirdly, embodiments of this application provide a method for fabricating an integrated sensing device, comprising the following steps:

[0021] The first reflector, the second reflector, and the microchannel cavity unit are assembled using plasma bonding or double-sided adhesive bonding; or

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

[0023] In some embodiments, the microchannel cavity unit can be fabricated using a commercial PDMS thin film through secondary machining, including photolithography, 3D printing, silicon etching, wet etching, and engraving.

[0024] Fourthly, embodiments of this application provide the application of an integrated sensing device in intracavity laser emission biomolecular analysis.

[0025] Fifthly, embodiments of this application provide the application of an integrated sensing device in the fluorescence lifetime analysis of intracavitary biomolecules.

[0026] The technical solution provided in this application integrates two optical elements, including a thicker glass-based dielectric mirror or an extremely thin optical glass, with a soft polymer interlayer pre-etched with flow channels, ultimately creating a stable optical microcavity through which liquids, cells, and viruses can pass. This can serve as a practical carrier for single-cell laser emission analysis and single-cell fluorescence lifetime research, with wide applications. Attached Figure Description

[0027] To more clearly illustrate the technical solutions in the embodiments of this application, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0028] Figure 1 This is a schematic diagram of the integrated sensing device for intracavity laser emission signal analysis provided in the embodiments of this application;

[0029] Figure 2 This is a schematic diagram of the structure of the microchannel cavity unit provided in the embodiments of this application;

[0030] Figure 3 This is a schematic diagram of the structure of an integrated sensing device for fluorescence signal testing provided in an embodiment of this application;

[0031] Figure 4 This is a schematic diagram of the integrated sensing device provided in this application embodiment for intracavity laser emission signal analysis. Detailed Implementation

[0032] To help those skilled in the art better understand the technical solutions of this application, the technical solutions in the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the description of the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0033] The terms "first," "second," etc., in the specification, claims, and accompanying drawings of this application are used to distinguish different objects, not to describe a specific order. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. For example, a process, method, software, product, or apparatus that includes a series of steps or units is not limited to the listed steps or units, but also includes steps or units not listed, or other steps or units inherent to such processes, methods, products, or apparatus.

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

[0035] Example 1

[0036] like Figure 1 The diagram shows the structure of the integrated sensing device for intracavity laser emission signal analysis provided in this application. It includes a first reflector, a microchannel cavity unit, and a second reflector. 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 method is described in detail below.

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

[0038] It is understood that the thickness of the first and second reflectors provided in this embodiment is set between 1 and 3 mm to better ensure that they are closer to rigid bodies and not easily deformed.

[0039] It should be noted that the integrated sensing device provided in the above embodiments can be used for laser emission testing. From top to bottom, the first reflector 110 is a square (25mm*25mm) optical glass with a high-reflectivity coating (dielectric coating). The reflector thickness is approximately 1-3mm; for example, the reflector used has a thickness of 3mm. The first reflector 110 has holes drilled at specific locations using mechanical drilling, with a hole diameter of 0.5-2mm.

[0040] Please see Figure 2 The diagram shows the structure of the microfluidic cavity unit provided in one embodiment, which is a microfluidic channel film. The microfluidic channel film includes PDMS (see reference). Figure 1 The PDMS film is made of silicone or SU8 photoresist or photocurable resin and has a thickness of 10-200um.

[0041] It is understood that the thickness of the microchannel cavity unit is determined by the thickness of a single cell, and the thickness of the microchannel cavity unit is selected between 10-200um to better meet the needs of practical applications.

[0042] Furthermore, the microchannel cavity unit is formed within the cavity between the reflective surfaces of the first and second reflective mirrors, which are coated with a high-reflectivity coating.

[0043] Please see Figure 2 The selected microfluidic channel membrane has a thickness of 50 μm and is cut using a custom die. The microfluidic channel membrane mainly consists of an inlet area, a detection area, and an outlet area. The inlet area is primarily used for liquid dispensing, and the dispensing methods include, but are not limited to, mechanical pumps (including pipettes), gravity dispensing, or mechanical valve dispensing. The detection area is a relatively wide and flat region, mainly used for sample detection, while the outlet area is mainly used for sample outflow. The sample automatically fills the microcavity and flows out through the outlet area. The samples used include fluorescent probes, free samples dissolved in aqueous or oil-phase buffer solutions such as nucleic acids and proteins, as well as micro / nanoscale biological samples that are roughly in a solid phase, such as viruses and cells.

[0044] Specifically, the integrated sensing device provided in the above embodiments of this application, from top to bottom, first consists of a square (25mm*25mm) quartz mirror with a high-reflectivity coating (dielectric coating). The mirror thickness is approximately 1-3mm, for example, the mirror used has a thickness of 3mm. The upper mirror has holes drilled at specific locations with a diameter of 0.5-2mm, for example, 1mm. The first mirror, the second mirror, and the microfluidic cavity unit are then assembled using plasma bonding or double-sided adhesive. The integrated sensing device provided in this embodiment can be used for laser emission testing.

[0045] It is understood that the integrated sensing device provided in this embodiment does not have special requirements for the structure of the reflecting surfaces of the first and second reflectors. In practice, it 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 high stability of the FP resonant cavity performance. The thickness of the microfluidic channel film ensures that the sample (cells, microspheres, etc.) in the detection area is a single layer, making it a practical carrier for single-cell laser emission analysis research.

[0047] Example 2

[0048] Please see Figure 3 The diagram below illustrates the structure of an integrated sensing device according to another embodiment, including a first optical glass, a microfluidic cavity unit, and a second optical glass. The thickness of the first and second optical glasses is 50-300 μm.

[0049] The specific implementation of the microchannel cavity unit provided in this embodiment can be found in Embodiment 1, and will not be repeated here.

[0050] It should be noted that, taking the device for fluorescence lifetime testing provided in the above embodiments as an example, both the top and bottom surfaces are made of low-fluorescence glass, which causes almost no interference to autofluorescence. Furthermore, the highly stable flow channel means that the sample molecule concentration within the test area is stable, which is more conducive to obtaining quantitative results. The device for fluorescence lifetime analysis is well-suited for fluorescence lifetime analysis equipment and its supporting imaging system.

[0051] Example 3

[0052] This application also provides a method for fabricating an integrated sensing device, comprising the following steps: assembling the first reflector, the second reflector, and the microchannel cavity unit by means of plasma bonding or double-sided adhesive bonding; or

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

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

[0055] Furthermore, the microfluidic channel film can be further processed by secondary machining of commercial PDMS films, such as photolithography, 3D printing, silicon etching, wet etching, or knife cutting.

[0056] It is understandable that when using materials with poor adhesion, such as SU8 and photocurable resin, to manufacture sandwich microchannels, the integrated assembly of the device can be achieved through stainless steel clamps, plastic clamps, double-sided adhesive, etc.

[0057] In this embodiment, the microfluidic channel membrane mainly consists of an inlet area, a detection area, and an outlet area. The inlet area is mainly used for liquid dispensing, and the dispensing methods include, but are not limited to, mechanical pumps (including pipettes), gravity dispensing, or mechanical valve dispensing. The detection area is a relatively wide and flat region, mainly used for sample detection, while the outlet area is mainly used for sample outflow. The sample automatically fills the microcavity and flows out through the outlet area. The samples used include fluorescent probes, free samples dissolved in aqueous or oil-phase buffer solutions such as nucleic acids and proteins, and micro / nanoscale biological samples that are roughly in a 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 inlet and outlet ports, and the thickness of the interlayer can all be changed. In the fabrication process, the thin-film PDMS can also be replaced with SU8 photoresist, double-sided adhesive, or other materials. In devices used for laser emission observation, the material, reflection wavelength, and limiting reflectivity of the dielectric reflective coating can all be adjusted according to actual needs. The thickness of the reflector substrate 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 method for fabricating the integrated sensing device provided in the above embodiments of this application integrates two optical elements, including a thicker glass-based dielectric mirror or an extremely thin optical glass, with a soft polymer interlayer pre-etched with flow channels. This ultimately produces a stable optical microcavity through which liquids, cells, and viruses can pass. It can serve as a practical carrier for intracavity laser emission spectroscopy biomolecular analysis, laser emission imaging analysis, fluorescence imaging of intracavity biological and molecular samples, fluorescence spectroscopy analysis, and fluorescence lifetime analysis. It has wide applications, a simple fabrication method, and is suitable for industrial production.

[0060] Please see Figure 4This diagram illustrates the integrated sensing device provided in the above embodiments of this application for intracavity laser emission signal analysis. The first reflector is 3mm thick, the microchannel cavity unit is 50μm thick, the second reflector is 3mm thick, and the high-reflectivity coating is 50nm thick. In this integrated sensing device, different concentrations of FITC fluorescent dye aqueous solutions are added to the microchannel cavity unit, and observation is performed using an OPO nanosecond pulsed laser (473nm) as the micropump light in a microscopic imaging system equipped with a spectrometer. Stable FP resonant cavity lasers (FSR approximately 2.7nm) can be observed in different regions within the observation area of ​​this device. Experiments tested the central region and the region near the edge; the laser threshold differences were small, demonstrating good stability and inter-regional consistency.

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

[0062] It should be understood that "at least one" in the embodiments of this application refers to one or more, and "more than one" refers to two or more. "And / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A alone, A and B simultaneously, or B alone, where A and B can be singular or plural. The character " / " generally indicates that the preceding and following related objects are in an "or" relationship. "At least one of the following" or similar expressions refer to any combination of these items, including any combination of single 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, and c can be single or multiple.

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

[0064] It should be noted that, for the sake of simplicity, the foregoing method embodiments are all described as a series of actions. However, those skilled in the art should understand that this application is not limited to the described order of actions, as some steps may be performed in other orders or simultaneously according to this application. Furthermore, those skilled in the art should also understand that the embodiments described in the specification are preferred embodiments, and the actions and modules involved are not necessarily essential to this application.

[0065] In the above embodiments, the descriptions of each embodiment have different focuses. For parts not described in detail in a certain embodiment, please refer to the relevant descriptions in other embodiments.

[0066] In the several embodiments provided in this application, it should be understood that the disclosed apparatus can be implemented in other ways. For example, the apparatus embodiments described above are merely illustrative; for instance, the division of the units described above is only a logical functional division, and in actual implementation, there may be other division methods. For example, multiple units or components may be combined or integrated into another system, or some features may be ignored or not executed. Furthermore, the coupling or direct coupling or communication connection shown or discussed may be through some interfaces; the indirect coupling or communication connection between devices or units may be electrical or other forms.

[0067] The embodiments of this application have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of this application. The description of the above embodiments is only for the purpose of helping to understand the method and core ideas of this application. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the ideas of this application. Therefore, the content of this specification should not be construed as a limitation of this application.

Claims

1. An integrated sensing device for intracavity laser emission signal analysis, characterized in that, The device includes a first reflector, a microfluidic cavity unit, and a second reflector arranged sequentially from top to bottom. The reflective surfaces of the first and second reflectors are coated with a high-reflectivity coating. The microfluidic cavity unit is a microfluidic channel film, which includes PDMS silicone or SU8 photoresist. The microfluidic channel film has an inlet area, a detection area, and an outlet area. The inlet area is used for adding liquid samples, the detection area is used for detecting samples, and the outlet area is used for sample outflow. The samples include free samples dissolved in aqueous or oil buffer solutions, or micro / nanoscale biological samples in solid phase. The dissolved aqueous phase includes fluorescent probes, nucleic acids, or proteins, and the solid micro / nanoscale biological samples include viruses or cells.

2. The integrated sensing device for intracavity laser emission signal analysis according to claim 1, characterized in that, The first and second reflectors are made of quartz.

3. The integrated sensing device for intracavity laser emission signal analysis according to claim 1, characterized in that, The first and second reflectors are optical glass substrates.

4. The integrated sensing device for intracavity laser emission signal analysis according to claim 2 or 3, characterized in that, The high reflectivity coating is a dielectric coating, and the thickness of the high reflectivity coating is 10-1000 nm.

5. The integrated sensing device for intracavity laser emission signal analysis according to claim 2 or 3, characterized in that, The thickness of the first reflector and the second reflector is 0.5-3mm.

6. The integrated sensing device for intracavity laser emission signal analysis according to claim 1, characterized in that, The thickness of the microchannel cavity unit is 10-200 μm.

7. A method for manufacturing an integrated sensing device as described in claim 1, characterized in that, Includes the following steps: The first reflector, the second reflector, and the microchannel cavity unit are assembled using plasma bonding or double-sided adhesive.

8. The method for preparing the integrated sensing device according to claim 7, characterized in that, The microchannel cavity unit is processed by secondary machining of commercial PDMS thin film, including photolithography, 3D printing, silicon etching, wet etching and engraving.

9. The application of the integrated sensing device according to claim 1 in intracavity laser emission spectroscopy biomolecular analysis.

10. An application of the integrated sensing device according to claim 1 in laser emission imaging analysis.

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