Integrated sensing device, preparation method therefor, and use thereof

Abstract

An integrated sensing device and a preparation method. Two optical elements, including a thick glass-based dielectric reflector provided with a liquid sample inlet and a liquid sample outlet, or extremely thin optical glass, are integrally bonded with a soft polymer interlayer in which a flow channel is formed in advance by etching, so as to finally generate a stable optical microcavity through which biological samples such as liquid samples and cells can pass. The device can serve as a practical carrier for single-cell laser emission analysis, single-cell fluorescence spectrum analysis and molecular fluorescence lifetime research, and is widely used.

Description

Integrated sensing device, preparation method and application thereof 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 application thereof. BACKGROUND

[0002] In biological analysis and cell biology research, people often need to analyze trace amounts of biochemical samples. For example, fluorescent probes, free samples in nucleic acid, protein, and other dissolved aqueous phase, oil phase buffer, and cell and other micro-nano scale biological samples in general solid phase. These studies often need to use supporting microfluidic sample processing devices and biological sensor devices that can amplify and modulate the information carried in the sample, so as to convert the microcosmic world into a signal that can be recognized, and complete sensitive optical analysis. Optical sensors convert the interaction between the analyte and the sensitive element into an optical signal for detection. Such sensors are not easily affected by external electromagnetic interference, can be detected remotely, and have a large amount of information detected because multiple signals can be transmitted through one channel.

[0003] Existing tests of this type often directly drop dyes on a glass slide (open surface), and it is difficult to effectively control the depth and thickness of the sample on the slide. In addition, some teams use closed PDMS flow channels to achieve similar needs, but PDMS polymer materials have strong spontaneous fluorescence properties, which can easily induce background fluorescence noise.

[0004] Existing tests of this type often use a parallelism control scheme in which two mirrors sandwich a small plastic bead (10-30 μL) in the middle. However, the stability between the mirror groups is poor. In addition, the quality factor of the area near the plastic bead and the plastic bead in the resonant cavity is significantly different, which is not conducive to the needs of precise quantitative analysis of cells in actual research.

[0005] SUMMARY

[0006] The embodiments of the present application provide an integrated sensing device, which can avoid the lack of stability and repeatability of existing devices.

[0007] In a first aspect, the embodiments of the present application provide an integrated sensing device, which comprises a first mirror, a microfluidic cavity unit and a second mirror arranged in sequence from top to bottom, and the reflective surfaces of the first mirror and the second mirror are coated with a high reflectivity coating.

[0008] In some embodiments, the first mirror and the second mirror are quartz or optical glass substrates.

[0009] In some embodiments, the high reflectivity coating is a dielectric coating, and the thickness of the high reflectivity coating is 10-1000 nm.

[0010] In some embodiments, the thickness of the first mirror and the second mirror is 0.5-3mm.

[0011] In some embodiments, the microfluidic channel cavity unit is a microfluidic channel film, which comprises PDMS silicone or SU8 photoresist or photocurable resin.

[0012] In some embodiments, the thickness of the microfluidic channel cavity unit is 10-200μm.

[0013] In some embodiments, the microfluidic channel film is provided with a sample inlet area, a detection area and a sample outlet area, the sample inlet area is used for liquid sampling, the detection area is used for sample detection, and the sample outlet area is used for sample outflow.

[0014] In some embodiments, the sample comprises dissolved water phase, free sample in oil phase buffer or solid phase micro-nano scale biological sample, the dissolved water phase comprises fluorescent probe or nucleic acid or protein, and the solid phase micro-nano scale biological sample comprises virus or cell.

[0015] In a second aspect, the embodiments of the present application provide an integrated sensing device for fluorescence signal testing, which comprises a first optical glass, a microfluidic channel cavity unit and a second optical glass arranged in sequence from top to bottom.

[0016] In some embodiments, the thickness of the first optical glass and the second optical glass is 50-300μm.

[0017] In some embodiments, the microfluidic channel cavity unit is a microfluidic channel film, which comprises PDMS silicone or SU8 photoresist or photocurable resin.

[0018] In some embodiments, the thickness of the microfluidic channel cavity unit is 10-200μm.

[0019] In some embodiments, the microfluidic channel film is provided with a sample inlet area, a detection area and a sample outlet area, the sample inlet area is used for liquid sampling, the detection area is used for sample detection, and the sample outlet area is used for sample outflow.

[0020] In some embodiments, the sample comprises dissolved water phase, free sample in oil phase buffer or solid phase micro-nano scale biological sample, the dissolved water phase comprises fluorescent probe or nucleic acid or protein, and the solid phase micro-nano scale biological sample comprises virus or cell.

[0021] In a third aspect, the embodiments of the present application provide a preparation method of the integrated sensing device, comprising the following steps:

[0022] The first mirror, the second mirror and the microfluid channel cavity unit are assembled by Plasma plasma bonding or double-sided adhesive paste.

[0023] The first optical glass, the microfluid channel cavity unit and the second optical glass are assembled by Plasma plasma bonding or double-sided adhesive paste.

[0024] In some embodiments, the microfluid channel cavity unit can be secondarily machined from a commercial PDMS film, and the secondary machining includes photolithography, 3D printing, silicon etching, wet etching and knife etching.

[0025] In a fourth aspect, the embodiments of the present application provide an application of the integrated sensing device in intracavity laser emission biomolecule analysis.

[0026] In a fifth aspect, the embodiments of the present application provide an application of the integrated sensing device in intracavity biomolecule fluorescence lifetime analysis.

[0027] The technical scheme provided by the present application integrates two optical elements, including a thick glass-based medium mirror or an extremely thin optical glass, with a soft polymer interlayer pre-engraved with a flow channel, to finally generate a stable optical microcavity through which liquid, cells, viruses and other samples can pass, which can be a practical carrier for single-cell laser emission analysis and single-cell fluorescence lifetime research, and has wide application. BRIEF DESCRIPTION OF DRAWINGS

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0029] FIG. 1 is a structural schematic diagram of an integrated sensing device for intracavity laser emission signal analysis provided by an embodiment of the present application;

[0030] FIG. 2 is a structural schematic diagram of a microfluid channel cavity unit provided by an embodiment of the present application;

[0031] FIG. 3 is a structural schematic diagram of an integrated sensing device for fluorescence signal testing provided by an embodiment of the present application;

[0032] FIG. 4 is a schematic diagram of an integrated sensing device for intracavity laser emission signal analysis provided by an embodiment of the present application. DETAILED DESCRIPTION

[0033] In order for those skilled in the art to better understand the technical solutions of the present application, the technical solutions in the embodiments of the present application will be described clearly and completely below in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the description of the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of the present application.

[0034] The terms "first", "second", and the like in the specification and claims of the present application and the above-described drawings are used to distinguish different objects, rather than to describe a specific order. In addition, the terms "include" and "have" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, software, product, or device 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 the process, method, product, or device.

[0035] Reference herein to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the present application. The appearance of the phrase in various places in the specification does not necessarily all refer to the same embodiment, nor does it necessarily refer to a separate or alternative embodiment in isolation from other embodiments. It is explicitly and implicitly understood by those skilled in the art that the embodiments described herein can be combined with other embodiments.

[0036] Embodiment 1

[0037] As shown in FIG. 1, the present application provides a structural schematic diagram of an integrated sensing device for intracavity laser emission signal analysis, which comprises a first mirror, a micro-channel cavity unit, and a second mirror. The reflecting surfaces of the first mirror and the second mirror 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 mode is described in detail below.

[0038] In the present embodiment, the first mirror and the second mirror are quartz or optical glass substrates or optical glass substrates, and the reflecting surfaces of the above-mentioned mirrors are coated with a high reflectivity coating. The high reflectivity coating is a dielectric coating, and the thickness of the high reflectivity coating is 10-1000 nm. The thickness of the first mirror and the second mirror is 1-3 mm.

[0039] It can be understood that the first mirror and the second mirror provided in the present embodiment have a thickness of 1-3 mm, which better ensures that it is closer to a rigid body and is not easily deformed.

[0040] 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, the first mirror 110 is a square (25mm*25mm) optical glass with a high reflectivity coating (dielectric coating), and the thickness of the mirror is about 1-3mm, for example, the thickness of the mirror used is 3mm. The first mirror 110 is mechanically punched at a specific position to form an aperture, and the aperture diameter is 0.5-2mm.

[0041] Referring to FIG. 2, the structure diagram of the microfluidic cavity unit provided in one of the embodiments is shown. The microfluidic channel film includes PDMS (see FIG. 1 for the PDMS film) silicone or SU8 photoresist or photocurable resin, and the thickness is 10-200um.

[0042] It can be understood that the thickness of the microfluidic cavity unit is determined by the thickness of the single-layer cell, and the thickness of the microfluidic cavity unit is selected to be between 10-200um, which better meets the actual application requirements.

[0043] Further, the microfluidic cavity unit is formed in the cavity between the reflecting surfaces of the first mirror and the second mirror coated with a high reflectivity coating.

[0044] Referring to FIG. 2, the thickness of the microfluidic channel film used is 50um, and the cutting is performed by a customized knife mold. The microfluidic channel film mainly includes a sample inlet area, a detection area, and a sample outlet area. The sample inlet area is mainly used for liquid sampling, and the sampling methods used include but are not limited to mechanical pumps (including pipettes), gravity sampling, or mechanical valve sampling, etc. The detection area is a relatively wide and flat area, mainly for sample detection, and the sample outlet area is mainly for sample outflow. The sample is automatically filled into the microcavity and flows out of the outside through the sample outlet area. The samples used include fluorescent probes, free samples in nucleic acid, protein, etc. dissolved in water phase, oil phase buffer, and virus, cell, etc. solid phase micro-nano scale biological samples.

[0045] Specifically, the integrated sensing device provided in the above embodiment, from top to bottom, first is a square (25mm*25mm) quartz mirror with a high reflectivity coating (dielectric coating), and the thickness of the mirror is about 1-3mm, for example, the thickness of the mirror used is 3mm. The mirror is mechanically punched at a specific position to form an aperture, and the aperture diameter is 0.5-2mm, for example, the aperture diameter is 1mm. The first mirror, the second mirror, and the microfluidic cavity unit are assembled by Plasma plasma bonding or double-sided adhesive paste. The integrated sensing device provided in the embodiment can be used for laser emission testing.

[0046] It can be understood that the integrated sensing device provided in the embodiment does not have special requirements for the structures of the reflection surfaces of the first mirror and the second mirror, and the reflection surfaces can be designed according to application needs in practice, for example, the reflection surfaces can be planar structures or concave mirror structures.

[0047] It should be noted that the PDMS cavity layer formed between the first mirror and the second mirror ensures the parallelism of the upper and lower mirrors to a great extent, and realizes the high stabilization of the FP resonant cavity performance. The thickness of the microfluid channel film can ensure that the samples (cells, microspheres, etc.) in the detection area are single-layer, and can be a practical carrier for single-cell laser emission analysis research.

[0048] Embodiment 2

[0049] Referring to FIG. 3, a structural schematic diagram of an integrated sensing device provided in another embodiment includes a first optical glass, a microfluid channel cavity unit, and a second optical glass. The thicknesses of the first optical glass and the second optical glass are 50-300 μm.

[0050] The specific implementation of the microfluid channel cavity unit provided in the embodiment can refer to Embodiment 1, which will not be described here. It should be noted that the device for fluorescence lifetime test provided in the above embodiment is taken as an example, the upper and lower surfaces are both taken by low-fluorescent glass, which almost does not cause interference to spontaneous fluorescence. Moreover, the flow channel is highly stable, which means that the sample molecule concentration in the test area is stable, and it is more conducive to obtain quantitative results. The device for fluorescence lifetime analysis can be well adapted to the fluorescence lifetime analysis equipment and the matching imaging system.

[0051] Embodiment 3

[0052] The application further provides a preparation method of the integrated sensing device, including the following steps: the first mirror, the second mirror, and the microfluid channel cavity unit are assembled by Plasma plasma bonding or double-sided adhesive paste.

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

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

[0055] Further, the microfluid channel film can be mechanically processed twice from a commercial PDMS film, for example, the commercial PDMS film can be mechanically processed twice by photoetching, 3D printing, silicon etching, wet etching, or knife etching.

[0056] It can be understood that when using SU8, light-cured resin and other materials with poor viscosity to manufacture the sandwich micro-channel, the integrated assembly of the device can be achieved by means of stainless steel clamps, plastic clamps, double-sided adhesive tape and the like.

[0057] In the embodiment, the microfluidic channel film mainly consists of a sample inlet area, a detection area and a sample outlet area. The sample inlet area is mainly used for liquid sampling, and the sampling methods include but are not limited to mechanical pump (including a pipette), gravity sampling or mechanical valve sampling and the like. The detection area is a relatively wide and flat area, mainly for sample detection, and the sample outlet area is mainly used for sample outflow. The sample is automatically filled into the microcavity, and flows out of the outside through the sample outlet area. The sample used includes fluorescent probes, free samples in nucleic acid, protein and other dissolved water phase, oil phase buffer, and virus, cell and other solid phase micro-nano scale biological samples.

[0058] It should be noted that the planar shape of the PDMS micro-channel in the micro-channel cavity unit, the size of the observation area, the size of the sample inlet and outlet, and the thickness of the sandwich can be changed. In the manufacturing process, the thin film PDMS can be replaced by SU8 photoresist, double-sided adhesive and other materials. In the device used for laser emission observation, the material of the medium reflective coating, the reflective wavelength and the limit reflectivity can be adjusted according to actual needs. The thickness of the mirror substrate glass (1-5mm) can also be adjusted according to needs. In the device used for fluorescence lifetime observation, the thickness and material of the upper and lower glass sheets can be adjusted according to actual needs (50-300μm).

[0059] The preparation method of the integrated sensing device provided by the above embodiment of the application integrates two optical elements, including a thick glass-based medium mirror or an extremely thin optical glass, with a soft polymer sandwich layer pre-engraved with a flow channel, to finally generate a stable optical microcavity through which liquid, cells, viruses and other samples can pass. It can be a practical carrier for laser emission spectrum biological molecule analysis, laser emission imaging analysis, fluorescence imaging of biological samples and molecular samples in the cavity, fluorescence spectrum analysis and fluorescence lifetime analysis, and has wide application, simple preparation method and suitability for industrial production.

[0060] Please refer to Fig. 4, which is a schematic diagram of the integrated sensing device provided by the above embodiment of the application for intracavity laser emission signal analysis, wherein the first mirror is 3 mm, the micro-channel cavity unit is 50 μm, the second mirror is 3 mm, and the thickness of the high reflectivity coating is 50 nm. In the integrated sensing device, different concentrations of FITC fluorescent dye aqueous solution are added in the micro-channel cavity unit, and the device is observed in a microscopic imaging system with a spectrometer under the micro-pumping of OPO nanosecond pulse laser (473 nm). In the observation area of the device, stable F-P resonant cavity laser (FSR is about 2.7 nm) can appear in different regions. In the experiment, the central region and the region close to the edge are tested, and the laser threshold difference is not large, which shows good stability and consistency between regions.

[0061] As shown in Fig. 4, the integrated sensing device provided by the application realizes the integration of the FP laser emission device for biomolecular analysis under relatively simple conditions through the fusion of various materials. The optical path of the FP resonant cavity is stabilized by the PDMS film produced by industrial production, and the stabilization and uniformity of the quality factor are realized. The thickness of the flow channel can ensure that the sample (cell) in the detection area is single-layer, which further improves the analysis accuracy. This invention not only makes the laser emission measurement result more easily reproducible, but also realizes the repeated use of the device without damaging the resonant cavity.

[0062] It should be understood that “at least one” in the embodiments of the application refers to one or more, and “multiple” refers to two or more. “And / or” describes the association relationship between the associated objects, which means that there can be three relationships, for example, A and / or B can represent the following three cases: A exists alone, A and B exist together, and B exists alone, wherein A and B can be singular or plural. The character “ / ” generally represents an “or” relationship between the front and rear associated objects. “At least one of the following” or similar expressions means any combination of these items, including any combination of single item or multiple items. For example, at least one of a, b, or c can represent a, b, c, a-b, a-c, b-c, or a-b-c, wherein a, b, and c can be single or multiple.

[0063] In addition, unless otherwise stated, the ordinal numbers “first”, “second”, etc. mentioned in the embodiments of the application are used to distinguish multiple objects, and are not used to limit the order, time sequence, priority or importance of the multiple objects.

[0064] It should be noted that, for the foregoing method embodiments, for the sake of simple description, they are all described as a combination of a series of actions, but those skilled in the art should know that the present application is not limited to the order of the actions described, because according to the present application, certain steps can be performed in other orders or simultaneously. Secondly, those skilled in the art should know that the embodiments described in the specification are all preferred embodiments, and the actions and modules involved are not necessarily necessary for the present application.

[0065] In the above embodiments, the description of each embodiment has its own focus, and the parts not described in detail in a certain embodiment can be referred to the related description of other embodiments.

[0066] In several embodiments provided in the present application, it should be understood that the disclosed device can be implemented by other means. For example, the device embodiments described above are only schematic, and the division of the above units is only a logical function division, and actual implementation can have another division manner, for example, a plurality of units or components can be combined or integrated into another system, or some features can be omitted or not executed. In addition, the coupling or direct coupling or communication connection between the displayed or discussed each other can be indirect coupling or communication connection through some interfaces, devices or units, and can be electrical or other forms.

[0067] The above describes the embodiments of the present application in detail, and the specific examples are applied to the principles and implementation modes of the present application. The above embodiment description 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, the specific implementation mode and application range will be changed, and the above description should not be understood as the limitation of the present application.

Claims

1. An integrated sensing device for intracavity laser emission signal analysis, characterized by, The first mirror, the micro-channel cavity unit and the second mirror are sequentially arranged from top to bottom, and the reflecting surfaces of the first mirror and the second mirror are coated with a high reflectivity coating.

2. The integrated sensing device for in-lumen laser emission signal analysis of claim 1, wherein, The first mirror and the second mirror are quartz or optical glass substrates.

3. The integrated sensing device for in-vivo laser emission signal analysis of claim 2, wherein, The high reflectivity coating is a dielectric coating, and the thickness of the high reflectivity dielectric coating is 10-1000 nm.

4. The integrated sensing device for in-vivo laser emission signal analysis of claim 2, wherein, The thickness of the first mirror and the second mirror is 0.5-3 mm.

5. The integrated sensing device for in-vivo laser emission signal analysis of claim 1, wherein, The micro-channel cavity unit is a micro-fluid channel film, and the micro-fluid channel film comprises PDMS silicone or SU8 photoresist or photocurable resin.

6. The integrated sensing device for in-vivo laser emission signal analysis of claim 1, wherein, The thickness of the micro-channel cavity unit is 10-200 μm.

7. The integrated sensing device for in-vivo laser emission signal analysis of claim 1, wherein, The micro-fluid channel film is provided with a sample inlet area, a detection area and a sample outlet area, the sample inlet area is used for adding liquid, the detection area is used for detecting the sample, and the sample outlet area is used for flowing out the sample.

8. The integrated sensing device for in-vivo laser emission signal analysis of claim 7, wherein, The sample comprises dissolved water phase, free sample in oil phase buffer or solid phase micro-nano scale biological sample, the dissolved water phase comprises fluorescent probe or nucleic acid or protein, and the solid phase micro-nano scale biological sample comprises virus or cell.

9. An integrated sensing device for fluorescence signal testing, characterized in that, The first optical glass, the micro-channel cavity unit and the second optical glass are sequentially arranged from top to bottom.

10. The integrated sensing device for fluorescence signal testing of claim 9, wherein, The thickness of the first optical glass and the second optical glass is 50-300 μm.

11. The integrated sensing device for fluorescence signal testing of claim 9, wherein, The micro-channel cavity unit is a micro-fluid channel film, and the micro-fluid channel film comprises PDMS silicone or SU8 photoresist or photocurable resin.

12. The integrated sensing device for fluorescence signal testing of claim 9, wherein, The thickness of the micro-channel cavity unit is 10-200 μm.

13. The integrated sensing device for fluorescence signal testing of claim 9, wherein, The micro-fluid channel film is provided with a sample inlet area, a detection area and a sample outlet area, the sample inlet area is used for adding liquid, the detection area is used for detecting the sample, and the sample outlet area is used for flowing out the sample.

14. The integrated sensing device for fluorescence signal testing of claim 13, wherein, The sample comprises dissolved water phase, free sample in oil phase buffer or solid phase micro-nano scale biological sample, the dissolved water phase comprises fluorescent probe or nucleic acid or protein, and the solid phase micro-nano scale biological sample comprises virus or cell.

15. A method of manufacturing the one-piece sensor device according to claim 1 or 9, characterized in that The method comprises the following steps: The first mirror, the second mirror and the micro-channel cavity unit are assembled by Plasma plasma bonding or double-sided adhesive paste; or The first optical glass, the micro-channel cavity unit and the second optical glass are assembled by Plasma plasma bonding or double-sided adhesive paste.

16. The method of claim 13, wherein the method further comprises: The micro-channel cavity unit can be mechanically processed by a commercial PDMS film, and the secondary mechanical processing comprises photoetching, 3D printing, silicon etching and wet etching.

17. Application of the integrated sensing device according to claim 1 in intracavity laser emission spectrum biomolecular analysis and laser emission imaging analysis.

18. Application of the integrated sensing device according to claim 9.

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