Interface microrheology detection device and method

Abstract

The present invention relates to the technical field of interface testing, and provides an interface microrheology detection device and method. Among them, the interface microrheology detection device includes a trough body, a glass slide, a dish body and a dish cover, the glass slide is connected to the trough body, and the glass slide and the trough body together form a groove; a stepped hole is provided at the bottom of the dish body; the stepped hole includes a small hole section and a large hole section, and the connection between the small hole section and the large hole section forms a connecting surface; the upper surface of the glass slide is connected to the connecting surface; and the dish cover is covered on the dish body. The interface microrheology detection device of the present invention has a groove, which can fix the solution of the small body phase, so that the immiscible liquid-liquid or gas-liquid two phases coexist in a stable upper and lower stratified state, ensure the stability of the interface position during the measurement process, and enable the interface microrheology detection to proceed smoothly. The interface microrheology detection method of the present invention realizes accurate particle tracing microrheology analysis of the interface layer by fluorescently marking the interfacial active component material and accurately positioning the interface layer.

Description

Technical Field

[0001] The invention relates to the technical field of interface testing, and in particular to an interface microrheology detection device and method. Background Art

[0002] Due to the particularity of the interface location, the current methods and equipment for in-situ and non-destructive detection of the interfacial microrheological properties of interfacial active ingredients such as surfactants, proteins, and polysaccharides are very limited, which restricts the exploration of the interfacial behavior and evolution mechanism of active ingredients.

[0003] Since the concept of particle tracer microrheology was proposed in the last century, it has been used to explore the local viscoelasticity and heterogeneity evolution during the formation of microgel networks and to determine the gel point. However, this technology has not yet been popularized in related research in the field of interfaces.

[0004] Traditional confocal micro-dishes cannot fix small bulk phase solutions. There are two problems that are not solved during interface measurement. The first is that the stability of the interface position cannot be guaranteed during the measurement process. The second is that it cannot ensure that the detected area is the interface layer rather than the bulk phase, which makes it impossible to carry out interface microrheology detection smoothly. Summary of the invention

[0005] The purpose of the present invention is to provide an interface microrheology detection device and method to solve the problem that the interface microrheology detection in the prior art cannot be carried out smoothly.

[0006] In order to achieve the above-mentioned purpose, according to a first aspect of the present invention, there is provided an interfacial microrheology detection device, comprising: a trough body; a glass slide, the glass slide is connected to the bottom of the trough body, the glass slide and the trough body together form a groove for carrying a liquid to be tested; a dish body, the dish body has a containing cavity, and the bottom of the dish body is provided with a stepped hole connected to the containing cavity; along the direction from top to bottom, the stepped hole comprises a small hole section and a large hole section that are connected to each other, and the connection between the small hole section and the large hole section forms a connecting surface; the glass slide is placed in the large hole section, the upper surface of the glass slide is connected to the connecting surface, and at least part of the trough body passes through the small hole section and extends into the containing cavity; and a dish cover, the dish cover is arranged on the dish body to close the dish body.

[0007] Furthermore, the trough body is arranged in a circular ring shape; the diameter of the groove is greater than or equal to 2 mm and less than or equal to 20 mm; the height of the groove is greater than or equal to 2 mm and less than or equal to 15 mm; the glass slide is arranged in a circular shape, and the diameter of the glass slide is greater than the outer diameter of the trough body; the diameter of the small hole section is smaller than the diameter of the glass slide, and the diameter of the large hole section is larger than the diameter of the glass slide; the outer diameter of the trough body is smaller than the diameter of the small hole section.

[0008] Furthermore, the trough body and the glass slide are fixedly connected; the glass slide and the dish body are detachably connected.

[0009] Furthermore, the material of the trough body is chromium, aluminum, zinc or polytetrafluoroethylene; the material of the glass slide is glass; the material of the dish body is glass, polyethylene, polystyrene or polypropylene; the material of the dish cover is glass, polyethylene, polystyrene or polypropylene.

[0010] Furthermore, the trough body and the glass slide are connected by α-ethyl cyanoacrylate; the glass slide and the dish body are connected by polyurethane or acrylate copolymer.

[0011] The present invention also provides an interface microrheology detection method, which is implemented by using the above-mentioned interface microrheology detection device. The interface microrheology detection method includes the following steps: preparing a particle tracer solution; preparing a mixed solution of an interfacial active substance and fluorescein; adding a sample to the groove of the tank body in the order of "phase 1-particle tracer solution-phase 2"; the mixed solution is used as one of phase 1 and phase 2, and the other of phase 1 and phase 2 is oil, water or gas selected according to the interface type; wherein the phase with a large density is used as phase 1, and the phase with a small density is used as phase 2; using the "3D" scanning mode of an inverted laser confocal device, after determining the position of the interface layer, a field of view is selected for video recording; according to the recorded video, a statistical analysis is performed on the particle motion trajectory on the computer side to obtain the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution and non-Gaussian coefficient microrheology indicators.

[0012] Furthermore, the particle tracer is polystyrene fluorescent particles, and the diameter of the particles is 0.1-10 μm.

[0013] Furthermore, the surfactant is protein, polysaccharide or polyphenol; the fluorescein is Nile blue, calcium fluorescent white, fluorescein isothiocyanate or rhodamine B.

[0014] Furthermore, the surfactant is lecithin or fatty acid glyceride; and the fluorescein is Nile red.

[0015] Furthermore, the combination of "phase 1-phase 2" in "phase 1-particle tracer-phase 2" includes "water-oil", "water-water", "oil-oil", "water-gas" or "oil-gas".

[0016] The interfacial microrheology detection device provided by the present invention has a groove, the size of the groove is relatively small, and the solution of the small phase can be fixed, so that immiscible liquid-liquid or gas-liquid two phases coexist in a stable upper and lower stratified state, thereby ensuring the stability of the interface position during the measurement process. The interfacial active substance will be adsorbed, rearranged, and conformationally evolved on this stable interface, so that the interfacial microrheology detection can be carried out smoothly.

[0017] The interfacial microrheology detection method provided by the present invention can fluorescently label the interfacial active substances on the basis of obtaining a stable interfacial layer. The fluorescently labeled interfacial active substances are adsorbed on the interfacial layer to ensure that the detected area is the interfacial layer rather than the bulk phase. At the same time, the "3D" scanning mode of the inverted laser confocal device is used to accurately locate the interfacial layer, thereby realizing accurate particle tracing microrheology analysis of the interfacial layer.

[0018] It has been found that the viscosity of pure water measured by the interfacial microrheology detection method provided by the present invention is close to the theoretical value.

[0019] The present application provides a stable interface microrheology detection device suitable for in-situ interface detection. The present application also provides an interface microrheology detection method with precise interface recognition means, which uses a fluorescent labeling method to perform precise interface recognition. The interface microrheology detection device and method provided by the present invention are highly accurate, simple and easy to operate, highly operable, and widely applicable, and can be applied to a variety of interfacial active components and different types of interfaces.

[0020] Based on the implementations provided in the above aspects, this application can also be further combined to provide more implementations. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0022] Figure 1 A schematic diagram of the three-dimensional structure of an interfacial microrheology detection device provided in an optional embodiment of the present invention;

[0023] Figure 2 An overall cross-sectional view of an interfacial microrheology detection device after assembly provided by an optional embodiment of the present invention;

[0024] Figure 3 A cross-sectional view of the disassembled structure of an interfacial microrheology detection device provided in an optional embodiment of the present invention;

[0025] Figure 4 It is a flow chart of the interfacial microrheology detection method of the present invention;

[0026] Figure 5 The position of the interface layer formed by 0.05% soybean seed coat polysaccharide at the interface formation time of 5 minutes, 30 minutes and 60 minutes and the particle tracking motion trajectory in the specific embodiment 2 of the present invention;

[0027] Figure 6The microrheological results of the interface layer formed by 0.05% soybean seed coat polysaccharide at the interface for 5 minutes, 30 minutes, and 60 minutes in the specific embodiment 2 of the present invention are: mean square displacement (A), viscoelastic modulus (B), Gaussian distribution (C), and non-Gaussian coefficient (D);

[0028] Description of Figure Numbers:

[0029] 1. trough body; 10. groove; 2. glass slide; 3. dish body; 30. stepped hole; 301. small hole section; 302. large hole section; 303. connecting surface; 4. dish cover. DETAILED DESCRIPTION

[0030] In order to enable those skilled in the art to better understand the scheme of the present invention, the technical scheme in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only embodiments of a part of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0031] like Figures 1 to 3 As shown, the interfacial microrheology detection device provided by the present invention includes: a trough body 1, a glass slide 2, a dish body 3 and a dish cover 4, the glass slide 2 is connected to the bottom of the trough body 1, and the glass slide 2 and the trough body 1 together form a groove 10 for carrying the liquid to be tested; the dish body 3 has a accommodating cavity, and the bottom of the dish body 3 is provided with a stepped hole 30 connected to the accommodating cavity; along the direction from top to bottom, the stepped hole includes a small hole section 301 and a large hole section 302 that are connected, and the connection between the small hole section 301 and the large hole section 302 forms a connecting surface 303; the glass slide 2 is placed in the large hole section 302, the upper surface of the glass slide 2 is connected to the connecting surface 303, and at least part of the trough body 1 passes through the small hole section 301 and extends into the accommodating cavity; the dish cover 4 is covered on the dish body 3 to close the dish body 3. Among them, the groove 10 is used to bind and stabilize the liquid to be tested and the interface; the glass slide 2 is used to carry the trough body 1 and the liquid to be tested, as well as the penetration of the fluorescent signal; the dish body 3 is used to carry and transfer the glass slide; the dish cover 4 is used for the penetration of the excitation light beam and to prevent the evaporation of the liquid.

[0032] In this way, the interfacial microrheology detection device provided by the present invention has a groove 10, which can fix the solution of the small body phase, so that the immiscible liquid-liquid or gas-liquid phases coexist in a stable upper and lower stratified state, ensuring the stability of the interface position during the measurement process. The interfacial active substance will be adsorbed, rearranged, and conformationally evolved on this stable interface, so that the interfacial microrheology detection can be carried out smoothly.

[0033] Optionally, the groove body 1 is arranged in a circular ring shape; the diameter of the groove 10 is greater than or equal to 2 mm and less than or equal to 20 mm; the height of the groove 10 is greater than or equal to 2 mm and less than or equal to 15 mm. In this way, the preparation process of the groove body 1 is simplified, and it is also ensured that the formed groove 10 can form a stable small-volume interface layer without hindering the transmission of optical signals.

[0034] Optionally, the tank body 1 may also be configured to have other regular shapes or irregular shapes.

[0035] Alternatively, if Figure 1 and Figure 2 As shown, the glass slide 2 is arranged in a circular shape, and the diameter of the glass slide 2 is larger than the outer diameter of the tank body 1. In this way, the circular glass slide 2 has no sharp corners, which is safer during the replacement process, and the bottom of the tank body 1 can be completely closed by the glass slide 2 to form a complete groove 10 structure.

[0036] Optionally, the diameter of the small hole section 301 is smaller than the diameter of the glass slide 2, and the diameter of the large hole section 302 is smaller than the diameter of the glass slide 2. In this way, the large hole section 302 can completely accommodate the glass slide 2 and ensure that the upper surface of the glass slide 2 can be connected to the connecting surface 303.

[0037] Optionally, the outer diameter of the trough body 1 is smaller than the diameter of the small hole section 301 , so that the trough body 1 can smoothly pass through the small hole section 301 and extend into the accommodating cavity of the dish body 3 .

[0038] Optionally, the shape and size of the vessel cover 4 are adapted to the shape and size of the vessel body 3 .

[0039] Alternatively, if Figure 2 and Figure 3 As shown, the upper surface of the outer peripheral wall of the dish body 3 has a plurality of recesses arranged at intervals, and the lower surface of the outer peripheral wall of the dish cover has a plurality of convexities arranged at intervals, and the plurality of convexities are arranged in one-to-one correspondence with the plurality of recesses.

[0040] Optionally, the tank body 1 and the glass slide 2 are fixedly connected. In this way, the groove 10 can be completely sealed to ensure that the interface is stable and leak-free during the detection process. In addition, in order to ensure the accuracy of each detection process, the tank body 1 and the glass slide 2 are defined as consumables and are not reused.

[0041] Optionally, the glass slide 2 and the dish body 3 are detachably connected. Since there will be liquid residue on the used glass slide 2 and the trough body 1, they cannot be reused. In this way, the glass slide 2 and the dish body 3 are detachably connected, and the only consumable parts are the glass slide 2 and the trough body 1, so that the dish body 3 can be reused, reducing resource waste.

[0042] Optionally, the material of the tank body 1 is chromium, aluminum, zinc or polytetrafluoroethylene. The above materials have slightly weaker hydrophobicity, which can weaken the depression of the water phase boundary and the bulge of the oil phase boundary, and maximize the flatness of the liquid surface and the interface layer.

[0043] Optionally, the material of the tank body 1 may also be other materials according to actual needs.

[0044] Optionally, the slide 2 is made of glass, which has good light transmittance, thus ensuring that the fluorescent signals of all particle tracers in the field of view can be accurately captured.

[0045] Optionally, the material of the dish body 3 is glass, polyethylene, polystyrene or polypropylene. The above materials are transparent, which is convenient for observing the morphology of the inner cavity sample, easy to clean, long service life, and convenient for repeated use.

[0046] Optionally, the material of the dish cover 4 is glass, polyethylene, polystyrene or polypropylene. The above materials are transparent and easy to penetrate the excitation light beam. In particular, although the light transmittance of polyethylene, polystyrene or polypropylene is slightly lower than that of glass, since the dish cover 4, in addition to sealing the dish body 3, only serves as a conducting medium for the excitation signal, it does not interfere much with the capture of the fluorescent signal of the tracer particles, and thus it can still be used as an alternative material.

[0047] Optionally, the tank body 1 and the glass slide 2 are connected by ethyl α-cyanoacrylate. Ethyl α-cyanoacrylate has strong adhesiveness and is difficult to debond, thus ensuring that a long-term stable groove with good sealing performance is formed between the tank body 1 and the glass slide 2.

[0048] Optionally, the lower surface of the trough body 1 and a portion of the upper surface of the glass slide 2 are bonded together by ethyl α-cyanoacrylate.

[0049] Optionally, the glass slide 2 and the dish body 3 are connected by polyurethane or acrylate copolymers. Polyurethane or acrylate copolymers can be quickly bonded, but the mechanical strength is slightly poor, ensuring the disassembly of the glass slide 2 and the dish body 3 and the high efficiency during the replacement process.

[0050] Optionally, part of the upper surface of the glass slide 2 is bonded to the connecting surface 303 of the dish body 3 by polyurethane or acrylate copolymer.

[0051] When the glass slide 2 and the dish body 3 are adhered by polyurethane or acrylate copolymer, the mechanical strength of the adhesion is weak and can be easily separated after ultrasonic treatment. Therefore, the only loss parts are the glass slide and the trough body 1, and the dish body 3 can be reused to reduce resource waste. At the same time, the trough body 1 and the glass slide 2 are connected by α-ethyl cyanoacrylate, and the two will not be separated during ultrasonic treatment. After use, the trough body 1 and the glass slide 2 can be taken out of the dish body 3 as a whole and thrown away.

[0052] like Figure 4As shown, the present invention also provides an interface microrheology detection method, which includes the following steps: preparing a particle tracer solution; preparing a mixed solution of an interfacial active substance and fluorescein; adding a sample to the tank body 1 in the order of "phase 1-particle tracer solution-phase 2"; the mixed solution is used as one of the phases 1 and 2, and the other of the phases 1 and 2 is oil, water or gas selected according to the interface type; wherein the phase with a large density is used as the phase 1, and the phase with a small density is used as the phase 2; using the "3D" scanning mode of an inverted laser confocal device, after determining the position of the interface layer, a field of view is selected for video recording; according to the recorded video, a statistical analysis is performed on the particle motion trajectory on the computer side to obtain the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution and non-Gaussian coefficient microrheology indicators.

[0053] In this way, the interface microrheology detection method provided by the present invention can accurately locate the interface layer by fluorescently labeling the interface active component material and applying the "3D" scanning mode of the inverted laser confocal device on the basis of obtaining a stable interface layer, and realize accurate particle tracing microrheology analysis of the interface layer. The pure water viscosity measured by the test is close to the theoretical value. This method has high accuracy, simplicity, strong operability, and wide applicability, and can be applied to a variety of interface active components and different types of interfaces.

[0054] When selecting the bulk phase 1 and the bulk phase 2 , preferably, the oil is pure oil, the water is pure water, and the gas is air; when one of the bulk phase 1 or the bulk phase 2 is air, no substance is added into the groove 10 .

[0055] The mixed solution is determined as phase 1 or phase 2 according to the properties of the surfactant and the interface type; the properties of the surfactant include fat solubility and water solubility; the interface types include "water-oil", "water-water", "oil-oil", "water-gas" or "oil-gas".

[0056] For example, when the surfactant is a water-soluble substance and the interface type is an oil-water interface, the mixed solution serves as phase 1 and phase 2 is pure oil; when the surfactant is a fat-soluble substance and the interface type is an oil-water interface, the mixed solution serves as phase 2 and phase 1 is pure water; when the surfactant is a fat-soluble substance and the interface type is an oil-gas interface, the mixed solution serves as phase 1 and phase 2 is air.

[0057] Optionally, Python software and IDL algorithm are used on the computer side to calculate the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution and non-Gaussian coefficient microrheological indicators based on Langevin and generalized Stokes-Einstein equations.

[0058] Optionally, the calculation of mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution and non-Gaussian coefficient microrheological indexes is realized by algorithms using running program packages and scripts of multiple existing software on the computer side.

[0059] Optionally, the particle tracer is a polystyrene fluorescent particle with a diameter of 0.1-10 μm. In this way, it is possible to avoid a weak fluorescence signal due to a particle tracer with a diameter that is too small, or an interference with the self-assembly behavior of the interfacial active substance at the interface due to a particle tracer with a diameter that is too large.

[0060] Optionally, the surface active substance is protein, polysaccharide or polyphenol; the fluorescein is Nile blue, calcium fluorescent white, fluorescein isothiocyanate or rhodamine B. The above fluorescein is a water-soluble dye, which can quickly dye water-soluble proteins, polysaccharides or polyphenols.

[0061] Optionally, the surfactant is lecithin or fatty acid glyceride, and the fluorescein is Nile red. Nile red is a fat-soluble dye that can quickly stain lecithin or fatty acid glyceride.

[0062] Optionally, the combination of "bulk phase 1-bulk phase 2" in "bulk phase 1-particle tracer-bulk phase 2" includes "water-oil", "water-water", "oil-oil", "water-gas" or "oil-gas". The detection device and method can detect the microrheological properties of all "liquid-liquid" and "liquid-gas" interface systems, and has a wide range of applications.

[0063] The present invention provides an interface microrheology detection device and method for particle tracer microrheology detection on an interface. In an optional embodiment of the present invention, a sample detection table consisting of a thin glass slide 2 and an annular trough 1 is first prepared, and then a sample is added thereto in the order of "bulk phase 1-particle tracer solution-bulk phase 2". Since the interfacial active components have been previously labeled with specific fluorescent substances, the position of the interfacial layer can be determined in the "3D" scanning mode of the laser confocal microscope, and then the movement video of the tracer particles at this position within a certain period of time is recorded, and the local viscosity, viscoelastic modulus and non-uniformity coefficient of the interfacial layer where the tracer is located are calculated based on a customized algorithm of Langevin and generalized Stokes-Einstein equations to obtain the microrheological properties of the interface. The method is simple to operate and has high accuracy, and is suitable for detecting the microrheological evolution mechanism of interfacial active components such as surfactants, proteins, polysaccharides, etc. at oil / water, gas / water, and gas / oil interfaces.

[0064] The interface microrheology detection device provided by the present invention allows immiscible liquid-liquid or gas-liquid two phases to coexist in a stable upper and lower layered state by setting the groove 10, and the interfacial active substance will be adsorbed, rearranged, and conformationally evolved on this stable interface, so that the interface microrheology detection can be carried out smoothly. This device solves the problem that the traditional confocal small dish cannot fix the small phase solution through simple modification. In addition, in this device, the glass slide and the dish body are adhered by polyurethane or acrylate, and the mechanical strength of the adhesion is weak, and it can be easily separated after ultrasonic treatment. Therefore, the loss parts are only the glass slide 2 and the groove body 1, which can realize the reuse of the dish body and reduce resource waste.

[0065] The interface microrheology detection method provided by the present invention can accurately locate the interface layer by fluorescently labeling the interface active component material and applying the "3D" scanning mode of the inverted laser confocal device on the basis of obtaining a stable interface layer, and realize accurate particle tracing microrheology analysis of the interface layer. The pure water viscosity measured by the test is close to the theoretical value. This method has high accuracy, simplicity, strong operability, and wide applicability, and can be applied to a variety of interface active components and different types of interfaces.

[0066] Several specific embodiments of the present invention are introduced below:

[0067] Example 1

[0068] Example 1 provides an interfacial microrheology detection device.

[0069] like Figure 1 As shown, the present invention provides an interfacial microrheology detection device, comprising: a trough body 1, used to constrain and stabilize the liquid to be tested and the interface; a glass slide 2, connected to the bottom of the trough body 1, used to carry the trough body 1 and the liquid to be tested, and to allow the penetration of fluorescent signals; a dish body 3, the bottom of which is connected to the glass slide 2, used to carry and transfer the glass slide; and a dish cover 4, buckled on the dish body 3, used to allow the penetration of the excitation light beam and prevent the evaporation of the liquid.

[0070] Specifically, in this embodiment, the material of the tank body 1 is aluminum, the size of the aluminum ring is 5 mm in diameter and 3 mm in height, and the dish body 3 and the glass slide 2 are bonded by acrylic copolymer. The material of the dish body 3 and the dish cover 4 is polyethylene.

[0071] Furthermore, the connection between the dish body 3 and the glass slide 2 can be separated after 5 minutes of ultrasonic treatment at 60-100W, and the glass slide 2 can be replaced after bonding again using an acrylic copolymer.

[0072] Example 2

[0073] Example 2 provides an interfacial microrheology detection method applied to an interfacial microrheology detection device.

[0074] Combination Figure 4 and an interface microrheology detection device. This embodiment provides a method for detecting the microrheological properties of microgels formed by soybean seed coat polysaccharides at an oil / water interface, comprising:

[0075] S1, prepare particle tracer solution;

[0076] In this step, polystyrene fluorescent particles with a diameter of 1 μm and an excitation wavelength of 540 nm are used as particle tracers. Specifically, a 0.01% polystyrene fluorescent particle dispersion is prepared using a mixture of isopropanol and deionized water at a ratio of 3:7 (v / v) as a diluent.

[0077] S2, preparing a mixed solution of the surfactant and fluorescein;

[0078] In this step, soybean seed coat polysaccharide and Nile blue were selected as the tested interfacial active substances and their corresponding fluorescent substances, respectively. Specifically, a 0.05% soybean seed coat polysaccharide solution was first prepared, and then Nile blue was added to the soybean seed coat polysaccharide solution to a final concentration of 3 μg / mL, and incubated for 12 hours under dark conditions to achieve Nile blue staining of soybean seed coat polysaccharide.

[0079] S3, adding samples to the tank 1 in the order of "bulk phase 1-particle tracer solution-bulk phase 2";

[0080] In this step, the 0.05% soybean seed coat polysaccharide solution labeled with Nile blue prepared in step S2 is used as "bulk phase 1"; the 0.01% polystyrene fluorescent particle dispersion prepared in step S1 is used as "particle tracer solution"; and pure soybean oil is used as "bulk phase 2".

[0081] Furthermore, 15 μL of 0.05% soybean seed coat polysaccharide solution was first added to the bottom of the tank 1, and then 5 μL of 0.01% polystyrene fluorescent particle dispersion was added, and after standing for about 1-2 minutes, 20 μL of soybean oil was added.

[0082] S4, using the “3D” scanning mode of the inverted laser confocal device, determine the position of the interface layer and select a suitable field of view for video recording;

[0083] In this step, the 540nm and 633nm excitation light sources in the inverted laser confocal device were turned on, and the appropriate z-axis range was set under a 20x microscope for 3D scanning and shooting to find the position of the interface microgel layer. After observing the soybean seed coat polysaccharide interface layer, the focal plane height was adjusted to the interface, and the field of view containing 80-100 polystyrene fluorescent particles was selected, and a 71s video recording (270 frames of images) was performed.

[0084] Furthermore, in order to explore the evolution of the microgel at the soybean seed coat polysaccharide interface, microrheological tests were performed at 5 minutes, 30 minutes, and 60 minutes after the interface was formed. The position of the interface layer and the imaging of the particle tracer at the three time points are shown in Figure 2. Figure 5 Shown on the left.

[0085] S5, statistical analysis of particle motion trajectories was performed on the computer according to a customized algorithm to obtain the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution, and non-Gaussian coefficient microrheological indicators.

[0086] In this step, the tracers in the captured images are first identified, their trajectories are counted, and drift removed in Python software to obtain the motion trajectories of all particles in each video, such as Figure 5 Shown on the right.

[0087] Furthermore, Python software was used to calculate the overall mean square displacement of all particles in each video, such as Figure 6 As shown in A, compared with the bulk solution of soybean seed coat polysaccharide, soybean seed coat polysaccharide will be enriched at the interface to form an elastic network, resulting in the particle tracer being subjected to viscous force, causing the diffusion coefficient and slope to decrease. The diffusion coefficient and viscosity corresponding to each curve are shown in the following table, proving that soybean seed coat polysaccharide will complete the formation of a microgel network at the interface within 30 minutes. In addition, to verify this experimental method, the viscosity of pure water measured under this condition was compared with the theoretical value of standard viscosity, and it was found that the experimental viscosity result was 0.8007mPa.s, which is consistent with the theoretical value of standard viscosity of pure water at 30°C, proving the accuracy and credibility of this device and method.

[0088] Solution 5 minutes 30 minutes 60 minutes Diffusion coefficient (μm2 / s) 0.4763 0.3611 0.3407 0.3363 Viscosity (mPa.s) 0.9174 1.2100 1.2827 1.2995

[0089] Further, using the IDL algorithm Figure 6 The mean square displacement data obtained by A is converted into the viscoelastic modulus, as Figure 6 As shown in Figure B, G, G', G" represent the total modulus, elastic modulus and viscous modulus of the interface layer, respectively. It can be seen from the figure that although the modulus of the soybean seed coat polysaccharide interface layer will increase with the extension of the interface formation time. However, due to the low concentration of soybean seed coat polysaccharide under this condition, the curves of 30 minutes and 60 minutes are similar, and the elastic modulus of the interface layer at all times is less than the viscous modulus, which once again proves that 0.05% soybean seed coat polysaccharide will complete the formation of the microgel network at the interface within 30 minutes and show viscous properties.

[0090] Furthermore, the IDL algorithm was used to calculate the Gaussian distribution of the tracer during the movement of the interfacial microgel network ( Figure 6 C) and non-Gaussian coefficients ( Figure 6D), analysis of the two images shows that the microgel formed by soybean seed coat polysaccharide at the interface is dynamically heterogeneous, and under this concentration condition, as the interface formation time increases, the size of the interface microgel network becomes narrower.

[0091] Example 3

[0092] Example 3 provides an interfacial microrheology detection method applied to an interfacial microrheology detection device.

[0093] Combination Figure 4 and an interface microrheology detection device. This embodiment provides a method for detecting the microrheological properties of microgels formed by soybean seed coat polysaccharides at an oil / water interface, comprising:

[0094] S1, prepare particle tracer solution;

[0095] In this step, polystyrene fluorescent particles with a diameter of 1 μm and an excitation wavelength of 540 nm are used as particle tracers. Specifically, a 0.01% polystyrene fluorescent particle dispersion is prepared using a mixture of isopropanol and deionized water at a ratio of 3:7 (v / v) as a diluent.

[0096] S2, preparing a mixed solution of the surfactant and fluorescein;

[0097] In this step, soybean seed coat polysaccharide and Nile blue were selected as the tested interfacial active substances and their corresponding fluorescent substances, respectively. Specifically, a 0.75% soybean seed coat polysaccharide solution was first prepared, and then Nile blue was added to the soybean seed coat polysaccharide solution to a final concentration of 3 μg / mL, and incubated for 12 hours in the dark to achieve Nile blue staining of soybean seed coat polysaccharide.

[0098] S3, adding samples to the tank 1 in the order of "bulk phase 1-particle tracer solution-bulk phase 2";

[0099] In this step, the 0.75% soybean seed coat polysaccharide solution labeled with Nile blue prepared in step S2 is used as "bulk phase 1"; the 0.01% polystyrene fluorescent particle dispersion prepared in step S1 is used as "particle tracer solution"; and pure soybean oil is used as "bulk phase 2".

[0100] Furthermore, 15 μL of 0.75% soybean seed coat polysaccharide solution was first added to the bottom of the tank 1, and then 5 μL of 0.01% polystyrene fluorescent particle dispersion was added, and after standing for about 1-2 minutes, 20 μL of soybean oil was added.

[0101] S4, using the “3D” scanning mode of the inverted laser confocal device, determine the position of the interface layer and select a suitable field of view for video recording;

[0102] In this step, the 540nm and 633nm excitation light sources in the inverted laser confocal device were turned on, and the appropriate z-axis range was set under a 20x microscope for 3D scanning and shooting to find the position of the interface microgel layer. After observing the soybean seed coat polysaccharide interface layer, the focal plane height was adjusted to the interface, and the field of view containing 80-100 polystyrene fluorescent particles was selected, and a 71s video recording (270 frames of images) was performed.

[0103] Furthermore, in order to explore the evolution process of soybean seed coat polysaccharide interface microgel, microrheological tests were performed at 5 minutes, 30 minutes, and 60 minutes after the interface was formed.

[0104] S5, statistical analysis of particle motion trajectories was performed on the computer according to a customized algorithm to obtain the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution, and non-Gaussian coefficient microrheological indicators.

[0105] In this step, the tracers in the captured images were first identified, their trajectories were counted, and their drifts were removed in Python software to obtain the motion trajectories of all particles in each video.

[0106] Furthermore, Python software was used to calculate the overall mean square displacement of all particles in each video. The diffusion coefficients and viscosities corresponding to the three time points are shown in the following table. It can be clearly seen that compared with Example 2, the microgel network viscosity formed by soybean seed coat polysaccharide at this concentration at the oil / water interface is higher and positively correlated with the formation time.

[0107] Solution 5 minutes 30 minutes 60 minutes Diffusion coefficient (μm2 / s) 0.1900 0.1766 0.1352 0.1016 Viscosity (mPa.s) 2.3001 2.4745 3.2323 4.3017

[0108] Furthermore, the obtained mean square displacement data were converted into viscoelastic modulus using the IDL algorithm.

[0109] Furthermore, the IDL algorithm was used to calculate the Gaussian distribution of the tracer during the movement of the interfacial microgel network.

[0110] Example 4

[0111] Example 4 provides an interfacial microrheology detection method applied to an interfacial microrheology detection device.

[0112] Combination Figure 4 and an interface microrheology detection device. This embodiment provides a method for detecting the microrheological properties of microgels formed by soy protein isolate at an oil / water interface, comprising:

[0113] S1, prepare particle tracer solution;

[0114] In this step, polystyrene fluorescent particles with a diameter of 1 μm and an excitation wavelength of 540 nm are used as particle tracers. Specifically, a 0.01% polystyrene fluorescent particle dispersion is prepared using a mixture of isopropanol and deionized water at a ratio of 3:7 (v / v) as a diluent.

[0115] S2, preparing a mixed solution of the surfactant and fluorescein;

[0116] In this step, soy protein isolate and Nile blue were selected as the tested interfacial active substances and their corresponding fluorescent substances, respectively. Specifically, a 0.20% soy protein isolate solution was prepared, and then Nile blue was added to the soy protein isolate solution to a final concentration of 3 μg / mL, and incubated for 12 hours in the dark to achieve Nile blue staining of the soy protein isolate.

[0117] S3, adding samples to the tank 1 in the order of "bulk phase 1-particle tracer solution-bulk phase 2";

[0118] In this step, the 0.20% soy protein isolate solution labeled with Nile blue prepared in step S2 is used as "bulk phase 1"; the 0.01% polystyrene fluorescent particle dispersion prepared in step S1 is used as "particle tracer solution"; and pure soybean oil is used as "bulk phase 2".

[0119] Furthermore, 15 μL of 0.20% soy protein isolate solution was first added to the bottom of the tank 1, and then 5 μL of 0.01% polystyrene fluorescent particle dispersion was added, and after standing for about 1-2 minutes, 20 μL of soybean oil was added.

[0120] S4, using the “3D” scanning mode of the inverted laser confocal device, determine the position of the interface layer and select a suitable field of view for video recording;

[0121] In this step, the 540nm and 633nm excitation light sources in the inverted laser confocal device were turned on, and the appropriate z-axis range was set under a 20x microscope for 3D scanning and shooting to find the position of the interface microgel layer. After observing the soy protein isolate interface layer, the focal plane height was adjusted to the interface, and the field of view containing 80-100 polystyrene fluorescent particles was selected, and a 71s video recording (270 frames of images) was performed.

[0122] S5, statistical analysis of particle motion trajectories was performed on the computer according to a customized algorithm to obtain the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution, and non-Gaussian coefficient microrheological indicators.

[0123] In this step, the tracers in the captured images were first identified, their trajectories were counted, and their drifts were removed in Python software to obtain the motion trajectories of all particles in each video.

[0124] Furthermore, Python software was used to calculate the overall mean square displacement of all particles in each video.

[0125] Furthermore, the obtained mean square displacement data were converted into viscoelastic modulus using the IDL algorithm.

[0126] Furthermore, the IDL algorithm was used to calculate the Gaussian distribution of the tracer during the movement of the interfacial microgel network.

[0127] Example 5

[0128] Example 5 provides an interfacial microrheology detection method applied to an interfacial microrheology detection device.

[0129] Combination Figure 4 and an interface microrheology detection device. This embodiment provides a method for detecting the microrheological properties of lecithin microgels formed at an oil / gas interface, comprising:

[0130] S1, prepare particle tracer solution;

[0131] In this step, polystyrene fluorescent particles with a diameter of 0.5 μm and an excitation wavelength of 540 nm are used as particle tracers. Specifically, a 0.01% polystyrene fluorescent particle dispersion is prepared using a mixture of isopropanol and deionized water at a ratio of 3:7 (v / v) as a diluent.

[0132] S2, preparing a mixed solution of the surfactant and fluorescein;

[0133] In this step, lecithin and Nile red were selected as the tested surfactants and their corresponding fluorescein, respectively. Specifically, a 0.20% lecithin soybean oil solution was prepared using soybean oil as a solvent, and then Nile red was added to the lecithin soybean oil solution to a final concentration of 3 μg / mL, and incubated for 12 hours in the dark to achieve Nile red staining of lecithin.

[0134] S3, adding samples to the tank 1 in the order of "bulk phase 1-particle tracer solution-bulk phase 2";

[0135] In this step, the 0.20% lecithin soybean oil solution labeled with Nile red prepared in step S2 is used as "bulk phase 1"; the 0.01% polystyrene fluorescent particle dispersion prepared in step S1 is used as "particle tracer solution"; and air is used as "bulk phase 2".

[0136] Furthermore, 15 μL of 0.20% lecithin soybean oil solution was first added to the bottom of the tank 1, and then 5 μL of 0.01% polystyrene fluorescent particle dispersion was added and allowed to stand for about 1-2 minutes.

[0137] S4, using the “3D” scanning mode of the inverted laser confocal device, determine the position of the interface layer and select a suitable field of view for video recording;

[0138] In this step, turn on the 540nm and 488nm excitation light sources in the inverted laser confocal device, set the appropriate z-axis range under the 20x microscope for 3D scanning and shooting, and find the position of the interface microgel layer. After observing the lecithin interface layer, adjust the focal plane height to the interface, select the field of view containing 80-100 polystyrene fluorescent particles, and record a 71s video (270 frames of images).

[0139] S5, statistical analysis of particle motion trajectories was performed on the computer according to a customized algorithm to obtain the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution, and non-Gaussian coefficient microrheological indicators.

[0140] In this step, the tracers in the captured images were first identified, their trajectories were counted, and their drifts were removed in Python software to obtain the motion trajectories of all particles in each video.

[0141] Furthermore, Python software was used to calculate the overall mean square displacement of all particles in each video.

[0142] Furthermore, the obtained mean square displacement data were converted into viscoelastic modulus using the IDL algorithm.

[0143] Furthermore, the IDL algorithm was used to calculate the Gaussian distribution of the tracer during the movement of the interfacial microgel network.

[0144] Examples 2 and 3 demonstrate that this detection method can be used to explore the effects of concentration and interface formation time on the microrheological properties of polysaccharide microgels formed at the oil / water interface; Example 4 introduces the specific steps of this detection method in exploring the microrheological properties of protein microgels formed at the oil / water interface; Example 5 introduces the specific steps of this detection method in exploring the microrheological properties of fat-soluble lecithin microgels formed at the oil / air interface. The above examples comprehensively demonstrate the feasibility of this scheme in exploring the microrheological properties of water-soluble or fat-soluble surfactants forming microgels at the oil / water interface or oil / air interface.

[0145] In a specific embodiment, the tank body 1 may also be made of chromium, zinc or polytetrafluoroethylene, and the surfactant may also be some other polysaccharides, proteins, lecithin, fatty acid glycerides, and the measurable interface types include any one of "water-oil", "water-water", "oil-oil", "water-gas" and "oil-gas", and examples are not given one by one here.

[0146] In combination with the above specific embodiments, it can be seen that the interface microrheology detection device and method provided by the present invention can achieve accurate particle tracing microrheology analysis of the interface layer on the basis of obtaining a stable interface layer. This method has high accuracy, is simple and easy to operate, has strong operability, and is widely applicable, and can be applied to a variety of interfacial active components and different types of interfaces.

[0147] The above are only specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art who is familiar with the technical field can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

[0148] Unless otherwise defined, the technical terms or scientific terms used in the present disclosure should be understood by people with ordinary skills in the field to which the present disclosure belongs. The "first", "second" and similar words used in the present disclosure do not indicate any order, quantity or importance, but are only used to distinguish different components. Similarly, similar words such as "one", "one" or "the" do not indicate quantity restrictions, but indicate that there is at least one. Similar words such as "include" or "comprise" mean that the elements or objects appearing before the word cover the elements or objects listed after the word and their equivalents, without excluding other elements or objects. Similar words such as "connect" or "connected" are not limited to physical or mechanical connections, but can include electrical connections, whether direct or indirect. "Up", "down", "left", "right" and the like are only used to indicate relative positional relationships. When the absolute position of the described object changes, the relative positional relationship may also change accordingly.

Claims

1. An interfacial microrheology detection device, characterized in that: include: Tank body (1); A glass slide (2), the glass slide (2) being connected to the bottom of the tank body (1), the glass slide (2) and the tank body (1) together forming a groove (10) for carrying the liquid to be tested; A dish body (3), the dish body (3) having a receiving cavity, the bottom of the dish body (3) being provided with a stepped hole (30) communicating with the receiving cavity; along a direction from top to bottom, the stepped hole comprises a small hole section (301) and a large hole section (302) communicating with each other, the connection between the small hole section (301) and the large hole section (302) forming a connecting surface (303); the glass slide (2) is placed in the large hole section (302), the upper surface of the glass slide (2) is connected to the connecting surface (303), and at least a portion of the trough body (1) passes through the small hole section (301) and extends into the receiving cavity; A dish cover (4), the dish cover (4) being arranged on the dish body (3); The tank body (1) and the glass slide (2) are connected via ethyl α-cyanoacrylate; The glass slide (2) and the dish body (3) are connected via polyurethane or acrylate copolymer.

2. The interfacial microrheology detection device according to claim 1, characterized in that: The tank body (1) is arranged in a circular ring shape; The diameter of the groove (10) is greater than or equal to 2 mm and less than or equal to 20 mm; The height of the groove (10) is greater than or equal to 2 mm and less than or equal to 15 mm; The glass slide (2) is arranged in a circular shape, and the diameter of the glass slide (2) is larger than the outer diameter of the tank body (1); The diameter of the small hole section (301) is smaller than the diameter of the glass slide (2), and the diameter of the large hole section (302) is larger than the diameter of the glass slide (2); The outer diameter of the groove body (1) is smaller than the diameter of the small hole section (301).

3. The interfacial microrheology detection device according to claim 1, characterized in that: The tank body (1) and the glass slide (2) are fixedly connected; The glass slide (2) and the dish body (3) are detachably connected.

4. The interfacial microrheology detection device according to claim 1, characterized in that: The material of the tank body (1) is chromium, aluminum, zinc or polytetrafluoroethylene; The material of the glass slide (2) is glass; The material of the dish body (3) is glass, polyethylene, polystyrene or polypropylene; The material of the dish cover (4) is glass, polyethylene, polystyrene or polypropylene.

5. An interfacial microrheology detection method, characterized in that: The interface microrheology detection method is implemented by using the interface microrheology detection device described in any one of claims 1 to 4, and the interface microrheology detection method comprises the following steps: Prepare particle tracer solution; preparing a mixed solution of a surfactant and fluorescein; Adding the sample into the groove (10) of the tank body (1) in the order of "bulk phase 1-particle tracer solution-bulk phase 2"; The mixed solution is used as one of the phase 1 and the phase 2, and the other of the phase 1 and the phase 2 is oil, water or gas selected according to the interface type; wherein the phase with a larger density is used as the phase 1, and the phase with a smaller density is used as the phase 2; Using the "3D" scanning mode of the inverted laser confocal device, determine the position of the interface layer and select the field of view for video recording; The particle motion trajectory is statistically analyzed on the computer based on the recorded video to obtain the mean square displacement, diffusion coefficient, viscosity, viscoelastic modulus, Gaussian distribution and non-Gaussian coefficient microrheological indicators.

6. The interfacial microrheology detection method according to claim 5, characterized in that: The particle tracer is polystyrene fluorescent particles, and the diameter of the particles is 0.1-10 μm.

7. The interfacial microrheology detection method according to claim 5, characterized in that: The surface active substance is protein, polysaccharide or polyphenol; The fluorescein is Nile blue, calcifluor white, fluorescein isothiocyanate or rhodamine B.

8. The interfacial microrheology detection method according to claim 5, characterized in that: The surface active substance is lecithin or fatty acid glyceride; The fluorescein is Nile red.

9. The interfacial microrheology detection method according to claim 5, characterized in that: The combination of "phase 1-phase 2" in "phase 1-particle tracer-phase 2" includes "water-oil", "water-water", "oil-oil", "water-gas" or "oil-gas".

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