CN109550528B - Multi-angle nondestructive observation method and device for internal channel of micro-fluidic chip - Google Patents

Multi-angle nondestructive observation method and device for internal channel of micro-fluidic chip Download PDF

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CN109550528B
CN109550528B CN201811534541.7A CN201811534541A CN109550528B CN 109550528 B CN109550528 B CN 109550528B CN 201811534541 A CN201811534541 A CN 201811534541A CN 109550528 B CN109550528 B CN 109550528B
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microfluidic chip
stainless steel
micro
internal channel
clamping device
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CN109550528A (en
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刘赵淼
王飓
逄燕
李梦麒
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Beijing University of Technology
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
    • G01B11/00—Measuring arrangements characterised by the use of optical techniques
    • G01B11/24—Measuring arrangements characterised by the use of optical techniques for measuring contours or curvatures
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/84—Systems specially adapted for particular applications
    • G01N21/88—Investigating the presence of flaws or contamination

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Abstract

The invention discloses a multi-angle nondestructive observation method and a device for an internal channel of a microfluidic chip, wherein the first part is a split type clamping device and consists of a left module and a right module; the second part is a main container which is provided with a split type clamping device and is used for containing liquid. In order to ensure that most laboratories only provided with a conventional optical microscope can observe the appearance of the internal channel of the microfluidic chip from multiple angles on the premise of not cutting and damaging the microfluidic chip, and the internal channel can be observed in a planar manner only through the upper side and the lower side of the top and the bottom. The morphological characteristics of the internal channel of the microfluidic chip can be observed through the cut rough plane on the premise that expensive equipment such as a confocal microscope and the like is not used, and the functions of the microfluidic chip are not damaged by cutting the microfluidic chip; the observation of multiple angles of the internal channel of the microfluidic chip can be realized, and a three-dimensional and comprehensive geometric image of the processed planar or non-planar microchannel can be obtained.

Description

Multi-angle nondestructive observation method and device for internal channel of micro-fluidic chip
Technical Field
The invention relates to a multi-angle nondestructive observation method and device for an internal channel of a microfluidic chip. The invention belongs to the technical field of microfluidics.
Background
Minute volume (10) for a long time-15～10-9L) the liquid, i.e. the micro-droplets, plays an important role in the industrial fields of petrochemical industry, food additives, pharmaceutical manufacturing, cosmetic preparation and the like. However, the traditional Top-down (Top-down) preparation method has difficulty in accurately controlling the components, sizes and fluxes of the micro-droplets, and limits the further development of micro-droplet-based engineering applications[1]. In recent years, a new method for continuously and rapidly generating micro-droplets with uniform size and controllable components is provided by a rapidly developed droplet microfluidic technology, and water-in-oil micro-droplets are formed mainly based on the interfacial instability effect between immiscible discrete phase and continuous phase fluids. The micro-droplets have the outstanding characteristics of small volume (which can be reduced to the picoliter to nanoliter magnitude), high flux (the generation frequency is up to thousands of hertz), high heat and mass transfer efficiency, no cross contamination among the micro-droplets, stable internal environment and the like, and are suitable for peopleThe method has wide application prospect in the biochemical fields of body tissue reconstruction, drug assembly, multiple emulsification, micro-nano material synthesis, DNA analysis and the like and the information processing field.
The existing related researches show that the micro-fluidic chip with the non-planar micro-channel characteristics is beneficial to the monodispersion generation of micro-droplets, the micro-droplets prepared by the method have the characteristic that the size is insensitive to the flow fluctuation caused by an injection device, and the generation method has been widely concerned in scientific research and industry at present. In the aspect of materials, the PDMS material has been widely applied to the preparation of microfluidic chips due to the advantages of low cost, simple preparation, good light transmittance and the like.
However, on the one hand, due to unavoidable machining errors during machining, variations in the channel dimensions that are difficult to estimate may be caused; on the other hand, since the fabrication process of the non-planar channel requires necessary manual operations, there is some uncertainty of the geometry, such as the application number: 201510379969.9. In order to correlate the correlation between geometry and performance of microfluidic chips, it is necessary to verify the specific dimensions of the channels used in the experiment by actual measurement. However, the PDMS channels have a cutting step in the manufacturing process, which results in a large roughness of the cut portion of the sidewall surface, and under the condition of not cutting the micro-channels, most of conventional optical microscopes in laboratories have difficulty in observing the shape and size of the specific channels through rough planes, and can only perform planar observation through smooth surfaces on the upper and lower sides, but cannot observe other planes from other angles, which causes great inconvenience in use in scientific research and production.
Disclosure of Invention
Based on the technical defects mentioned in the background technology, the invention aims to enable most laboratories only provided with a conventional optical microscope to observe the appearance of the internal channel of the microfluidic chip from multiple angles on the premise of not cutting and damaging the microfluidic chip, and not only can carry out planar observation through the upper side and the lower side of the top and the bottom. The invention provides a multi-angle nondestructive observation method and a multi-angle nondestructive observation device for an internal channel of a microfluidic chip.
In order to achieve the above object, the present invention provides a clamping device capable of rotating a microfluidic chip at multiple angles, which can help the method provided by the present invention to be better implemented. This device mainly is formed by two parts equipment, and its characteristic includes:
the first part is split type clamping device for the centre gripping and the rotation of micro-fluidic chip, and preferred, the first part comprises left side module and right side module, and the left side module includes left threaded rod 1, left side cylinder cover 2, and the long stainless steel of left side presss from both sides 3, and the right side module includes the short stainless steel of right side presss from both sides 4, right side cylinder cover 5 and right threaded rod 6. Further, in the left module, the main body part of the left cylindrical sleeve 2 is an acrylic cylindrical rod, a small hole is axially formed in the acrylic cylindrical rod and used for leading in a left threaded rod 1, and a part of the outer wall surface of the left threaded rod 1 is cut into a plane and is attached to the plane of the middle section of the left long stainless steel clamp 3; the left long stainless steel clamp 3 is a three-section metal sheet made of stainless steel, and round heads are arranged at the tail ends of two ends of the left long stainless steel clamp 3 and used for clamping a microfluidic chip. The structure of right side module and left side module is similar mutually, and the main part of right side cylinder cover 5 is ya keli cylinder pole, and inside axial has the aperture to be used for letting in right threaded rod 6, and the outer wall has partly to cut for the plane be used for with the laminating of the interlude plane of the short stainless steel clamp 4 in right side, and the short stainless steel clamp 4 in right side is the syllogic sheetmetal of being made by the stainless steel, and the both ends end that the short stainless steel clamp 4 in right side has the button head to be used for pressing from both sides and get micro-fluidic chip.
The second part is a main container which is provided with a split type clamping device and is used for containing liquid. The main body container comprises an acrylic transparent cavity 7. The left module and the right module are symmetrically arranged on two sides of the transparent acrylic cavity 7; particularly, four angles of transparent appearance chamber 7 of ya keli all have protruding stand, and the upper portion of protruding stand all has the aperture, and the aperture is used for with assembly left threaded rod 1 or right threaded rod 6. The bottom of the transparent acrylic cavity 7 has good smoothness and flatness due to the use of acrylic materials, so that the subsequent optical observation of the microfluidic chip inside the transparent acrylic cavity is facilitated.
The three-section metal sheet refers to a clamping device structure consisting of three planes.
In addition, the invention also provides a method for better implementing multi-angle nondestructive observation of the internal channel of the microfluidic chip by virtue of the clamping device capable of realizing multi-angle rotation of the microfluidic chip, which is characterized by comprising the following steps:
(1) preferably, the material of the microfluidic chip is observed by using a refractometer, so that the refractive index corresponding to the material used by the microfluidic chip to be observed is obtained.
(2) Preferably, glycerol aqueous solutions with different concentration ratios are prepared, and refractive indexes corresponding to the glycerol aqueous solutions with different concentrations are respectively measured by using a refractometer, so as to obtain the concentration value of the group of glycerol aqueous solutions closest to the refractive index of the microfluidic chip material. The concentration value is adjusted by properly adding water or glycerol, so that the refractive index of the glycerol aqueous solution is consistent with that of the material of the microfluidic chip, and the concentration value of the glycerol aqueous solution at the moment is recorded.
(3) Preferably, the outlet and inlet of the microfluidic chip to be observed are connected with a conduit to prevent the liquid drops from infiltrating into the channel. And fixing the micro-fluidic chip to be observed by using a split type clamping device, and adjusting the angle to be observed.
(4) Preferably, the split type clamping device and the microfluidic chip are placed on an object stage of an optical microscope, a glycerol aqueous solution with a prepared concentration is poured into a main container of the clamping device for realizing multi-angle rotation of the microfluidic chip, and the microfluidic chip is immersed.
(5) Preferably, the optical microscope is used for observing the internal channel of the microfluidic chip to be measured to obtain an experimental image.
Compared with the prior art, the invention has the following beneficial effects.
1. The invention provides a multi-angle nondestructive observation method and device for an internal channel of a microfluidic chip. By using the method, the morphological characteristics of the internal channel of the microfluidic chip can be observed through the cut rough plane without using expensive equipment such as a confocal microscope and the like and without cutting the microfluidic chip to destroy the functions of the microfluidic chip;
2. the invention provides a multi-angle nondestructive observation method and device for an internal channel of a microfluidic chip. The method and the device are simple, convenient and quick to implement. The method and the device can realize the observation of multiple angles of the internal channel of the microfluidic chip, obtain the three-dimensional and comprehensive geometric morphology image of the processed planar or non-planar microchannel, and help the professional scientific research personnel and engineers verify and evaluate the performance corresponding to the geometric dimension of the processed microchannel;
drawings
In order to more clearly illustrate the technical solution in the implementation of the present invention, the following briefly describes the attached drawings that are needed and applied in the embodiments. It is clear that the described figures represent only a part of an embodiment of the invention and not all embodiments, and that a person skilled in the pertinent art will be able to derive other design solutions and design solution figures of similar characteristics from these figures without inventive effort.
FIG. 1a is a left module of a split type clamping device provided in the multi-angle nondestructive observation method and device for the internal channel of the microfluidic chip according to the present invention;
FIG. 1b is a right module of a split type clamping device provided in the multi-angle nondestructive observation method and device for the internal channel of the microfluidic chip according to the present invention;
FIG. 2 is a main container part provided in the multi-angle nondestructive observation method and apparatus for internal channels of microfluidic chips according to the present invention;
FIG. 3 is a multi-angle nondestructive observation method and apparatus for internal channels of microfluidic chips according to the present invention, which is a clamping apparatus capable of multi-angle rotation of microfluidic chips and provided by a split type clamping apparatus and a main container;
FIG. 4 is a schematic diagram of a method and apparatus for multi-angle nondestructive observation of internal channels of a microfluidic chip according to an embodiment of the present invention, in which a solution having a refractive index matching the refractive index of the microfluidic chip is used to immerse the microfluidic chip so as to allow a microscope to clearly observe the microchannel;
FIG. 5a is a side view of a micro-channel with a bottom of a cambered surface obtained by an inverted optical microscope and a camera without using the method of the present invention in the embodiment of the multi-angle nondestructive observation method and apparatus for an internal channel of a microfluidic chip of the present invention.
FIG. 5b is a side view of a micro-channel with a bottom of a cambered surface obtained by an inverted optical microscope and a camera using the method of the present invention in an embodiment of the multi-angle nondestructive observation method and apparatus for an internal channel of a microfluidic chip of the present invention.
Detailed Description
The idea, specific results and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments and the accompanying drawings to fully understand the objects, features and effects of the present invention. It is to be understood that the drawings illustrate only some embodiments of the invention and not all embodiments of the invention, and that other embodiments may be devised by those skilled in the art without the use of the inventive faculty, and that the invention may be embodied in other specific forms and arrangements of similar features. All technical features in the invention can be interactively combined with other inventive features on the premise of not conflicting with each other.
In order to describe the method of the present invention in more detail, so that one skilled in the art can easily implement the method and obtain the effects of the present invention, the figures mentioned in the following examples have been enlarged with necessary characteristic details, and it is obvious that the sizes of the actual microfluidic chip product are very small.
(1) Manufacturing a PDMS micro-fluidic chip with a micro-channel at the bottom of the cambered surface:
step (1) is referred to application number: 201510379969.9, a PDMS microfluidic chip with a micro-channel at the bottom of a cambered surface is manufactured. Specifically, a PDMS (polydimethylsiloxane) main agent and a coagulant are uniformly mixed according to a ratio of 10:1, then the reagent is placed in a normal-temperature vacuum environment for about 40-60 minutes until all bubbles are separated out, a part of the reagent is reserved for standby, the rest is poured onto a silicon chip containing a T-shaped micro-channel male die and a square male die, and the silicon chip is placed in an oven at the temperature of 80 ℃ for about 1 hour to be solidified. And after the PDMS is solidified, the PDMS is taken off from the silicon wafer template, and a main body part of the microfluidic chip with a complete T-shaped microchannel structure and a bottom plate part with a square groove are cut. And respectively punching a through hole at the outlet and inlet position of the main body part of the micro-fluidic chip with the complete T-shaped micro-channel structure and two opposite angles of the bottom plate with the square groove.
Putting a clean blank silicon wafer on a centrifugal spin coater, pouring liquid PDMS in a well-proportioned ratio into the center of the silicon wafer, starting the spin coater, throwing a PDMS reagent to form a liquid film attached to the silicon wafer, and putting the silicon wafer with a liquid film in an oven to solidify the PDMS film to form a solid elastic film.
And punching the inlet and outlet of the main body part of the PDMS microfluidic chip containing the T-shaped microchannel groove by using a puncher. And (3) processing the surface containing the channel structure in the main body part of the chip and the film surface on the silicon wafer for 3-5 seconds by using a corona machine processor, and then bonding the surface and the film surface. The bonded silicon wafer (with the chip body structure thereon) was placed on a hot plate at 80 ℃ and heated for about 15 minutes. And lightly scratching the edge of the chip main body structure by using a blade, and taking down the chip main body structure to obtain the PDMS micro-channel containing the single-sided film. And (3) processing the other surface of the film by using a corona machine, bonding the other surface of the film on the PDMS bottom plate part with the square groove, placing the whole bonded microfluidic chip on a hot plate at the temperature of 80 ℃, and heating for 20 minutes.
Slowly injecting the liquid PDMS with the prepared proportion from the punched small hole of the bottom plate with the square groove, so that the film in contact with the groove bulges and deforms towards one side of the groove of the micro-channel, slowly extracting a part of PDMS after the PDMS is filled, so that the film on the bottom surface of the micro-channel inclines towards one side of the square groove, placing the micro-channel on a heating plate after the micro-channel is stabilized, and heating the micro-channel at 80 ℃ for 15 minutes to solidify the liquid PDMS, thereby obtaining the PDMS micro-fluidic chip with the micro-channel with the bottom of the cambered surface.
(2) Preparation of glycerol aqueous solution with refractive index matched with PDMS:
uniformly mixing a PDMS (polydimethylsiloxane) main agent and a coagulant according to a ratio of 10:1, and then placing the reagent in a normal-temperature vacuum environment for about 40-60 minutes until all bubbles are separated out. The refractive index was measured by pouring the liquid PDMS in the prepared ratio on a refractometer to obtain a refractive index value of about 1.405. Preparing glycerol aqueous solutions with different concentration ratios, respectively measuring the corresponding refractive indexes of the glycerol aqueous solutions with different concentrations by using a refractometer to obtain the concentration value of the glycerol aqueous solution group closest to the refractive index of the microfluidic chip material, adjusting the concentration value by properly adding water or glycerol to ensure that the refractive index of the glycerol aqueous solution is close to 1.405, and recording the concentration value of the glycerol aqueous solution at the moment, wherein the concentration value is about 55% w/w.
(3) Side-view shooting of PDMS micro-fluidic chip with micro-channel at bottom of cambered surface:
since the fabrication of the arc-shaped bottom surface of the microchannel requires a part of manual operation, it is difficult to obtain the quantified geometrical size of the arc surface without observing it. Therefore, the clamping device which is shown in fig. 3 and can realize multi-angle rotation of the microfluidic chip is placed on an object stage of a conventional optical inverted microscope by observing the clamping device through the method provided by the invention, and the manufactured PDMS microfluidic chip with the micro-channel at the bottom of the cambered surface is fixed by using the split type clamping device shown in fig. 1a and 1b, so that the side wall surface of the PDMS microfluidic chip is parallel to the object stage, and the cambered surface characteristic of the PDMS microfluidic chip can be observed from the side surface.
And (3) adjusting the focal length and shooting a conventional optical inverted microscope to obtain a side view image which is shown in figure 5a and is obtained by shooting the microchannel with the cambered bottom directly through the rough cutting surface without using the method provided by the invention. And connecting the outlet and the inlet of the micro-fluidic chip to be observed with a conduit to prevent the liquid drops from infiltrating the inside of the channel. Leading 55% w/w glycerol aqueous solution into the main container shown in FIG. 2, the immersed part of the PDMS microfluidic chip with the micro-channel at the bottom of the arc surface is nearly transparent in 55% w/w glycerol aqueous solution due to the refractive index, and continuing to add 55% w/w glycerol aqueous solution and immersing the microfluidic chip as shown in FIG. 4. Focusing and shooting are carried out on a conventional optical inverted microscope, and a side-view image with a cambered bottom micro-channel obtained by shooting by using the method provided by the invention is obtained as shown in figure 5 b.
The preferred embodiments of the present invention have been specifically described above. It should be emphasized that the invention is not limited to the embodiments described, and that those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the invention, and those having similar features are included in the scope defined by the claims of the present application.

Claims (3)

1. The utility model provides a multi-angle nondestructive observation device of micro-fluidic chip inner channel which characterized in that: the device is mainly assembled by two parts;
the first part is a split type clamping device and is used for clamping and rotating the microfluidic chip, the first part consists of a left side module and a right side module, the left side module comprises a left threaded rod (1), a left side cylindrical sleeve (2) and a left side long stainless steel clamp (3), and the right side module comprises a right side short stainless steel clamp (4), a right side cylindrical sleeve (5) and a right threaded rod (6); in the left side module, the main body part of the left side cylindrical sleeve (2) is an acrylic cylindrical rod, a small hole is axially formed in the acrylic cylindrical rod and used for leading in a left threaded rod (1), and one part of the outer wall surface of the left threaded rod (1) is cut into a plane and is attached to the plane of the middle section of the left side long stainless steel clamp (3); the left long stainless steel clamp (3) is a three-section metal sheet made of stainless steel, and round heads are arranged at the tail ends of two ends of the left long stainless steel clamp (3) and used for clamping a microfluidic chip; the structure of the right module is similar to that of the left module, the main part of the right cylindrical sleeve (5) is an acrylic cylindrical rod, a small hole is formed in the inner axial direction and used for leading in a right threaded rod (6), one part of the outer wall surface is cut into a plane and used for being attached to the middle section plane of the right short stainless steel clamp (4), the right short stainless steel clamp (4) is a three-section metal sheet made of stainless steel, and round heads are arranged at the tail ends of two ends of the right short stainless steel clamp (4) and used for clamping a microfluidic chip;
the second part is a main container which is provided with a split type clamping device and contains liquid; the main body container comprises an acrylic transparent cavity (7); the left module and the right module are symmetrically arranged on two sides of the transparent acrylic cavity (7); four angles of the transparent acrylic containing cavity (7) are provided with protruding upright columns, the upper parts of the protruding upright columns are provided with small holes, and the small holes are used for assembling the left threaded rod (1) or the right threaded rod (6).
2. The multi-angle nondestructive observation device for the internal channel of the microfluidic chip according to claim 1, wherein: the three-section metal sheet refers to a clamping device structure consisting of three planes.
3. The method for multi-angle nondestructive observation of the internal channel of the microfluidic chip by using the device of claim 1 is characterized in that: comprises the following steps of (a) carrying out,
(1) observing the material of the microfluidic chip by using a refractometer to obtain the refractive index corresponding to the material used by the microfluidic chip to be observed;
(2) preparing glycerol aqueous solutions with different concentration ratios, and respectively measuring the refractive indexes corresponding to the glycerol aqueous solutions with different concentrations by using a refractometer to obtain the concentration value of the group of glycerol aqueous solutions closest to the refractive index of the microfluidic chip material; adjusting the concentration value by properly adding water or glycerol to ensure that the refractive index of the glycerol aqueous solution is consistent with that of the material of the microfluidic chip, and recording the concentration value of the glycerol aqueous solution at the moment;
(3) connecting an outlet and an inlet of a micro-fluidic chip to be observed with a guide pipe to prevent liquid drops from infiltrating the inside of the channel; fixing the micro-fluidic chip to be observed by using a split type clamping device, and adjusting the angle to be observed;
(4) placing the split type clamping device and the microfluidic chip on an objective table of an optical microscope, pouring a glycerol aqueous solution with a prepared concentration into a main container of the clamping device for realizing multi-angle rotation of the microfluidic chip, and immersing the microfluidic chip;
(5) and observing the internal channel of the micro-fluidic chip to be measured by using an optical microscope to obtain an experimental image.
CN201811534541.7A 2018-12-14 2018-12-14 Multi-angle nondestructive observation method and device for internal channel of micro-fluidic chip Active CN109550528B (en)

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CN101923045A (en) * 2010-05-07 2010-12-22 重庆大学 Micro-fluidic chip observing platform and dual-visual field micro-fluidic chip observing system
CN103386345A (en) * 2013-08-07 2013-11-13 苏州扬清芯片科技有限公司 Micro-fluidic chip clamp
CN105013550A (en) * 2015-07-09 2015-11-04 清华大学深圳研究生院 Micro-fluidic chip fixture and micro-fluidic chip
CN108801980A (en) * 2018-03-30 2018-11-13 北京工业大学 Microfluid refractive index and concentration measurement system based on digital holographic microscopy and method

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010041230A2 (en) * 2008-10-10 2010-04-15 Cnrs-Dae Microfluidic integrated device for sample processing
CN101923045A (en) * 2010-05-07 2010-12-22 重庆大学 Micro-fluidic chip observing platform and dual-visual field micro-fluidic chip observing system
CN103386345A (en) * 2013-08-07 2013-11-13 苏州扬清芯片科技有限公司 Micro-fluidic chip clamp
CN105013550A (en) * 2015-07-09 2015-11-04 清华大学深圳研究生院 Micro-fluidic chip fixture and micro-fluidic chip
CN108801980A (en) * 2018-03-30 2018-11-13 北京工业大学 Microfluid refractive index and concentration measurement system based on digital holographic microscopy and method

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