TWI507803B - Dielectric particle controlling chip, method of manufacturing the same and method of controlling dielectric particles - Google Patents

Description

Dielectric particle handling wafer and its manufacturing method and manipulation dielectric Particle method

The invention relates to a method for controlling a dielectrophoretic wafer and a dielectric particle, in particular to a control wafer and a dielectric particle manipulation method for controlling a dielectric particle by a dielectrophoretic force and an alternating current permeation.

At present, in the field of microfluidic wafers, there are many techniques for controlling dielectric particles such as cells, microorganisms, and dielectric particles, and methods such as fluid flow, magnetic force, electrophoresis, electrodynamics, and dielectrophoresis are generally used. Although it is easier to control the particle mixing by fluid flow, the object to be controlled is the movement of the liquid, and there may still be repulsive force between the dielectric particles, so the dielectric particles are transported by the liquid flow to promote the dielectric particles. When the reaction is contacted with each other, or when the dielectric particles are uniformly mixed with the fluorescent material or the calibration substance, the disturbance caused by the fluid flow force in the micro flow channel does not necessarily drive the dielectric particles to contact each other to overcome the repulsive force, and The mixing effect is poor due to the poor perturbation effect, or a longer mixing reaction time is required. Because the dielectrophoretic force is low in particle destructiveness and can directly manipulate the particles themselves for movement, Good handling, so it is a good way to control the particles.

Another technique for manipulating particles is to use AC electroosmotic flow, which mainly utilizes the difference in the size of the liquid eddy current caused by the asymmetric AC electroosmotic flow between the large and small electrodes, so that the liquid force flows from the small electrode to the large electrode and transmits. Particles, although the use of AC electroosmotic flow can effectively drive the liquid to drive the particle flow transmission, but the current design must consider the size of the large and small electrodes, and the location of the stagnation point generated by the large and small electrodes, and requires a considerable number of electrodes to match Control, process technology is complicated.

Accordingly, it is an object of the present invention to provide a wafer that is convenient to fabricate and that utilizes dielectrophoretic forces to interact with alternating current electroosmotic flow to manipulate dielectric particles.

Another object of the present invention is to provide a method of fabricating the above wafer.

It is still another object of the present invention to provide a method of transporting, mixing and collecting dielectric particles by means of the above wafer.

Thus, the dielectric particle handling wafer of the present invention comprises a wafer body defining a reaction space having an opening facing upward to accommodate a dielectrophoretic liquid, and a first electrode layer and a finger spaced apart from the one of the wafer body a forked second electrode layer, and a dielectric layer. The first electrode layer and the second electrode layer are respectively located at a bottom edge of the reaction space, the first electrode layer has a first connecting portion, and a plurality of first finger electrode portions respectively extending outward from the first connecting portion The second electrode layer has a second connecting portion, and a plurality of second finger electrode portions extending outward from the second connecting portion, the first finger electrical portions The pole portions and the second finger electrode portions are staggered and arranged at intervals. The dielectric layer is made of a dielectric material and is disposed to be fixed to the wafer body by covering the first electrode layer and the second electrode layer.

Thus, the method for fabricating a dielectric particle handling wafer of the present invention comprises the steps of: (a) coating a substrate with an interdigitated first electrode layer and a finger-shaped second electrode layer, the first The electrode layer has a plurality of strip-shaped first interdigitated electrode portions, the second electrode layer having a plurality of strip-shaped second interdigitated electrode portions, and the first interdigitated electrode portions and the same The two-finger electrode portions are arranged in a staggered arrangement with each other; (b) a top surface of the substrate is covered with a dielectric layer composed of a dielectric material and covering the first electrode layer and the second electrode layer; and (c) Forming a layer on the top surface of the substrate to define a layered body covering the first electrode layer and the second electrode layer and accommodating the reaction space of the dielectrophoresis liquid.

Therefore, the method for controlling dielectric particles of the present invention is suitable for controlling the dielectric particles by the above-mentioned dielectric particles, and comprises the steps of: (a) injecting a dielectrophoresis liquid containing dielectric particles into the reaction space; (b) applying an alternating current to the first electrode layer and the second electrode layer, and the alternating current applied to the first electrode layer and the second electrode layer has a phase difference of 180°, so that two adjacent first fingers The electrode portion and the second finger electrode portion cooperate to generate a positive dielectrophoretic force field for attracting the dielectric particles, and cooperate with the positively above to generate a driving medium to the inward and inward direction from the two long sides thereof An alternating current seepage field that flows in the direction of the first connection portion and the second connection portion that are electrically connected from the end thereof.

The effect of the invention: through the first fork electrode layer and the first The top surface of the two electrode layer is covered with a dielectric layer, and the positive dielectric field strength generated by the first electrode layer and the second electrode layer is reduced by the presence of the dielectric layer, so that the first electrode is The AC electroosmotic force field generated by the layer and the second electrode layer can be relatively developed, and the dielectric particles can be effectively manipulated through the interaction of the positive dielectrophoretic force field and the AC electroosmotic force field.

3‧‧‧chip body

30‧‧‧Reaction space

31‧‧‧Substrate

32‧‧‧Layered body

4‧‧‧First electrode layer

41‧‧‧First finger fork electrode

42‧‧‧First connection

5‧‧‧Second electrode layer

51‧‧‧Second finger fork electrode

52‧‧‧Second connection

520‧‧‧ Radial gap

6‧‧‧Dielectric layer

7‧‧‧Three-dimensional microstructure

71‧‧‧Upper convex

72‧‧‧ recessed

800~802‧‧‧ arrow

Other features and effects of the present invention will be apparent from the following description of the drawings, wherein: FIG. 1 is a perspective, cross-sectional view of a first preferred embodiment of a dielectric particle handling wafer of the present invention; Figure 3 is a side cross-sectional view of the first preferred embodiment; Figure 4 is a flow chart showing the manufacturing steps of the dielectric particle handling wafer of the present invention; and Figure 5 is a dielectric diagram of the present invention. FIG. 6 is a partial top plan view of the first preferred embodiment, schematically illustrating the direction of action of the AC electroosmotic force field generated above the recess; FIG. 7 is a view similar to FIG. The two adjacent alternating current electroosmotic flow fields drive the dielectrophoretic liquid to generate a swirling flow over an upper convex portion; FIG. 8 is a first preferred embodiment of the first preferred embodiment for concentrating and fixing the biotin modified latex particles by a control mode (1). FIG. 9 is a microscopic image of the first preferred embodiment driving the biotin-modified latex particles along the lower concave portions through the manipulation mode (2); FIG. 10 is a first preferred embodiment of the microscopic image. Formula (3) driven by biotin A modified microscopic image of the mixed latex particles in the center of the upper convex portions; FIG. 11 is a first preferred embodiment for driving the biotin-modified latex particles and the streptavidin-modified fluorescent particles to be rotated and mixed by the manipulation mode (3). Microscopic image of the reaction; FIG. 12 is a microscopic image of the first preferred embodiment for driving a rotational mixing reaction between S. aureus and a fluorescent antibody through a manipulation mode (3); FIG. 13 is a view of the dielectric particle manipulation wafer. In the case where the three-dimensional microstructure is not provided, when the sinusoidal alternating current is applied, the microscopic image of the fluorescent particles is rotated and mixed between the adjacent first and second finger electrode portions; FIG. 14 is the dielectric The particle manipulation wafer drives the square wave alternating current without applying the three-dimensional microstructure, and drives the fluorescent particles to be concentrated on the microscopy directly above the first finger electrode portion and above the second finger electrode portion. Figure 15 is a top plan view of a second preferred embodiment of the dielectric particle handling wafer of the present invention; Figure 16 is a partial enlarged view of Figure 15 and schematically illustrates the direction of action of the AC electroosmotic flow field; and Figure 17 It is the second preferred embodiment that collects the fluorescent dielectric particles in a microscopic image above a first connection.

As shown in Figures 1, 2 and 3, a first preferred embodiment of the dielectric particle handling wafer of the present invention can be used to manipulate the transport, mixing and collection concentration of dielectric particles in the dielectrophoretic fluid. The dielectric particles may be latex particles or biological particles such as cells, bacteria and yeast, but are implemented. When the dielectric particles are not limited to the above types.

The dielectric particle handling wafer comprises a wafer body 3, and a first electrode layer 4, a finger-shaped second electrode layer 5, a dielectric layer 6, and a solid layer laminated on the wafer body 3. Microstructure 7. The wafer body 3 has a substrate 31, and an annular layered body 32 covering the top surface of the substrate 31 and cooperating with the substrate 31 to define an opening-facing reaction space 30. The reaction space 30 is for injection. Dielectrophoresis fluid. In the present embodiment, the layered body 32 is made of PDMS (polydimethyl siloxane), but in practice, the wafer body 3 is provided with a plurality of openings into the reaction space 30, and the method is not The inventive focus of the present invention is therefore not described in detail, and is not limited to the above types.

It should be noted that since the reaction space 30, the first electrode layer 4, the second electrode layer 5, the dielectric layer 6, and the structure of the three-dimensional microstructure 7 are relatively small, for convenience, the figure The proportion of each component is an enlarged schematic view of the original structure. When implemented, the size of the components is not limited by the ratio shown in the figure.

The first electrode layer 4 and the second electrode layer 5 are fixed to the top surface of the substrate 31 by a microelectromechanical process, and are located at the bottom edge of the reaction space 30. The first electrode layer 4 has a plurality of strip-shaped first interdigitated electrode portions 41 extending left and right and having a front-rear interval, and a strip first electrically extending between the left ends of the first interdigitated electrode portions 41. Connection portion 42. The second electrode layer 5 is formed in a bilaterally symmetrical shape with the first electrode layer 4, and has a plurality of strip-shaped second finger electrode portions 51 extending forward and backward and extending left and right, and an electrical connection to the second finger a second connecting portion 52 between the right ends of the electrode portions 51, etc. The first interdigitated electrode portion 41 and the second interdigitated electrode portions 51 are arranged in a staggered arrangement in parallel with each other in parallel.

The dielectric layer 6 is covered and fixed on the top surface of the substrate 31 covering the first electrode layer 4 and the second electrode layer 5, and is located above the first electrode layer 4 and the second electrode layer 5, so that the The first electrode layer 4 and the second electrode layer 5 are not in contact with the dielectrophoretic liquid in the reaction space 30.

In this embodiment, the dielectric layer 6 is made of a dielectric material, such as a photoresist or a tantalum oxide. In this embodiment, the SU-8 photoresist is taken as an example, but in practice, the dielectric material is used. Not limited to the above categories.

The three-dimensional microstructure 7 is overlaid on the dielectric layer 6 covering the dielectric layer 6, and the cross section is in the form of a continuous undulating wave. The three-dimensional microstructure 7 has a plurality of strip-shaped upper protrusions 71 and a plurality of strip-shaped lower recesses 72 extending longitudinally left and right along the first and second finger-tip electrode portions 41, respectively. The upper convex portions 71 are formed in a crest shape and are respectively located above the gap between the adjacent first and second finger electrode portions 41 and 51, and the lower concave portions 72 are in a trough shape. The first finger electrode portion 41 and the second finger electrode portion 51 are directly above.

In the present embodiment, the first finger electrode portion 41 and the second finger electrode portion 51 have a width of 50 μm, and the distance between the adjacent first and second finger electrode portions 41 and the second finger electrode portion 51 is 30 μm. The thickness of the dielectric layer 6 is 2 mm, the height of the upper convex portions 71 of the three-dimensional microstructure 7 is 12 mm, and the height of the lower concave portions 72 is 2 mm. However, the dimensions of the first electrode layer 4, the second electrode layer 5, the dielectric layer 6, and the three-dimensional microstructure 7 are not limited thereto.

As shown in Figures 1, 3 and 4, the method of manufacturing the dielectric particle handling wafer comprises the following steps:

Step (1) coating the first electrode layer 4 and the second electrode layer 5 on the substrate 31. A conductive material is coated and fixed on the top surface of the substrate 31 through a microelectromechanical process to form the first electrode layer 4 and the second electrode layer 5. In this embodiment, the material of the first electrode layer 4 and the second electrode layer 5 is ITO (Indium Tin Oxide), but the implementation is not limited thereto. Since the first electrode layer 4 and the second electrode layer 5 are fabricated by a microelectromechanical process and are not the focus of the present invention, they will not be described in detail.

Step (2) molding the dielectric layer 6. Applying a dielectric material to the top surface of the substrate 31 to cover the first electrode layer 4 and the second electrode layer 5, and only exposing a small first connecting portion 42 and a small second connecting portion 52 for guiding Receive a signal. In this embodiment, the dielectric material is SU-8 photoresist, and SU-8 is diluted with a photoresist developing solution, and then coated on the top surface of the substrate 31, and is dried and shaped.

Step (3) Forming the three-dimensional microstructure 7 includes the following sub-steps:

Step (3-1) coating the photosensitive hardening material on the top surface of the dielectric layer 6, and correspondingly located above the first electrode layer 4 and the second electrode layer 5.

Step (3-2) applies an alternating current to the first electrode layer 4 and the second electrode layer 5 to induce deformation of the photosensitive hardening material to produce a three-dimensional microstructure. An alternating current is applied to the second finger electrode portions 41 and the second finger electrode portions 51 through the first connecting portion 42 and the second connecting portion 52, respectively. Electrically attracting the photosensitive hardening material to the first finger electrode portions 41 and the non-uniform electric field distribution formed by the matching of the first finger electrode portions 41 and the second finger electrode portions 51 When the edge of the second finger electrode portion 51 is the strongest electric field, a relatively high crest-like convex portion 71 is concentratedly formed above the two adjacent first and second finger electrode portions 41 and 51, and Forming a relatively lower trough-like recess 72 directly above the first finger electrode portion 41 and the second finger electrode portions 51, and further, the first finger electrode portions 41 and the same A three-dimensional microstructure 7 having a cross-section continuous high and low undulations is formed between the two-finger electrode portions 51.

In this embodiment, in the present example, the photosensitive hardening material used in the stereoscopic microstructure 7 is a UV-curable adhesive (UV adhesive), and the alternating current applied to the first electrode layer 4 and the second electrode layer 5 is square. The wave is 800Vpp, 30kHz, and the two alternating currents have a phase difference of 180°, and the larger the applied voltage is, the larger the shape variable of the photosensitive dielectric material is, that is, the higher the convex portions 71 are, so the same can be applied by modulation. The waveform, voltage and/or frequency of the alternating current are adjusted to produce a high and low undulating shape of the three-dimensional microstructure 7, which is not limited to the above-mentioned alternating current conditions. However, when implemented, the photosensitive hardening material may be replaced by other photosensitive materials of photohardening properties, such as infrared light curing glue or halogen light curing adhesive, etc., and alternating current conditions applied to the first electrode layer 4 and the second electrode layer 5 Can be adjusted accordingly.

The step (3-3) causes the stereo microstructure 7 to be photosensitive and hardened. When the alternating current is applied to the first electrode layer 4 and the second electrode layer 5, the stereo microstructure 7 is irradiated with UV light to be photohardened and shaped.

Step (4) forming a layered annular layer on the substrate 31 32. Forming a layered body 32 surrounding the three-dimensional microstructure 7 on the top surface of the substrate 31 by PDMS, so that the layered body 32 cooperates with the substrate 31 to define an opening face upward and covering the reaction space of the three-dimensional microstructure 7 30. Since the manner in which a reaction space 30 is formed on the substrate 31 is numerous and is not the focus of the present invention, it is not limited to the above.

As shown in Figures 1, 3 and 5, when the dielectric particles are manipulated by the dielectric particles of the present invention for manipulation of dielectric particles, for example, mixing of two dielectric particles, mixing of dielectric particles and dielectrophoresis liquid, or The transmission and collection of electric particles, etc., include the following steps:

Step (1) A dielectrophoresis liquid containing dielectric particles is added to the reaction space 30. According to the purpose of the dielectric particle manipulation to be performed, the dielectric particles to be mixed and mixed are added to the dielectrophoresis solution and added to the reaction space 30, or a dielectric particle and the dielectric particles are contained. The reagents of the mixed reaction are added to the reaction space 30 together, or a sample containing the dielectric particles to be concentrated is added to the reaction space 30, but when implemented, the reaction space 30 is placed for manipulation of the dielectric particles. The type is not limited to the above.

In step (2), an alternating current is applied to the first electrode layer 4 and the second electrode layer 5 to start manipulation of the dielectric particles.

In the present invention, an alternating current is applied to the first electrode layer 4 and the second electrode layer 5, and the alternating current applied to the first electrode layer 4 and the second electrode layer 5 has a phase difference of 180°. The method of driving the first finger electrode portions 41 and the second finger electrode portions 51 to cooperate with the wave-like upper convex portions 71 to generate a positive dielectrophoretic force field, and An AC electroosmotic force field is respectively generated above the trough-like recess 72.

Since the first and second finger electrode portions 41 and the second finger electrode portions 51 are arranged in a staggered manner, the AC current is generated by the recess 72 of the first finger electrode portion 41 above the first finger electrode portion 41. The force field is an example for explanation.

The first finger electrode portion 41 has a second finger electrode portion 51 on the front and rear sides of the first finger electrode portion 41, and the end of the first finger electrode portion 41 is adjacent to the second connecting portion 52, so the first finger electrode The portion 41 is surrounded by the two second finger electrode portions 51 and the second connecting portion 52. Therefore, the alternating current seepage force field generated by the first finger electrode portion 41 in the upper lower recess portion 72 thereof may have The front and rear opposing forces from the upper and lower convex portions 71 on the front and rear sides of the lower concave portion 72, and the force from the end of the first finger electrode portion 41 toward the left side toward the first connecting portion 42, as shown in FIG. As shown in FIG. 800, the net force of the AC electroosmotic force field causes the dielectrophoretic liquid above the recess 72 to flow toward the left toward the first connecting portion 42, as indicated by an arrow 801 in FIG. Similarly, the AC electroosmotic force field generated in the recess 72 above the second finger electrode portion 51 drives the dielectrophoretic solution to flow to the right in the direction of the second connecting portion 52. That is, when the alternating electroosmotic flow field drives the flow of the dielectrophoretic liquid, the flow direction of the dielectrophoretic liquid of the crest-like concave portions 72 on the adjacent sides of each of the crest-like upper convex portions 71 is reversed in the left and right directions. Therefore, the direction in which the dielectric particles are driven to move is also reversed.

The positive dielectrophoretic force field and the alternating current electroosmosis due to changes in parameters such as signal waveform, frequency and/or voltage magnitude of the applied alternating current The size of the flow field will also change accordingly, so the parameters of the signal waveform, frequency and/or voltage of the alternating current can be modulated to control the movement mode of the dielectric particles and the flow mode of the dielectrophoretic liquid to achieve the desired control. purpose.

The change of the signal waveform of the alternating current affects the magnitude of the square root voltage (Vrms) of the alternating current, and the magnitude of the Vrms directly affects the positive medium generated by the first finger electrode portion 41 and the second finger electrode portion 51. The size of the electrophoretic force field and the AC electroosmotic flow field. Under the same Vpp and frequency AC conditions, the square wave: sine wave: the Vrms ratio of the triangular wave is 6:3:2, so the sine wave alternating current should produce the same positive dielectrophoretic force field and alternating current electroosmotic force field size as the square wave alternating current. When the sine wave voltage (Vpp) is at least twice the square wave AC. Changing the alternating current frequency will affect the relative size of the positive dielectrophoretic force field and the alternating current electroosmotic force field. Under the application of square wave alternating current, the positive dielectric force generated will be strong enough to reduce the square wave alternating current frequency to below 10 kHz. The AC electroosmotic flow field will be relatively present.

By adjusting the alternating current waveform, frequency and/or voltage applied by the modulation, and adjusting the relative size of the positive dielectrophoretic force field and the alternating current electroosmotic force field, the following dielectric particle manipulation modes are generated, and the manipulation modes are alternated. Implementation to achieve the desired control objectives: Manipulation mode (1): Adsorption and immobilization of dielectric particles using a positive dielectrophoretic force field. When the positive dielectrophoretic force field is much larger than the alternating current electroosmotic force field and is sufficient to attract the dielectric particles, that is, the alternating current electroosmotic force field is very weak or almost absent, since the positive dielectrophoretic force field is the strongest The area is located at the first finger electrode portion 41 and the second finger electrode portion At the end of 51, the dielectric particles are respectively concentrated and fixed to the lower recesses 72 at the ends of the first finger electrode portions 41 and the second finger electrode portions 51.

Referring to Figure 6, the control mode (2): the AC electroosmotic flow field is used to control the flow of the dielectrophoretic liquid to drive the dielectric particles to move to collect the dielectric particles. When the AC electroosmotic flow field is sufficient to overcome the attraction force of the positive dielectrophoretic force field, the dielectric particles dispersed in the reaction space 30 can be attracted downward to the top surface of the stereo microstructure 7 through the positive dielectrophoretic force field. The alternating current seepage force field of the lower recess 72 drives the dielectrophoretic liquid on the front and rear sides to drive the dielectric particles to flow directly above the first and second finger electrode portions 41 or the second finger electrode portion 51. The first connecting portion 42 or the second connecting portion 52 connected in the direction flows, and the dielectric particles are moved to the upper portion of the first connecting portion 42 and the second connecting portion 52 to be concentrated by the flow of the dielectrophoretic liquid.

Referring to Figure 7, the control mode (3): the interaction between the positive dielectrophoretic force field and the alternating dielectrophoretic force field is used to drive the dielectrophoretic fluid to generate a swirling flow, which drives the dielectric particles to aggregate and produce a rotational mixing phenomenon. By causing the positive dielectrophoretic force field and the alternating current electroosmotic flow fields to be apparently generated, the dielectric particles are respectively attracted to the top of the crest-like upper convex portions 71 by a positive dielectrophoretic force field, but The dielectric particles attract the positioning. At the same time, the AC electroosmotic force field of the concave portion 72 under the front and rear adjacent sides of each of the upper convex portions 71 respectively drives the dielectrophoretic liquid to flow to the left and reverse to the right.

The two adjacent alternating current electroosmotic force fields in the opposite direction form a balance point of force in the central region of the upper convex portion 71, and drive the dielectrophoretic liquid at the balance point of the force to generate a maximum swirling flow, as indicated by arrow 802. Show, Thereby, the dielectric particles above the upper convex portion 71 are brought to the central region to concentrate and generate a mixing phenomenon of rotational displacement, which can quickly and effectively promote a mixed reaction between the dielectric particles and the dielectrophoresis liquid, for example, the dielectric particles and The fluorescent reagent in the dielectrophoresis solution is combined.

Hereinafter, an experimental example in which the dielectric microparticle manipulation wafer and the manipulation method actually manipulate the dielectric microparticles will be described.

Experimental Example (1) Concentration of transmission of dielectric particles.

The dielectric particles used were latex particles surface-modified with biotin (biotin), the particle size of the latex particles was 4 μm, and 300 mM sucrose solution (conductivity 5 S/cm) was used as the dielectrophoresis solution. The latex particles modified with biotin were formulated to a concentration of 107 particles/ml, and 40 μl of the latex particle sample was injected into the reaction space 30.

As shown in FIG. 8, sinusoidal alternating current (120 Vpp, 100 kHz) is applied to the first electrode layer 4 and the second electrode layer 5, and the biotin-modified latex particles are respectively adsorbed by the above-described phenomenon of the manipulation mode (1). The upper convex portion 71 is concentrated and fixed to a portion above the ends of the first finger electrode portion 41 and the second finger electrode portion 51.

Then, the frequency of the sine wave alternating current (120Vpp, 5kHz) is lowered, the positive dielectrophoretic force field is lowered, and the alternating current electroosmotic force field is relatively increased, and the latex particles modified with biotin are driven by the above control mode (2). The electrophoretic liquid driven by the AC electroosmotic force field above the concave portion 72 flows, and the dielectric particles are respectively carried over the lower concave portions 72, and are moved along the longitudinal direction of the lower concave portions 72 to the first connection portions on the left and right sides. 42 and the upper area of the second connecting portion 52 are concentrated as shown in FIG.

Next, the applied alternating current waveform is changed to a square wave. In the case of applying square wave alternating current (120Vpp, 100kHz), the positive dielectrophoretic force field will be much larger than the alternating current electroosmotic force field, and the above control mode (1) will be generated. In other cases, the latex particles modified with the biotin are all adsorbed and fixed to the portions of the upper convex portion 71 located above the ends of the first and second finger electrode portions 41 and 51.

However, when the square wave AC frequency is reduced to 3 kHz, the positive dielectrophoretic force field will decrease, and the AC electroosmotic force field will become significantly larger, which will result in the manipulation of the above control mode (3), and the latex particles will be Focusing on the central mixing of the upper convex portions 71, the mixing is as shown in FIG.

Experimental Example (2) Mixed reaction of two different dielectric particles.

20 μl of the above latex particle sample and 20 μl of the sample containing the streptavidin-modified fluorescent particles (particle size 0.1 mm) were simultaneously added to the reaction space 30, and the mixing test was carried out in the above-described manipulation mode (3). The sample containing the streptavidin-modified fluorescent particles (particle diameter: 0.1 mm) was set to a concentration of 107 particles/ml using a 300 mM sucrose aqueous solution as a dielectrophoresis solution.

When a square wave alternating current (120 Vpp, 3 kHz) is applied, the latex particles are concentrated to rotate at the central portion of the upper convex portion 71, and mixed with the fluorescent particles in the solution, and can be reacted in a short time (5 min). A sufficient intensity of fluorescence is obtained, as shown in FIG.

Experimental Example (3) The biological dielectric particles were subjected to a fluorescent cursor setting.

As shown in Figure 12, this experimental example is directly based on biological dielectric micro The manipulation of the granules is described. The bio-dielectric particles used are Staphylococcus aureus (Accession No. BCRC14957), the particle size is about 1 μm, and the substance involved in the mixed reaction is fluorescent-labeled anti-protein A (IgG) 20 g. /ml, and Staphylococcus aureus was formulated to a concentration of 109 particles/ml with a 300 mM sucrose solution (conductivity of 5 S/cm) as a dielectrophoresis solution. 20 μl of S. aureus samples and 2.5 ml of IgG antibody samples were simultaneously added to the reaction space 30, and square wave alternating current (120 Vpp, 3 kHz) was applied to drive Staphylococcus aureus and Anti-protein by the above control mode (3). The A (IgG) mixed reaction bond and can generate sufficient fluorescence intensity within 5 min. Therefore, it can be seen that the method has the advantages of high speed and high efficiency for the mixed manipulation of the biological particles.

According to the above experimental examples, the dielectric layer 6 is covered over the interdigitated first electrode layer 4 and the second electrode layer 5, and the dielectric layer 6 is covered with a continuous high and low undulating three-dimensional microstructure 7 The dielectric layer 6 and the three-dimensional microstructure 7 can be effectively utilized to reduce the intensity of the positive dielectrophoretic force field generated by the first electrode layer 4 and the second electrode layer 5, thereby reducing the phenomenon that the dielectric particles are directly adsorbed and fixed. The alternating current seepage force field generated by the first electrode layer 4 and the second electrode layer 5 can be conveniently manipulated to control particle movement. The structure of the upper convex portion 71 and the lower concave portion 72 of the three-dimensional microstructure 7 can be further matched, so that the dielectric particle manipulation wafer of the present invention can further pass the alternating current waveform, frequency and/or voltage level applied by the modulation. The method further modulates the relative size of the positive dielectrophoretic force field and the alternating current electroosmotic force field generated by the first electrode layer 4 and the second electrode layer 5, thereby generating different control modes and conveniently switching according to usage requirements. Control mode is one A new dielectric particle manipulation method and wafer structure design for integrated dielectrophoresis and AC electroosmotic manipulation modes.

However, in the implementation, it is not necessary to provide the three-dimensional microstructure 7 , and the alternating current waveform, frequency and voltage applied to the first electrode layer 4 and the second electrode layer 5 can be modulated to adjust the position. The relative magnitude relationship between the positive dielectrophoretic force field and the alternating current electroosmotic force field generated by the first finger electrode portion 41 and the second finger electrode portion 51 can also produce the effects of the above three control modes.

As shown in Figures 13 and 14, in the case where a stereoscopic structure is not used, in the case of applying a sine wave (120 Vpp, 30 kHz), the particle size can be driven by the positive dielectrophoretic force field and the alternating current electroosmotic force field. The 1 mm fluorescent particles are rotated between the first finger electrode portions 41 and the second finger electrode portions 51. When the applied alternating current is changed to a square wave (120 Vpp, 3 kHz), the 1 mm fluorescent particles are driven to move directly above the first finger electrode portions 41 and directly above the second finger electrode portions 51. Gather and rotate the mix.

As shown in FIG. 15 and FIG. 16 , the second preferred embodiment of the dielectric particle handling wafer of the present invention is different from the first preferred embodiment in that the three-dimensional microstructure is not disposed in the embodiment, and the first electrode layer is 4. The structural design of the second electrode layer 5 and the dielectric layer 6 is different from that of the first preferred embodiment. For convenience of description, only the differences between the present embodiment and the first preferred embodiment will be described below.

In this embodiment, the first electrode layer 4 and the second electrode layer 5 are designed to be radially inner and outer, and the first electrode layer 4 has a circular shape. The first connecting portion 42 and the plurality of first finger electrode portions 41 extending radially outward from the first connecting portion 42 are radially distributed. The second electrode layer 5 has an annular second connecting portion 52 spaced around the periphery of the first electrode layer 4, and a plurality of radially distributed radially inward from the inner edge of the second connecting portion 52. The second finger electrode portion 51 protrudes from the connecting portion 42, and the first finger electrode portions 41 and the second finger electrode portions 51 are arranged in a staggered arrangement. The second connecting portion 52 has a radial through hole 520. The first finger electrode portion 41 has a width that is radially outward, and the second finger electrode portions 51 are radially inward. The first finger electrode portion 41 and the second finger electrode portion 51 are equidistantly spaced apart, and one of the first finger electrode portions 41 extends radially outwardly. Through the radial gap 520.

In this embodiment, the first connecting portion 42 has a diameter of 800 μm, and the first finger electrode portion 41 extends outward from the periphery of the first connecting portion 42 to have a length of 1.1 mm and a width of 60 μm. The second connecting portion 52 The radius is 1.7 mm, the distance between the two adjacent first finger electrode portions 41 and the second finger electrode portion 51 is 30 μm, and the distance between the first finger electrode portions 41 and the second connecting portion 52 is 100 μm. The second finger electrode portion 51 and the first connecting portion 42 are spaced apart by 300 μm.

When the dielectric particle manipulation wafer of the present embodiment is used, the specific dielectric particles in the sample can be collected by the manipulation mode of the manipulation mode (2) of the first embodiment. The mode of use of this embodiment will be described below by way of an experimental example.

Experimental Example (4) Concentrated fluorescent dielectric particles were collected.

The fluorescent dielectric particles used were FluoSpheres® Polystyrene Microspheres (particle size 1.0 μm), Orange Fluorescent (540/560), and a concentration of 10 x 107 beads/mL. After the fluorescent dielectric particle sample is injected into the reaction space 30, a sine wave alternating current (120 Vpp, 5 kHz) is applied to the first electrode layer 4 and the second electrode layer 5, respectively, and the two-string alternating current has a phase difference of 180°. Under the voltage and frequency conditions of the sinusoidal alternating current, the flow of the dielectrophoresis liquid driven by the alternating current electroosmotic force directly above the first finger electrode portion 41 and the second finger electrode portions 51 will drive The electric particles are respectively displaced toward the first connecting portion 42 and the second connecting portion 52, and the distance between the first finger electrode portion 41 and the second connecting portion 52 is smaller than the second finger electrode portions 51 and The first connecting portions 42 are spaced apart from each other, so that the AC electroosmotic force field radially inward of the first finger electrode portions 41 is relatively larger than the AC electroosmotic force field radially outward of the second finger electrode portions 51. Therefore, the flow of the dielectrophoretic liquid above the first finger electrode portion 41 is relatively large, and most of the fluorescent dielectric particles are radially inwardly directed from all sides to the first connecting portion 42 to be concentrated and concentrated.

After applying AC power for 60 seconds, a fluorescent intensity sufficient for interpretation can be obtained above the first connecting portion 42, as shown in FIG. It can be shown that even in the case of extremely low dielectric particle content, the present invention can quickly collect and concentrate the dielectric particles dispersed in the sample in a fixed area, which can greatly improve the applicable dielectric particle concentration limit. And has excellent sensitivity. Since biological particles such as cells, bacteria, and fungi are common dielectric particles that can be manipulated by dielectrophoretic force, the method can also be applied to the collection and concentration of biological particles in a sample.

In addition, the first finger electrode portion 41 and the second electrode electrode portion 51 are further extended by the first electrode layer 4 and the second electrode layer 5 designed by the annular and radial structures. The length of the way to expand the collection range of dielectric particles, can be used to collect and concentrate a wide range of dielectric particles, is a new design of dielectric particle collection and concentration.

It should be noted that, in addition to the dielectric layer 6 alone, the continuous undulating three-dimensional microstructure (not shown) may be additionally formed on the dielectric layer 6 to enable the peaks. The upper convex portion is located above the gap between the two adjacent first finger electrode portions 41 and the second finger electrode portion 51, and the valley-shaped lower concave portions are respectively located directly above the first finger electrode portions 41. Directly above the second finger electrode portions 51, the positive dielectrophoretic force field strength can be further attenuated, and the above various control modes can be more conveniently performed by the modulation of the positive dielectrophoretic force field and the alternating current electroosmotic force field.

In summary, the design of the transparent layer 7 is covered by the first electrode layer 4 and the second electrode layer 5, and the layer 3 is coated on the dielectric layer 6. The dielectric layer 6 and the presence of the stereoscopic microstructure 7 reduce the positive dielectrophoretic force field intensity generated by the first electrode layer 4 and the second electrode layer 5, so that the first electrode layer 4 and the second electrode The AC electroosmotic force field generated by the combination of layer 5 can be relatively developed, and the dielectric particles can be effectively manipulated through the interaction between the positive dielectrophoretic force field and the AC electroosmotic force field, for example, the micro dielectric in the concentrated dielectrophoresis liquid is collected. particle. Further, the three-dimensional microstructure design of the high-low undulation of the three-dimensional microstructure 7 and the modulation of the waveform, frequency and/or voltage of the alternating current applied by the modulation may be performed to adjust the first electrode layer 4 and the second electrode layer 5 Produced The relative size relationship between the positive dielectrophoretic force field and the AC electroosmotic flow field enables the dielectric particle manipulation wafer of the present invention to be used for transmitting mixed dielectric particles in addition to transmitting dielectric particles, and can be used for two different media. The mixing reaction of the electric particles and the mixing reaction of the dielectric particles with other substances in the dielectrophoresis liquid can achieve the desired mixing effect in a short time, and have excellent mixing efficiency. Therefore, the dielectric particle manipulation wafer of the present invention can be further combined with other detection devices, and can be widely applied to the detection and analysis of biological particles or other dielectric particles in the food, environmental and medical fields, and is quite convenient and practical, so that the object of the present invention can be achieved. .

The above is only the preferred embodiment of the present invention, and the scope of the present invention is not limited thereto, that is, the simple equivalent changes and modifications made by the patent application scope and patent specification content of the present invention, All remain within the scope of the invention patent.

3‧‧‧chip body

30‧‧‧Reaction space

31‧‧‧Substrate

32‧‧‧Layered body

4‧‧‧First electrode layer

41‧‧‧First finger fork electrode

42‧‧‧First connection

5‧‧‧Second electrode layer

51‧‧‧Second finger fork electrode

52‧‧‧Second connection

7‧‧‧Three-dimensional microstructure

71‧‧‧Upper convex

72‧‧‧ recessed

Claims (22)

A method for fabricating a dielectric particle manipulation wafer, comprising the steps of: (a) coating a substrate with a first interdigitated first electrode layer and an interdigitated second electrode layer, the first electrode layer having a first connecting portion, and a plurality of spaced strip-shaped first interdigitated electrode portions, the second electrode layer having a second connecting portion, and a plurality of spaced strip-shaped second interdigitated electrode portions, The first finger electrode portions are respectively extended from the first connecting portion toward the second connecting portion, and the second finger electrode portions are respectively extended from the second connecting portion toward the first connecting portion. And the first finger electrode portions and the second finger electrode portions are staggered in parallel with each other; (b) the top surface of the substrate is covered with a dielectric material and covers the first electrode layer and a dielectric layer of the second electrode layer; and (c) forming a layer on the top surface of the substrate to cooperate with the substrate to define a first electrode layer and the second electrode layer, and accommodating the dielectrophoresis liquid a layered body of the reaction space. The method for fabricating a dielectric particle manipulation wafer according to claim 1, further comprising the step of: d) coating a top surface of the dielectric layer with a three-dimensional microstructure having a continuous upper and lower undulations, the three-dimensional microstructure having And a plurality of convex portions and lower concave portions extending along the first and second finger electrode portions, wherein the upper convex portions are in a crest shape, and respectively located at two adjacent first fingers Above the gap between the fork electrode portion and the second finger electrode portion, the lower recesses are in a trough shape and are located above the first finger electrode portions and above the second finger electrode portions. The method for manufacturing a dielectric particle manipulation wafer according to claim 2, wherein the step (d) comprises the steps of: (d1) coating a top surface of the dielectric layer with a photosensitive hardening material; (d2) respectively for the first Applying an alternating current to the electrode layer and the second electrode layer, and using the non-uniform electric field distribution between the first interdigitated electrode portion and the second interdigitated electrode portion to induce the photohardening material to have a strong electric field strength The first finger electrode portions are concentrated above the edge regions of the second finger electrode portions, and the three-dimensional microstructures are formed on the top surface of the dielectric layer; and (d3) the first electrode layer and the first electrode layer When the second electrode layer is applied with an alternating current, the three-dimensional microstructure is photohardened and shaped. The method for manufacturing a dielectric particle handling wafer according to claim 1, wherein the first electrode layer of the step (a) further has a first connecting portion, and the second electrode layer further has a second a connecting portion, the first finger electrode portions are respectively extended from the first connecting portion toward the second connecting portion, and the second finger electrode portions are respectively connected to the first connecting portion from the second connecting portion The direction of the portion is extended, and the first finger electrode portions and the second finger electrode portions are staggered in parallel with each other. The method for manufacturing a dielectric particle manipulation wafer according to claim 1, wherein the first electrode layer and the second electrode layer of the step (a) are radially inner and outer spacers, and the first electrode The layer further has a first connecting portion, the second electrode layer further has a second connecting portion spaced around the first connecting portion, the first finger electrode portions are radially distributed from the first connecting portion Radially outwardly projecting toward the second connecting portion, the second finger electrode portions are radially distributed from the second connecting portion Extending inward toward the first connecting portion. The method of fabricating a dielectric particle manipulation wafer according to claim 5, wherein the second connecting portion has a radial notch, and wherein a first interdigitated electrode portion extends radially outward through the radial notch. The method for manufacturing a dielectric particle manipulation wafer according to claim 5, wherein the width of the second finger electrode portions in step (a) is gradually narrowed radially inward, and respectively adjacent to the first The one-finger electrode portions maintain the same pitch. The method of manufacturing a dielectric particle handling wafer according to claim 5, wherein the distance between the end of the second finger electrode portion and the first connecting portion is greater than the end of the first finger electrode portion The pitch of the second connecting portion. The method of fabricating a dielectric particle manipulation wafer according to claim 1, wherein the dielectric material constituting the dielectric layer in the step (b) is a photoresist or a tantalum oxide. The method of manufacturing a dielectric particle manipulation wafer according to claim 3, wherein the photosensitive hardening material is selected from the group consisting of ultraviolet light curing glue, infrared light curing glue or halogen light curing glue. A dielectric particle manipulation wafer comprises: a wafer body defining a reaction space for opening an opening facing up to accommodate a dielectrophoretic liquid; a finger-shaped first electrode layer and an interdigitated second electrode layer spaced apart Provided on the wafer body, respectively located at the bottom edge of the reaction space, the first electrode layer has a first connection portion, and a plurality of respectively from the first connection a first finger electrode portion extending outwardly, the second electrode layer having a second connecting portion, and a plurality of second finger electrode portions extending outward from the second connecting portion, the first finger The electrode portion and the second finger electrode portions are staggered and arranged at intervals; and a dielectric layer is formed of a dielectric material and covers the first electrode layer and the second electrode layer to be fixed on the wafer Ontology. The dielectric particle manipulation wafer of claim 11, further comprising a three-dimensional microstructure covering the top surface of the dielectric layer and located above the first electrode layer and the second electrode layer, the three-dimensional microstructure cross section is The upper and lower undulations have a plurality of upper convex portions and a plurality of lower concave portions extending along the first finger electrode portions and the second finger electrode portions, and the upper convex portions are in a crest shape, and Separatingly located above the gap between the two adjacent first and second finger electrode portions, and the lower concave portions are in a trough shape, respectively located directly above the first finger electrode portions and the first The two-finger electrode portion is directly above. The dielectric particle manipulation wafer according to claim 11 or 12, wherein the first finger electrode portions and the second finger electrode portions are staggered and arranged in parallel with each other. The dielectric particle manipulation wafer of claim 11 or 12, wherein the first electrode layer and the second electrode layer are radially inner and outer spacers, and the second connection portion is annular and spaced around The first connecting portion is radially outward, and the first finger electrode portions are radially distributed from the first connecting portion to the second connecting portion radially outward, and the second finger electrode portions Radially distributed from the second connecting portion to the radial direction The inner portion protrudes toward the first connecting portion. The dielectric particle handling wafer of claim 14 wherein the second attachment portion has a radial indentation and wherein a first interdigitated electrode portion extends radially outwardly through the radial indentation. The dielectric particle manipulation wafer according to claim 14, wherein the width of the second finger electrode portions is gradually narrowed radially inwardly, and is respectively maintained adjacent to the adjacent first finger electrode portions. The same spacing. The dielectric particle handling wafer of claim 14, wherein a distance between the end of the second finger electrode portion and the first connecting portion is greater than a distance between the end of the first finger electrode portion and the second connecting portion . The dielectric particle handling wafer of claim 11, wherein the dielectric material constituting the dielectric layer is selected from the group consisting of photoresist or tantalum oxide. The dielectric particle manipulation wafer according to claim 12, wherein the stereoscopic microstructure is composed of a photosensitive hardening material selected from the group consisting of ultraviolet light curing glue, infrared light curing glue or halogen light curing glue. . A method for controlling dielectric particles, which is suitable for handling dielectric particles by using a dielectric particle manipulation wafer according to any one of claims 11 to 19, and comprising the steps of: (a) dielectrophoresis containing dielectric particles a liquid is injected into the reaction space; and (b) an alternating current is applied to the first electrode layer and the second electrode layer, and an alternating current applied to the first electrode layer and the second electrode layer has a phase difference of 180°. Two adjacent first finger electrode portions and second finger fork The electrode portion cooperates between the two to generate a positive dielectrophoretic force field for attracting the dielectric particles, and cooperates with the upper portion thereof to generate a driving medium to flow from the opposite sides thereof toward the inner side, and from the end thereof An alternating current seepage field that is electrically connected to the first connecting portion and the second connecting portion. The method for controlling dielectric particles according to claim 20, wherein the step (b) is such that the positive dielectrophoretic force field is smaller than the alternating current electroosmotic force field, so that the dielectric particles are respectively subjected to the adjacent alternating current electroosmotic flow force. The position-driven dielectrophoretic liquid is driven to be displaced directly above the first finger electrode portion and the second finger electrode portions, and along the first finger electrode portions and the second finger electrodes The minister shifts the concentration toward the first connecting portion and the second connecting portion, respectively. The method for controlling dielectric particles according to claim 20, wherein the step (b) is: respectively, attracting the dielectric particles to the adjacent first finger electrodes and the first electrode by the positive dielectrophoretic force field a relative flow of the dielectrophoresis liquid driven by the two alternating current electroosmotic force places generated above the two adjacent first and second finger electrode portions and opposite to the second finger electrode portion And generating a swirling flow above the gap between the two adjacent first and second finger electrode portions to drive the gap between the two adjacent first and second finger electrode portions The dielectric particles are concentrated and rotationally displaced toward the vortex flow of the dielectrophoresis liquid.