Lateral and vertical electrophoresis method for micro-nano biological and metabolite sensors and actuators using two electric field intensities

Abstract

A lateral and vertical DEP method for micro-nano biological and metabolite sensors and actuators using two electric field intensities. The method specifically comprises the following steps: S1, before a metal etching process, performing resist plasma etching to form a resist profile at a side wall, wherein a high-pressure baseline formulation comprises oxygen, nitrogen, and argon in a ratio of 1:4:140 at a pressure of 1600 mT and a radiofrequency power of 1300 watts, used to generate a resist with a pre-designed angle; S2, performing metal etching, wherein a baseline instruction comprises chlorine, boron trichloride, and argon in a ratio of, but not limited to, 1:0.4:0.2, at a pressure of 8 mT, with a source power preferably of 1200 watts and a bias power preferably of 175 watts; and S3, performing metal profiling to measure the remaining thickness. The method enables a lateral attractive force of PDEP at Y in a medium.

Description

A lateral and vertical electrophoresis method for micro- and nano-bio- and metabolite sensors and actuators using two-strength electric fields Technical Field

[0001] The present invention relates to the field of micro- and nano-organisms and metabolite technologies, and in particular to a lateral and vertical DEP method for micro- and nano-organism and metabolite sensors and actuators using two-strength electric fields. Background Art

[0002] Dielectrophoresis (DEP) is a particle displacement method that exploits its dielectric properties. In this technique, a sinusoidal, time-varying, and spatially inhomogeneous electric field is used to manipulate the position of particles due to their dielectric properties. DEP is capable of focusing, translating, and capturing a wide range of materials (such as environmental, biological, and clinical analytes) in fluid suspension media, as well as characterizing, purifying, and enriching them.

[0003] The development of contactless, label-free, and label-free manipulation studies by integrating dielectrophoretic microelectrodes into lab-on-a-chip systems has further revealed the potential applications of dielectrophoresis in nano- and micro-machines. Indirect physical contact or contactless particle motion has numerous applications in areas such as drug discovery and delivery, as well as disease screening, separation, and biological sample analysis. This is primarily due to the ability to eliminate any attendant contact damage and associated issues, compared to direct physical contact.

[0004] In fact, it becomes even more challenging when the size of the moving object is a few micrometers or nanometers, which cannot be handled through direct physical contact. Therefore, the advantage of this non-contact particle movement method is that it can eliminate the effects caused by physical contact. For these reasons, movement through indirect physical contact using the properties of dielectrophoresis has been proposed. Other technologies such as fluorescence-activated cell sorting (FACS), magnetic-activated cell sorting (MACS), and field-flow fractionation (FFF) can also be used for the movement and separation of particles, especially cells, but DEP-based particle and cell separation using the force of dielectric polarization (FDEP) has better reliability and performance in terms of sensitivity and selectivity. Technical issues

[0005] Most dielectrophoresis (DEP) electrode designs and research currently on the market focus on a single, higher-field electrode. Consequently, a single, higher-field DEP microelectrode generates a high electric field gradient only from the top edge of the microelectrode, resulting in low efficiency for manipulation, separation, and fractionation in the medium via vertical attraction and repulsion of the target's NDEP and PDEP. However, less attention has been paid to combining two higher-field DEP electrodes to generate lateral and vertical motion of target or non-target particles through uniform and directional DEP forces.

[0006] In view of the above situation, a method for performing lateral and longitudinal dielectrophoretic particle separation by utilizing two relatively high intensity electric field points is required. Technical Solutions

[0007] The primary objective of this invention is to overcome the shortcomings of the prior art by providing a lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two-strength electric fields. This method enables more efficient manipulation, separation, and classification of target and non-target particles in a medium through the lateral attractive force of the PDEP in the Y axis and the vertical repulsive force of the NDEP in the Z axis.

[0008] The technical solution adopted by the present invention to achieve its technical purpose is:

[0009] A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two-strength electric fields is summarized. The method includes: SiO2, Ti / TiN and Al / Si / Cu addition steps, photolithography steps of photoresist coating, exposure and development, and photoresist profile removal steps by lateral etching of the photoresist slope, metal etching and photoresist removal.

[0010] The specific steps include:

[0011] S1. Performing resist plasma etching to form a resist profile at the sidewalls prior to the metal etching process, wherein a high-pressure baseline recipe including oxygen (O2), nitrogen (N2), and argon (Ar) in a ratio of 1:4:140 at a pressure of 1600 mT and a radio frequency (RF) power of 1300 watts is used to produce a pre-designed angle resist;

[0012] S2. Perform metal etching, wherein the baseline command includes chlorine (Cl2), boron trichloride (BCL3) and argon (Ar) in a ratio of but not limited to 1:0.4:0.2, a pressure of 8 mT, a source power of preferably 1200 W, and a bias power of preferably 175 W;

[0013] S3, measuring the remaining thickness of the metal profiling measurement;

[0014] Among them: the process based on inductively coupled plasma (ICP) introduces chemical and physical etching mechanisms, and the etching quality is controlled through four-step etching process conditions;

[0015] The four-step etching process conditions include:

[0016] Breakthrough etching (BE), controlled by time;

[0017] Main etch (ME) controlled by endpoint detection;

[0018] Over-etch (OE), which is controlled by the timing of the etching process;

[0019] Residue removal etch (RRE), which is controlled by the timing of the etching process.

[0020] Preferably, the oxygen (O 2 ) gas is a main gas for resist etching, the nitrogen (N 2 ) gas is a buffer gas to maintain a higher chamber pressure, and the argon (Ar) gas is used to increase plasma density.

[0021] Preferably, the chlorine gas (CL2) is a chlorine-based gas, which serves as a primary etchant for plasma-free aluminum, and in parallel CL2 is also a primary etchant for resist.

[0022] Preferably, the breakthrough etching (BE) is performed to remove the native oxide and resist remaining after the photolithography process.

[0023] Preferably, the main etch (ME) is to remove bulk aluminum.

[0024] Preferably, the over-etching (OE) and the residue removal etching (RRE) are for removing the remaining aluminum on the thicker aluminum area and removing the aluminum residue, respectively.

[0025] Preferably, the timing selection in the breakthrough etch (BE), over-etch (OE) and residue removal etch (RRE) processes is based on visual inspection through a microscope for color monitoring, a critical dimension scanning electron microscope (CDSEM) for critical dimension measurement, and a transmission scanning electron microscope (FESEM) for cross-sectional views.

[0026] Preferably, the method further uses a microfluidic channel with one input and three outputs (left, right and middle microfluidic channels) to separate target particles and non-target particles into two different locations at the top surface of the electrode and between the two conical electrodes.

[0027] Preferably, the method includes a lateral separation DEP force, which is a PDEP attractive force in the Y-axis from between the two electrodes to the top electrode surface, and a separation yield of the particle lateral motion synchronized with the capillary in the X-axis to the electrode ends (the ends of the left and right outlet microfluidic channels).

[0028] Preferably, the method includes a vertical separation DEP force, which is a NDEP repulsive force from the top electrode surface to between the two electrodes in the X-axis, and a separation yield of the particle vertical motion (end of the middle microfluidic channel) synchronized with the capillary in the X-axis.

[0029] Preferably, two electric fields of different strengths provide separation and fractionation in the medium through lateral attraction of PDEP in the Y-axis, vertical repulsion of target and non-target particles by NDEP in the Z-axis, and flow collection from one input microchannel with three output microchannels using capillary forces in the X-axis. Beneficial effects

[0030] Compared with the prior art, the beneficial effects of the present invention are: the lateral and vertical DEP method of micro-nano biological and metabolite sensors and actuators using two-intensity electric fields provides two-point higher-intensity electric field dielectrophoresis microelectrodes to generate higher electric field gradients from the top and bottom edges of the microelectrode, so as to more effectively operate, separate and classify target and non-target particles in the medium through the lateral attraction of PDEP at the Y axis and the vertical repulsion of NDEP on the Z axis.

[0031] Furthermore, the lateral and vertical DEP methods for micro-nano bio- and metabolite sensors and actuators using two strength electric fields also provide yield collection using capillary forces at the X-axis flow of one input microchannel with three output microchannels. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] 1-7 are schematic diagrams illustrating the working principle of the present invention for lateral and vertical dielectrophoretic particle separation using two relatively high-intensity electric field points.

[0033] FIG8(ai) is a schematic diagram illustrating the steps of fabricating a microelectrode array for dielectrophoresis (DEP)-based applications according to the present invention.

[0034] FIG9 is a schematic diagram showing a resist angle of about 16-20 degrees generated by lateral etching in a resist bevel process according to the present invention.

[0035] FIG. 10 is a schematic diagram showing an aluminum tilt angle of about 65-70 degrees obtained by the present invention for the resist angle after the aluminum etching process. Best Mode for Carrying Out the Invention

[0036] A lateral and vertical DEP approach for micro-nano bio- and metabolite sensors and actuators using two electric field intensities is presented. This invention presents a novel structure of a microelectrode array with two higher-intensity electric field points for dielectrophoresis (DEP)-based particle manipulation. The microelectrode array is fabricated using a complementary metal-oxide-semiconductor (CMOS) technology process, employing state-of-the-art techniques for forming the slope profile of the resist and the metal etching process. The introduced structure exhibits more effective electric field gradients and asymmetric distribution of the electric field for particle manipulation, separation, and classification in both lateral and vertical directions. Furthermore, based on electric field analysis, the two points of the higher-intensity electric field microelectrode with a sidewall profile angle of 20° generate higher electric field gradients from the top and bottom edges of the microelectrode, resulting in more effective manipulation, separation, and classification.

[0037] As a result, target or non-target particles are separated at two different locations: on top of the electrode surface and between the two electrodes. Separation occurs at the top of the electrode due to the lateral attraction and the PDEP lateral force on the y-axis. Simultaneously, separation between the two electrodes generates a vertical force on the x-axis due to the NDEP vertical repulsion. Using a single input channel with a tree output channel, left and right output channels with PDEP lateral forces on the y-axis, and a center output channel with NDEP vertical forces on the x-axis, the yield of target or non-target particles separated at two different locations is achieved. Modes for Carrying Out the Invention

[0038] In the description of the present invention, it should be noted that the terms "center," "upper," "lower," "left," "right," "vertical," "horizontal," "inner," and "outer," etc., indicating orientations or positional relationships, are based on the orientations or positional relationships shown in the accompanying drawings, or are the orientations or positional relationships in which the inventive product is typically placed when in use. These terms are intended solely to facilitate the description of the present invention and to simplify the description, and are not intended to indicate or imply that the device or component referred to must have a specific orientation, be constructed, or operate in a specific orientation. Therefore, they should not be construed as limitations on the present invention. Furthermore, the terms "first," "second," and "third," etc., are used solely to distinguish descriptions and should not be construed as indicating or implying relative importance.

[0039] In the description of the present invention, it should also be noted that, unless otherwise expressly specified or limited, the terms "disposed," "installed," "connected," and "connected" should be understood in a broad sense. For example, they may refer to fixed connections, detachable connections, or integral connections; they may refer to mechanical connections or electrical connections; they may refer to direct connections or indirect connections through an intermediate medium; and they may refer to internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on the specific circumstances.

[0040] To make the objectives, technical solutions, and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and examples. However, it should be understood that the specific embodiments described herein are merely illustrative of the present invention and are not intended to limit the scope of the present invention. In addition, in the following description, descriptions of well-known structures and technologies are omitted to avoid unnecessary confusion of the present invention.

[0041] 1-10 , which may be viewed alone or in any combination thereof, the lateral and vertical DEP methods for micro-nano bio- and metabolite sensors and actuators using two-strength electric fields will now be described in more detail with reference to the accompanying drawings.

[0042] In an embodiment of the present invention, the method includes using a machine to separate target particles and non-target particles into two different locations at the top surface of the electrode and between two conical electrodes, a microfluidic channel at an input end, and three ends of an output configuration, a left microfluidic channel, a right microfluidic channel, and a middle microfluidic channel.

[0043] The method includes the lateral separation DEP force, which is the PDEP attractive force in the Y-axis from between the two electrodes to the top electrode surface, and the separation yield of the particle lateral motion synchronized with the capillary in the X-axis to the electrode ends (the ends of the left and right outlet microfluidic channels).

[0044] The method also includes the vertical separation DEP force, which is the NDEP repulsive force from the top electrode surface to the two electrodes in the X-axis, and the separation yield of the particle vertical motion synchronized with the capillary in the X-axis (the end of the middle microfluidic channel).

[0045] 1-7 illustrate the working principles of lateral and vertical dielectrophoretic particle separation using two higher intensity electric field points according to an embodiment of the present invention;

[0046] Figure 1 illustrates the plane of particle motion in three dimensions, with two-point-strength electric fields, preferably at the bottom and top edges of a conical electrode. Thus, the multiple descending arrows indicate the PDEP attractive force to the two-point-strength electric field on the y-axis, transverse to the direction of motion. The opposite direction represents the NDEP repulsive force to the two-point-strength electric field on the z-axis, perpendicular to the direction of motion. It should be understood that the capillary fluid flow direction is along the x-axis. It is important to note that all motion in the x, y, and z axes occurs simultaneously.

[0047] Figure 2 shows the mechanism of separation by lateral attraction in the Y-axis direction and vertical repulsion in the Z-axis direction, and the pre-lateral and vertical motions of target or non-target particles are shown in Figure 2;

[0048] 3 and 4 show the lateral and vertical motion of target or non-target particles, while the lateral and vertical motion of target or non-target particles is shown in FIG2 .

[0049] FIG5 shows a column illustrating lateral and vertical motion of target or non-target particles according to a preferred example of the present invention;

[0050] FIG6 shows the integration of a microfluidic microchannel with one input microchannel and three output microchannels according to a preferred embodiment of the present invention. Thus, the yield is collected using capillary force on the X-axis flow of one input microchannel and three output microchannels;

[0051] FIG. 7 shows two points where particles are exposed to a higher intensity electric field according to a preferred example of the present invention.

[0052] In an embodiment of the present invention, a microelectrode array with two high-intensity electric field points for dielectrophoresis (DEP) is fabricated using complementary metal-oxide-semiconductor (CMOS) processing technology. This begins with the deposition of 1.15 μm of silicon oxide (SiO2) on a silicon substrate via plasma-enhanced chemical vapor deposition (PECVD). Physical vapor deposition (PVD) is then used to deposit thin layers of 0.053 μm titanium / titanium nitrite (Ti / TiN) with thicknesses of 60 nm / 30 nm as an adhesion layer. Following the Ti / TiN deposition, an aluminum / silicon / copper (Al / Si / Cu) layer with a thickness of at least 4.0 μm is deposited using a preferred, but not limited to, ratio of 98 / 1 / 1 wt % by PVD. It should be understood that a photoresist layer with a thickness of 4.0 μm, including ultraviolet (UV) curing to harden the photoresist, is applied to transfer the square array design onto the Al / Si / Cu layer.

[0053] Figure 8 (ac) shows the addition steps of SiO2, Ti / TiN, and Al / Si / Cu;

[0054] Figure 8 (d-f) shows the photolithography steps of photoresist coating, exposure, and development;

[0055] Figure 8(gi) shows the steps of photoresist profile removal by lateral etching to achieve photoresist bevel, metal etching, and photoresist removal.

[0056] Before the final step of aluminum / silicon / copper etching, after the photolithography process, the following steps are performed to form the resist bevel profile.

[0057] In a preferred embodiment, the first step includes a resist plasma etching process. Therefore, prior to the metal etching process, resist plasma etching is used to shape the resist profile at the sidewalls, thereby enabling precise control of the microelectrode profile angle. Photoresist etching at the top of the photoresist sidewalls is a technique that smoothes the photoresist based on a chemical etching reaction mechanism. It should be understood that various desired resist angles can be achieved by appropriately controlling parameters such as the gases used and their ratios, pressure, radio frequency (RF) power, and temperature.

[0058] During fabrication, a high-pressure baseline recipe consisting of oxygen (O2), nitrogen (N2), and argon (Ar) in a ratio of 1:4:140, at a pressure of 1600 mT and an RF power of 1300 W, was used to produce resists with pre-designed angles. Oxygen (O2) served as the primary gas for resist etching, while N2 served as a buffer gas to maintain a high chamber pressure, and Ar was added to increase plasma density. It should be noted that modifying the etch time significantly affected the angle of the photoresist profile and was used to alter the profile. This photoresist profile could be replicated during the metal etching process.

[0059] Furthermore, a metal etching process is performed, with the chamber system configuration and gas composition specified for anisotropic etching based on chemical and physical etching mechanisms. For this metal etching process, the baseline instructions include chlorine (Cl2), boron trichloride (BCl3), and argon (Ar) gases in a preferred ratio of, but not limited to, 1:0.4:0.2, a pressure of 8 mT, a preferred source power of 1200 watts, and a preferred bias power of 175 watts. Therefore, Cl2, a chlorine-based gas, serves as the primary etching gas for aluminum without the need for plasma. Cl2 is also the primary etchant for resist. Excessive Cl2 in the chemical reaction results in a low selectivity between aluminum and resist.

[0060] It should be noted that the metal etching process is the most challenging step in standard CMOS technology. Therefore, BCl3 is used as a heavy molecule for physical bombardment, while argon (Ar) is primarily used for residue removal. Therefore, an inductively coupled plasma (ICP) process based on both chemical and physical etching mechanisms was introduced. To control etching quality, a new four-step etching process was introduced. Therefore, the first step is a time-controlled breakthrough etch (BE), and the second step is a main etch (ME) controlled by endpoint detection. The third and fourth steps are the overetch (OE) and residue removal etch (RRE), respectively, both of which are controlled by the etching process timing. It should be understood that BE is performed to remove native oxide and resist remaining after the photolithography process, and ME is performed to remove bulk aluminum. The main reasons for using OE and RRE are to remove residual aluminum in thick areas and to remove aluminum residue, respectively. It should be understood that the timing of the BE, OE, and RRE processes was based on visual inspection by microscope for color monitoring, critical dimension scanning electron microscope (CDSEM) for critical dimension measurement, and field emission scanning electron microscope (FESEM) for cross-sectional views.

[0061] After establishing the etch process instructions, the final step in etch process monitoring is to measure the remaining silicon dioxide (SiO2) thickness. Using an aluminum etch recipe, based on etch rate testing for each material, aluminum selectivity and resist selectivity are optimized to be 1:1. Lateral etching using the resist bevel process results in a resist angle of approximately 16 degrees.

[0062] The tapered profile photoresist before the metal etching process is shown in Figure 9. After the aluminum etching process, an aluminum tilt angle of approximately 65-70 degrees is obtained as the resist angle;

[0063] FIG10 clearly shows the tapered cross-section dielectrophoresis electrode.

[0064] In this embodiment of the present invention, based on electric field analysis and experimental work, two points of a high-intensity electric field microelectrode with a 20° sidewall profile angle generate higher electric field gradients from the top and bottom edges of the microelectrode, enabling more efficient manipulation, separation, and fractionation in the medium in both lateral and vertical directions. The device was tested with three engineered particles of varying diameters. The first three minutes at the slowest flow rate were considered to demonstrate the effects of FDEP on various particles. The goal was to capture 10 μm particles in the region between the microelectrodes and release 1 and 3 μm particles from this region. By selecting an appropriate frequency between the crossover frequencies for 10 μm engineered particles and other particles, the DEP force required to actuate and capture the desired particles can be applied. The crossover frequency (fxo) for 10 μm is approximately 25 kHz, while the crossover frequencies for 3 and 1 μm particles are 380 and 1315 kHz, respectively. Therefore, a DEP signal frequency of 200 kHz was selected for the 10 μm isolation experiment. At this frequency, 10 μm particles experience a repulsive NDEP force in the vertical direction along the Z axis, while other particles experience a lateral PDEP attraction in the Y axis. Consequently, 1 and 3 μm particles are attracted to the highest electric field region at the top electrode surface. 10 μm particles are repelled from the highest electric field regions on the left and right sides, located between the two higher-intensity electric field profile electrodes.

[0065] It should be understood that the application of the novel two-point high-field DEP electrode can be applied to lab-on-a-chip systems for active filtration, such as circulating tumor cell applications and renal albumin filtration (artificial kidney). Therefore, the application of the two-point high-field DEP electrode in lab-on-a-chip can potentially eliminate the traditional design of the lateral one-point high-field DEP electrode, labeling technology, biomarkers, and passive filters (such as pore filters).

[0066] It should also be understood that the two-point higher-intensity electric field dielectrophoresis electrode can also be applied to biomedical and bioMEMS systems, such as circulating tumor cell (CTC) prognosis or renal albumin filtration (artificial kidney). The two-point higher-intensity electric field dielectrophoresis electrode manufacturing process has been miniaturized in a lab-on-a-chip system. The two-point higher-intensity electric field dielectrophoresis electrode in the lab-on-a-chip system is more compact, reliable, and easier to operate.

[0067] The working principle and specific usage process of the lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two high-intensity electric field points:

[0068] Dielectrophoretic microelectrodes provide a versatile platform for manipulating, separating, and characterizing a variety of biological entities, including cells, bacteria, proteins, viruses, and metabolites;

[0069] The method includes performing a resist plasma etch prior to a metal etching process to shape a resist profile at a sidewall, wherein a high-pressure baseline recipe includes using oxygen (O 2 ), nitrogen (N 2 ), and argon (Ar) gases in the following ratios of 1:4:140 at a pressure of 1600 mT and a radio frequency (RF) power of 1300 watts to produce a pre-designed angle resist;

[0070] Performing metal etching, wherein the baseline command includes chlorine (Cl2), boron trichloride (BCL3) and argon (Ar) in a ratio of but not limited to 1:0.4:0.2, a pressure of 8 mT, a source power of preferably 1200 W, and a bias power of preferably 175 W;

[0071] Measure the remaining thickness of silicon dioxide (SiO2);

[0072] Among them, the process based on inductively coupled plasma (ICP) introduces chemical and physical etching mechanisms, and the etching quality is controlled through four-step etching process conditions;

[0073] Among them, the four-step etching process conditions include:

[0074] Time-controlled breakthrough etch (BE); main etch (ME) controlled by endpoint detection; overetch (OE), controlled by the timing of the etch process; residue removal etch (RRE), controlled by the timing of the etch process;

[0075] Two-strength electric fields provide separation and fractionation in the medium through lateral attraction by PDEP in the Y axis, vertical repulsion of target and non-target particles by NDEP in the Z axis, and flow collection from one input microchannel with three output microchannels using capillary force in the X axis.

[0076] It should be noted that although the above embodiments have been described herein, this does not limit the scope of patent protection of the present invention. Therefore, based on the innovative concept of the present invention, changes and modifications to the embodiments described herein, or equivalent structures, equivalent processes, or equivalent functional transformations made using the contents of the present invention's specification and drawings, and direct or indirect application of the above technical solutions to other related technical fields, are all included in the scope of protection of the present invention's patent. Industrial Applicability

[0077] It should be noted that this invention demonstrates that the concept of two higher-intensity electric fields can be further explored in functional activities based on dielectrophoretic forces to transport, accumulate, separate, and characterize micron / nanoscale particles. Using two-point higher-intensity electric field dielectrophoretic microelectrodes, particles can be manipulated, separated, and classified both horizontally and vertically. Compared to conventional single-point high-intensity electric field dielectrophoretic microelectrodes, which can only manipulate, separate, and classify particles in the vertical direction, this invention can improve non-contact operation methods, with high sensitivity, high selectivity, and label-free detection of target particles for analytical applications.

[0078] It should be noted that the present invention provides improved process handling, separation, and fractionation performance with higher efficiencies exceeding 90% in the application of two-point high-field DEP electrodes. Therefore, the use of a two-point high-field DEP electrode profile is preferred due to the advantages of PDEP and NDEP in terms of particle motion in both the lateral and vertical directions under dielectrophoretic forces (FDEP). In terms of active filters, the two-point high-field DEP electrode is a non-contact filter with self-cleaning capabilities that does not affect process handling, separation, and fractionation, achieving efficiencies exceeding 90%. The two-point high-field DEP electrode profile was chosen because it enables process handling, separation, and fractionation with higher efficiencies exceeding 90%, making it advantageous for biomedical applications such as circulating tumor cell prognosis or renal albumin filtration.

Claims

1. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two-strength electric fields, characterized in that, The specific steps include: S1. performing resist plasma etching to form a resist profile at the sidewalls prior to a metal etching process, wherein a high-voltage baseline recipe including oxygen, nitrogen, and argon in a ratio of 1:4:140 at a pressure of 1600 mT and a RF power of 1300 watts is used to produce a pre-designed angle resist; S2, performing metal etching, wherein the baseline instruction includes chlorine, boron trichloride and argon, the ratio of which is but not limited to 1:0.4:0.2, the pressure is 8 mT, the source power is preferably 1200 watts, and the bias power is preferably 175 watts; S3, measuring the remaining thickness of metal profiling; Among them: the process based on inductively coupled plasma introduces chemical and physical etching mechanisms, and the etching quality is controlled through four-step etching process conditions; The four-step etching process conditions include: Breakthrough etching, controlled by time; Main etch controlled by endpoint detection; Overetching, which is controlled by the timing of the etching process; Residue removal etching, which is controlled by the timing of the etching process.

2. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The oxygen gas is a main gas for resist etching, the nitrogen gas is a buffer gas to maintain a higher chamber pressure, and the argon gas is used to increase plasma density.

3. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The chlorine gas is a chlorine-based gas that is used as a primary etchant for plasma-free aluminum.

4. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The breakthrough etching is to remove the native oxide and resist remaining after the photolithography process.

5. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The main etch is to remove bulk aluminum.

6. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The over-etching and residue removal etching are to remove the remaining aluminum on the area where the aluminum is thicker and to remove the aluminum residue, respectively.

7. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The timing of the breakthrough etch, over-etch and residue removal etch processes is based on visual inspection by microscope for color monitoring, critical dimension scanning electron microscope for critical dimension measurement and emission scanning electron microscope for cross-sectional view.

8. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The method also uses a microfluidic channel with a one-input and three-output configuration to separate target particles and non-target particles into two different locations at the top surface of the electrode and between two conical electrodes.

9. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The method includes lateral separation DEP forces, which are PDEP attractive forces in the Y-axis from between the two electrodes to the top electrode surface, and separation yields synchronized with the capillary lateral motion of particles in the X-axis to the electrode ends.

10. A lateral and vertical DEP method for micro-nano bio- and metabolite sensors and actuators using two strength electric fields according to claim 1, characterized in that: The method includes a vertical separation DEP force, which is a NDEP repulsive force from the top electrode surface to between the two electrodes in the X-axis, and a separation yield of the particle vertical motion synchronized with the capillary in the X-axis.

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