阅读说明：本技术 具有dep阵列的微流体设备 (Microfluidic device with DEP array ) 是由 希瑟·莫顿 洛塔尔·施密德 爱德华多·博阿达 于 2019-09-12 设计创作，主要内容包括：一种微流体设备,包括多个微流体通道和多个对应的介电电泳(DEP)电极阵列,每个微流体通道被布置成将流体引导到DEP电极阵列上,使得在使用中,靶微粒通过DEP电极阵列操纵。该设备还包括用于将设备连接到交流电源的第一连接点和第二连接点,每个DEP电极阵列的第一输入端经由第一导体连接到第一连接点,并且每个DEP电极阵列的第二输入端经由第二导体连接到第二连接点。在每个电极的第一输入端与第一连接点之间的第一导体的电阻以及在每个电极的第二输入端与第二连接点之间的第二导体的电阻基本上比所连接的电极阵列的总电阻小至少一个数量级。(A microfluidic device comprising a plurality of microfluidic channels and a plurality of corresponding Dielectrophoresis (DEP) electrode arrays, each microfluidic channel being arranged to direct fluid onto a DEP electrode array such that, in use, target particles are manipulated by the DEP electrode array. The device further comprises a first connection point and a second connection point for connecting the device to an alternating current power supply, the first input of each array of DEP electrodes being connected to the first connection point via a first conductor and the second input of each array of DEP electrodes being connected to the second connection point via a second conductor. The resistance of the first conductor between the first input of each electrode and the first connection point and the resistance of the second conductor between the second input of each electrode and the second connection point are substantially at least one order of magnitude less than the total resistance of the connected array of electrodes.)

1. A microfluidic device comprising a plurality of microfluidic channels and a plurality of corresponding Dielectrophoresis (DEP) electrode arrays, each microfluidic channel being arranged to direct fluid onto a DEP electrode array, the device further comprising at least one first connection point and at least one second connection point for connecting the device to an alternating current power supply, a first input of each DEP electrode array being connected to the first connection point via a first conductor and a second input of each DEP electrode array being connected to the second connection point via a second conductor, wherein:

the resistance of the first conductor between the first input of each electrode and the first connection point and the resistance of the second conductor between the second input of each electrode and the second connection point are substantially at least one order of magnitude less than the total resistance of the connected array of electrodes.

2. The microfluidic device of claim 1, wherein each DEP electrode array is associated with only one microfluidic channel.

3. The microfluidic device of claim 1 or 2, wherein the DEP electrode arrays are electrically connected in parallel.

4. The microfluidic device of any one of claims 1 to 3, wherein the first and second connection points are the only connection points on the device.

5. The microfluidic device according to any one of the preceding claims, wherein the DEP electrode arrays each comprise an array of interdigitated electrodes (IDEs).

6. The microfluidic device of claim 5, wherein each IDE array comprises a first set of electrodes comprising 5 to 40 electrodes interdigitated with a second set of electrodes comprising 5 to 40 electrodes.

7. The microfluidic device of claim 5 or 6, wherein each IDE array is about 2mm to 8mm long.

8. The microfluidic device of claim 7, wherein each IDE array is operable to generate an average electric field of about 80kV/m RMS.

9. The microfluidic device according to any one of the preceding claims, wherein the DEP electrode arrays each have a resistance of 1.6-2.4k Ω.

10. The microfluidic device according to any one of the preceding claims, wherein the device is a microfluidic cartridge insertable into a corresponding analytical device.

11. A microfluidic device according to any one of the preceding claims comprising at least two arrays of DEP electrodes.

12. The microfluidic device according to any one of the preceding claims, wherein the first and second conductors comprise electrically conductive leads.

13. The microfluidic device of claim 12, wherein the conductive leads comprise a conductive material deposited on a substrate.

14. The microfluidic device of claim 13, wherein each of the conductive leads comprises one or more internal gaps free of conductive material to increase an edge length of the conductive lead.

15. The microfluidic device of claim 13 or 14, wherein the electrically conductive material comprises gold, nickel platinum, or aluminum.

16. The microfluidic device according to any one of the preceding claims, wherein the target microparticle is a target cell.

Technical Field

The present invention relates to microfluidic devices, and to microfluidic devices that use Dielectrophoresis (DEP) to selectively manipulate target particles.

Background

Dielectrophoresis (DEP) is a well-known phenomenon that can be used to selectively move and/or manipulate particles based on their dielectric properties. The particles will move in the direction of the field gradient (positive DEP) or in the opposite direction (negative DEP).

In particular, DEP arrays (including, for example, interdigitated electrode arrays) can be adapted based on their geometry and the voltage and frequency of the power supply connected to the array to selectively manipulate certain particular particles from a fluid flowing through the DEP array.

In principle, DEP offers a promising particle selection mechanism for microfluidic diagnostic applications, where, for example, a fluid sample is processed to identify and analyze fluid-borne pathogen particles or particles associated with pathogens. Examples include analyzing a fluid sample of a patient to identify the presence and number of mycobacterium tuberculosis cells to diagnose and assess the severity of tuberculosis infection.

Generally, to produce a commercially viable point-of-care (POC) microfluidic diagnostic device, the device must be able to provide accurate diagnosis within an acceptable period of time. The constraint typically dictates a minimum flow rate of the sample through the device in order to make a diagnosis from the sample within an acceptable time period. To achieve the required sample flow levels in a commercially acceptable size and cost device, it is often necessary to provide multiple parallel processing channels, each performing the same diagnostic process (e.g., isolating particles of interest).

Unfortunately, many technical problems arise when attempting to implement DEP-based particle manipulation systems in microfluidic devices that include multiple parallel processing channels. For example, the DEP electrode array must be carefully "tuned" to ensure that the electric field it generates targets the desired microparticles. However, devices comprising multiple DEP electrode arrays, while necessary for commercial implementation, are difficult to implement because differences within the device tend to cause the DEP electrode arrays to behave differently from one another (e.g., capture target particles at inconsistent rates). Such performance differences beyond a certain tolerance level can lead to unreliable diagnostic results.

Furthermore, implementing microfluidic devices that include multiple DEP electrode arrays requires relatively high levels of power to be supplied to the device to drive the electrodes. To avoid unacceptable levels of power dissipation due to heating, it is therefore necessary to increase the amount of conductive material connecting the DEP electrode array to the power supply. However, on the scale and geometry typically associated with commercially practical microfluidic POC devices (e.g. disposable cartridges inserted into analytical devices), the conductive material may peel away from the substrate surface due to poor adhesion of the conductive material, and merely increasing the amount of conductive material may be difficult.

It is an object of certain embodiments of the present invention to at least partially address the above disadvantages of the prior art.

Summary of The Invention

According to a first aspect of the present invention, there is provided a microfluidic device comprising a plurality of microfluidic channels and a plurality of corresponding dielectrophoresis DEP electrode arrays. Each microfluidic channel is arranged to direct fluid onto the DEP electrode array. The device further comprises a first connection point and a second connection point for connecting the device to an alternating current power supply, the first input of each array of DEP electrodes being connected to the first connection point via a first conductor, and the second input of each array of DEP electrodes being connected to the second connection point via a second conductor. The resistance of the first conductor between the first input of each electrode and the first connection point and the resistance of the second conductor between the second input of each electrode and the second connection point are substantially at least one order of magnitude less than the total resistance of the connected array of electrodes.

Where the DEP electrode array utilises positive DEP (pdep), the microfluidic channel is arranged such that, in use, the target particles are held or attracted towards the DEP electrode array.

Optionally, each DEP electrode array is associated with only one microfluidic channel.

Optionally, the DEP electrode arrays are electrically connected in parallel.

Optionally, the first connection point and the second connection point are the only connection points on the device.

Optionally, the DEP electrode arrays each comprise an array of interdigitated electrodes (IDEs).

Optionally, each IDE array includes a first set of 5 to 40 electrodes interdigitated with a second set of 5 to 40 electrodes.

Optionally, each IDE array is about 2mm to 8mm in length and about 2.7-3.0mm in width. In some cases, an auxiliary IDE array is provided that is 1.7mm in width.

Optionally, each IDE array consists of 50 μm wide fingers, spaced apart by 50 μm.

Alternatively, each IDE array may be operated at a peak-to-peak voltage of about 12V (4V RMS) to generate an average electric field of about 80kV/m RMS.

Optionally, the DEP electrode arrays each have a resistance of 1.6-2.4k Ω. This is for an electrode having a conductivity of 150-220. mu.S/cm and a width of 2mm and a length of 4 mm.

The DEP electrode array resistance depends on the conductivity of the solution. This will vary greatly for different samples or sample preparation methods. It also depends on the width of the IDE array.

Optionally, the device is a microfluidic cartridge insertable into a corresponding analytical device.

Optionally, the apparatus comprises at least 2 DEP electrode arrays.

More commonly, the apparatus comprises more than 2 DEP electrode arrays

The preferred number of DEP electrodes depends to some extent on the sample and the cells of interest. For example, when testing sputum samples, it may be advantageous to have more than 2 DEP electrodes to allow adequate flux flow-through — however, in the presence of a large number of cells of interest, it is feasible to use only two DEP electrodes.

Optionally, the first conductor and the second conductor comprise electrically conductive leads.

Optionally, the conductive leads comprise a conductive material deposited on the substrate.

Optionally, each of the conductive leads includes one or more internal gaps free of conductive material to increase an edge length of the conductive lead.

Optionally, the conductive material comprises gold, platinum or aluminum. The conductive material may also include multiple layers, with an upper surface layer of gold, platinum, or aluminum, and an optional lower substrate layer of, for example, nickel or tantalum. A lower substrate layer is typically included to improve adhesion, and for an upper layer, biocompatible materials are typically used. For example, the conductive material may be 5um nickel with a thin gold layer on top.

Optionally, the target microparticle is a target cell.

According to certain aspects of the present invention, there is provided a technique for arranging a microfluidic device comprising a plurality of DEP electrode arrays connected generally in parallel and powered by a common pair of connection points. The technique specifically recognizes that if, for each electrode array, the total resistance between a connection point and the corresponding DEP electrode array input is an order of magnitude less than the total resistance of the connected DEP electrode array as calculated by the following equation;

any differences in DEP operation due to resistance changes to and from the connection point to the electrode input remain within acceptable levels.

Reference to X being "an order of magnitude less than Y" means that X is less than or equal to 1/10 of Y.

By this recognition, it is possible to provide a microfluidic device comprising, for example, a plurality of DEP electrode arrays arranged in parallel and advantageously powered by a single set of connection points, with operational consistency of the DEP electrode arrays not being below an acceptable level. Without this particular knowledge, unacceptable levels of non-uniformity are expected to occur in microfluidic devices of this configuration, or the amount of conductive material would need to be higher than necessary for the conductors, thereby increasing the cost and size of the apparatus.

Various other features and aspects of the present invention are defined in the claims.

Brief Description of Drawings

Embodiments of the invention will now be described, by way of example only, with reference to the accompanying schematic drawings in which similar elements are provided with corresponding reference symbols, and in which:

FIG. 1 provides a simplified schematic of an apparatus for providing alternating current to a plurality of electrode arrays for use in a microfluidic device to selectively capture target particles using Dielectrophoresis (DEP);

FIG. 2 provides a simplified schematic of a portion of a microfluidic device including a plurality of microfluidic channels;

FIG. 3 provides a diagram depicting an equivalent circuit diagram of the device of FIG. 1, including three electrode arrays connected in parallel and powered by a single pair of power pads (supply pads);

FIG. 4 provides a graph depicting an ideal situation, wherein the resistance of the conductive leads is zero;

FIG. 5 provides a schematic diagram illustrating a portion of a conventional conductive lead; a portion of a conventionally modified conductive lead, as well as portions of conductive leads according to some embodiments of the present invention, and portions of conductive leads according to some alternative embodiments of the present invention; and

fig. 6 provides an exploded view of a microfluidic cartridge that may be arranged in accordance with embodiments of the present invention.

Detailed Description

Fig. 1 provides a simplified schematic of an apparatus for providing alternating current to a plurality of electrode arrays for use in a microfluidic device to selectively capture target particles using Dielectrophoresis (DEP). The example described uses positive dep (pdep).

The device 101 comprises a first electrode array 102, a second electrode array 103 and a third electrode array 104. Each electrode array includes an array of interdigitated electrodes (IDEs). In a typical implementation, more electrode arrays (e.g. 32 parallel channels, each with its own electrode array (2 mm width per channel) may be used to provide a suitable footprint for a POC device), but for clarity only three electrode arrays are shown in fig. 1.

Although the figures show an array of linear interdigitated electrodes, it will be appreciated that arrays having different physical configurations may be used, for example interdigitated spiral electrodes may be used.

The first electrode array 102, the second electrode array 103, and the third electrode array 104 are connected to a connection point provided by a first power supply pad 105 and another connection point provided by a second power supply pad 106 via conductive leads. In use, the power supply pad is connected to an ac power supply 107.

The first electrode array 102, the second electrode array 103, and the third electrode array 104 are connected in parallel. More specifically, the first power supply pad 105 is connected to a first input terminal of the first electrode array 102, a first input terminal of the second electrode array 103, and a first input terminal of the third electrode array 104 via a first conductive lead 108. The first conductive lead 108 includes: a main branch 108a common to all electrode arrays; a first sub-branch 108b, the first sub-branch 108b being connected to a first input of the first electrode array 102; a second sub-branch 108c, the second sub-branch 108c being connected to the first input terminal of the second electrode array 103; and a third sub-branch 108d, the third sub-branch 108d being connected to the first input of the third electrode array 104. The second power supply pad 106 is connected to a second input terminal of the first electrode array 102, a second input terminal of the second electrode array 103, and a second input terminal of the third electrode array 104 via a second conductive lead 109. The second conductive lead 109 includes: a main branch 109a common to all electrode arrays; a first sub-branch 109b, the first sub-branch 109b being connected to the second input terminal of the first electrode array 102; a second sub-branch 109c, the second sub-branch 109c being connected to a second input of the second electrode array 103; and a third sub-branch 109d, the third sub-branch 109d being connected to the second input of the third electrode array 104.

In use, the apparatus shown in figure 1 forms part of a microfluidic device. The operation of such a device is described in more detail with reference to fig. 2.

Fig. 2 provides a simplified schematic of a portion of a microfluidic device comprising a plurality of microfluidic channels 201, 202, 203. Each microfluidic channel passes over a corresponding electrode array 204, 205, 206. As described above, the electrode arrays are each electrically connected in parallel, and are connected to the first power supply pad 207 and the second power supply pad 208 via the first conductive lead and the second conductive lead. The region through which each microfluidic channel passes over the corresponding electrode array (referred to as the "DEP bed") is where the target particles are attracted and held during operation. At the point where the channel intersects the DEP bed, the DEP beds 209, 210, 211 typically have a larger width than the microfluidic channel-this ensures that all the sample passes through the electrodes (it does not flow around the outside).

In operation, an alternating current is applied to the power supply pads 207, 208. An alternating current propagates through the first and second electrically conductive leads, establishing an alternating electric field at each DEP bed 209, 210, 211. A fluid containing target particles flows through each microfluidic channel. Assuming that the voltage and frequency of the alternating current are properly selected, as the target particles pass through each DEP bed, they are attracted to the electrodes and held within the DEP bed due to dielectrophoresis.

In one example, the device is arranged to target and focus mycobacterium tuberculosis cells for visualization and further processing. In this arrangement, the electrode array typically has 20 pairs of electrode fingers, with the array having a bed width of 2.7mm intersecting a channel of 2mm and a total length of about 4mm (the width of each electrode finger and the spacing between the fingers is 50 microns), and the AC power supply provides a peak-to-peak voltage of about 12V (4V RMS) at a frequency of 10MHz, which generates an average electric field of 80kV/m RMS.

When looking at mycobacterium tuberculosis, the sample fluid is typically the sputum of a human subject (although it will be understood that a variety of cells may be targeted, and other biological sample materials may be used, including resuspended swab material, blood, plasma, saliva, etc.) that has been pre-treated, such as diluted with buffer and desalted.

In this way, processing and/or analysis of the fluid may be performed. For example, if each target microparticle is labeled with a fluorescent label, the number of target microparticles captured or retained in each DEP bed after a predetermined amount of flow time can be assessed by visual inspection using optical equipment such as a microscope (not shown).

Alternatively, or in addition, once the fluid containing the target particles has passed through the apparatus and the target particles are held in the DEP bed, a second fluid may be passed through the microfluidic channel while the alternating current is reduced or turned off. Thus, the target particles are released into a second fluid, which may be flushed into another chamber (not shown) for further processing or analysis or eluted from the apparatus. This provides a more concentrated sample for subsequent processing such as lysis, PCR (polymerase chain reaction) and nucleic acid detection.

For accurate analysis of target particles, especially to perform analyses involving estimation of target particle volume (volume) in a fluid sample, it is important that the DEP bed must operate consistently. That is, the dielectrophoresis is uniform so that each DEP bed attracts and holds the target particles to the same extent. For consistent operation of each DEP bed, it is important to ensure that the electric field differences produced by each electrode array are minimized.

As can be understood with reference to fig. 2, assuming that only one set of power supply pads is used, the electrode array gradually moves away from the first power supply pad and the second power supply pad from left to right.

The further the electrode is from the power supply pad, the further the distance the alternating current must travel through the conductive material before reaching the electrode. Therefore, as the distance from the electrode to the power supply pad increases, the total resistance between the power supply pad and the electrode gradually increases. This difference in resistance means that the voltage at each electrode array, and hence the electric field generated at each DEP bed, is different. Thus, the dielectrophoresis of each DEP bed is different so that the target particles are not attracted and retained to the same extent.

This is illustrated in the equivalent circuit shown in fig. 3, which is the equivalent circuit of the device shown in fig. 1, comprising three electrode arrays connected in parallel and powered by a single pair of power supply pads.

Fig. 3 shows that the resistance between the power supply pads for the first electrode array is different from the resistance between the power supply pads and the second electrode array, resulting in a voltage difference (and thus a difference in electric field strength) at each electrode array.

More specifically, fig. 3 shows the resistance (R) of the first electrode array_IDE1) Resistance (R) of the second electrode array_IDE2) And resistance (R) of the third electrode array_IDE3). Fig. 3 also shows the resistance (R) of the portion of the first conductive lead 108 between the first power supply pad 105 and the first input terminal of the first electrode array_L1) (ii) a The "additional" resistance (R) of the portion of the first conductive lead 108 between the first power supply pad 105 and the first input of the second electrode array_L2) And a further "additional" resistance (R) of the portion of the first conductive lead 108 between the first power supply pad 105 and the first input of the third electrode array_L3)。

Fig. 3 also shows the resistance (R) of the portion of the second conductive lead 109 between the second power supply pad 106 and the second input terminal of the first electrode array_L4) (ii) a The "additional" resistance (R) of the portion of the second conductive lead 109 between the first power supply pad 105 and the second input of the second electrode array_L5) And a further "additional" resistance (R) of the portion of the second conductive lead 109 between the second power supply pad 106 and the second input of the third electrode array_L6)。

As can be appreciated with reference to fig. 3, although the electrode arrays are connected in parallel, the voltage on each electrode will be different for a given voltage input Vac due to the difference in the total amount of resistance between each electrode input and the power supply pad. That is to say that the position of the first electrode,V_IDE1greater than V_IDE2And V is_IDE2Greater than V_IDE3。

To make V_IDE1、V_IDE2And V_IDE3The difference between them is minimized, and it is necessary to reduce the resistance of the conductive leads as much as possible (e.g., R in fig. 3)_L1、R_L2、R_L3、R_L4、R_L5And R_L6). Indeed, in an ideal implementation, the conductive leads (e.g., R in FIG. 3)_L1、R_L2、R_L3、R_L4、R_L5And R_L6) Will be zero. In such an ideal case, the equivalent circuit of fig. 3 would simply become the resistance of the three electrode arrays in parallel (as shown in fig. 4), and in this case the voltage (and hence the electric field) across the electrode arrays would be the same.

Therefore, it is desirable to reduce the resistance of the conductive leads as much as possible. This may be achieved by increasing the conductivity of the conductive leads, which for any given conductive material may be achieved by increasing the width or thickness of the conductive material forming the conductive leads. However, in implementations where size is a critical factor (e.g., point of care (POC) microfluidic devices), the surface area available for deposition of conductor material is greatly constrained. This same size constraint typically prevents each electrode array from being powered by its own set of power supply pads, which would otherwise be an alternative means of ensuring that a consistent voltage is applied across each electrode array. Increasing the thickness of the lead is also undesirable due to the resulting increase in production cost.

Connecting the electrode arrays in parallel ensures that if one electrode fails, the remaining electrodes will continue to function. Also, by connecting DEP electrodes in parallel, the electrode resistance of each individual DEP electrode does not itself become correlated with the "lead-in" of the downstream electrode.

According to certain embodiments of the present invention, it has been found that for implementations in which multiple electrode arrays are connected in parallel (thus making a single set of power supply pads available to power the electrodes), if the total resistance of each conductive lead to an electrode is approximately ten times less than the total resistance of each electrode, the difference between the electric fields at each electrode array will be sufficiently reduced to ensure an acceptable consistent level of operation.

Referring to fig. 1, the resistance between the point a and the point B is the total resistance of the conductive leads from the first power supply pad 105 to the first input terminal of the first electrode array 102, the resistance between the point B and the point C is the resistance of the first electrode array 102, and the resistance between the point C and the point D is the total resistance of the conductive leads from the second power supply pad 106 to the second input terminal of the first electrode array 102. Similarly, the resistance between the point a and the point E is the total resistance of the conductive leads from the first power supply pad 105 to the first input terminal of the second electrode array 103, the resistance between the point E and the point F is the resistance of the second electrode array 103, and the resistance between the point F and the point D is the total resistance of the conductive leads from the second power supply pad 106 to the second input terminal of the second electrode array 103. Similarly, the resistance between the point a and the point G is the total resistance of the conductive leads from the first power supply pad 105 to the first input terminal of the third electrode array 104, the resistance between the point G and the point H is the resistance of the third electrode array 104, and the resistance between the point H and the point D is the total resistance of the conductive leads from the second power supply pad 106 to the second input terminal of the third electrode array 104.

Thus, according to some embodiments of the present invention, the differences between the electric fields generated at the first electrode array 102, the second electrode array 103 and the third electrode array 104 are sufficiently reduced to ensure an acceptable consistent operating level under the following circumstances: such asThe calculated total resistance at all connected electrode arrays 102 (resistance between B and C), 103 (resistance between E and F), and 104 (resistance between G and H) is at least about ten times greater than the resistance between point a and point B + the resistance between point C and point D and the resistance between point a and point E + the resistance between point F and point D and the resistance between point a and point G + the resistance between point H and point D.

Methods of producing devices with appropriate resistance ratios are known to those skilled in the art, and the resistance balance may be modeled at the design stage and/or Quality Control (QC) tests may be performed to check the resistance ratios.

As described herein, in certain embodiments, a suitable resistance ratio may be provided by controlling the geometry and/or cross-sectional area of the conductive leads.

In other embodiments, suitable resistivity may be provided by selecting the materials comprising the conductive leads and/or the electrode array based on the resistivity of these materials, instead of or in addition to controlling the geometry and/or cross-sectional area of the conductive leads. In such embodiments, the material of the conductive leads has a lower resistivity than the material of the electrode array.

It should be understood that different combinations of materials for the conductive leads and the electrode array may be used to provide suitable resistance ratios.

Examples of suitable materials for the conductive leads include gold, silver, aluminum, beryllium, titanium, or chromium.

Examples of suitable materials for the electrode array include aluminum, beryllium, tungsten, zinc, nickel, titanium, or platinum.

In certain embodiments, the electrically conductive leads are comprised of gold and the electrode array is comprised of platinum. Advantageously, the combination may provide a suitable resistance ratio while also improving oxidation resistance.

In certain embodiments, a non-oxidizing protective layer may be provided to cover the conductive leads and/or the electrode array to reduce or prevent oxidation.

In certain embodiments, the conductive leads and/or electrode arrays may be composed of more than one layer of material to form a "composite" conductive lead or electrode array. For example, the conductive leads and/or the electrode array may be composed of multiple layers made of different materials.

In one embodiment, the base layer and the top layer are composed of a first material, and the intermediate layer located between the base layer and the top layer is composed of a second material different from the first material.

In another embodiment, the base layer and the top layer are composed of different materials. The base layer may be composed of titanium or chromium to promote adhesion to the substrate. The base layer may be about 5-10nm in depth. The top layer may be comprised of aluminum or gold.

In the above embodiments, the cross-sectional area and/or thickness of each layer may be independently selected to provide a suitable resistance ratio.

The level of consistency acceptable across the DEP electrode will vary depending on the sample being studied and the desired efficacy and efficacy of the current test. Preferably, an acceptable level of uniformity across the array of different electrodes is a field strength difference between the electrodes of less than 25%, more preferably a field strength difference between the electrodes of less than 10%, more preferably a field strength difference between the electrodes of less than 1%.

Too much difference may result in inefficient capture or manipulation of cells in at least some of the channels, and sometimes excessive heat generation in some of the channels.

In addition to the above requirements to ensure consistent DEP bed operation, it is also desirable to reduce the resistance of the conductive leads to minimize energy consumption due to heating of the conductive leads and to maximize energy consumption in the electric field at the DEP bed. Particularly considering the relatively high power dissipation involved in driving multiple electrode arrays (a device comprising 32 electrode arrays can be expected to consume about 1W of power). A particular problem that needs to be solved with multiple DEP arrays is that heating of the sample can cause serious problems, as the sample is particularly concentrated in DEP arrays in the middle of the cassette, which is difficult to cool.

Conductive leads used in conventional electronic applications on a scale similar to that of microfluidic devices (e.g., integrated circuits) are typically too small to provide the desired resistance levels. In other words, the conductivity of such conductive leads is too low. In other applications, a fairly thick conductive layer and/or wider leads are used to achieve acceptable levels of conductivity.

In order to minimize the resistance of the conductive leads connecting the electrode array to the power supply pads, the width and/or thickness area of the conductive material forming the conductive leads must be increased as much as possible. In general, it is necessary to increase the width of the conductive leads beyond the size typically used in electronic applications having similar scales.

However, on the scale of typical microfluidic geometries and using conventional techniques of lift-off lithography, in which a conductive material is bonded/deposited on a substrate, such as a polymer substrate (e.g., acrylic or polypropylene) or a glass substrate, to form conductive leads, increasing the surface area of the conductive leads beyond a certain size increases the likelihood that the conductive material will lift off the substrate. This is because, if the ratio of the surface area of the conductive material to the length of its outer edge is below a threshold value, the surface tension of the conductive material at the edge of the conductor lead becomes greater than the adhesion of the conductive material to the substrate. Typically, the ratio of the surface area of the conductive material to the length of its outer edge is reduced by providing an internal gap to the conductor leads to increase the amount of edge. Preferably, the internal gap is an elongate gap which results in a conductive material having a plurality of elongate sections or portions (this configuration has the added benefit that if one of the elongate sections has a manufacturing defect, this does not result in a complete failure.

According to an embodiment of the invention, the conductive leads are arranged such that both the conductor material surface area and the conductor lead edge length are maximized. More specifically, the conductive leads include an internal gap that is free of conductive material to increase the ratio of the outer edge of the conductive lead to its total surface area. This reduces the surface tension of the conductor material and increases the adhesion at the edge of the conductive lead, thereby reducing the likelihood of delamination.

Fig. 5 depicts this concept.

Fig. 5 provides a schematic diagram illustrating a portion of a conventional conductive lead 501 used in electronics on a scale similar to a particular microfluidic application. For certain microfluidic applications, for example, those including multiple electrode arrays, particularly those including multiple electrode arrays driven by a single set of power pads, the surface area of the conductive leads may not provide sufficient conductive material to provide the desired level of conductivity.

Fig. 5 shows a portion of a conventional modified conductive lead 502 that provides a desired level of conductivity due to the addition of conductive material. However, the edge of the conductive lead 502 risks peeling off from the substrate due to the increase in surface tension.

Fig. 5 also shows a portion of the conductive leads 503, 503a, according to a particular embodiment of the present invention, wherein the portion of the conductive leads 503, 503a provides a desired level of conductivity due to an increased amount of conductive material, but includes internal gaps 504, 504a that are devoid of conductive material to increase the ratio of the length of the outer edges of the conductor leads to the total surface area thereof. It can also be seen that the inner gaps 504, 504a can have varying shapes.

In the embodiment shown in fig. 1, the conductive leads 107a, 107b, 107c, 108a, 108b, 108c connecting the electrodes to the power supply pads are provided by a plurality of connected parallel conductive leads separated by gaps. Providing a plurality of connected conductive leads introduces an internal gap within the conductive leads and thereby creates an overall edge length of the conductive leads, which thus reduces the risk of peeling of the conductive leads from the substrate.

It will be appreciated that any suitable shape and size of gap may be used. For example, in some cases, the gap may be a square cut rather than parallel elongated sections.

Fig. 6 provides a schematic view of a microfluidic device according to a particular embodiment of the present invention.

More specifically, fig. 6 provides an exploded view of a microfluidic cartridge 601 comprising a first substrate layer 602 (typically comprising polypropylene). As described above, on the substrate layer 602 is the conductive layer 603, and the conductive layer 603 includes an electrode array, a conductive lead, and a pair of power supply pads. In particular, as described above, the electrode array, the conductive leads and the power supply pads are arranged such that the resistance of the conductive leads between the input end of each electrode and the power supply pad is substantially at least an order of magnitude less than the total resistance of the connected electrode array. Further, in certain embodiments, as described above, the conductive leads include an internal gap that is free of conductive material to increase the ratio of the outer edge of the conductive lead to its total surface area.

Above the electrically conductive layer is a microfluidic channel layer 604, the microfluidic channel layer 604 comprising microfluidic channels arranged to direct fluid onto the electrode array, which forms a DEP bed in which target particles are attracted and held.

The microfluidic channel layer 604 extends over the portion of the conductive layer comprising the electrode array, but exposes the power supply pads 605a, 605b for connection to an AC power supply. The microfluidic channel layer 604 includes: an inlet port 606 through which a fluid to be analyzed is driven; and a corresponding outlet port 607 through which fluid that has passed through the cartridge exits.

In use, the microfluidic cartridge is inserted into a point-of-care analysis device that receives a fluid sample, performs any necessary pre-processing steps (e.g., changing the viscosity, salt content (ionic strength) of the fluid, adding dyes or fluorophores, etc.), and then drives the fluid (via suitable microfluidic pumping as is well known in the art) into the inlet port 606. At the same time, insertion of the microfluidic cartridge brings the AC power supply into contact with the power supply pads 605a, 605 b. The AC power supply provides an alternating current at a predetermined frequency and voltage that, given the geometry of the electrodes, causes the target particles to be retained within the DEP bed as fluid flows through the microfluidic cartridge. The cartridge holder may also provide a degree of cooling to the cartridge.

As described above, in one mode of operation, once the analysis device drives the fluid sample all the way through the microfluidic cartridge, the analysis device drives a second fluid (e.g., purified water) through the microfluidic cartridge and turns off the AC power supply. In this way, target particles are released from the DEP bed via the outlet port 607 and exit the cassette, and can be collected in a collection chamber. Analysis of the target particles in the collection chamber can then be performed by an analytical device.

It will be appreciated that one possible implementation of a microfluidic device according to a particular embodiment of the present invention is described above.

While specific embodiments are described above, it should be understood that in a simple embodiment, there is no complex chip for a separate point of care (POC) device. The cartridge or chip is a self-contained microfluidic chip device that flows the sample through multiple channels, the cells of interest are manipulated by the DEP electrodes such that they are held on or near the DEP electrodes, and then the DEP electrodes can be simply viewed under a microscope to see if the cells have been held and are or are not present.

Similarly, in more complex variations, microfluidics can be combined in a single cartridge to allow for concentration and additional processing, such as lysis (lysis), PCR, molecular detection, and the like.

The electrode array has both resistance (due to the conductivity of the solution) and reactance (the electrode fingers act as capacitors), while the leads have only resistance. Throughout this document we refer to the electrode array as "resistive" because the reactance of the electrode array need not be considered in the above embodiments. In fact, for "typical" solution conductivity, and at 10MHz, the electrode reactance in terms of voltage drop is negligible. However, if the sample conductivity decreases or the frequency increases significantly, it may become significant. In this regard, the term "resistance" should be understood to mean the magnitude of the AC impedance.

All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and/or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations and/or steps in which at least some of such features are mutually exclusive. Each feature disclosed in this specification (including any accompanying claims, abstract and drawings) may be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise. Thus, unless expressly stated otherwise, each feature disclosed is one example only of a generic series of equivalent or similar features. The invention is not restricted to the details of the foregoing embodiments. The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

With respect to the use of substantially any plural and/or singular terms herein, those having skill in the art can translate from the plural to the singular and/or from the singular to the plural as is appropriate and/or applicable. Various singular/plural permutations may be expressly set forth herein for the sake of clarity.

It will be understood by those within the art that, in general, terms used herein, and especially in the appended claims, are generally intended as "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "includes" should be interpreted as "includes but is not limited to," etc.). It will be further understood by those within the art that if a specific number of an introduced claim recitation is intended, such an intent will be explicitly recited in the claim, and in the absence of such recitation no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed to imply that the introduction of a claim recitation by the indefinite articles "a" or "an" limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases "one or more" or "at least one" and indefinite articles such as "a" or "an" (e.g., "a" and/or "an" should be interpreted to mean "at least one" or "one or more"); the same holds true for the use of definite articles used to introduce claim recitations. In addition, even if a specific number of an introduced claim recitation is explicitly recited, those skilled in the art will recognize that such recitation should be interpreted to mean at least the recited number (e.g., the bare recitation of "two recitations," without other modifiers, means at least two recitations, or two or more recitations).

It will be understood that various embodiments of the present disclosure have been described herein for purposes of illustration, and that various modifications may be made without departing from the scope of the disclosure. Therefore, the various embodiments disclosed herein are not intended to be limiting, with the true scope being indicated by the following claims.