A method including a digital microfluidic (DMF) system, a DMF cartridge, and integrated optical fiber sensing.

Abstract

A cartridge for use with an instrument for performing fluid measurements, an instrument for operating the cartridge, and a method for manufacturing and using the cartridge. [Solution] The present invention is a cartridge for use with an instrument for performing fluid measurement, the cartridge comprising: a digital microfluidic substrate having a plurality of electrowetting electrodes that operate to perform droplet operations on liquid droplets in a droplet operation gap; a top plate separated from the digital microfluidic substrate to form a droplet operation gap and having an opening for injecting liquid into the droplet operation gap; and a fiber assembly including an optical fiber probe having a sensing end that protrudes into the droplet operation gap and is positioned in close proximity to one or more electrowetting electrodes.

Description

Cross - reference to related applications

[0001] This application claims priority to U.S. Patent Application No. 62 / 964,424, filed on January 22, 2020 which is incorporated herein by reference.

Technical Field

[0002] This disclosure relates to digital microfluidic (DMF) systems, DMF cartridges, and methods of using integrated optical fiber sensing.

Background Art

[0003] Digital microfluidic systems are useful for handling and manipulating droplets for various lab - on - a - chip applications. Optical sensing techniques are commonly used in digital microfluidic systems, for example, for detecting analytes within droplets. Simple fiber optic sensors have been described for use in detecting light in digital microfluidic systems. However, there is a need in the art to enhance the optical fiber sensing capabilities for performing complex analyses using digital microfluidics.

Summary of the Invention

Problems to be Solved by the Invention

[0004] This disclosure provides a cartridge for use with an apparatus for performing fluid measurements, an apparatus for operating the cartridge, and methods of fabricating and using the cartridge. This disclosure also includes a fiber assembly useful for assembling the cartridges of the disclosure.

Means for Solving the Problems

[0005] In one embodiment, the cartridge performs droplet manipulation on liquid droplets within the droplet manipulation gap. A digital microcontroller with multiple electrowetting electrodes that operate to perform the operation. Contains chlorofluid. The cartridge also contains a set of two or more electrowetting electrodes. This may include an optical fiber probe that enters the droplet manipulation gap adjacent to the torch, As a result, droplets located on any electrode among the set of two or more electrodes will fall into the optical fiber. You can make contact with the robe.

[0006] In another embodiment, the cartridge controls the liquid droplets within the droplet operation gap. A digital machine with multiple electrowetting electrodes that operate to perform the operation. A microfluidic substrate and a digital microfluidic substrate to form a droplet operation gap A top plate that is separated and has an opening for injecting liquid into the droplet operation gap. The cartridge also enters the droplet operation gap and has one or more Optical fiber with a sensing end positioned in close proximity to the electrowetting electrode. It may include a fiber assembly with a probe.

[0007] In some embodiments, two optical fiber probes enter the droplet manipulation gap. Arranged in close proximity to the above set of electrowetting electrodes, two or more of the electrodes A droplet located on any electrode in the set can come into contact with the optical fiber probe. The probe can contain a ligand. The droplet in the cartridge is electrolytic. The contact electrode can be controlled to reach the optical fiber probe. Low viscosity Using oil or other filler material (e.g., filler fluid), gaps around the droplet It can fill gaps.

[0008] This disclosure also provides a method for performing an assay. The method involves a photopharmaceutical containing a ligand. The steps include preparing the Iber probe, which has a volume of less than approximately 1000 nL, and the ligand. The steps involve preparing droplets containing a sample that has potential affinity for the probe, and then testing the droplets with a probe. The end of the probe is brought into contact with the droplet, and the droplet is vibrated without the droplet leaving contact with the probe. It can be equipped with a step. In some cases, the oscillating range is approximately 0. The frequency is approximately 5 to 15 Hz, or approximately 4 to 10 Hz. In some cases, the volume of the droplet is approximately 90 Less than 0 nL, or less than approximately 800 nL, or less than approximately 700 nL, or less than approximately 600 nL It is full, or less than approximately 500 nL, or less than approximately 400 nL. In some cases, the volume of the droplet The product is between 200 and 400 nL.

[0009] In some cases, the fiber optic probe contains multiple ligands, and the droplet contains multiple samples. In some cases, the method involves preparing multiple optical fiber probes and multiple droplets. The step involves bringing each of the multiple droplets into contact with the corresponding optical fiber probe, and the The step of vibrating each of the plurality of droplets that are in contact with the corresponding optical fiber probe. and, including. In another embodiment, for a series of assays, a single droplet is used in a certain process. It can be transported from one probe to another.

[0010] In some cases, the vibration is transmitted via electrowetting electrodes. Vibration is for droplet manipulation of a droplet actuator or an electro-wetting cartridge. It is performed through an electro-wetting electrode in the gap. In some cases, the optical fiber probe is aligned such that its end is adjacent to the droplet manipulation electrode. In some cases, the optical fiber probe is aligned such that its end is close to the edge of the droplet manipulation electrode. In some cases, the vibration is approximately orthogonal to a line extending along the length of the optical fiber probe. In some cases, the vibration substantially coincides with a line extending along the length of the optical fiber probe. In some cases, the vibration is multi-directional. In some cases, the vibration is multi-directional within a plane parallel to a line extending along the length of the optical fiber probe. In some cases, the vibration is performed using an elongated droplet that is, for example, 2X, 3X, 4X or longer than that, where X is the number of electro-wetting electrodes used to elongate the droplet. In some cases, the assay is selected from the following, namely, molecular library screening assay, binding kinetics assay, affinity determination assay, binding site mapping assay, competition analysis assay, specificity determination assay, and characterization of antibody binding, and combinations of the above. In some cases, the assay generates a response curve.

[0011] In some cases, the assay is selected from the following, namely, molecular library screening assay, binding kinetics assay, affinity determination assay, binding site mapping assay, competition analysis assay, specificity determination assay, and characterization of antibody binding, and combinations of the above. In some cases, the assay generates a response curve. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The gist of the invention has been explained above using general terminology, but the following may not necessarily be drawn to scale. I will now explain the attached drawings that are not included. [Figure 1] A block diagram of an embodiment of a DMF system including integrated optical fiber sensing is shown. [Figure 2] Figure 2A shows an example of a DMF cartridge related to a DMF device, and also shows an example of an optical fiber interface. Figure 2B illustrates an example of a DMF cartridge related to a DMF device, and an example of an optical fiber interface. [Figure 3] This is a side view of a portion of an embodiment of a DMF cartridge in a DMF system, with an optical fiber probe introduced into the gap from above. [Figure 4] This is a side view of a portion of an embodiment of a DMF cartridge in a DMF system, with an optical fiber probe introduced into the gap from below. [Figure 5] This is a side view of a portion of an embodiment of a DMF cartridge in a DMF system, where an optical fiber probe is introduced into the gap from the side. [Figure 6] This is a side view of a portion of an embodiment of a DMF cartridge in a DMF system, where an optical fiber probe is introduced into the gap and optical sensing operations are performed in reflective mode. [Figure 7A] A side view of a portion of an embodiment of a DMF cartridge for a DMF system is shown, in which the sensor surface is located at the tip of an optical fiber probe. [Figure 7B] A side view of a portion of an embodiment of a DMF cartridge for a DMF system is shown, in which the sensor surface is located at the tip of an optical fiber probe. [Figure 8] A side view of a portion of an embodiment of a DMF cartridge for a DMF system is shown, in which one or more optical elements are provided at the tip of an optical fiber probe. [Figure 9]A side view of an embodiment of a DMF cartridge for a DMF system, including both a primary optical measuring device and a secondary optical measuring device, is shown. [Figure 10] A flowchart illustrating an example of how to use a DMF system and / or DMF cartridge, including integrated optical fiber sensing, is shown. [Figure 11] An example of a sample droplet that vibrates back and forth on the droplet manipulation electrode and is in contact with the optical fiber probe is illustrated. [Figure 12A] An oblique view from the top of an exemplary instance of a DMF cartridge including integrated fiber sensing is shown. [Figure 12B] An oblique view from the bottom of an exemplary instance of a DMF cartridge containing integrated fiber sensing is shown. [Figure 13A] An exploded view from the top of an exemplary instance of a DMF cartridge containing integrated fiber sensing is shown. [Figure 13B] An exploded view from the bottom of an exemplary instance of a DMF cartridge containing integrated fiber sensing is shown. [Figure 14A] Figures 14A and 14B illustrate one of several embodiments of the fiber assembly of a DMF cartridge. [Figure 14B] One of several embodiments of the fiber assembly for a DMF cartridge is illustrated. [Figure 15] This document shows an example of the process for prototyping fiber sensing. [Figure 16] An example of an electron microscope image of a nanoparticle sensor surface is shown. [Figure 17] Figures 17A and 17B show plots of exemplary test results in which the affinity of protein A and IgG was determined using fiber-optic-based surface plasmon resonance detection within a DMF device. [Modes for carrying out the invention]

[0013] The present invention relates to a digital microfluidic (DMF) system, a DMF cartridge, and a collection This relates to a method including integrated optical fiber sensing.

[0014] DMF systems and DMF cartridges, for example, the droplet operation of DMF cartridges. This may include an optical fiber probe directly inserted into the gap. DMF system For example, the components include a DMF cartridge, one or more illumination sources, and one or more optical measuring devices. and may include a controller. The DMF cartridge is a DMF cartridge This may further include an optical fiber probe directly inserted into the droplet manipulation gap. .

[0015] Several mechanisms, DMF systems, DMF cartridges, and methods are described below. The tip of the optical fiber probe is directly inserted into the droplet handling gap of the DMF cartridge. To provide an optical fiber probe.

[0016] Several mechanisms, DMF systems, DMF cartridges, and methods are described below. Optical fiber plugs inserted through the top, bottom and / or sides of the DMF cartridge They may offer a robe.

[0017] In some embodiments, the present disclosure relates to the droplet operation gap of a DMF cartridge. We propose a single optical fiber probe that can function as a conduit for light entering and exiting the sensing region. To provide.

[0018] The droplets used in the assays disclosed herein are much smaller in volume than in many conventional assays. For example, less than approximately 1000 nL, or less than approximately 900 nL, or less than approximately 800 nL, or less than approximately 700 nL, or less than 600 nL, or less than approximately 500 nL, or approximately 40 Less than 0 nL, or less than approximately 300 nL, or less than approximately 200 nL, or less than approximately 100 nL It could be full.

[0019] The droplets used in the SPR assay described herein are different from those used in many conventional SPR assays. Small amounts, for example, less than approximately 1000 nL, or less than approximately 900 nL, or approximately 800 Less than nL, or less than approximately 700 nL, or less than 600 nL, or less than approximately 500 nL, Or less than approximately 400 nL, or less than approximately 300 nL, or less than approximately 200 nL, or approximately It may be less than 100 nL.

[0020] The droplets used in the SPR biomolecular interaction assays disclosed herein are similar to those used in many conventional SPR assays. Much smaller amounts than those used in somatic molecular interaction assays, for example, less than about 1000 nL, or Less than approximately 900 nL, or less than approximately 800 nL, or less than approximately 700 nL, or 600 nL Less than L, or less than approximately 500 nL, or less than approximately 400 nL, or less than approximately 300 nL, Or it may be less than approximately 200 nL, or less than approximately 100 nL. SPR biomolecular interaction Examples of assays include molecular library screening assays and binding reaction rate assays. I. Affinity determination assay, binding site mapping assay, competitive analysis assay, specificity determination assay. Examples include assays and characterization of antibody binding.

[0021] In some embodiments, the present disclosure provides a way to supply excitation light to an optical fiber probe. One or more light sources and light emitted from the same optical fiber probe are received and processed. It may provide one or more optical measuring devices for this purpose.

[0022] In some embodiments, the present disclosure uses a free-space optical system to supply excitation light. One or more light sources and one or more optical fibers for receiving and processing emitted light. A probe and can be provided. Similarly, in some embodiments, the present disclosure provides optical fiber One or more illumination sources to supply excitation light coupled to a bar probe, and free-space optics It may provide one or more optical measuring devices for receiving and processing emitted light. ru.

[0023] In some embodiments, the present disclosure directly connects the droplet operation gap of the DMF cartridge. An indirectly inserted optical fiber probe, in which the optical sensing operation is in reflection mode, transmission mode Optical fiber probes, which can occur in both reflection and transmission modes. To provide.

[0024] In some embodiments, the present disclosure relates to the droplet operation gap of a DMF cartridge. At the tip of the optical fiber probe, other components and / or elements (e.g., sensing It provides layers, optical elements.

[0025] Furthermore, this disclosure relates to a DMF system in which integrated optical fiber sensing is provided. And provides instructions on how to use the DMF cartridge.

[0026] Figure 1 shows a block diagram of an embodiment of the DMF system 100 including integrated optical fiber sensing. This is a diagram. The DMF system 100 is, for example, a plasmon resonance (P) system for the analysis of a sample. This could be a R) system and / or a localized surface plasmon resonance (LSPR) system. Analysis, for example, involves the detection, identification, quantification, or measurement of a sample, and / or the combined reaction. This can refer to the interaction between the substance and the specimen, such as kinetic and thermodynamic interactions. An example specimen is... This may include small molecules, proteins, peptides, atoms, and ions, but these may include Not limited to. For example, using the DMF system 100, ligators to macromolecules such as receptors can be used. The rate of the bond reaction can be measured.

[0027] The DMF system 100 also uses biolayer interferometry (BLI) or monochromatic reflection. These can be configured as reflectance interferometer sensors such as ctometry (SCORE). Ingress sensors use technologies such as chemical vapor deposition (CVD) or physical vapor deposition (PVD), For thin film coatings of zinc sulfide, titanium dioxide, magnesium fluoride, silicon dioxide, etc. Using common materials, light is produced along with the ultimate metallic layer of plasmonic metals such as gold. It is deposited onto the tip of the fiber. For example, the ligand can be immobilized on the tip of the fiber. The wavelength of light reflected from the sensor depends on the presence or absence of a sample interacting with the ligand. This causes a shift or change in the interference pattern. Analysis is, for example, the detection of a sample. Identification, quantification, or measurement, and / or other interactions between a sample and other substances, such as binding reaction rates. This can mean an interaction. Examples of samples include small molecules, proteins, peptides, atoms, and may include, but is not limited to, ions. For example, using DMF system 100 This method can be used to measure the binding kinetics of ligands to macromolecules such as receptors. In this state, the light reflected by the BLI fiber sensor exposed to the sample is the sample This can be compared to light reflected by a BLI fiber sensor that is not exposed to the light.

[0028] The DMF system 100 can also be configured as a fluorescence measurement system. The sensor can be an unmodified sensor, or a fluorescent substance (fluorophore) can be used. It can be fixed to the tip of the fiber. In another example, the tip of the fiber is quenched By means of, or by energy transitions such as Forster resonance energy transition, fluorescence It can be coated using a coating that modifies the luminescence properties of the material. Alternatively, multiple fluorescent substances can be used in combination with each other simultaneously. The fluorescence intensity and / or lifetime can be measured. It may be either in the solution or captured at the tip of the optical fiber. The fluorescence may be specific to the sample, or it may be bound to the sample, or Potential for generating fluorescent materials using small molecules, quantum dots, and other interacting materials. There are also analyses, for example, the detection, identification, quantification, or measurement of a sample, and / or This can refer to interactions between the sample and other substances, such as binding reaction rates. Examples of samples include: This may include, but is not limited to, small molecules, proteins, peptides, atoms, and ions. No. For example, using the DMF system 100, ligands can be bound to macromolecules such as receptors. The reaction rate can be measured.

[0029] The DMF system 100 combines fiber optic-based sensors with digital microfluidics. It is characterized by integration. For example, the DMF system 100 uses the DMF cartridge 110. The DMF cartridge 110 may include, for example, droplet merging, splitting, dispensing, and dilution. A droplet actuator device that provides DMF functionality for generally performing droplet manipulation such as the above. It could be. One application of these DMF functions is sample preparation. Furthermore, the DMF function is used in other processes such as flushing during waste removal or execution. It may be used. The DMF cartridge 110 is coupled to the optical fiber probe 130. This may include an onboard sensing area 158 in the DMF cartridge 110. A single optical fiber probe 130 enters and exits the onboard sensing area 158. It can function as a conduit for light. The optical fiber probe 130 is one optical fiber or May include bundles of multiple optical fibers. DMF cartridge 110, onboard sensing Details of region 158 and the optical fiber probe 130 are shown below in Figures 3 to 9. It will be explained.

[0030] In various embodiments, the sensing end of the optical fiber probe 130 is It is aligned adjacent to the droplet handling electrode 120. In various embodiments, optical fiber - The end of the probe 130 is aligned so that it is close to the edge of the droplet handling electrode 120. In various embodiments, the end of the optical fiber probe 130 is such that the end is capable of handling droplets. It is aligned so as to be in a straight line with the edge of the electrode 120.

[0031] The DMF system 100 further includes a controller 150 and a DMF interface 15 2. It may include an illumination source 154 and an optical measuring device 156. Controller 1 50 includes a DMF cartridge 110, an illumination source 154, and an optical measuring device 156, etc. It can be electrically coupled to various hardware components of the DMF system 100. For example The controller 150 then connects to the DMF cartridge via the DMF interface 152. It can be electrically coupled to 110, where the DMF interface 152 is, for example For the DMF cartridge 110, a plug-connectable connector is provided for mechanical and electrical connection. It can be used as an interface. DMF cartridge 110, controller 150 The DMF interface 152, illumination source 154, and optical measuring device 156 are, This also forms the DMF device 105.

[0032] Controller 150 is used, for example, in general-purpose computers, dedicated computers, and personal computers. To be used as a computer, microprocessor, or other programmable data processing device. The controller 150 can store, interpret, and / or implement software instructions. It provides processing capabilities for rows and other data, and controls the overall operation of the DMF system 100. This is useful. Controller 150 controls the data and / or power aspects of these devices. It can be configured and programmed to control the following. For example, controller 150 This is achieved by activating / deactivating the electrodes of the DMF cartridge 110. It controls the operation of droplets inside. Generally, the controller 150 controls any of the DMF system 100. It can be used for various functions. For example, the controller 150 can be used by the printer manufacturer itself DMF cartridges can be checked in a similar way to how to check the ink cartridges of a particular brand. Controller 150 can be used to authenticate 110, and DMF cartridge It can be used to verify that the 110 has not expired, and control The LA150 operates by executing a protocol for its purpose, thereby replacing the DMF cartridge It can be used to check the cleanliness level of 110, and so on.

[0033] The controller 150 has one or more input interfaces that connect the processing unit to the input device. - May include a face. The input interface allows the user of the DMF system 100 to... This allows commands to be communicated to the processor. Such an example command is One is the execution of program code. Input devices include keyboards and mouse devices. , voice activation systems, touchscreens, and / or other suitable systems known to those skilled in the art It can take the form of a device.

[0034] In some embodiments, the controller 150 provides a graphical user interface. One or more output interfaces (such as GUIs) that connect processing units to output devices. - May include faces. This allows the DMF system 100 to process various experimental results and other information. It becomes possible to communicate the results of the operation to the user. Software instructions are controllers It can be stored in the RA150 memory unit, and conventional semiconductor random access memory (RAM) or other forms of memory known in the art may include, and / or The software instructions are for hard drives, USB drives, and read / write CDs. One or more such as ROM, DVD, tape drive, flash drive, optical drive, etc. These instructions can be stored in the form of program code on a computer-readable storage medium. It is executed in response to the interaction between the DMF system 100 and the user via an input device. obtain.

[0035] In some embodiments, the DMF cartridge 110 is a capacitive feedback system. This may include sensing. In other words, signals from a volume sensor that can detect the position and volume of a droplet. This is the number. Furthermore, in other embodiments, instead of capacitive feedback sensing, In addition to or in addition to the above, the DMF cartridge 110 optically analyzes the position and volume of droplets. It may include a camera that takes measurements, and these measurements trigger the controller 150. This allows the droplet to be rerouted to the appropriate location.

[0036] In some embodiments, the DMF cartridge 110 has a heating zone (not shown) This may include various sample preparation steps and assays that benefit from temperature control. It will be understood that this is incorporated herein by reference. U.S. Article 8,658,11 As described in No. 1, thermal control is generally done in the following three ways, namely, (1 (2) Thermal control of the entire DMF cartridge 110, and whether it is in contact with the controlled area or The DMF cartridge 110 uses a heater located in close proximity to the controlled area. (3) Thermal control of a certain region, and using a heater integrated into the DMF cartridge 110 The heat of a certain area of ​​the DMF cartridge 110 or the entire DMF cartridge 110 is used. Control (e.g., within a substrate including electrode paths or arrays, and / or on a top substrate) When this happens, it can be obtained in the manner of the upper circuit board of the DMF cartridge 110. Combinations of the methods described above are also possible. An example of heating technology is the one adjacent to the cartridge. Examples include heater bars attached to the device and heaters integrated into the cartridge itself. It can be done.

[0037] In the integrated heater method, thermal control is directly integrated into the DMF cartridge 110. The element can be used to create and control temperature zones. Thermal control element (heating and (and / or cooling) the lower and / or upper boards of the DMF cartridge 110. If present, and if integrated on the bottom and / or top surface of either substrate It is possible, or it can be integrated within the structure of either substrate, or placed between substrates. It is possible. In some cases, the thermal control element is electronically coupled to the controller 150. Furthermore, it can be controlled. A heat zone can be created using a separate heating element. Therefore, it can function as a separate heat zone within the DMF cartridge 110. Depending on the configuration, multiple steps in analysis, such as sample preparation and thermal cycling, are possible. This is performed simultaneously at different temperatures in different thermal zones of the DMF cartridge 110. This requires different temperatures. For example, droplet manipulation via electrowetting In use, droplets are physically transported or travel back and forth between thermal zones of different fixed temperatures. This allows for the implementation of a thermal cycle for amplification reactions.

[0038] In one embodiment, the heater of the heat zone is formed using a thin conductive film. This is possible. Suitable thin film examples include Pt heater wire and transparent indium oxide. One example is (ITO). In one embodiment, the substrate of the DMF cartridge 110 Metal vias (e.g., copper) are used.

[0039] The temperature of the heat zone can also be controlled using thermocouples for temperature regulation. Using a device, the temperature of the heat zone is measured and the temperature readings are supplied to the controller. The roller can be used to precisely control the temperature of the relevant heat zone.

[0040] The DMF device 105 can be connected to a network. For example, controller 15 0 can communicate with networked computer 160 via network 162. The networked computer 160 is, for example, any centralized server or It can be used as a cloud server. Network 162 is, for example, the internet. A local area network (LAN) or wide area network to connect to It can be designated as WAN.

[0041] In the DMF system 100, the illumination source 154 and the optical measuring device 156 are DMF Onboard sensing area 158 of cartridge 110 and / or fiber optic probe It can be positioned in relation to B130. The lighting source 154 is not limited to the following. However, for example, white light-emitting diodes (LEDs), halogen bulbs, arc lamps, incandescent lamps, These include lasers, which can be used as light sources in the visible range (wavelength 400-800 nm). The illumination source 154 is not limited to a white light source. The illumination source 154 is a DMF system 100 This allows for any useful color of light. The illumination source 154 is a DMF cartridge 110 Excitation light 132 is supplied to the onboard sensing region 158.

[0042] The optical measuring device 156 is used, for example, to obtain a reading of light intensity. It can be an optical transducer device. The optical measuring device 156 is, for example, a charge-coupled Composite devices, photodetectors, spectrometers, photodiode arrays, cameras, or any of the above. This can be combined with a single light source 15. The DMF system 100 is not limited to four and one optical measuring device 156. Any detection operation in the DMF system 100 and / or DMF cartridge 110 To support this, multiple illumination sources 154 and / or multiple optical measuring devices 156 It can include. The optical measuring device 156 is onboard the DMF cartridge 110. The emitted light 134 from the sensing region 158 is received and processed. Therefore, DMF The single optical fiber probe 130 of cartridge 110 is located in the onboard sensing area. Both excitation light 132 to 158 and emitted light 134 from the onboard sensing region 158 It can function as a conduit for that purpose.

[0043] The components of the DMF system 100 and / or DMF device 105 are DMF cartridges. Onboard sensing area 158 and / or fiber optic probe 13 of the ledge 110 It can be optically coupled to 0, and can also be decoupled. Optical coupling / decoupling is, for example, used in optical fiber connectors. This can be a fiber coupler and / or a free-space optical coupler.

[0044] Conventional methods for measuring optical stimuli from DMF devices use free-space optics to capture the stimuli. To capture. In this context, the limitations of free-space optics are mainly due to stray light and optical loss, which affect performance. This is due to a relatively lower performance. The decrease in performance is due to the lens, phi Using optical components such as routers and other similar optical components These are often overcome by [other methods], but these components reduce the cost of the system. To finish. Compared to conventional methods, the integration of optical fiber sensors into the DMF is slightly This has advantages. For example, the integration of fiber optic-based sensors and digital microfluidics. The advantages of the DMF system 100, which features the following, are, but are not limited to, local optics To enable surveying techniques, and low-cost methods to achieve high optical performance (for example, low cost To provide a reduction in the number of components, and to improve the signal, light within the system It may be possible to limit losses and limit the collection of stray light to reduce noise.

[0045] In the DMF system 100, the optical fiber probe 130 is the optical fiber probe 1 The tip of 30 is directly located in the droplet handling gap of the DMF cartridge 110. It is integrated into the DMF cartridge 110 (see Figures 3 to 9). In this way, the optical fiber The tip of the Iber probe 130 is directly in contact with the droplets being processed by the DMF cartridge 110. They can interact with each other. Therefore, the tip of the optical fiber probe 130 is onboard. A sensing area 158 is formed, where the DMF cartridge 110 is onboard sensing The droplet can be moved between the sizing region 158 and the DMF system 100 and / Alternatively, the DMF cartridge 110 contains one optical fiber probe 130 and one O Not limited to the onboard sensing area 158. This is merely an example. DMF Stem 100 and / or DMF cartridge 110 can connect to any number of fiber optic cables. It may include a range 130 and / or an onboard sensing area 158. Furthermore, one or more The optical fiber probe 130 above can be provided in the form of a fiber assembly 125. It is possible (see Figures 14A and 14B). Further details of the embodiment of the optical fiber probe 130 are shown below with reference to Figures 3 to 9. It is done and explained.

[0046] Figures 2A and 2B show an example of the optical fiber interface of DMF device 10. An embodiment of the DMF cartridge 110 related to 5 is shown. In this embodiment, DMF Cartridge 110 can be dropped into a compatible DMF device 105 such as DMF device 105. It can be designed to do so. In this example, the DMF device 105 is a DMF cartridge It includes a recessed area 164 for receiving the ledge 110, i.e., the DMF cartridge. 110 is sized to fit into the recessed area 164 of the DMF device 105. In this manner, the DMF cartridge 110 is fluidly coupled to the DMF device 105, optically coupled, and / or electrically coupled. This embodiment involves coupling the cartridge to the device. This automatically aligns the optical fiber elements of the cartridge with the optical fiber elements of the device. This is an example of various embodiments. In one embodiment (not shown), alignment is performed on the corresponding devices. A post or opening that fits into the post or DMF fiber interface The opening can be increased by various similar methods that are apparent to those skilled in the art.

[0047] For example, regarding optical coupling, the optical fiber probe 130 of the DMF cartridge 110 is This can include one optical fiber or a bundle of multiple optical fibers. The fiber can be used in multi-mode, single-mode, or a combination of the two (multiple colors) It may have a cladding layer and / or a DMF cartridge 110. This allows one or more optical fibers to be coupled to the DMF device 105, It may include one or more interfaces. An optical interface is, for example, This includes optical fiber connectors, optical fiber couplers, and / or free-space optical couplers. This is possible. For example, Figures 2A and 2B show that the DMF cartridge 110 is connected to the DMF device 1 When loaded into the recessed area 164 of 05, the optical fiber inside the DMF cartridge 110 The outer end of the probe 130 is connected to, for example, the illumination source 154 and / or the optical measuring device 1 This shows that one or more optical fibers 166 connected to 56 can be nearly in a straight line. The system and cartridge are connected to the DMF cartridge when the DMF cartridge is connected to the device. To ensure that the optical connection between the fiber element and the cartridge is optically coupled Alignment elements can be included. For example, DMF cartridge 110 is connected to DMF device 10 When inserted into 5, the DMF device 105 connects the fiber and D in the DMF cartridge 110. Automatic alignment step to maximize coupling efficiency between the fiber and the MF device 105 This can be done to electrically and optically couple the cartridge to the device. It should be understood that a wide variety of mechanisms are possible.

[0048] Figures 3, 4, and 5 show the DMF cartridge 11 of the DMF system 100 shown in Figure 1. The image shows a side view of an embodiment in part of 0, where the optical fiber probe 130 is located above It is introduced into the gap from the front, bottom, and sides. The DMF cartridge 110 is a droplet controller. It includes a lower substrate 112 and an upper substrate 114 separated by a gap 116. Yes, it is possible. The droplet operation gap 116 is a filling such as a gas or liquid that does not mix well with the droplet. A filler fluid that does not significantly interfere with the desired analytical process, and which can be used to fill a room with the filler fluid. Yes, it is possible. In one embodiment, the filler fluid is a silicone oil or hexadecane, etc. It is a low-viscosity oil. Furthermore, the droplet operation electrode 120 (for example, electrowetting) The arrangement of the electrodes can be provided on the lower substrate 112. DMF cartridge 11 0 can include any line or path of the droplet handling electrode 120.

[0049] In one embodiment, the lower substrate 112 is almost transparent to white light (or any colored light). It can be made of a material that exhibits brightness. For example, the lower substrate 112 can be glass, plastic, etc. or formed from polymers known as thermoplastic elastomers (TPEs). This is possible. In another example, the lower board 112 is a nearly transparent printed circuit board (PCB). Alternatively, the PCB may include holes or openings that allow light to pass through. Similar to the lower substrate 112, the upper substrate 114 is resistant to white light (or any color light). It can be formed from a material that exhibits transparency. For example, the upper substrate 114 can be glass, plastic It can be formed from stick or TPE. Furthermore, the inner surface of the upper substrate 114 is transparent Light conductive layer (e.g., indium tin oxide (ITO), poly(3,4-ethylenedioxymethyl) Offen polystyrene sulfonate (PEDOT:PSS), or other similar clear Alternatively, a conductive layer 118 such as an opaque (e.g., non-transparent) conductive coating may be used. In other embodiments, all areas of the DMF cartridge 110 It does not need to include a nearly transparent substrate and / or a coating or layer. For example, The substrate and / or coating or layer are transparent, translucent, and It does not have to be transparent or opaque. In some cases, such a substrate does not require a transparent layer. This would be inappropriate if it is a free-space optical probe. This is because it does not use transparent light. Fiber probes can be used, which provides flexibility in the selection of substrates and manufacturing technologies. This disclosure is advantageous in that it increases the benefits of this disclosure.

[0050] "top", "bottom", "over", "under" The terms "in" and "on" refer to the upper base of a DMF cartridge. Regarding the relative positions of the components of the DMF cartridge, such as the relative positions of the board and the lower substrate, This is used throughout the specification. The DMF cartridge is related to its orientation in space. It will be understood that it functions regardless.

[0051] In the DMF cartridge 110, the droplet operation gap 116 allows any droplet operation The liquid of interest, for example, but not limited to, liquid reagents, buffers, sample fluids, etc. This can be used as a space for processing. The height of the gap can be, for example, several hundred microns. This can be done. The droplet manipulation electrode 120 manipulates droplets via electrowetting. It can be used to carry out operations. "Droplet operations" refers to... This refers to arbitrary droplet manipulation on a digital microfluidic device or cartridge. For example, droplet operations include loading droplets into digital microfluidic devices and source Dispensing one or more droplets from a single droplet, splitting one droplet into two or more droplets, separation or division Dividing, transporting a droplet from one place to another in any direction, transporting two or more droplets Combining or joining into a single droplet, droplet dilution, droplet mixing, droplet stirring, droplet deformation, the location of the droplet Fixed-position retention, droplet incubation, droplet heating, droplet evaporation, droplet cooling, droplet disposal, DM Droplet transport from cartridge F 110, other droplet operations described herein, and / or This can be any combination of the operations described above. Furthermore, the droplet operation gap To control the temperature of the process occurring at 116, a Peltier heat pump is used. A temperature control element (not shown), such as an at pump, is combined with the DMF cartridge 110. It can be used.

[0052] Furthermore, Figures 3, 4, and 5 show the electrowetting method (for example, droplet handling electrode 1 The DMF cartridge 110 is described as one that manipulates droplets via (using 20). However, this is merely an example. In other embodiments, the droplets may be disposed of in other ways, for example. , but not limited to, optical methods, magnetic methods, thermal capillary methods, surface acoustic wave methods, and Other methods such as dielectrophoresis and other electrical methods, as well as any combination thereof Through this, it can be operated within the DMF cartridge 110.

[0053] Referring to Figure 3, this is the DMF cartridge 110 of the DMF system 100. As shown, the optical fiber probe 130 is introduced into the droplet manipulation gap 116 from above. For example, passing through the upper substrate 114, almost perpendicular to the plane of the DMF cartridge 110 (Yes). The sample droplet 140 is in the droplet manipulation gap 116 and a specific droplet manipulation electrode. It is located above 120. The tip of the optical fiber probe 130 is inside the sample droplet 140. / Directly interacts with sample droplet 140, thereby enabling onboard sensing region 1 58 can be formed. The DMF cartridge 110 is transmitted via droplet operation to the onboard sensor It can be used to move droplets between the spooling region 158 and the optical fiber. Lobe 130 is directed directly into the droplet manipulation gap 116 and along the path of the sample droplet 140. By accumulating these, localized optical survey techniques become possible. Figure 3 shows the optical fiber probe. This shows excitation light 132 delivered to the sample droplet 140 via bulb 130. Furthermore, The light 134 emitted from the sample droplet 140 is ejected via the optical fiber probe 130. In this case as well, the optical fiber probe 130 uses one optical fiber or multiple optical fibers It can contain bundles of fiber.

[0054] Referring to Figure 4, the DMF cartridge 110 of the DMF system 100 is shown. The optical fiber probe 130 is introduced into the droplet manipulation gap 116 from below (for example) (Passing through the lower substrate 112, it is approximately perpendicular to the plane of the DMF cartridge 110.) .

[0055] Referring to Figure 5, the DMF cartridge 110 of the DMF system 100 is shown. The optical fiber probe 130 is introduced into the droplet manipulation gap 116 from the side (for example) Between the lower substrate 112 and the upper substrate 114, relative to the plane of the DMF cartridge 110 (They are almost parallel.)

[0056] Referring now to Figure 6, an example of a DMF cartridge 110 of the DMF system 100. This is a view showing that the optical fiber probe 130 is introduced into the droplet manipulation gap 116, and the optical sensor The single operation is performed in reflection mode. Figure 6 shows the optical fiber probe shown in Figure 5. The configuration of the 130 is shown, but the optical fiber probe 130 is shown in Figures 4 and 5. The configuration is equally applicable to operation in reflection mode.

[0057] Figure 6 shows that excitation light 132 is transmitted from the illumination source 154 via the optical fiber probe 130 to the sun. This indicates that it has been delivered to the pull droplet 140. Furthermore, from the sample droplet 140 The emitted light 134 is projected onto the optical measurement device 156 via the optical fiber probe 130. This configuration of the DMF system 100 and DMF cartridge 110 is for the optical properties of liquids. In a manner that enables reflection sensing of properties (for example, the optical properties of sample droplet 140), D Provides an integrated optical fiber along with the MF cartridge. Optical fiber probe 13 0 is the DMF cartridge 110, which also includes other components necessary for digital microfluidics. This is only a part of it.

[0058] In this case as well, the illumination source 154 can be one or more light sources. The light source is monochromatic. Alternatively, it can be made pleochroic. These light sources are not limited to the following, but include light-emitting diodes. Examples include LEDs, lasers, incandescent light sources, fluorescent light sources, or any combination thereof. The light source can be modulated so that its intensity can be adjusted very quickly. Furthermore, it may include one or more sensors, such as a photodetector, to adjust the brightness. The light source may also include additional filters to ensure the quality of the incident light. Furthermore, A light source can be a combination of multiple individual light-emitting elements. These elements are the same Alternatively, it can become active at different wavelengths.

[0059] In this case as well, the optical measuring device 156 may be one or more optical sensors. Yes, it is possible. The optical measuring device 156 can be, for example, a narrowband sensor or a wideband sensor. The optical measuring device 156 converts the optical signal into an electronic signal that can be further processed. The optical measuring device 156 also has multiple sensors sensitive to various parts of the light spectrum. A spectrometer can be used. This is a spectrometer that is sensitive to various parts of the electromagnetic spectrum. Alternatively, it can be in the form of multiple individual sensors. The optical measuring device 156 is best It may include additional optical filters necessary to ensure the performance of the optical measuring device 1. 56 is an additional configuration that enables control of the illumination source 154 using closed-loop control such as a correlator. It can contain constituent elements.

[0060] During operation, excitation light 132 from the illumination source 154 passes through the optical fiber probe 130. The optical fiber probe 130 is used to probe the droplet being investigated (e.g., sample droplet 140). The incident light is guided to the fiber. The emitted light 134 is directed to the same core (co) of the fiber, depending on the system configuration. Reflected back from the droplet along the (re) or along different cores. The 130 can have one or more cores. These cores may be, for example, It can be in single mode or multi-mode (or any combination of the two). The reflected emitted light 134 is coupled to the optical measuring device 156.

[0061] The DM shown in Figures 3, 4, and 5 is performing optical sensing operations in reflection mode. The configuration of the optical fiber probe 130 inside the F cartridge 110 is as follows, however Specific techniques for droplet characterization, not limited to these, namely, (1) Reflectance spectroscopy such as infrared spectroscopy, (2) Variable path length spectroscopy, (3) Reading of enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction Absorbance spectroscopy, such as that used for reading PCR results. (4) Raman spectroscopy, (5) Photoelectron spectroscopy such as laser-induced destruction spectroscopy and atomic emission spectroscopy, (6) Fluorescence spectroscopy, such as that used for reading ELISA and PCR (7) Turbidity measurement, (8) Time-resolved spectroscopy such as photon correlation spectroscopy and fluorescence correlation spectroscopy, (9) Microscale thermophoresis, This enables the use of [this feature].

[0062] Figure 7A is a side view of an example of a DMF cartridge 110 of the DMF system 100. The sensor surface 142 is located at the tip of the optical fiber probe 130 within the droplet operation gap 116. It is placed at the end.

[0063] Various technologies are known to be useful in the manufacture of sensors using optical fibers. In one embodiment, this disclosure relates to Non-Patent Document 1 (Jeong, Hyeon-Ho & Erdene, Norov & Le e, Seung Ki & Jeong, Dae & Park, Jae-Hyoung. (2011) Fabrication of fiber-optic l ocalized surface plasmon resonance sensor and its application to detect antibody -antigen reaction of interferon-gamma. Optical Engineering. 50. 124405-124405. 1 The technology described in 0.1117 / 1.3662418. will be utilized. Non-patent document 2 (Proll, G., Mark ovic, G., Steinle, L., & Gauglitz, G. (2009). Reflectometric Interference Spectr. oscopy. Methods in Molecular Biology: Biosensors and Biodetection, 503, 167-178 See also (doi: 10.1007 / 978-1-60327-567-5_8). Jeonget al. The entire disclosure by Proll et al. is incorporated herein by reference.

[0064] The sensor surface 142 converts, for example, the characteristics of a droplet or a sample in a droplet into an optical response. An optical layer that can perform the following can be formed. For example, the characteristics converted to the optical response are: Factors such as the presence of the sample, temperature, temperature changes, and changes in the properties of the sample or solution can be cited.

[0065] In one example, the sensor surface 142 may use an interference filter. The interference filter is true It can be deposited on the fiber using empty deposition techniques. The interference filter is monochromatic intensity or This allows for the measurement of bonding to the surface using phase measurement or multicolor spectroscopy.

[0066] One example of a technique for manufacturing filters useful for optical fiber probes in the present disclosure is non Patent Document 3 (Proll G., Markovic G., Steinle L., Gauglitz G. (2009) Reflectometric Interference Spectroscopy. In: Rasooly A., Herold KE (eds) Biosensors and Bio detection. Methods in Molecular Biology TM It was published in vol. 503 (Humana Press), The entire disclosure is incorporated herein by reference.

[0067] In another embodiment, the sensor surface 142 may be an interference film that can be deposited using wet chemistry. It may be a tar or an optical material. The optical material may include, but is not limited to, metal nanoparticles. No. Using properties such as surface plasmon resonance or surface-enhanced Raman spectroscopy, droplets and The sample in the liquid and / or droplet can be analyzed. Another example of the sensor surface 142 is lithography. The technology may include the deposition of nanostructured surfaces. Examples of materials include metals (e.g.) Plasmonic nanoparticles), glass (e.g., diffraction elements), plastics (e.g., nanoimpedances) Examples include linted diffraction elements. Furthermore, the sensor surface 142 receives the excitation light 132. This can take the form of a dielectric microsphere, which is used to focus the light into a smaller region.

[0068] The optical sensor surface 142 allows the sample to bind from the solution to the surface in response to the amplified signal. It may include a layer involved in the process. For example, this layer may be deposited on the optical sensor surface 142. This layer can also be used to enhance the optical signal from the sample at the tip of the fiber. It can be used without the optical sensor surface 142 as a layer for concentrating the sample. In one embodiment, the layer is a gel matrix that promotes an increase in bonding sites. In another embodiment, The layer is a porous material that promotes an increase in surface density of the sample. Bonding sites (e.g., sparse Increase the aqueous site or the specific binding site (e.g., antibody / antigen or aptama) Various chemical enhancements for adding (- / sample) are known to those skilled in the art. Porous materials are In some cases, this may involve polymer brushes, gels, nanoparticles, or other high-surface-area structures. These can be typical organic materials. These porous organic media include polydimethylsiloxane, diviny This can be a synthetic polymer such as polybenzene or polyethylene glycol. In another embodiment, Porous media can be of biological origin, such as cellulose, chitin, or collagen.

[0069] As an alternative, the porous medium could be made from practically inorganic materials such as nanostructured carbon, silica, or titania. Porous media can be made by combining these materials arbitrarily, and the unique properties of the specimen By utilizing their chemical properties, their surface concentrations can be increased.

[0070] In another example, the sensor surface 142 is an LSPR sensor functionalized with one or more capture molecules. It can be a sensor layer. The LSPR sensor layer has metal nanostructures and / or one of the layers is metal. It can be composed of a certain multilayer nanostructure. Examples of metals commonly used in LSPR sensors include Examples include nanoparticles of gold, silver, platinum, palladium, and copper. In one example, the captured molecule is It is a ligand immobilized on the surface of the LSPR sensor layer. For example, as a ligand, Proteins, antibodies, antigens, or aptamers are carboxyl, NTA, or strepto Using Vidin surface adhesion chemistry, metal nanoparticles can be attached to the sensor layer. In this example, Gand is one of the two binding partners, and the other binding partner is Sample This is the target sample 144 in droplet 140.

[0071] In another example, the sensor surface 142 is a gel matrix containing fluorescein Any pH-sensitive optical layer can be used. The optical properties of these pH-sensitive gels are determined by the occurrence of these properties. It is related to the pH of the environment. As a result, the optical signal readout is for sample droplet 140. It can change based on pH.

[0072] In another example, the sensor surface 142 may have a heat-sensitive layer such as a thermochromic coating, or an oxidation coating. It could even be an LSPR sensor coated with a passivation layer such as silicon. The optical properties of sensors are strongly correlated with the temperature of the environment in which they are placed. The optical signal can vary as a function of the temperature of the sample droplet 140.

[0073] The sensor surface 142 is at the tip of the optical fiber probe 130 inside the DMF cartridge 110 The configurations at the ends, as shown in Figure 7A, are as follows, but are not limited to these: Specific characterization techniques, namely, (1) Monochromatic reflectometry, (2) Reflection such as biolayer interferometry and monochromatic reflectometry (SCORE) Type interferometry, (3) Surface-enhanced Raman spectroscopy, (4) Surface plasmon resonance, (5) Diffraction optics measurement, (6) Solid-phase microextraction (in combination with optical readout method), (7) Temperature measurement; This makes it possible.

[0074] Figure 7B shows the tip of the optical fiber probe 130 including the nanoparticle sensor surface 143. This is a side view of an example of a DMF cartridge 110 of the MF system 100. Exemplary nano Details of the particle sensor surface 143 are shown below with reference to Figure 15.

[0075] Referring now to Figure 8, an embodiment of the DMF cartridge 110 of the DMF system 100. This is a side view, and one or more optical elements 148 are located within the droplet operation gap 116. It is provided at the tip of the Iver probe 130. One or more optical elements 148 can be used individually, and In addition to the sensor surface 142 shown in Figure 7, one or more optical elements 148 may be provided. Using this method, light is directed into the sample droplet 140 and / or the sensor surface 142, and / or, outside, can be coupled more effectively. One or more optical elements 148 are not limited to However, this includes the following, namely, (1) Tapered optical fibers such as conical tapers, (2) U-shaped optical fiber, (3) Lenses such as hemispherical lenses, ball lenses, GRIN lenses, and aspherical lenses , (4) Prisms such as ATR prisms and Kretschmann prisms, (5) Angled polishes such as side polishes and wedge polishes, (6) Diffusion element, (7) Reflections such as those from Fabry-Perot etalons and transflection cavities. Cavity, It can include...

[0076] Figure 9 shows the same model except with the addition of a second illumination source 154' and a second optical measuring device 156'. A DMF cartridge of the DMF system 100, which is configured in much the same way as shown in Figure 6. This is a side view of embodiment 110. In this embodiment, the illumination source 154' and / or optical measurement The constant device 156' can operate independently or in conjunction with the illumination source 154 and the optical measuring device 156. It can be used in combination with a coupled optical fiber probe 130.

[0077] Similar to optical measuring device 156, optical measuring device 156' includes a photodetector, a camera, It can take the form of an optical measurement device such as a spectrometer or hyperspectral imager. Device 156' can perform time-resolved measurements. Furthermore, from the optical measurement device 156' The input can be used to control illuminator 154 and / or illuminator 154'.

[0078] The presence of the second illumination source 154' and the second optical measuring device 156' allows for various operating modes. This makes it possible. For example, using a first illumination source 154 and a first optical measuring device 156 The optical sensing operation is performed entirely in reflection mode via the optical fiber probe 130. It is possible to perform both illumination and collection using the optical fiber probe 130. In another example, using the first illumination source 154 and the second optical measuring device 156', light The sensing operation can be performed in transmission mode. That is, the optical fiber probe 130 Illumination is performed using the first illumination source 154, but an external second optical measuring device 156' is used. And collect. In yet another example, a second illumination source 154' and a first optical measuring device 15 Using 6, the light sensing operation can again be performed in transmission mode, i.e., external second illumination. Illumination is performed using light source 154', but optical fiber probe 130 and first optical measurement device Collect using Vice 156.

[0079] Referring to Figure 10, we see a DMF system including integrated optical fiber sensing. Flowchart of an example of using 100 and / or DMF cartridge 110 200 This is a diagram. Method 200 may include, but is not limited to, the following steps.

[0080] Step 210 includes a DMF system and / or DMF with integrated RI sensing. Prepare the cartridge. For example, the droplet operation gap 116 of the DMF cartridge 110. DMF system 100 and / or D including an integrated optical fiber probe 130 Prepare the MF cartridge 110 as shown in Figures 1 to 9, for example.

[0081] In step 215, the droplets to be processed are transported to the sensing area of ​​the DMF cartridge. For example, referring to Figures 3 to 9, the sample droplet 140 to be processed is a droplet Using the operation, the DMF cartridge 110 is transported to the onboard sensing area 158. However, here the onboard sensing area 158 is the optical fiber probe 130 It's at the tip.

[0082] In step 220, the optical sensing operation uses integrated optical fiber sensing. This is carried out within the MF cartridge. For example, referring to Figures 6 to 9, the optical sensor The operation involves the droplet operation gap 116 of the DMF cartridge 110, the illumination source 154, and Using the integrated optical fiber probe 130 on the optical measurement device 156, the DMF cart This is carried out at Ridge 110. During the operation, excitation light 132 from illumination source 154 is used in the optical field It passes through the Iver probe 130 and heads towards the sample droplet 140 being investigated, and into it. Next, the emitted light 134 is directed from the sample droplet 140 to the optical fiber probe 130. The light is reflected along the way and returns to the optical measuring device 156, and the optical measuring device 156 processes it. To capture the optical reading, for example, the optical sensing within the DMF cartridge 110 The operation can occur in reflection mode, as shown in Figures 6, 7, and 8. In another embodiment... The optical sensing operation within the DMF cartridge 110 is as shown and explained in Figure 9. This can occur in reflective mode, transmitted mode, or both reflective and transmitted modes.

[0083] In certain embodiments, the droplet is vibrated while it is in contact with the sensor. It may be useful. For example, in one embodiment, the first reaction electrode and the second reaction electrode are They are activated alternately, inducing droplet vibration between the first and second reaction electrodes, S This induces droplet movement on the PR sensor surface. In one embodiment, the first reaction electrode The vibration of the droplet between the first and second reaction electrodes is linear. In one embodiment, the vibration is approximately 0 The range is approximately 0.5 to 15 Hz. In another embodiment, the vibration range is approximately 4 to 10 Hz. It is enclosed.

[0084] In another embodiment, the SPR sensor surface is positioned between three or more reaction electrodes. Three or more reaction electrodes are activated alternately, inducing droplet vibration between the three or more reaction electrodes. This induces droplet movement on the SPR sensor surface. In another embodiment, three or more The vibration of the droplet between the reaction electrodes is circular.

[0085] In one embodiment, the electrode set moves droplets from one electrode to the next within the set. The vibrations caused by this process keep the droplet in contact with the sensing tip of the sensor. It is positioned relative to the sensing tip of the sensor.

[0086] Figure 11 shows a device that vibrates back and forth on the droplet manipulation electrode 120 and is connected to the optical fiber probe 130. An example of 140 2X sample droplets in contact is shown. In Figure 11, the droplets are generally... A line along the path of the droplet manipulation electrode 120, along the length of the optical fiber probe 130. It is moving in a direction perpendicular to (not shown). Within the scope of this disclosure, other types of movement For example, it coincides with a line (not shown) along the length of the optical fiber probe 130. It will be understood that movement in various directions is possible. Also, vibrations cause the droplet to elongate. and may include shortening. Furthermore, although the image shows a 2x stretch, the stretching can be 3x, 4x, It will be understood that it may be or more (X = used to extend the droplet) (Number of activated electrodes).

[0087] Figures 12A and 12B show DMF cartridge 1 including integrated fiber sensing. Each of the 10 exemplary instances is shown with a top perspective view and a bottom perspective view. For example, Figures 12A and 12B show the upper substrate 114 of the DMF cartridge 110, Examples of fiber optic assembly 125 and optical fiber core 146 are shown.

[0088] The fiber assembly 125 includes fibers extending from the first end edge of the assembly, and It can be configured as a set of fibers extending from the second end edge of the assembly. When the senburi 125 is assembled together with the DMF cartridge 110, the upper circuit board 114 and The droplet operation gap 116 between the lower substrate 112 and the optical fiber probe is sealed, and the optical fiber probe is It extends within the cap. Ideally, the fiber is electrowetting one or more times. An electrode 120, which interacts with one or more electrowetting electrodes 120. The droplet comes into contact with the end of the optical fiber probe, one or more electrowetting electrons It extends in close proximity to electrode 120. For assays requiring droplet vibration, the electrode's... The set vibrates by moving a droplet from one electrode to the next within the set. Keep the probe's sensing tip in contact with the sensor's sensing tip. They can be positioned opposite each other.

[0089] Figures 13A and 13B show top disassembled portions of an embodiment of the DMF cartridge 110, respectively. The figure and the lower exploded view are shown. The upper substrate 114 contains buffer, reagents and samples. Includes an opening for depositing reagents, buffers and samples within the MF cartridge 110. This enables the use of a multichannel pipette to introduce the liquid into the DMF cartridge 110. To achieve this, the openings are spaced apart. The footprint of the DMF cartridge 110 is It can be optimized for use in equipment used with well plates. The lower substrate 112 is A printed circuit board with patterned electrodes inside to control the operation of a droplet Yes. The lower substrate 112 is hydrophobic to optimize the electrowetting force of the DMF. It has a sex coating. The upper substrate 114 is made of an injection-molded polymer such as polycarbonate, acrylic, or cyclic olefin copolymer. The upper substrate 114 is made of an injection-molded polymer such as polycarbonate, acrylic, or cyclic olefin copolymer. To enhance its conductivity, a conductive coating such as indium tin oxide is applied to the upper substrate 114. The upper substrate 114 is sealed to the lower substrate 112 using an elastomer adhesive such as silicone. The fiber is introduced into the cartridge through a V-groove in the upper substrate 114 that helps align the fiber. The above elastomer adhesive also seals around the fiber to prevent it from entering the gap between the upper substrate 114 and the lower substrate 112. The fiber assembly 125 is fixed to the upper substrate 114 using any of adhesives, fasteners, ultrasonic welding, heat staking, or other fixing methods. The fiber assembly 125 is fixed to the upper substrate 114 using any of adhesives, fasteners, ultrasonic welding, heat staking, or other fixing methods. The fiber assembly 125 is fixed to the upper substrate 114 using any of adhesives, fasteners, ultrasonic welding, heat staking, or other fixing methods.

[0090] Figures 14A and 14B are various views of an embodiment of the fiber assembly 125 of the DMF cartridge 110. Figure 14A shows an exploded view and a cross-sectional view of the fiber assembly 125. Figure 14B shows a perspective view of the fiber assembly 125. The fiber assembly 125 may include two main components: a plurality of optical fibers 180 (i.e., those forming the optical fiber probe 130 and the fiber holder 182). The fiber assembly 125 may include two main components: a plurality of optical fibers 180 (i.e., those forming the optical fiber probe 130 and the fiber holder 182). The fiber holder 182 also includes two alignment holes 184, one at each end. To manufacture the fiber assembly 125, the optical fibers 180 are cut to a certain length and adhered to the fiber holder 182. The optical fibers 180 are adhered to the fiber holder 182. The optical fibers 180 extend from one side (the sensor end) of the fiber holder 182 by about 20 To manufacture the fiber assembly 125, the optical fibers 180 are cut to a certain length and adhered to the fiber holder 182. The optical fibers 180 extend from one side (the sensor end) of the fiber holder 182 by about 20 It extends by mm and extends approximately 5 mm from the opposite side (the mating surface 186 of the device). The river was cut with a ruby ​​scribe to make it flush with the mating surface 186 of the instrument, and then polished. Obtain. If the optical fiber 180 is cleave along its length, the other end (se The sensor end is polished. Then, the sensor end forms an optical fiber probe 130. Further processing is performed to achieve this. For example, the processing involves applying nanoparticles to the tip of the sensor end. This may include coating.

[0091] In summary, referring again to Figures 1 to 14B, the DMF system 100, DMF Cartridge 110 and / or method 200 are disposable cartridges (e.g., D Optical fibers (for example, optical fiber plugs) directly integrated into the MF cartridge 110. Using lobe 130), localized optical survey techniques are enabled. Compared to conventional methods, The integration of fiber optic sensors and digital microfluidics in the DMF system 100 This enables localized optical survey techniques and low-cost methods for achieving high optical performance. To provide a method (for example, by reducing the number of components for low cost) and to improve the signal, This limits light loss within the stem and restricts stray light collection for low noise. [Examples]

[0092] [Example 1] Referring to Figure 15, this is the actual process 300 for fabricating the sensor fiber. This is an example. Process 300 involves (1) stripping the fiber to a predetermined length and cleaving it. (1) Strip and cleave fibers to length (2) Secure the fibers to the jig (3) Step of depositing the sensor on the tip of the fiber, and (4) Optical properties This may include, but is not limited to, steps to test the integrity of the fiber and its coupling properties. It is not determined. This is an optical fiber for inserting into the droplet operation gap of the DMF cartridge. This is one example of how to fabricate and prepare a bar probe.

[0093] [Example 2] This experiment uses a fiber-optic-based surface plasmon resonance detection method within a DMF device. Using this, the common protein-protein interaction between protein A and IgG The purpose was to determine compatibility. The experiment involved autonomous processing and small-scale operation of a DMF device. The advantages of this, as well as the low background and limited configuration requirements of fiber optic detection. We demonstrated the advantages of both primeness and alignment.

[0094] [material] A 125 μm diameter optical fiber is cleaved and processed to deposit gold nanoparticles at the tip of the fiber. The material was deposited, and then the surface of these nanoparticles was finished with carboxyl groups. In summary, The optical fibers were thoroughly cleaned and oxidized. An example of oxidation is sodium hydroxide. Exposure to highly basic solutions such as sodium, ozone exposure, or oxygen plasma exposure Next, a self-organizing interface layer is deposited, and gold is attached to the sensor. Combine. In this example, mercapto such as (3-mercaptopropyl)trimethoxysilane. Silane compounds were used. Finally, the optical fiber was fitted with gold nanoparticles of the desired size and shape. The material was immersed in a solution containing the material. In this case, gold nanoparticles with a diameter of 10 nm were used. After compounding, thiol compounds were used to express functional groups for future chemistry. Various compounds The object can be used. For example, a compound having a thiol group on one side and a hydroxy group on the opposite side will express a carboxyl surface for future chemical reactions. 3-Mercapto-1-propanol was used. For example, FIG. 16 shows details of an example of the nanoparticle sensor surface 143 ( see FIG. 7B). FIG. 16 shows an electron micrograph of the tip of the obtained optical fiber probe and shows an optical fiber cladding 145, an optical fiber core 146, and a nanoparticle coating 147.

[0095] After surface finishing, the sensor was inserted into the vial of the DMF cartridge so that the tip was aligned with the edge of the electrode. Each channel of the collected data references various optical fiber sensors.

[0096] The DMF cartridge was loaded with 2 centistokes of polydimethylsiloxane to prepare an oil environment for the DMF device. The wells of the DMF device were filled with 100 μL of pH 7.4 phosphate buffered saline (PBS), 35 μL of ethanolamine, 35 μL of pH 1. 5 glycine + hydrochloric acid (HCl), 35 μL of protein A (the ligand used). Also, the cartridge was loaded with 8 μL of 1 0% glycerol in PBS and 16% glycerol in PBS for fiber sensitivity calibration, 8 μL of 1-ethyl-3-(-3-dimethylaminopropyl) carbodi imide hydrochloride (EDC) and 8 μL of N-hydroxysuccinimide (NHS) to activate the carboxyl surface of the sensor, and 8 μL of 900 nM IgG for analysis. However, only 2 μL is required for this protocol. For all reagents, 0. to stabilize the interface between oil and water. It contained 1% Tween-20 surfactant.

[0097] As shown in Figure 11, the droplet was vibrated while in contact with the probe. Using a setting electrode, the droplet is extended to a 2X droplet while maintaining contact with the probe. The device was moved back and forth across three electrodes at 10 Hz for 10 minutes.

[0098] [method] The entire experiment was performed automatically by the DMF cartridge. First, the sensor sensitivity was set. Calibration was performed using glycerol. This procedure involves the following steps: (1) 700 nL of PBS was introduced for baseline, (2) Measure the refractive index shift of 700 nL of 10% glycerol. (3) Rinse the sensor with 700 nL of PBS. (4) Measure the refractive index shift of 700 nL of 16% glycerol, and (5) Rinse the sensor with 700 nL of PBS. It was something.

[0099] The above evaluates the degree of signal shift caused by a known refractive index. Next, the cartridge The system's coupling reaction rate is measured automatically. This involves the following steps: Please note that many steps were carried out in parallel with other steps for the purpose of optimization. . (1) Step of adjusting the sensor with 700 nL of PBS, (2) A step of washing the sensor with 700 nL of glycine-HCl, (3) Rinse the fibers with 700 nL PBS. (4) 350 nL of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Mix 350 nL of unit N-hydroxysuccinimide with the sensor in each channel. - A step of activating the carboxyl surface with a mixture, (5) Rinse the fibers with 700 nL PBS. (6) 700 nL of protein A solution is introduced into each sensor. This allows the protein The step involves immobilizing protein A on the sensor surface and having it function as a ligand for the protein A antibody. , (7) Rinse the fibers with 700 nL PBS. (8) The step of preparing an IgG sample by the following method, namely, (a) Collect 350 nL of 900 nM IgG and dilute it with 700 nL of PBS. The above formulations were completely mixed, and one half was divided into 700 nL of mixture to obtain a 300 nM sample. Create, (b) Take the remaining 350 nL from 8a and dilute it with 700 nL of PBS. The mixture was then divided into 700 nL portions, with 350 nL reserved for a 100 nM sample. Dilute further with drops, and (c) Repeat 8b to produce samples of 33 nM, 11 nM, and 3.67 nM. Complete. Discard the remaining 350nL drop. By doing so, the step of creating an IgG sample, (9) Introduce 700 nL of IgG sample into various sensors. IgG and protein The meeting with A is measured in real time, and (10) Rinse the fiber with 700 nL PBS. IgG from protein A A step to measure dissociation in real time.

[0100] [Results and Analysis] Using the steps described above, response curves were created for each IgG sample. These curves are: First, adjust the sensitivity to refractive index shift obtained from glycerol correction, and then The amount of protein A immobilized on the sensor measured during the experiment was adjusted. Results This is shown in plot 400 in Figure 17A. Plot 400 is the response curve A The following are shown: B, C, D, E, and F. Legend 410 shows that the response curve A is an affinity of 0 nM. Correspondingly, response curve B corresponds to an affinity of 3.67 nM, and response curve C corresponds to an affinity of 11 nM. The response curve D corresponds to an affinity of 33 nM, the response curve E corresponds to an affinity of 100 nM, and the response curve F This supports affinity of 3000 nM.

[0101] A 1:1 reaction rate model was applied to and fitted to this data. The process shown in Figure 17B is shown. As shown in 405, this is consistent with measurements of the same sample on other instruments, non This consistently yields results of close correlation and a measured affinity of 2nM. However, other instruments Unlike the machine, this entire experiment, including sample preparation, involves cross-contamination with pre- or post-samples. It was performed autonomously with disposable fluid cartridges without any risk. Furthermore, in the experiment, It consumes only 350 nL of sample solution, dramatically reducing the volume compared to other systems. Ta.

[0102] This invention is implemented using hardware, software, or a combination thereof. It can be implemented in one or more computer systems or other processing systems. Therefore, the present invention relates to one or more computers capable of performing the functions described herein. To plan the system.

[0103] "preferably," "commonly," and "typically" Terms such as "cally)" are used to limit the scope of the claimed embodiment or to specify a particular embodiment. This means that the features are important or essential to the structure or function of the claimed embodiment. These terms are not used herein for the purpose of [specific purposes]. Intended to highlight alternative or additional features that may or may not be used in the context. It is.

[0104] The term "substantially" is used herein to mean any quantitative comparison, value, or measurement. To express the degree of inherent uncertainty that may arise from the fixed or other expressions, and to address the problem The quantitative representation of the object changes from the reference without altering its basic function. It is used to express the degree to which something is possible.

[0105] The terms "a," "an," and "the" are used in this application, including in the claims. When used in this context, it refers to "one or more." Therefore, for example, "target" A reference to "a subject" is inappropriate when the context is clearly the opposite (for example, multiple objects). Includes multiple objects, excluding those listed above.

[0106] "to include, to provide ("comprise", "comprises", & "comprising")", and "to include The term "include", "includes", "including" is non-restrictive. This is intended, and the descriptions of items in the list may be replaced or added to the listed items. This does not exclude other similar items that may be subject to similar treatment.

[0107] All publications referenced in the above specification are incorporated herein by reference. .

[0108] Various modifications and variations of the disclosed methods, compositions, and uses of the present invention are within the scope of the present invention. And without deviating from the spirit of this, by considering the scope of this disclosure and claims, This will become clear to the industry. The present invention relates to a particular preferred aspect or embodiment. Although disclosed herein, the claimed invention is not excessively limited to such specific embodiments. Please understand that it should not be limited.

Claims

1. A cartridge for use with the device, Multiple elements that operate to perform droplet manipulation on liquid droplets within a droplet manipulation gap. A digital microfluidic having a crowetting electrode, and The droplet handling gasket adjacent to two or more sets of electrowetting electrodes An optical fiber probe that enters the tubing, and any electrode from a set of two or more electrodes A droplet located on top of the optical fiber probe can come into contact with the optical fiber probe. Robe and, A cartridge equipped with these features.

2. A cartridge for use with the device, Multiple elements that operate to perform droplet manipulation on liquid droplets within a droplet manipulation gap. A digital microfluidic substrate having a crowetting electrode, Separated from the digital microfluidic substrate to form a droplet operation gap, and the A top plate having an opening for injecting liquid into a droplet operation gap, One or more of the electrowetting electrodes enter the droplet manipulation gap. A fiber optic probe having a sensing end positioned in close proximity to the fiber optic Swertia and A cartridge equipped with these features.

3. In the cartridge according to claim 1 or 2, light entering the droplet operation gap The fiber probe is positioned in close proximity to two or more sets of electrowetting electrodes. The droplets are arranged such that they are located on any electrode among the set of two or more electrodes, and the photophoton A cartridge that can come into contact with the Iber probe.

4. In the cartridge according to any one of claims 1 to 3, the probe is a ligand A cartridge containing this.

5. The cartridge according to any one of claims 1 to 4, further comprising the optical fiber - A droplet whose contact with the probe can be controlled by the electrowetting electrode. Prepare, cartridge.

6. In the cartridge according to any one of claims 1 to 5, the gap is further A cartridge containing low-viscosity oil for filling.

7. In the cartridge according to any one of claims 1 to 6, the top plate is It has two or more grooves or openings, each containing an optical fiber from the fiber assembly. A cartridge for aligning probes.

8. In the cartridge according to any one of claims 1 to 7, the sensing end is A cartridge containing the surface of a particle sensor.

9. A method for performing an assay, The steps include preparing an optical fiber probe containing a ligand, Its volume is less than approximately 1000 nL, and it has potential affinity for the ligand. The steps include preparing droplets containing the sample, The droplet is brought into contact with the end of the probe, and the droplet is moved away from contact with the probe. The steps include: Vibrating the droplet without stopping, Methods that include...

10. A method according to claim 9, wherein the vibration range is approximately 0.5 to approximately 15 Hz.

11. The method according to claim 9 or 10, wherein the vibration range is about 4 to about 10 Hz, 。

12. In the method according to any one of claims 9 to 11, the volume of the droplet is approximately 900 nL. A method that is less than.

13. In the method according to any one of claims 9 to 11, the volume of the droplet is approximately 800 nL. A method that is less than.

14. In the method according to any one of claims 9 to 11, the volume of the droplet is approximately 700 nL. A method that is less than.

15. In the method according to any one of claims 9 to 11, the volume of the droplet is approximately 600 nL. A method that is less than.

16. In the method according to any one of claims 9 to 11, the volume of the droplet is approximately 500 nL. A method that is less than.

17. In the method according to any one of claims 9 to 11, the volume of the droplet is approximately 400 nL. A method that is less than.

18. In the method according to any one of claims 9 to 17, further, The signal from the droplet is measured via the aforementioned optical fiber probe, and a computer is used. The step of calculating a response curve from the signal. Methods that include...

19. In the method according to any one of claims 9 to 18, the optical fiber probe is multiple A method comprising several ligands, wherein the droplets contain multiple samples.

20. In the method according to any one of claims 9 to 19, the preparation operation is further multi The steps include preparing a number of optical fiber probes and a plurality of droplets, and the plurality of droplets The steps of bringing it into contact with the corresponding optical fiber probe, and the corresponding optical fiber A method comprising the step of vibrating each of the plurality of droplets in contact with a probe.

21. In the method according to any one of claims 9 to 20, the vibration is electrowet A method performed via a junction electrode.

22. In the method according to claim 21, each optical fiber probe has an electro-weave end. A method of aligning the electrodes adjacent to the setting electrodes.

23. In the method according to claim 21 or 22, each optical fiber probe has an end that A method of aligning the electrodes so as to be close to the edge of the octowetting electrode.

24. In the method according to any one of claims 9 to 23, the vibration is a digital micro Through the electrowetting electrode in the droplet operation gap of the fluid (DMF) cartridge The method by which something is done.

25. In the method according to any one of claims 9 to 24, the vibration is the optical fiber A method that is approximately perpendicular to the line extending along the length of the probe.

26. In the method according to any one of claims 9 to 24, the vibration is the optical fiber A method that closely coincides with a line extending along the length of the probe.

27. A method according to any one of claims 9 to 24, wherein the vibration is multidirectional, 。

28. In the method according to claim 27, the vibration is along the length of the optical fiber probe. A method that is multidirectional within a plane parallel to the extending line.

29. In the method according to any one of claims 9 to 28, the vibration is performed by elongated liquid A method performed using drops.

30. The method according to claim 29, wherein the elongated droplet is a 2X droplet.

31. The method according to claim 29, wherein the elongated droplet is a 3X droplet.

32. In the method according to any one of claims 9 to 31, the assay is, in other words, Molecular library screening assays, binding reaction rate assays, affinity determination assays SEX, binding site mapping assays, thermodynamic investigations, sandwich assays, competitive analysis assays Specificity determination assays, antibody binding characterization, and combinations thereof, A method to be selected from.

33. In the DMF system, the system is It is a cartridge, Multiple units that operate to perform droplet operations on liquid droplets within a droplet operation gap. Digital microfluidic with lectrowetting electrodes, The droplet handling gear adjacent to two or more sets of electrowetting electrodes An optical fiber probe that enters the pit, and any of the two or more sets of electrodes A droplet located above the electrode can come into contact with the optical fiber probe, probe, A cartridge equipped with, The electrowetting electrode and the optical fiber probe are operationally coupled The controller and An illumination source arranged to illuminate one or more droplets within the droplet operation gap, Optically coupled to the optical fiber probe, and receiving signals from the optical fiber probe. Optical measuring devices positioned to be trusted, A DMF system equipped with this feature.

34. A digital microfluidic (DMF) cartridge, A first substrate having one or more electrowetting electrodes for performing droplet operations. and, A second substrate offset from the first substrate, A droplet operation gap defined between the first substrate and the second substrate, and one or more The upper electrowetting electrode controls the liquid droplet within the droplet manipulation gap. A droplet operation gap that operates to carry out the operation, A sensing region within the droplet manipulation gap for contacting the liquid droplet, Excitation light is supplied from the first illumination source to the sensing region, and from the sensing region, the first An optical fiber probe that supplies emitted light to an optical measurement device, A DMF cartridge equipped with this feature.

35. In the DMF cartridge according to claim 34, the sensing area is the optical fiber A DMF cartridge including a sensor surface positioned on the terminal surface portion of an Iber probe.

36. In the DMF cartridge according to claim 34 or 35, the sensing area is The liquid droplets are brought into contact with the sensing area by one or more electrowetters. A DMF cartridge is positioned within the droplet operation gap adjacent to the ing electrode.

37. In the DMF cartridge according to any one of claims 34 to 36, the optical fiber The bar probe penetrates the first substrate and the sensing area is located within the droplet manipulation gap. The DMF cartridge is used to position the cartridge.

38. In the DMF cartridge according to any one of claims 34 to 37, the optical fiber The bar probe penetrates the second substrate and the sensing area is located within the droplet manipulation gap. The DMF cartridge is used to position the cartridge.

39. In the DMF cartridge according to any one of claims 34 to 38, the optical fiber The bar probe is substantially parallel to the first substrate and the second substrate, and the first substrate and the second substrate The droplet enters the droplet manipulation gap between the plate and the sensor. A DMF cartridge for arranging the area.

40. In the DMF cartridge according to any one of claims 34 to 39, the first substrate Alternatively, at least one of the second substrates introduces liquid into the droplet operation gap. A DMF cartridge having at least one opening for...

41. In the DMF cartridge according to any one of claims 34 to 40, the sensing The area contains the ligand, and is a DMF cartridge.

42. In the DMF cartridge according to any one of claims 34 to 41, the sensing The area includes the nanoparticle sensor surface, and is a DMF cartridge.

43. In the DMF cartridge according to any one of claims 34 to 42, the optical fiber The bar probe is a DMF cartridge that includes one or more optical elements in the terminal surface portion.

44. In the DMF cartridge according to any one of claims 34 to 43, the droplet operation The gap contains a filler material that does not mix with the liquid droplets, and is a DMF cartridge.

45. In the DMF cartridge according to any one of claims 34 to 44, the first substrate and the second substrate allows excitation light from the second illumination source to pass through the first substrate and sense The light reaches the region, and the emitted light from the sensing region passes through the second substrate for the second optical measurement. A DMF cartridge containing a material transparent enough to allow access to the device. 。

46. A digital microfluidic (DMF) system, A DMF cartridge according to any one of claims 34 to 45, Excitation light is supplied to the sensing area of ​​the DMF cartridge via an optical fiber probe. to supply, one or more light sources, The light emitted from the sensing region is received and processed via the optical fiber probe. and one or more optical measuring devices, A DMF system equipped with this feature.

47. In the DMF system according to claim 48, further, To control the execution of droplet manipulation by one or more electrowetting electrodes, A controller that can communicate with one or more electrowetting electrodes in an operable manner. A DMF system equipped with this feature.

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