Multimodal measurement device, system, and method

Abstract

A multimodal measurement device comprises a split analyte sensor layer, a seal layer above the sensor layer, configured to divide the sensor layer into two or more detection zones, including a detection channel associated with each detection zone, a membrane holding layer above the seal layer, comprising a holder for a nanoporous membrane chip, and delivery channels fluidly connecting two or more surfaces of the chip to the two or more detection zones of the sensor layer via the detection channels, and a cap layer above the membrane holding layer, comprising a plurality of application channels including input and output ports, wherein the application channels are fluidly connected to chip via the delivery channels of the membrane holding layer. Related systems and methods are also disclosed.

Description

MULTIMODAL MEASUREMENT DEVICE, SYSTEM, AND METHODCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. provisional application No. 63 / 639,786 filed on April 29, 2024, incorporated herein by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with government support under TR003281, and AG088071 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND OF THE INVENTION

[0003] The use of photonic biosensors to measure refractive light index changes in a sample is well known. The detected change in refractive light of a sample provides for the detection of analytes. Some known optical structures of a biosensor cause a refractive index change as a result of binding of an analyte to an optical surface and / or reagent, which creates a detectable change in an optical resonance frequency. These known biosensors provide high sensitivity and label free detection of desired analytes.

[0004] Typically, detection elements of biosensors are created on silicon substrates using traditional silicon-based nanoscale manufacturing processes, such as complementary metal- oxide-semiconductor ("CMOS") fabrication processes. The use of silicon-based fabrication processes creates biosensor detection elements for integrated photonics that have exceptional optical and biochemical characteristics. For example, silicon-based fabrication processes enable precise and / or intricate optical structures of a detection element to be manufactured in silicon or silicon nitride. The optical structures include, for instance, ring resonators, spiral waveguides,grating couplers, and Mach-Zehnder Interferometers ("MZI"). These structures generally require near defect-free optical paths to ensure results are not affected by material impurities or structure defects.

[0005] While silicon-based processes provide precise biosensor detection elements on a substrate, known biosensors typically have costly fluid and light interconnections. For example, optical fiber bonding of input and output optics is typically needed for interfacing with light paths of a detection element. Further, many biosensors have complex active fluid delivery mechanisms to bring a sample into contact with the detection element. Oftentimes, a fluid sample is pulled or pushed to a detection element using external pumps that control sample volume and flow rate through the biosensor. This light and fluid interconnect complexity increases the cost of instrumentation, and the biosensors themselves.

[0006] Thus, there is a need in the art for improved multiplex photonic biosensors and related methods.SUMMARY OF THE INVENTION

[0007] Some embodiments of the invention disclosed herein are set forth below, and any combination of these embodiments (or portions thereof) may be made to define another embodiment.

[0008] In one aspect, a multimodal measurement device comprises: a split analyte sensor layer; a seal layer above the sensor layer, configured to divide the sensor layer into two or more detection zones, including a detection channel associated with each detection zone; a membrane holding layer above the seal layer, comprising a holder for a nanoporous membrane chip, and delivery channels fluidly connecting two or more surfaces of the chip to the two or more detection zones of the sensor layer via the detection channels; and a cap layer above the membrane holding layer, comprising a plurality of application channels including input and output ports, wherein the application channels are fluidly connected to chip via the delivery channels of the membrane holding layer.

[0009] In one embodiment, the detection channels, delivery channels, and application channels comprise microfluidic channels.

[0010] In one embodiment, the cap layer further comprises a purge valve.

[0011] In one embodiment, the device is configured to detect two or more analytes simultaneously.

[0012] In one embodiment, the sensor layer comprises a photonic integrated circuit (PIC) disposed directly on a substrate, optically coupled to a light source and a photodetector.

[0013] In one embodiment, the substrate comprises at least one of a cassette, a slide, a membrane, a fibrous substrate, or a test card.

[0014] In one embodiment, the photonic integrated circuit comprises: an input coupler; at least two output couplers; at least one waveguide optically connecting the input coupler to the output couplers; and a plurality of detection elements disposed as clusters along the at least one waveguide.

[0015] In one embodiment, the detection element clusters are positioned in the two or more detection zones.

[0016] In one embodiment, wherein the detection element clusters are separated by at least 700 pm.

[0017] In one embodiment, the detection elements include at least one capture molecule.

[0018] In one embodiment, the input coupler is aligned with an optical input port, and the at least two output couplers are aligned with an optical output port.

[0019] In one embodiment, the detection elements include at least one of a ring resonator, a double ring resonator, a cylindrical resonator, a spherical resonator, a spiral waveguide, a Vernier filter, a photonic crystal, and a Mach-Zehnder Interferometer ("MZI").

[0020] In one embodiment, the at least one waveguide comprises a silicon nitride waveguide.

[0021] In one embodiment, the at least one waveguide splits into a plurality of branches from the input coupler to the at least two output couplers.

[0022] In one embodiment, each detection element cluster is positioned on separate branches.

[0023] In one embodiment, the cap layer comprises polydimethylsiloxane (PDMS).

[0024] In one embodiment, the membrane holder layer comprises silicone.

[0025] In one embodiment, the chip is configured to hold two or more cell cultures on its surfaces.

[0026] In one embodiment, the chip is configured to hold a different cell culture on each of its surfaces.

[0027] In another aspect, a multimodal measurement device comprises: a photonic integrated circuit (PIC) disposed directly on a substrate, optically coupled to a light source and a photodetector; a seal layer above the PIC, configured to divide the PIC into first and second detection zones, including a first detection channel associated with the first detection zone, and a second detection channel associated with the second detection zone; a membrane holding layer above the seal layer, comprising a holder for a nanoporous membrane chip, a top delivery channel fluidly connecting the top surface of the chip to the first detection zone of the PIC via the first detection channel, and a bottom delivery channel fluidly connecting the bottom surface of the chip to the second detection zone of the PIC via the second detection channel; and a cap layer above the membrane holding layer, comprising: a top application channel including input and output ports, fluidly connected to the top surface of the chip via the top delivery channel; and a bottom application channel including input and output ports, fluidly connected to the bottom surface of the chip via the bottom delivery channel.

[0028] In one embodiment, the first and second detection channels, top and bottom delivery channels, and top and bottom application channels comprise microfluidic channels.

[0029] In one embodiment, the top application channel further comprises a purge valve.

[0030] In one embodiment, the chip is configured to hold a first cell culture on its top surface, and a second cell culture different from the first on its bottom surface.

[0031] In another aspect, a multimodal measurement method comprises: providing the device as described above; applying a fluid to the chip via the cap layer; and measuring the presence or quantity of an analyte from the chip via the PIC.

[0032] In another aspect, a photonic integrated circuit (PIC) comprises: a substrate; an input coupler on the substrate; at least two output couplers on the substrate; at least one waveguide on the substrate optically connecting the input coupler to the output couplers; and a plurality of detection elements disposed as clusters on the on the substrate along the at least one waveguide.

[0033] In one embodiment, the detection element clusters are separated by at least 700 pm.

[0034] In one embodiment, wherein the detection elements include at least one capture molecule.

[0035] In one embodiment, the detection elements include at least one of a ring resonator, a double ring resonator, a cylindrical resonator, a spherical resonator, a spiral waveguide, a Vernier filter, a photonic crystal, and a Mach-Zehnder Interferometer ("MZI").

[0036] In one embodiment, the at least one waveguide comprises a silicon nitride waveguide.

[0037] In one embodiment, the at least one waveguide splits into a plurality of branches from the input coupler to the at least two output couplers.

[0038] In one embodiment, wherein each detection element cluster is positioned on separate branches.BRIEF DESCRIPTION OF THE DRAWINGS

[0039] The foregoing purposes and features, as well as other purposes and features, will become apparent with reference to the description and accompanying figures below, which are included to provide an understanding of the invention and constitute a part of the specification, in which like numerals represent like elements, and in which:

[0040] Fig. 1 shows top and perspective views of an exemplary multimodal measurement device in accordance with some embodiments.

[0041] Fig. 2 shows a cutaway view of an exemplary multimodal measurement device in accordance with some embodiments.

[0042] Fig. 3 shows a cutaway view of an exemplary multimodal measurement device in accordance with some embodiments.

[0043] Fig. 4A shows cutaway views of an exemplary multimodal measurement device in accordance with some embodiments.

[0044] Fig. 4B shows exemplary apical and basal channel pathways of an exemplary multimodal measurement device in accordance with some embodiments.

[0045] Fig. 5 shows an exploded view of an exemplary multimodal measurement device in accordance with some embodiments.

[0046] Fig. 6 shows an exploded view of an exemplary multimodal measurement device in accordance with some embodiments.

[0047] Fig. 7 shows an exemplary photonic integrated circuit (PIC) sensor for use in an exemplary multimodal measurement device in accordance with some embodiments.

[0048] Figs. 8A-8B show exemplary PIC sensors for use in an exemplary multimodal measurement device in accordance with some embodiments.

[0049] Figs. 9A-9D show an exemplary experimental multimodal measurement device in accordance with some embodiments.

[0050] Figs. 10A-10B show exemplary manufacturing schematics for an experimental device in accordance with some embodiments.

[0051] Figs. 11A-11F show exemplary details of a manufactured device in accordance with some embodiments.

[0052] Figs. 12A-12B show exemplary manufacturing schematics for an experimental device in accordance with some embodiments.

[0053] Figs. 13A-13B show exemplary assembly jigs in accordance with some embodiments.

[0054] Fig. 14 shows an exemplary experiment schematic in accordance with some embodiments.

[0055] Fig. 15 shows an exemplary experimental schematic in accordance with some embodiments.

[0056] Fig. 16 shows exemplary experimental results in accordance with some embodiments.

[0057] Fig. 17 shows exemplary experimental results in accordance with some embodiments.DETAILED DESCRIPTION OF THE INVENTION

[0058] It is to be understood that the figures and descriptions of the present invention have been simplified to illustrate elements that are relevant for a clearer comprehension of the present invention, while eliminating, for the purpose of clarity, many other elements found inmultimodal measurement devices, systems, and methods. Those of ordinary skill in the art may recognize that other elements and / or steps are desirable and / or required in implementing the present invention. However, because such elements and steps are well known in the art, and because they do not facilitate a better understanding of the present invention, a discussion of such elements and steps is not provided herein. The disclosure herein is directed to all such variations and modifications to such elements and methods known to those skilled in the art.

[0059] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, exemplary methods and materials are described.

[0060] As used herein, each of the following terms has the meaning associated with it in this section.

[0061] The articles "a" and "an" are used herein to refer to one or to more than one ( / .e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0062] "About" as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, and ±0.1% from the specified value, as such variations are appropriate.

[0063] Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Where appropriate, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within thatrange, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0064] The terms "patient," "subject," "individual," and the like are used interchangeably herein, and refer to any animal amenable to the systems, devices, and methods described herein. The patient, subject or individual may be a mammal, and in some instances, a human.

[0065] The terms "fluid" or "liquid medium" are used herein to refer to a substance which is in the form of a liquid at ambient temperature or room temperature. Non-limiting examples of fluid include: water, blood, blood serum, body fluids, cell media, or the like, and any combination thereof.

[0066] The term "membrane" is used herein to refer to a film capable of performing separations. The separation may be absolute (i.e., non-permeable membrane), selective (i.e., semi-permeable membrane), or limited (i.e., permeable membrane).

[0067] Referring now in detail to the drawings, in which like reference numerals indicate like parts or elements throughout the several views, in various embodiments, presented herein are multimodal measurement devices, systems, and methods.

[0068] Disclosed herein is a layered microfluidic device that can perform multimodal measurements, such as for example to recapitulate the epidermis and dermis of human skin while allowing continuous monitoring of both apical and basal analytes of interest simultaneously.

[0069] Referring now to Figs. 1-6, an exemplary multimodal measurement device 100 is shown. In some embodiments, a multimodal measurement device 100 comprises a split analyte sensor layer 101, a seal layer 102 above the sensor layer 101, a membrane holding layer 104 above the seal layer 102, and a cap layer 107 above the membrane holding layer 104. In some embodiments, the device 100 is configured to detect two or more analytes simultaneously.

[0070] In some embodiments, the seal layer 102 is configured to divide the sensor layer 101 into two or more detection zones 103. In some embodiments, the seal layer 102 includes a detection channel associated with each detection zone 103.

[0071] In some embodiments, the membrane holding layer 104 comprises a holder for a nanoporous membrane chip 105, and delivery channels 106 fluidly connecting two or more surfaces of the chip 105 to the two or more detection zones 103 of the sensor layer 101 via the detection channels. In some embodiments, the membrane holder layer 104 comprises silicone. In some embodiments, the chip 105 is configured to hold two or more cell cultures on its surfaces (e.g., one or more cells of a first cell type positioned or cultured on a first surface of the membrane, and one or more cell types of a second cell type positioned or cultured on a second surface of the membrane). In some embodiments, the chip 105 is configured to hold a different cell culture on each of its surfaces.

[0072] In some embodiments, the cap layer 107 comprises a plurality of application channels 108 including input and output ports, wherein the application channels 108 are fluidly connected to chip 105 via the delivery channels 106 of the membrane holding layer 104. In some embodiments, the cap layer further includes a purge valve 109 which may be used to create and control an air-liquid interface (ALI). In some embodiments, the cap layer 107 comprises polydimethylsiloxane (PDMS). In some embodiments, portions of the application channels 108, delivery channels 106, and / or detection channels / zones 103 can be combined to form an apical channel path 198 and / or basal channel path 199.

[0073] In some embodiments, the sensor layer 101 comprises a photonic integrated circuit (PIC) 110 disposed directly on a substrate and is optically coupled to a light source and a photodetector. In some embodiments, the substrate comprises at least one of a cassette, a slide, a membrane, a fibrous substrate, or a test card.

[0074] In some embodiments, the detection channels and / or detection zones 103, delivery channels 106, and / or application channels 108 comprise microfluidic channels.

[0075] In some embodiments, the seal layer 102 and membrane holding layer 104 comprise a silicone and adhesive double channel seal, a holder for a nanoporous membrane chip 105 (such as those manufactured by SiMPore), and an adhesive membrane chip seal. The overall goal of these layers is to support and incorporate the nanoporous membrane chip 105 such that cells grow effectively and are in close proximity to the sensor for higher resolution realtime measurements. In some embodiments, the device is configured for any suitable biological material including any suitable genetic cell types, populations, and / or cultures. In some embodiments, the device is configured to grow a first biological material on the apical side of the membrane and second biological material on the basal side of the membrane. Analytes secreted apically by the first biological material will flow through the membrane holding layer directly over the right sensing bank on the PIC surface. Analytes secreted by the second biological material will be flown directly over the PIC without the need to reroute the flow pathway. To allow for differential sensing, the sensing compartments are kept separate as shown in Figs. 2-3.

[0076] In some embodiments, the device is configured for epidermal keratinocytes growth on the apical side of the membrane and dermal fibroblasts growth on the basal side of the membrane. Analytes secreted apically by the keratinocytes will flow through the membrane holding layer directly over the right sensing bank on the PIC surface. Analytes secreted by the fibroblasts will be flown directly over the PIC without the need to reroute the flow pathway. To allow for differential sensing, the epidermal and dermal analyte compartments are kept separate as shown in Figs. 2-3.

[0077] In some embodiments, the cap layer 107 comprises an adhesive top channel and polydimethylsiloxane (PDMS) cap. The purpose of this component is to form a pathway for analyte to flow across the apical side of the membrane and allow for cannula to interface with the device to deliver fluid via pressure-driven flow without leaking. The cap layer 107 may include any suitable number of channels, including one, two, three, or any number greater than one. The device 100 can be configured to use any suitable number of pumps, including one, two, three, or any number greater than one, and these pumps may be any suitable pump typeincluding, but not limited to, pressure driven pumps and peristaltic pumps. In some embodiments, the device 100 is configured to have a flow rate in the range of 5 pL / min to 200 pL / min.

[0078] As shown the PDMS cap is also equipped with a purge valve 109 in the apical channel. This valve provides an exit point for fluid that has filled the top channel to allow for the development of an air-liquid interface. For example, in certain instances, the valve allows for the development of an air-liquid interface for the epidermal keratinocytes while still allowing for the recapitulated skin to be fed and maintained basal ly from fluid filling the bottom channel. For epidermal keratinocytes to differentiate properly, the keratinocytes must be apically exposed to air to recapitulate conditions seen in vivo.

[0079] Further details of the figures follow. Fig. 1 shows a top down and a perspective view of a fully assembled device 100. In some embodiments, the device 100 is about 10 mm by 20 mm by 2 mm. The membrane window and left basal ring resonator (RR) sensing bank on the PIC can be seen through the left half of the apical channel. Note the 90-degree right turn made by the left half of the apical channel.

[0080] Fig. 2 shows a cross section of the example device showing the left half of apical channel, apical channel outlet, air-liquid interface (ALI) purge valve, and basal channel inlet and outlet.

[0081] Fig. 3 shows another cross section of the example device. At this plane, the device comprises a transfer pathway between the left half of the apical channel, through the membrane chip holding layer, and across the right RR sensing bank on the PIC surface. Compared to Fig. 2, Fig. 3 takes a cross section in the same plane but moved closer to the glass slide overhang.

[0082] Fig. 4A shows additional front-side (top row) and back-side (bottom row)cross- sectional views of the device 100.

[0083] Fig. 4B shows a rendering of apical channel 198 and basal channel 199 pathways within the device and how the channels interact with the nanoporous membrane 105 and PIC 110. Arrows show inlet and outlet flow direction.

[0084] Fig. 5 shows an exploded view of an example device 100. In total this device is made of 12 layers, each cut and stacked individually. In some embodiments, the device comprises multiple sublayers. As shown in Fig. 5, sublayer 1 comprises a PDMS cap, sublayer 2 comprises an adhesive channel (left half of apical channel), sublayers 3 and 4 comprise a membrane window seal of silicone (3) and adhesive (4), sublayer 5 comprises a silicone membrane chip holder, sublayers 6 and 7 comprise a double channel seal of adhesive (6) and silicone (7), sublayer 8 comprises an adhesive double channel (basal channel and right half of apical channel), sublayer 9 comprises a PET PIC seal and partitioner, sublayer 10 comprises a silicone PIC holder, sublayer 11 comprises an adhesive (underneath) PIC seal, and sublayer 12 comprises a glass slide. The split analyte sensor layer 101 comprises sublayers 10-12, the seal layer 102 comprises sublayers 8-9, the membrane holding layer 104 comprises sublayers 3-7, and the cap layer 107 comprises sublayers 1-2.

[0085] Fig. 6 shows an exploded view of the device where one-sided PET is the only adhesive sealing layer used in the device. In total this device is made of 10 layers, each cut and stacked individually. In some embodiments, the device comprises multiple sublayers. As shown in Fig. 6, sublayer 1 comprises a PDMS cap, sublayer 2 comprises an adhesive channel (left half of apical channel), sublayer 3 comprises a PET membrane window seal, sublayer 4 comprises a silicone membrane chip holder, sublayer 5 comprises a PET double channel seal, sublayer 6 comprises an adhesive double channel (basal channel and right half of apical channel), sublayer 7 comprises a PET PIC seal and partitioner, sublayer 8 comprises a silicone PIC holder, sublayer 9 comprises an adhesive (underneath) PIC seal, and sublayer 10 comprises a glass slide. The split analyte sensor layer 101 comprises sublayers 8-10, the seal layer 102 comprises sublayers 6-7, the membrane holding layer 104 comprises sublayers 3-5, and the cap layer 107 comprises sublayers 1-2. This device design can streamline prototype building andmanufacturing as there are 10 layers instead of 12. Although only a difference of two layers, a significant amount of assembly time is saved.

[0086] Manufacturing can be simplified using the one-sided adhesive PET layer (sublayer 9 in Fig. 5) to replace the double silicone and adhesive layers. As adhesive bound to adhesive can be prone to leakage, silicone layers of the exact same footprint can be added to prevent leaking and form a tight seal. However, prototypes have shown that the one-sided adhesive PET can also work as a seal for both above (sublayers 3 and 4 in Fig. 5) and below the membrane chip (sublayers 6 and 7 in Fig. 5). With such substitutes, the device design layer stack would be as shown in Fig. 6.

[0087] Referring now to Figs. 7-8, exemplary split analyte sensor layers 101 are shown. In some embodiments the split analyte sensor layer 101 comprises a photonic integrated circuit (PIC) 110. In some embodiments, the PIC 110 includes at least one first (input) coupler 111 such as a grating coupler and / or edge coupler, at least two second (output) couplers 112 such as grating couplers and / or edge couplers, at least one waveguide 113 between the first coupler 111 and the second couplers 112, and a plurality of detection elements and / or detection element clusters 114 disposed along the at least one waveguide 113. In some embodiments, the at least one first (input) coupler 111 is aligned with an optical input port. In some embodiments, the at least two second (output) couplers 112 are aligned with one or more optical output ports. In some embodiments, the detection element clusters 114 are positioned in the two or more detection zones associated with the detection channels 115. In some embodiments, the detection element clusters are separated by at least 100 pm, 300 pm, 500 pm, or 700 pm.

[0088] In some embodiments, the at least one detection element 114 can include at least one capture molecule. In some embodiments, the capture molecule binds to, or is otherwise configured to, capture analytes, such as proteins, antibodies, peptides, nucleic acids, antigens, cytokines, chemokines, and any other suitable substance.

[0089] In some embodiments the at least one detection element 114 includes at least one of a ring resonator, a double ring resonator, a cylindrical resonator, a spherical resonator, aspiral waveguide, photonic crystal, a Mach-Zehnder Interferometer ("MZI"), and any other suitable design or combinations thereof.

[0090] In some embodiments, the at least one waveguide 113 comprises a silicon nitride waveguide. In some embodiments, the at least one waveguide 113 splits into a plurality of branches from the first coupler 111 to the at least two second couplers 112. In some embodiments, the at least one detection element 114 is positioned on one of the plurality of branches. In some embodiments, each detection element cluster 114 is positioned on separate branches. In some embodiments, the at least one detection element 114 is positioned to contact a fluid sample within the sample detection channel 115.

[0091] As shown, for example in Fig. 7, the PIC 110 sensor may include two clusters of photonic sensor elements (ring resonators (RR)) is able to interface with two microfluidic channels simultaneously. The left sensing RR bank is outlined in black, while the right sensing RR bank is outlined in red.

[0092] Additional example PIC layouts are shown in Fig 8, including PIC layouts for edge- coupled designs (A, B, and C) and grating coupled designs (D and E). Every layer of the device could remain the same as described above except for layers 6, 7, and 8 of the embodiment of Fig. 6, and layers 8, 9, and 10 of the embodiment of Fig. 5. Since the PIC partitioning / sealing layer would need to change, the subsequent double channel layer would need to be changed to allow for adequate sensor exposure for both apical and basal channels to be achieved.

[0093] In some embodiments, the PIC 110 comprises a rectangular prism or cuboid shape. In some embodiments, the PIC 110 has a length between 1000 and 10000 pm, a width between 1000 and 10000 pm and a height between 700 pm and 800 pm. In some embodiments, an array of PICs 110 may be connected to the substrate for multiplex applications. In some embodiments, the PIC 110 can include 0 to 100, 1 to 20, 2 to 10, 2 or more, 5 or more, 8 or more, or any suitable number of detection elements and / or detection clusters 114.

[0094] In some embodiments, a PIC 110 with zero detection elements and / or detection clusters 114 may be included in an array. In some embodiments, the PIC 110 may be used as a reference for light calibration and / or adjustment.

[0095] In some embodiments, the at least one detection element and / or detection cluster 114 has an extinction ratio greater than 5 dB under aqueous cladding. In some embodiments, each detection element and / or detection cluster 114 has a unique extinction ratio. In some embodiment, the spectral footprint of individual detection element and / or detection cluster 114 sharing the same bus waveguide may be made distinguishable by altering the quality factor and / or extinction ratio, thus enabling the identification of control / experimental rings. In some embodiments, intentional reduction of the quality factor and / or extinction ratio broadens (or lessens the depth of) resonance dips in the spectrum, effectively labeling the ring in question based on the peak shape.

[0096] In some embodiments, the sensor layer 101 comprises a supportive glass substrate, a double-sided adhesive layer for sealing underneath the photonic integrated circuit (PIC) sensor 110, a silicone PIC sensor holder, a one-sided silicone adhesive polyethylene terephthalate (PET) seal for above the PIC to divide the sensor into two separate sensing detection zones 103, and an adhesive double channel to incorporate flow within the two discrete compartments. The overall goal is to incorporate a partitioned PIC 110 into the device 100 such that it can directly interface with the membrane 105 and two separate microfluidic channels (apical and basal). Use of this partitioned PIC 110 significantly simplifies instrumentation needed for optical input / output (I / O) relative to the use of individual PICs in each microfluidic channel.

[0097] In some embodiments, the sensor layer 101 includes any suitable sensor types such as field-effect transistors (FET), bipolar junction transistors (BJT), and / or other electronic sensors.

[0098] Although the one device design integrates the novel PIC design shown in Fig. 7, the modular aspect of the device allows different PIC geometries to easily be swapped out with the current PIC design. Figs. 8A-8B show various exemplary PIC geometries that could be includedin the device including edge-coupled PICs where light input / output (I / O) occurs by bringing a fiber array close to the edge of the chip, and face-coupled PICs where light I / O occurs on the face via a grating coupler or similar. In the example shown in Fig. 8B, while the PIC chip is not specifically designed for use in the above-described microfluidic implementation, it can still be utilized with ring resonator banks 1-4 are exposed to the first channel, and banks 13-16 to the second channel, while banks 5-12 are inaccessible.

[0099] Overall, the device 100 differs from other microfluidics-based organ-on-a-chip devices as it is not only able to deliver real-time biosensing data of signaling occurring in situ but can simultaneously record soluble secretions from two biologically relevant compartments of the model. As such, specific design choices for the PIC layout were made specifically with discrete portioning capabilities in mind while still allowing the same sensor I / O to be used. A different solution to the need for dual apical and basal sensing could be to add additional sensors to the device. However, the ability of the device to sense two compartments of the microfluidic device without adding additional sensors allows the device to have true 'real-time' sensing capabilities (one does not have to realign the device to a different I / O configuration) and keep manufacturing costs at a minimum. In the context of modeling human skin, the device 100 can output pertinent data regarding both epidermal and dermal signaling cascades. Such information is vital to discover drugs with a higher efficacy or improve the current understanding of disease processes within the skin.

[0100] This platform and device can be used to study any tissues or areas of interest in the body of a subject that could be associated with any apical or basal polarity. Broadly speaking, any barrier tissues found in vivo can be modeled and further studied using the disclosed device. Examples of other biological applications include the blood-brain barrier, blood-retina barrier, blood-mammary gland barrier, liver sinusoid, air-lung interface, osteomyelitis / bone infection, glomerular capsule, and vascular-tendon interface such as human tendon-on-chip (hToC).

[0101] One aspect of the present invention relates to partitioning various cell types on either side of a membrane (e.g., a nanoporous membrane, a nanoporous membrane chip, nanoporous membrane chip 105), and creating microenvironments on each side of the 1membrane for the respective cell type(s), such that analytes secreted on either side of the membrane may be collected and analyzed separately (e.g., differential sensing). For example, in some embodiments, one or more cell of at least a first cell type are positioned or cultured in a first microenvironment formed on a first side or surface of the membrane (in some examples referred to as an apical side of the membrane), and one or more cells of at least as second cell type are positioned or cultured in a second microenvironment formed on a second side or surface of the membrane (in some examples referred to as a basal side of the membrane). For example, but without limitation, in some embodiments, device 100 comprises epidermal keratinocytes cultured on the first side of the membrane with a first microenvironment comprising an air liquid interface with one or more gases, and fibroblasts cultured on the second side of the membrane with a second microenvironment comprising one or more fluids such as cell culture media configured to feed and maintain the one or more cells. It should be appreciated that in this example the one or more fluids introduced to the second microenvironment may pass through the membrane and feed and maintain the one or more cells cultured on the first side of the membrane, in addition to feeding and maintain the one or more cells cultured on the second side of the membrane.

[0102] In some embodiment, the one or more cell types may comprise any known cell type known by one of ordinary level of skill in the art. For example, but without limitation, the one or more cell types may comprise: mammalian cells, plant cells, stem cells, organoids, subject- derived cells or organoids, skin cells, collagen cells, erythrocytes, epidermal cells, epidermal keratinocytes, vascular cells, fibroblasts, hepatocytes, lymphocytes, endothelial cells, muscle cells, myocytes, nerve cells, epithelial cells, erythrocytes, neurons, glial cells, bone cells, plant cells, dermal fibroblasts, and any primary cell or any combinations thereof.

[0103] It should be appreciated that the microenvironments created on each surface of the membrane may be supplied or cultured with any known culturing methods, materials, gases, fluids, known by one of ordinary level of skill in the art in order to create a suitable microenvironment for the one or more cell types. For example, but without limitation, the microenvironments may comprise any of: biomimetic conditions, recapitulated cell specificconditions or environments, hypoxic / anoxic conditions, hypercarbia / hypercapnia conditions, stimulation conditions, air, water, fluid, air-liquid interface, gas, cell culture media, buffering fluid, inflammatory cytokines, bacteria supernatants, lipopolysaccharide (LPS), or any other cellular insult factor where the cells are stimulated with any insult factor / molecule, or any combinations thereof.

[0104] In some embodiments, a biological sample of a subject is obtained and prepared for analysis. The sample may be any suitable cytological sample. In some embodiments, the sample is derived from a solid tissue sample or biopsy sample. For example, in some embodiments, the sample is a suspension of cells collected with a brush biopsy, scrape biopsy, or needle biopsy. In some embodiments, the sample comprises cytology samples collected from fine needle aspiration samples, or bodily fluids. In some embodiments, the sample may be obtained from a region or area of interest on a subject. For example, in some embodiments, the sample may be obtained from an area of interest that may or may not be suspected of having a disease or disorder (e.g., malignant or benign tissue). In some embodiments, the sample is processed prior to analysis. For example, the sample may be processed to permeabilize and fix the cells contained therein. However, in some embodiments, processing of the sample is not necessary.

[0105] In some embodiments, a multimodal measurement method comprises providing the device 100 as described herein, applying a fluid to the chip 105 via the cap layer 107, application channels 108, delivery channels 106, and detection channels / zones 103, and measuring the presence or quantity of an analyte from the chip 105 via the PIC 110 of the sensor layer 101.

[0106] The disclosed devices and methods may be used to replicate or recapitulate one or more disease states of tissue or cells, wherein one or more analytes measured from the cells may indicate the presence or absence of a disease or disorder. In another embodiment, one or more treatments may be applied to the tissues or cells, wherein the efficacy or potency of the treatment may be measured based on the one or more measured analytes. For example, in some embodiments, various skin disorders may be interrogated with experimental therapies in order to analyze the efficacy or potency of the treatment.

[0107] In one example device 100 includes epidermal cells (epidermal keratinocytes) on the first surface of the membrane, and dermal cells (dermal fibroblasts) on the second surface of the membrane. In certain embodiments, the presence, amount, concentration and / or the change thereof of one or more measured analytes (e.g., cytokines such as IL-33) from one or both sides of the membrane can be used to determine the presence of a disease state, monitor progression of a disease state and / or evaluate the efficacy of a treatment condition.EXPERIMENTAL EXAMPLES

[0108] The invention is now described with reference to the following Examples. These Examples are provided for the purpose of illustration only and the invention should in no way be construed as being limited to these Examples, but rather should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

[0109] Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the present invention and practice the claimed methods. The following working examples therefore specifically point out exemplary embodiments of the present invention and are not to be construed as limiting in any way the remainder of the disclosure.

[0110] Referring now to Figs. 9A-9D, an exemplary experimental device was built for testing purposes. Fig. 9A shows red dye being loaded into the top application channel after filling both the apical and basal channels with water. Since the apical channel outlet is sealed, and the air-liquid interface (ALI) purge valve is open, red dye remains only in the left half of the apical channel (slight diffusion occurring directly after the ALI purge valve canula. Fig. 9B shows red dye is removed from the apical channel after pushing air through the ALI purge valve. The apical channel outlet remained clamped. Fig. 9C shows that once the ALI purge valve is closed and the apical channel outlet is opened, red dye can flow uninterrupted through the full length of the apical channel. Slight backfilling is observed in the ALI purge valve tubing due to an improperly placed clamp. Fig. 9D shows after closing the apical channel outlet, flowing waterthrough the apical channel inlet and out the unplugged ALI purge valve allows for the red dye to be cleared from the left half of the apical channel once again.

[0111] Mass manufacturability of the device was then explored. To test the manufacturability a device built with component pieces as opposed to being built by individual layers as described above to increase assembly speed and repeatability. In a first example (Figs. 10A-10B), a 3-component device was explored. Described herein the three components depicted are referred to as Cl (M6-12), C2 (M4, 5), and C3 (Ml, 2, 3) according to their stacking order during manufacturing ('M' stands for material). Each individual layer and subsequent component layer combination is shown in the schematic of Fig. 10B. Minor changes were implemented based on testing, including the pressure sensitive adhesive (PSA) being switched from an acrylic to a silicone to both improve bonding between layers and imaging clarity, improving assembly by switching to a 0.005" adhesive.

[0112] Minor issues with the first run of manufactured components are detailed below and have since been addressed. For example, the PIC did not reliably fit in the PIC holding compartment of Cl, the membrane chip did not reliably fit in the membrane chip holding compartment of Cl (Fig. 11A), and the blue lining on the front and back of Cl ended up sticking to, and therefore stretching and bowing, the adhesive double channel layer preventing proper adhesive and changing channel geometry (Fig. 11B).

[0113] A few workarounds were developed to make the first round of manufactured parts usable. A first idea of using a 3D resin printed PIC 'plug' (Fig. 11C) was relatively easy to make at scale, but did not form a reliable seal, fit poorly, and did not include a PIC making filling the basal and apical channels with dye the only feasible proof-of-concept test. The second option was to use a razor blade to remove the extra acrylic and make the PIC holding space larger (Fig. 11D). While this method was better than the first option and about 50% more effective, the fit was still poor, and the small acrylic shavings interfered with the sealing of the PIC. The most effective method involved using a razor blade to fit in between the PIC holding layer and adhesive seal to remove the acrylic PIC holding layer as built and replace it with an in-house silicone PIC holding layer. This method was around 95% effective, but still took a lot of patience,was time intensive, and was an all-around cumbersome process that requires a steady hand.These manufacturing issues were addressed, and devices are now able to be produced at scale and tested with minimal hand assembly (Fig. 11D).

[0114] In a second example to further address the abovementioned issues a new component-based layer stack was adopted (Figs. 12A-12B). In this design, the third layer (the PIC holding layer) was now an additional component bringing the total number of manufactured components to 4. While not pictured explicitly in the above schematic, the blue linings surrounding now C2 (previously Cl) were also changed to make them more rigid, further preventing contact with the double channel adhesive layer and solving the issue of bowing in the adhesive channel layer of C2.

[0115] Referring now to Figs. 13A-13B, to assemble the device with the manufactured components, two different assembly jigs were designed and 3D printed out of resin. One jig was used to ensure the four components are placed completely flush to each other. With the assembly jig the alignment of the channels are guaranteed and therefore the PIC is guaranteed to be parallel to the edge of the device. This is imperative to the optical alignment capabilities of the Split PIC device. The second jig ensures that the Split PI C is placed parallel to the edge of the glass slide on the underside of the SplitPIC. If the SplitPIC is straight against the edge of the glass, all that is required to make sure the PIC is straight on the stage is to keep the glass slide straight.

[0116] Various tests were performed on the SplitPIC device, including testing that the device can maintain two discrete sensing channels with uniform resonance shifts as shown in Fig. 14, where the experiment confirmed that sensing could occur in one channel (apical) without cross-talk in the other channel (basal). This experiment was a simple bulk refractive index measurement, in which varying concentrations of sucrose were used.

[0117] Another experiment (Fig. 15) was performed to confirm that the apical and basal channels experience similar protein sensing profiles. In this experiment, the membrane was blocked to prevent cross talk. The results are shown in Fig. 16. Fig. 17 shows human umbilical vein endothelial cells (HUVECs) seeded within the experimental device.

[0118] The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention.

Claims

CLAIMSWhat is claimed is:

1. A multimodal measurement device, comprising: a split analyte sensor layer; a seal layer above the sensor layer, configured to divide the sensor layer into two or more detection zones, including a detection channel associated with each detection zone; a membrane holding layer above the seal layer, comprising a holder for a nanoporous membrane chip, and delivery channels fluidly connecting two or more surfaces of the chip to the two or more detection zones of the sensor layer via the detection channels; and a cap layer above the membrane holding layer, comprising a plurality of application channels including input and output ports, wherein the application channels are fluidly connected to chip via the delivery channels of the membrane holding layer.

2. The device of claim 1, wherein the detection channels, delivery channels, and application channels comprise microfluidic channels.

3. The device of any of claims 1-2, wherein the cap layer further comprises a purge valve.

4. The device of any of claims 1-3, wherein the device is configured to detect two or more analytes simultaneously.

5. The device of any of claims 1-4, wherein the sensor layer comprises a photonic integrated circuit (PIC) disposed directly on a substrate, optically coupled to a light source and a photodetector.

6. The device of claim 5, wherein the substrate comprises at least one of a cassette, a slide, a membrane, a fibrous substrate, or a test card.

7. The device of any of claims 5-6, wherein the photonic integrated circuit comprises: an input coupler; at least two output couplers; at least one waveguide optically connecting the input coupler to the output couplers; and a plurality of detection elements disposed as clusters along the at least one waveguide.

8. The device of claim 7, wherein the detection element clusters are positioned in the two or more detection zones.

9. The device of any of claims 7-8, wherein the detection element clusters are separated by at least 700 pm.

10. The device of any of claims 7-9, wherein the detection elements include at least one capture molecule.

11. The device of any of claims 7-10, wherein the input coupler is aligned with an optical input port, and the at least two output couplers are aligned with an optical output port.

12. The device of any of claims 7-11, wherein the detection elements include at least one of a ring resonator, a double ring resonator, a cylindrical resonator, a spherical resonator, a spiral waveguide, a Vernier filter, a photonic crystal, and a Mach-Zehnder Interferometer ("MZI").

13. The device of any of claims 7-12, wherein the at least one waveguide comprises a silicon nitride waveguide.

14. The device of any of claims 7-13, wherein the at least one waveguide splits into a plurality of branches from the input coupler to the at least two output couplers.

15. The device of claim 14, wherein each detection element cluster is positioned on separate branches.

16. The device of any of claims 1-15, wherein the cap layer comprises polydimethylsiloxane (PDMS).

17. The device of any of claims 1-16, wherein the membrane holder layer comprises silicone.

18. The device of any of claims 1-17, wherein the chip is configured to hold two or more cell cultures on its surfaces.

19. The device of any of claims 1-18, wherein the chip is configured to hold a different cell culture on each of its surfaces.

20. A multimodal measurement device, comprising: a photonic integrated circuit (PIC) disposed directly on a substrate, optically coupled to a light source and a photodetector; a seal layer above the PIC, configured to divide the PIC into first and second detection zones, including a first detection channel associated with the first detection zone, and a second detection channel associated with the second detection zone; a membrane holding layer above the seal layer, comprising a holder for a nanoporous membrane chip, a top delivery channel fluidly connecting the top surface of the chip to the first detection zone of the PIC via the first detection channel, and a bottom delivery channel fluidly connecting the bottom surface of the chip to the second detection zone of the PIC via the second detection channel; anda cap layer above the membrane holding layer, comprising: a top application channel including input and output ports, fluidly connected to the top surface of the chip via the top delivery channel; and a bottom application channel including input and output ports, fluidly connected to the bottom surface of the chip via the bottom delivery channel.

21. The device of claim 20, wherein the first and second detection channels, top and bottom delivery channels, and top and bottom application channels comprise microfluidic channels.

22. The device of any of claims 20-21, wherein the top application channel further comprises a purge valve.

23. The device of any of claims 20-22, wherein the chip is configured to hold a first cell culture on its top surface, and a second cell culture different from the first on its bottom surface.

24. A multimodal measurement method, comprising: providing the device of claim any of claims 1-23; applying a fluid to the chip via the cap layer; and measuring the presence or quantity of an analyte from the chip via the PIC.

25. A photonic integrated circuit (PIC), comprising: a substrate; an input coupler on the substrate; at least two output couplers on the substrate; at least one waveguide on the substrate optically connecting the input coupler to the output couplers; anda plurality of detection elements disposed as clusters on the on the substrate along the at least one waveguide.

26. The device of claim 25, wherein the detection element clusters are separated by at least 700 pm.

27. The device of any of claims 25-26, wherein the detection elements include at least one capture molecule.

28. The device of any of claims 25-27, wherein the detection elements include at least one of a ring resonator, a double ring resonator, a cylindrical resonator, a spherical resonator, a spiral waveguide, a Vernier filter, a photonic crystal, and a Mach-Zehnder Interferometer ("MZI").

29. The device of any of claims 25-28, wherein the at least one waveguide comprises a silicon nitride waveguide.

30. The device of any of claims 25-29, wherein the at least one waveguide splits into a plurality of branches from the input coupler to the at least two output couplers.

31. The device of claim 30, wherein each detection element cluster is positioned on separate branches.

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