Multi-modal surface plasmon resonance detector with additively manufactured liquid-cooled frame for measuring molecular interactions

Abstract

A multi-modal surface plasmon resonance detector for measuring molecular interactions is provided herein. The multi-modal surface plasmon resonance detector includes a detector frame manufactured by metal additive manufacturing, the detector frame comprising integrated conformal cooling passages. The multi-modal surface plasmon resonance detector includes a prism providing a plurality of optical faces for a plurality of optical paths. The multi-modal surface plasmon resonance detector includes a first detection path configured for surface plasmon resonance detection. The multi-modal surface plasmon resonance detector includes a second detection path configured for surface plasmon resonance imaging. The multi-modal surface plasmon resonance detector includes a sensor chip interface configured to receive a sensor chip. The multi-modal surface plasmon resonance detector includes a liquid cooling system configured to circulate liquid through the integrated conformal cooling passages.

Description

Docket No.: INST-01-PCTMULTI-MODAL SURFACE PLASMON RESONANCE DETECTOR WITH ADDITIVELY MANUFACTURED LIQUID-COOLED FRAME FOR MEASURING MOLECULAR INTERACTIONSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority from U.S. Provisional Application Serial No.63 / 742,398 filed on January 6, 2025, and U.S. Provisional Application Serial No. 63 / 742,399 filed on January 6, 2025, each of which is incorporated herein by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] Not Applicable.INTRODUCTION

[0003] Surface plasmon resonance (SPR) biosensing has become a widely adopted technique for label -free detection and characterization of biomolecular interactions. The technique exploits optical phenomena occurring at metal-dielectric interfaces where incident light at specific angles excites collective oscillations of free electrons in thin metal films, creating evanescent fields that extend into sample media. Changes in refractive index within these evanescent fields caused by molecular binding events shift the resonance condition, enabling real-time monitoring of association and dissociation kinetics without requiring fluorescent or radioactive labels.

[0004] Commercial SPR instrumentation has evolved along two distinct technological trajectories. Traditional SPR systems, exemplified by the Biacore product line (Cytiva, Marlborough, MA), employ angular interrogation with focused illumination and photodiode array or line camera detection to achieve high sensitivity measurements. These systems ty pically monitor 2-8 interaction zones and achieve noise floors of approximately 0.03 RU RMSD, enabling detection of small molecule binding events. SPR imaging systems, including the Sierra SPR-24 / 32 Pro (Bruker, Billerica, MA) and the LSA platform (Carterra, Salt Lake City, UT), employ wide-field illumination and two-dimensional camera detection to monitor arrays of 24-384 or more interaction zones simultaneously, though with reduced sensitivity' of approximately 0.5 RU RMSD.

[0005] The thermal sensitivity of SPR measurements presents ongoing challenges for instrument design. Refractive index varies with temperature at approximately 10-4Docket No.: INST-01-PCTRIU / °C for aqueous solutions (Homola, J., "Surface Plasmon Resonance Sensors for Detection of Chemical and Biological Species," Chem. Rev. 2008, 108, 462-493), requiring temperature stability of ±0.001 °C to achieve noise floors below 0.01 RU. Commercial systems address thermal management through thermoelectric cooler (TEC) assemblies with forced air cooling, resulting in instrument footprints of 3,800-4,500 cm3 for the detector assembly alone and acoustic signatures of 54-58 dB(A) during operation.

[0006] Detector frame construction in existing SPR instruments typically employs multi-component assemblies comprising 15-25 precision-machined parts including base plates, optical mounting blocks, TEC housings, heat sinks, fan mounting brackets, thermal interface plates, and various alignment components. Each component requires individual manufacturing, inspection, and assembly, with tolerances of ±0.01-0.05 mm for optical alignment. Assembly requires skilled technicians, specialized fixtures, and labor times of 8-1 hours per detector. This manufacturing complexity contributes to instrument costs of $300,000-500,000 for traditional SPR systems and $600,000-800,000 for imaging systems, with annual service contracts of $30,000-50,000.

[0007] Liquid cooling approaches have been applied in high-performance optical systems, as described in Mici et al., "Optomechanical Performance of 3D-Printed Mirrors with Embedded Cooling Channels," SPIE Proceedings 2015, where metal additive manufacturing enabled creation of optical mirror substrates with internal cooling passages. However, application of such approaches to SPR biosensing geometries, where thermal management, optical alignment, and biochemical interface requirements intersect, presents distinct challenges not addressed in the optical engineering literature.

[0008] Multi-wavelength SPR detection has been explored as a means to extend dynamic range and improve sensitivity. U.S. Patent No. 6,862,094 to Nikitin et al. describes a two-dimensional imaging SPR apparatus using multiple wavelengths for simultaneous detection, enabling tracking of both sides of the SPR dip and extending the measuring range. However, such systems operate in a single detection modality and do not address the tradeoff between high-sensitivity detection and high-throughput array imaging within a single instrument.

[0009] Prism geometries for SPR coupling have been described in various configurations. U.S. Patent Application Publication No. 2019 / 0360932 describes an SPR imaging system using a hemispherical prism with a high numerical aperture lens for achieving high optical resolution and wide SPR angle range. U.S. Patent No. 8,149,411 describes a sensor unit using curved transparent structures to eliminate the need for refractiveDocket No.: INST-01-PCTindex matching materials. These approaches address specific optical coupling challenges but do not provide for integration of multiple detection modalities within a single compact instrument.

[0010] Prism geometries for SPR detection have been explored in various configurations. U.S. Patent Application Publication No. 2019 / 0360932 describes an SPR imaging system using a hemispherical prism with a high numerical aperture lens for achieving high optical resolution and wide SPR angle range. U.S. Patent No. 8,149,411 describes a sensor unit using curved transparent structures to eliminate the need for refractive index matching materials. These approaches address specific optical coupling challenges but do not provide for integration of multiple detection modalities within a single compact instrument.

[0011] Fragment-based drug discovery applications place particular demands on SPR instrumentation. Detection of weak binding interactions with equilibrium dissociation constants from 100 pM to 10 mM between small molecule fragments (molecular weight less than 300 Da) and protein targets requires high protein immobilization densities of 5,000 RU or greater (Erlanson, D.A., et al., "Twenty Years On: The Impact of Fragments on Drug Discovery," Nat. Rev. Drug Discov. 2016, 15, 605-619). Screening campaigns testing hundreds of compounds per surface require regeneration conditions that presen e protein activity across multiple cycles, a challenge that current harsh chemical regeneration approaches do not adequately address.

[0012] Engineered photoswitchable proteins, including OptoNanobodies with LOV2 domain insertions (Gil, A.A., et al., "Optogenetic Control of Protein Binding Using Light-Switchable Nanobodies," Nat. Commun. 2020, 11, 4044), offer potential for light-controlled binding affinity that could enable gentle surface regeneration. Characterization of such proteins requires real-time monitoring of affinity changes during light activation, a capability not provided by existing SPR instruments that lack integrated photochemistry paths.

[0013] The separation of high-sensitivity detection and array imaging capabilities into distinct instrument platforms creates workflow inefficiencies for researchers requiring both capabilities. Screening large numbers of molecular interactions in imaging mode followed by detailed kinetic characterization of hits in high-sens i ti x i t mode requires sample transfer between instruments, increasing experimental time and introducing potential for sample degradation or contamination. Instrument costs exceeding $1 million for combined capabilities limit accessibility for academic laboratories and smaller research organizations.Docket No.: INST-01-PCT

[0014] Additive manufacturing technologies, including selective laser melting (SLM) and direct metal laser sintering (DMLS). enable fabrication of complex internal geometries in thermally conductive metal alloys that would be impractical using conventional subtractive machining. Conformal cooling channel designs have been applied in injection mold tooling to improve thermal uniformity and cycle times. Application of such manufacturing approaches to optical biosensor instrumentation presents opportunities for integration of thermal management, structural support, and optical mounting features that have not been fully explored in the SPR biosensing field.SUMMARY

[0015] The present disclosure addresses the technical problem of providing a compact SPR instrument that integrates both high-sensitivity detection and imaging modes within a single device, with a liquid-cooled, additively manufactured frame achieving temperature stability of ±0.003°C at the sensing surface, reduced acoustic signature compared to fan-cooled systems, and consolidated manufacturing complexity. According to an aspect of the present disclosure, a multi-modal surface plasmon resonance detector for measuring molecular interactions is provided. The multi-modal surface plasmon resonance detector includes a detector frame manufactured by metal additive manufacturing, the detector frame comprising integrated conformal cooling passages. The multi-modal surface plasmon resonance detector includes a prism providing a plurality of optical faces for a plurality of optical paths. The multi-modal surface plasmon resonance detector includes a first detection path configured for surface plasmon resonance detection. The multi-modal surface plasmon resonance detector includes a second detection path configured for surface plasmon resonance imaging. The multi-modal surface plasmon resonance detector includes a sensor chip interface configured to receive a sensor chip. The multi-modal surface plasmon resonance detector includes a liquid cooling system configured to circulate liquid through the integrated conformal cooling passages. The multi-modal surface plasmon resonance detector operates by circulating temperature-controlled liquid through the integrated conformal cooling passages to maintain thermal stability at a sensing surface while enabling independent operation of the first detection path and the second detection path through the plurality of optical faces of the prism. The first detection path provides surface plasmon resonance detection with sensitivity of 0.05 RU RMSD or less suitable for kinetic characterization of molecular interactions, while the second detection path provides surface plasmon resonance imaging detection suitable for high-throughput screening of arrays ofDocket No.: INST-01-PCTmolecular interactions. The pyramidal geometry of the prism enables orthogonal optical paths that share a common sensing surface without optical interference, allowing users to switch between detection modes without mechanical reconfiguration. In working examples, the multi-modal surface plasmon resonance detector achieved temperature stability’ of ±0.003°C at the sensing surface during continuous operation, with the first detection path providing sensitivity of 0.03 RU RMSD and the second detection path enabling simultaneous monitoring of multiple interaction zones across the sensing surface.

[0016] According to other aspects of the present disclosure, the multi-modal surface plasmon resonance detector may include one or more of the following features. The detector frame may be manufactured from a thermally conductive metal alloy. The thermally conductive metal alloy may comprise AlSilOMg aluminum alloy. The thermally conductive metal alloy may comprise a copper alloy. The thermally conductive metal alloy may comprise titanium. The thermally conductive metal alloy may comprise stainless steel. The integrated conformal cooling passages may be formed during the metal additive manufacturing. The integrated conformal cooling passages may have a diameter in a range of 2 mm to 4 mm. The integrated conformal cooling passages may follow a serpentine channel pattern. The integrated conformal cooling passages may follow a parallel channel pattern. The integrated conformal cooling passages may be routed within 5 mm to 10 mm of the prism. The integrated conformal cooling passages may comprise teardrop-shaped crosssections configured for printing without removable supports. The plurality of optical paths may comprise orthogonal optical paths. The second detection path may be orthogonal to the first detection path. The first detection path may be configured for surface plasmon resonance detection providing sensitivity of 0.05 RU RMSD or less. The sensor chip interface may be configured to receive the sensor chip at a top face of the prism. The liquid cooling system may be configured to circulate temperature-controlled liquid through the integrated conformal cooling passages. The liquid cooling system may be configured to maintain a temperature stability of ±0.003 °C at a sensing surface. The liquid cooling system may be configured to operate over a temperature range of 4°C to 40°C. The liquid cooling system may be configured to circulate the temperature-controlled liquid at a flow rate in a range of 150 mL / min to 250 mL / min. The liquid cooling system may provide a heat transfer coefficient in a range of 2000 W / m2K to 5000 W / m2K. The prism may comprise a pyramidal prism. The prism may be fabricated from BK7 glass. The prism may be fabricated from SF 10 glass. The first detection path may comprise a near-infrared light source operating at a wavelength of 850 nm (e.g., in a range of 840 nm to 860 nm). The first detection pathDocket No.: INST-01-PCTmay comprise a light source having a power in a range of 5 mW to 10 mW. The first detection path may comprise a line camera detector. The second detection path may comprise a two-dimensional camera having at least 5 megapixels. The first detection path may be configured to provide a sensitivity of 0.05 RU RMSD or less. The second detection path may be configured to provide a sensitivity of 0.5 RU RMSD. The multi-modal surface plasmon resonance detector may further comprise a photochemistry path configured to deliver ultraviolet light at a wavelength of 365 nm (e.g., in a range of 350 nm to 380 nm) to the sensor chip interface.

[0017] According to another aspect of the present disclosure, a liquid-cooled detector frame for optical biosensing is provided. The liquid-cooled detector frame includes a monolithic frame manufactured by metal additive manufacturing. The liquid-cooled detector frame includes integrated conformal cooling passages formed within the monolithic frame. The liquid-cooled detector frame includes a coolant inlet and a coolant outlet fluidically connected to the integrated conformal cooling passages. The liquid-cooled detector frame includes mounting features for optical components. The liquid-cooled detector frame operates by receiving temperature-controlled coolant through the coolant inlet, routing the coolant through the integrated conformal cooling passages that are positioned proximate to sensing zones and optical component mounting regions, and returning the coolant through the coolant outlet. The integrated conformal cooling passages provide heat transfer coefficients of 2000-5000 W / m2K in turbulent flow regimes, enabling precise temperature regulation across the sensing region and optical mounting areas. The monolithic frame consolidates structural support, thermal management, and optical mounting features into a single component, reducing manufacturing complexity compared to traditional detector assemblies that require 15-25 precision-machined components. In working examples, the liquid-cooled detector frame achieved temperature variation of less than 0.005°C between optical component mounting points during continuous 8-hour operation, with the compact detector volume of 576 cm3representing a reduction compared to traditional TEC-fan assemblies requiring 3,800-4,500 cm3.

[0018] According to other aspects of the present disclosure, the liquid-cooled detector frame may include one or more of the following features. The monolithic frame may be manufactured from a thermally conductive metal alloy. The thermally conductive metal alloy may comprise AlSilOMg aluminum alloy having athermal conductivity in a range of 130 W / m K to 180 W / m K. The thermally conductive metal alloy may comprise a copper alloy having a thermal conductivity in a range of 350 W / m K to 400 W / m K. The integratedDocket No.: INST-01-PCTconformal cooling passages may be formed during the metal additive manufacturing. The integrated conformal cooling passages may be routed around a sensing zone and optical component mounting regions. The mounting features may comprise a prism mounting region, a light source mounting region, and a detector mounting region. The integrated conformal cooling passages may have a diameter in a range of 2 mm to 4 mm. The integrated conformal cooling passages may comprise teardrop-shaped cross-sections having a self-supporting geometry with an apex angle of less than 45 degrees. The integrated conformal cooling passages may be spaced 5 mm to 10 mm from a prism interface. The liquid-cooled detector frame may further comprise a temperature sensor embedded in the monolithic frame proximate to the sensing zone. The monolithic frame may be configured to interface with a thermoelectric cooler module positioned external to the monolithic frame. The mounting features may comprise precision-machined surfaces having a tolerance of ±0.01 mm. The monolithic frame may have a resonant frequency greater than 200 Hz. The monolithic frame may have a detector volume of 576 cm3. The integrated conformal cooling passages may be configured to provide unified thermal management for a light-emitting diode and a camera. The integrated conformal cooling passages may comprise a camera cooling passage routed proximate to the detector mounting region. The integrated conformal cooling passages may comprise heat transfer enhancement structures. The monolithic frame may be configured to produce an acoustic signature of 38 dB(A) to 40 dB(A) during operation. The liquid-cooled detector frame may further comprise quick-disconnect fittings at the coolant inlet and the coolant outlet for connection to an external chiller. The integrated conformal cooling passages may comprise a first set of cooling passages and a second set of cooling passages configured for multi-zone temperature control. The monolithic frame may be manufactured by selective laser melting. The integrated conformal cooling passages may comprise self-supporting geometries having tapered upper surfaces configured for printing without removable supports. The monolithic frame may integrate structural support, thermal management, and optical mounting features in a single component. The monolithic frame may be configured to maintain a temperature variation of less than 0.005 °C between optical component mounting points. The integrated conformal cooling passages may be routed around a prism interface, optical mounting points, and a sample staging area. The monolithic frame may comprise fluidic ports integrated into the frame structure. The integrated conformal cooling passages may follow a three-dimensional path traversing different depths within the monolithic frame. The monolithic frame may be configured to remove heat from electronic subsystems connected to a same coolant loop.Docket No.: INST-01-PCT

[0019] According to another aspect of the present disclosure, a method for detecting molecular interactions using a multi-modal surface plasmon resonance detector is provided. The method includes circulating liquid through integrated conformal cooling passages in a detector frame to maintain thermal stability at a sensing surface. The method includes operating in a first detection mode using a first optical path for surface plasmon resonance detection. The method includes operating in a second detection mode using a second optical path for surface plasmon resonance imaging detection. The method includes switching between the first detection mode and the second detection mode. The method enables users to screen large numbers of molecular interactions in the second detection mode, identify hits from the screening results, and then validate the identified hits in the first detection mode with kinetic characterization providing sensitivity of 0.05 RU RMSD or less, all within a single device without sample transfer or instrument switching. The circulating liquid maintains thermal stability at the sensing surface, which is particularly advantageous for surface plasmon resonance detection where temperature-induced refractive index drift can compromise measurement accuracy. In working examples, the method was used to characterize small molecule binding to carbonic anhydrase II, with the resulting rate constants matching published values for these interactions. In prophetic examples, the method enables automated antibody screening w orkflow s where 96 antibody variants are screened in the second detection mode followed by kinetic validation ith sensitivity of 0.05 RU RMSD or less of top hits in the first detection mode, completing the entire workflow in 6-8 hours with minimal user intervention.

[0020] According to other aspects of the present disclosure, the method may include one or more of the following features. The detector frame may be additively manufactured. Circulating liquid may comprise circulating temperature-controlled liquid through the integrated conformal cooling passages. The first optical path may be configured for surface plasmon resonance detection providing sensitivity of 0.05 RU RMSD or less. The second optical path may be orthogonal to the first optical path. Switching between the first detection mode and the second detection mode may comprise switching via software control without mechanical reconfiguration. The method may further comprise screening a plurality of molecular interactions in the second detection mode to identify hits. The method may further comprise validating identified hits in the first detection mode following screening in the second detection mode. The method may further comprise operating the first detection mode and the second detection mode concurrently. The method may further comprise activating a photochemistry path to deliver ultraviolet light to the sensing surface during surface plasmonDocket No.: INST-01-PCTresonance monitoring. The photochemistry path may deliver ultraviolet light at a wavelength of 365 nm (e.g., in a range of 350 nm to 380 nm) for activating photoactivatable crosslinkers. The method may further comprise monitoring photocoupling dynamics in real-time using the first detection mode during ultraviolet activation. Circulating the temperature-controlled liquid may comprise maintaining a temperature stability of ±0.003°C at the sensing surface. Circulating the temperature-controlled liquid may comprise transitioning from 25°C to 37°C in less than 10 minutes. The method may further comprise screening an array of molecular interactions in the second detection mode, wherein the array comprises microarray spots having a diameter in a range of 100 pm to 500 pm. The method may further comprise extracting kinetic parameters comprising an association rate constant, a dissociation rate constant, and an equilibrium dissociation constant from data obtained in the first detection mode. The method may further comprise characterizing antibody-antigen binding interactions. The method may further comprise performing epitope binning using the second detection mode. The method may further comprise integrating with nucleic acid programmable protein array technology to express proteins directly on the sensing surface. The method may further compnse activating photoactivatable coupling chemistry using the photochemistry path while monitoring coupling efficiency using the first detection mode. The first detection mode may provide a sensitivity7of 0.05 RU RMSD or less. The second detection mode may provide a sensitivity of 0.5 RU RMSD. The method may further comprise visualizing fluid flow patterns across the sensing surface using the second detection mode. The method may further comprise detecting small molecules having a molecular weight of less than 500 Da binding to proteins. The method may further comprise performing fragment-based drug discovery screening. The method may further comprise regenerating the sensing surface using photo-triggered release of captured molecules. The method may further comprise monitoring binding kinetics at a plurality of array spots simultaneously using the second detection mode. The method may further comprise operating the first detection mode at a frame rate in a range of 100 Hz to 1000 Hz. The method may further comprise operating the second detection mode at a frame rate in a range of 1 Hz to 500 Hz. The method may further comprise maintaining the sensing surface at a temperature of 25°C or 37°C for biological studies.

[0021] According to another aspect of the present disclosure, a method for manufacturing a detector frame for optical biosensing is provided. The method includes fabricating a monolithic frame using metal additive manufacturing. The method includes forming integrated conformal cooling passages within the monolithic frame. The methodDocket No.: INST-01-PCTincludes post-processing mounting surfaces of the monolithic frame. The method enables creation of complex internal geometries including conformal cooling passages routed around sensing zones and optical component mounting regions that cannot be economically produced through conventional machining. The integrated conformal cooling passages are formed during the metal additive manufacturing process, enabling three-dimensional routing paths that pass within 5-10 mm of all thermal zones while avoiding interference with optical mounting features and structural supports. The post-processing of mounting surfaces provides precision-machined features suitable for optical component mounting with tolerances of ±0.01 mm. In working examples, the method produced a detector frame in a single additive manufacturing build with a print time of 6-12 hours followed by 2-4 hours of post-processing for support removal and surface machining, consolidating functions of 15-25 precision-machined components into a single monolithic part.

[0022] According to other aspects of the present disclosure, the method for manufacturing may include one or more of the following features. Fabricating the monolithic frame may comprise fabricating from a thermally conductive metal alloy. Forming the integrated conformal cooling passages may comprise forming during the metal additive manufacturing. Post-processing the mounting surfaces may comprise post-processing to optical tolerances. Fabricating the monolithic frame may comprise selective laser melting. The thermally conductive metal alloy may comprise AlSilOMg aluminum alloy. The thermally conductive metal alloy may comprise a copper alloy. Forming the integrated conformal cooling passages may comprise forming teardrop-shaped passage cross-sections configured for printing without removable supports. Forming the integrated conformal cooling passages may comprise forming self-supporting geometries. Fabricating the monolithic frame may comprise a print time in a range of 6 hours to 12 hours. Postprocessing the mounting surfaces may comprise a post-processing time in a range of 2 hours to 4 hours. Post-processing the mounting surfaces may comprise machining mounting surfaces to a tolerance of ±0.01 mm. Forming the integrated conformal cooling passages may comprise forming passages having a diameter in a range of 2 mm to 4 mm. Forming the integrated conformal cooling passages may comprise routing the passages within 5 mm to 10 mm of a prism interface region. Forming the integrated conformal cooling passages may comprise forming a serpentine channel pattern. Forming the integrated conformal cooling passages may comprise forming a parallel channel pattern. The method may further comprise integrating structural support features, thermal management features, and optical mounting features in the monolithic frame as a single component. The method may further compriseDocket No.: INST-01-PCTforming heat transfer enhancement structures within the integrated conformal cooling passages. The method may further comprise forming a coolant inlet port and a coolant outlet port integrated into the monolithic frame. The method may further comprise forming a temperature sensor cavity within the monolithic frame proximate to a sensing zone. Forming the integrated conformal cooling passages may comprise forming passages that traverse different depths within the monolithic frame. The method may further comprise forming a camera cooling passage routed proximate to a detector mounting region. The method may further comprise forming a prism mounting region, a light source mounting region, and a detector mounting region in the monolithic frame. The monolithic frame may consolidate functions of a plurality of precision-machined components into a single part. The method may further comprise removing support structures from the integrated conformal cooling passages during post-processing. The monolithic frame may be configured to achieve a resonant frequency greater than 200 Hz for optical stability. The method may further comprise forming a first set of cooling passages and a second set of cooling passages for multi-zone temperature control. The monolithic frame may be configured to provide unified thermal management for optical components and electronic subsystems.

[0023] According to another aspect of the present disclosure, a multi-modal surface plasmon resonance detector for measuring molecular interactions is provided. The multimodal surface plasmon resonance detector includes a detector frame manufactured by metal additive manufacturing, the detector frame comprising integrated conformal cooling passages routed proximate to a prism interface and around a sensing zone. The multi-modal surface plasmon resonance detector includes a prism providing a plurality of optical faces for a plurality of optical paths. The multi-modal surface plasmon resonance detector includes a first detection path configured for surface plasmon resonance detection, the first detection path comprising a light source, optical elements, and a detector. The multi-modal surface plasmon resonance detector includes a second detection path configured for surface plasmon resonance imaging detection, the second detection path comprising a light source, optical elements, and a camera. The multi-modal surface plasmon resonance detector includes a photochemistry path configured to deliver ultraviolet light to a sensor surface. The multimodal surface plasmon resonance detector includes a sensor chip interface configured to receive a sensor chip. The multi-modal surface plasmon resonance detector includes a liquid cooling system configured to circulate temperature-controlled liquid through the integrated conformal cooling passages. The multi-modal surface plasmon resonance detector includes a microfluidic flow cell positioned above the sensor chip for delivery of samples to the sensorDocket No.: INST-01-PCTsurface. The detector frame integrates structural support, thermal management, and optical mounting features. The first detection path and the second detection path are configured for mode switching without mechanical reconfiguration. The multi-modal surface plasmon resonance detector operates by circulating temperature-controlled liquid through the integrated conformal cooling passages to maintain thermal stability while enabling independent or concurrent operation of the first detection path, the second detection path, and the photochemistry path through the plurality of optical faces of the prism. The orthogonal optical path architecture enables the first detection path to provide kinetic measurements with sensitivity of 0.05 RU RMSD or less while the second detection path provides array -based imaging, with the photochemistry path enabling in-situ activation of photoactive chemistries during surface plasmon resonance monitoring. In working examples, the multi-modal surface plasmon resonance detector was used to visualize fluid flow patterns during fluidics development using the second detection path while collecting kinetic data with sensitivity of 0.05 RU RMSD or less using the first detection path, demonstrating the capability7for concurrent multi-modal operation. In prophetic examples, the photochemistry7path enables real-time monitoring of photoactivatable coupling chemistry, eliminating the need for unstable NHS / EDC reagents and providing precise temporal control over surface immobilization.

[0024] In another aspect, a multi-modal surface plasmon resonance detector for measuring molecular interactions includes a detector frame manufactured by metal additive manufacturing, the detector frame comprising integrated conformal cooling passages routed proximate to a prism interface and around a sensing zone. The detector includes a prism providing a plurality of optical faces for a plurality of optical paths, a first detection path configured for surface plasmon resonance detection comprising a light source, optical elements, and a detector, and a second detection path configured for surface plasmon resonance imaging detection comprising a light source, optical elements, and a camera. The detector further includes a photochemistry path configured to deliver ultraviolet light to a sensor surface, a sensor chip interface configured to receive a sensor chip, a liquid cooling system configured to circulate temperature-controlled liquid through the integrated conformal cooling passages, and a microfluidic flow cell positioned above the sensor chip for delivery of samples to the sensor surface. The detector frame integrates structural support, thermal management, and optical mounting features, and the first detection path and the second detection path are configured for mode switching without mechanical reconfiguration.

[0025] In another aspect, a multi-modal surface plasmon resonance detector forDocket No.: INST-01-PCTmeasuring molecular interactions includes a prism providing a plurality of optical faces for a plurality of optical paths, a first detection path configured for surface plasmon resonance detection comprising a first light source, first optical elements, and a first detector, and a second detection path configured for surface plasmon resonance imaging detection comprising a second light source, second optical elements, and a two-dimensional camera. The first detection path and the second detection path are configured to interrogate a common sensing surface through different optical faces of the prism, and the first detection path and the second detection path are configured for mode switching without mechanical reconfiguration.

[0026] In another aspect, a multi-modal surface plasmon resonance detector for measuring molecular interactions includes a pyramidal prism having a plurality of optical faces arranged to enable a plurality of orthogonal optical paths addressing a common sensing surface. The detector includes a first detection path configured for surface plasmon resonance detection providing sensitivity of 0.05 RU RMSD or less, the first detection path comprising a light source and a detector arranged along a first diagonal pair of optical faces of the pyramidal prism. The detector includes a second detection path configured for surface plasmon resonance imaging detection comprising a light source and a two-dimensional camera arranged along a second diagonal pair of optical faces of the pyramidal prism orthogonal to the first diagonal pair, and a sensor chip interface configured to receive a sensor chip at a top face of the pyramidal prism. The orthogonal arrangement of the first detection path and the second detection path enables simultaneous or sequential interrogation of the common sensing surface without optical interference.

[0027] In another aspect, a multi-modal surface plasmon resonance detector for measuring molecular interactions includes a prism providing a plurality of optical faces for a plurality of optical paths, a first detection path configured for surface plasmon resonance detection, and a second detection path configured for surface plasmon resonance imaging detection, the second detection path being orthogonal to the first detection path. The detector includes a photochemistry path configured to deliver light at a wavelength suitable for activating photoactivatable functional groups to a sensing surface, and a sensor chip interface configured to receive a sensor chip. The first detection path, the second detection path, and the photochemistry path are configured for independent or concurrent operation.

[0028] In another aspect, a method for detecting molecular interactions using a multi-modal surface plasmon resonance detector includes operating in a first detection mode using a first optical path through a prism for surface plasmon resonance detection, andDocket No.: INST-01-PCToperating in a second detection mode using a second optical path through the prism for surface plasmon resonance imaging detection, wherein the second optical path is different from the first optical path. The method includes switching between the first detection mode and the second detection mode via software control without mechanical reconfiguration of optical components, wherein the first detection mode and the second detection mode interrogate a common sensing surface.

[0029] In another aspect, a method for detecting molecular interactions includes screening a plurality of molecular interactions in a surface plasmon resonance imaging mode using a first optical path through a prism to identify hits, switching to a surface plasmon resonance detection mode using a second optical path through the prism without mechanical reconfiguration, and validating identified hits in the surface plasmon resonance detection mode with sensitivity of 0.05 RU RMSD or less. The screening and validating are performed using a common sensing surface within a single device.

[0030] In another aspect, a method for detecting molecular interactions includes monitoring a sensing surface using surface plasmon resonance detection through a first optical path of a prism, activating a photochemistry path to deliver ultraviolet light at a wavelength in a range of 350 nm to 380 nm to the sensing surface while continuing to monitor the sensing surface, and detecting photocoupling dynamics in real-time using the surface plasmon resonance detection. The photochemistry path is configured to activate photoactivatable crosslinkers on the sensing surface without requiring chemical activation reagents.DRAWINGS

[0031] Those of skill in the art will understand that the drawings, described below, are for illustrative purposes only. The drawings are not intended to limit the scope of the present teachings in any way.

[0032] FIG. 1 illustrates a method for detecting molecular interactions using a multi-modal surface plasmon resonance detector.

[0033] FIG. 2 depicts a method for manufacturing a detector frame for optical biosensing.

[0034] FIG. 3 shows a method for detecting molecular interactions with an optional photochemistry path.

[0035] FIG. 4 presents a system workflow for a multi-modal surface plasmon resonance detector.Docket No.: INST-01-PCT

[0036] FIG. 5 depicts a multi-modal surface plasmon resonance detector.

[0037] FIG. 6 illustrates a liquid cooling system and detector frame for thermal management.

[0038] FIG. 7 depicts a detector frame manufactured using metal additive manufacturing.

[0039] FIG. 8 illustrates a cutaway view of the detector frame of FIG. 7.

[0040] FIG. 9 depicts a cross-sectional view of the detector frame showing internal cooling via geometry.

[0041] FIG. 10 presents a cutaway view of the detector frame revealing cooling vias and heat transfer structures.

[0042] FIG. 11 illustrates a cutaway view of the detector frame showing a coolant flow path.

[0043] FIG. 12 depicts a cross-sectional view of the detector frame showing internal cooling via geometry.

[0044] FIG. 13 illustrates a plan view of the detector frame showing cooling via network and structural features.

[0045] FIG. 14 depicts a multi-modal detector assembly with orthogonal optical paths.

[0046] FIG. 15 illustrates a multi-modal detector assembly with a pyramidal prism structure.

[0047] FIG. 16 depicts a prototype multi-modal detector assembly.DETAILED DESCRIPTION

[0048] All patents, applications, published applications and other publications cited herein are incorporated by reference in their entirety. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary7skill in the art to which the invention belongs.

[0049] Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of this invention. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure. In the event that there is a plurality of definitions for a term herein, those in this section prevail unless stated otherwise. Headings used herein are for organizational purposes only and in no way limit the invention described herein.Docket No.: INST-01-PCT

[0050] Abbreviations and Definitions

[0051] To facilitate understanding of the invention, a number of terms and abbreviations as used herein are defined below as follows:

[0052] Surface Plasmon Resonance (SPR): As used herein, the term "surface plasmon resonance" or "SPR" refers to an optical phenomenon occurring at a metal-dielectric interface where incident light at a specific angle excites collective oscillations of free electrons (surface plasmons) in a thin metal film, creating an evanescent field extending approximately 200 nm into the sample medium, wherein changes in refractive index within this evanescent field caused by molecular binding events shift the resonance condition, enabling label-free detection and quantification of biomolecular interactions.

[0053] Detector Frame: As used herein, the term "detector frame" refers to a structural component manufactured by metal additive manufacturing that provides mechanical support for optical components, thermal management through integrated cooling passages, and precision mounting features for a prism, light sources, detectors, and associated optical elements, wherein the detector frame consolidates functions that would otherwise require 15-25 separate precision-machined components in traditional detector assemblies.

[0054] Metal Additive Manufacturing: As used herein, the term "metal additive manufacturing" is broadly defined to include manufacturing processes that build three-dimensional metal structures layer-by-layer from metal powder or wire feedstock, including but not limited to selective laser melting (SLM), direct metal laser sintering (DMLS), electron beam melting (EBM), and laser powder bed fusion, wherein such processes enable creation of complex internal geometries including conformal cooling passages that cannot be economically produced through conventional subtractive machining methods.

[0055] Integrated Conformal Cooling Passages: As used herein, the term "integrated conformal cooling passages" refers to internal channels formed within a monolithic frame during metal additive manufacturing, wherein the passages follow three-dimensional routing paths that conform to the geometry of sensing zones, optical component mounting regions, and structural features, and wherein the passages are configured to circulate temperature-controlled liquid to provide heat transfer coefficients in a range of 2000 W / m2K to 5000 W / m2K in turbulent flow regimes.

[0056] Thermally Conductive Metal Alloy: As used herein, the term "thermally conductive metal alloy" refers to metal alloys suitable for additive manufacturing that exhibit thermal conductivity sufficient for effective heat transfer in optical biosensing applications, including but not limited to AlSilOMg aluminum alloy having a thermal conductivity in aDocket No.: INST-01-PCTrange of 130 W / m K to 180 W / m K, copper alloys having a thermal conductivity in a range of 350 W / m K to 400 W / m K. titanium alloys, and stainless steel alloys.

[0057] Pyramidal Prism: As used herein, the term "pyramidal prism" refers to an optical element fabricated from high-refractive-index glass such as BK7 or SF10 that provides a plurality' of geometrically distinct optical faces enabling independent, simultaneous interrogation of a shared sensing surface using different detection modalities through orthogonal optical paths without optical interference between the paths. Suitable pyramidal prisms may be obtained from commercial optical component suppliers including Edmund Optics (Barrington, NJ), Thorlabs (New ton. NJ), Newport Corporation (Irvine, CA), and Optosigma Corporation (Santa Ana, CA), or may be custom fabricated to specification by precision optical manufacturers such as Esco Optics (Oak Ridge, NJ), Sydor Optics (Rochester, NY), or Shanghai Optics (Irvine, CA). BK7 and SF10 optical glass substrates suitable for prism fabrication are available from Schott AG (Mainz, Germany) and Ohara Corporation (Branchburg, NJ).

[0058] Prism Geometries. Prism geometries for SPR detection have been explored in various configurations. Hemisphencal prisms provide wide angular ranges but introduce optical distortions that complicate image analysis. Dove prisms enable compact single-axis optical paths but limit the range of accessible incident angles. Pyramidal prism geometries provide multiple optical faces that could potentially enable independent optical paths addressing a common sensing surface, but integration of such geometries with thermal management and multi-modal detection has not been reported in the literature known to the inventors.

[0059] First Detection Path: As used herein, the term "first detection path" refers to an optical path configured for traditional high-sensitivity surface plasmon resonance detection, comprising a light source, polarizing optics, focusing elements, and a detector optimized for narrow-field detection, wherein the first detection path is configured to provide a sensitivity' of 0.05 RU RMSD or less, suitable for kinetic characterization of molecular interactions including detection of small molecules having a molecular weight of less than 500 Da binding to proteins.

[0060] Second Detection Path: As used herein, the term "second detection path" refers to an optical path configured for surface plasmon resonance imaging detection, comprising a light source with wide-field optics and a two-dimensional imaging camera having at least 5 megapixels, wherein the second detection path is configured to capture the entire sensing surface simultaneously for monitoring arrays of interaction zones with aDocket No.: INST-01-PCTsensitivity of approximately 0.5 RU RMSD, suitable for high-throughput screening applications.

[0061] Photochemistry Path: As used herein, the term "photochemistry path" refers to an optical path positioned to deliver ultraviolet or visible light in a wavelength range of 300 nm to 700 nm, and particularly at 365 nm (e.g., in a range of 350 nm to 380 nm), to a sensor surface for in-situ activation of photoactive chemistries including photoactivatable crosslinkers such as diazirines and benzophenones, wherein the photochemistry path operates independently of or concurrently with the first detection path and the second detection path.

[0062] Temperature Stability: As used herein, the term "temperature stability " refers to the degree of temperature variation at a sensing surface during continuous operation, wherein a temperature stability’ of ±0.003°C indicates that the temperature at the sensing surface varies by no more than 0.003°C from a setpoint temperature during operation.

[0063] Sensor Chip Interface: As used herein, the term "sensor chip interface" refers to a mounting surface configured to receive a sensor chip comprising a thin glass substrate coated with a thin metal film and functionalized with bioactive surface chemistry, wherein the sensor chip is pressed against a top face of a prism using an optical matching interface to ensure optical continuity7for surface plasmon resonance excitation. Suitable sensor chips may be obtained from commercial suppliers including Cytiva (Marlborough, MA) under the Biacore sensor chip product line, Nicoya Lifesciences (Kitchener, Ontario, Canada). Reichert Technologies (Buffalo. NY), Bio-Rad Laboratories (Hercules. CA) under the ProteOn sensor chip product line, Bruker (Billerica, MA) under the Sierra SPR sensor chip product line, and Xantec Bioanalytics (Dusseldorf, Germany). Gold-coated glass substrates suitable for custom sensor chip fabrication are available from Platy pus Technologies (Madison, WI), Ssens BV (Enschede, Netherlands), and EMF Corporation (Ithaca, NY). Index matching fluids and optical coupling compounds for the optical matching interface are available from Cargille Laboratories (Cedar Grove, NJ) and Thorlabs (Newton. NJ).

[0064] Liquid Cooling System: As used herein, the term "liquid cooling system" refers to a closed-loop system comprising a thermoelectric cooler module, a centrifugal pump, a coolant reservoir, and a temperature controller, configured to circulate temperature-controlled liquid through integrated conformal cooling passages at a flow rate in a range of 150 mL / min to 250 mL / min, wherein the liquid cooling system is configured to maintain temperature stability at a sensing surface and to operate over a temperature range of 4°C to 40°C. Suitable thermoelectric cooler modules may be obtained from commercial suppliersDocket No.: INST-01-PCTincluding TE Technology (Traverse City, MI), Laird Thermal Systems (Durham, NC), Ferrotec (Santa Clara, CA), and CUI Devices (Lake Oswego, OR). Centrifugal pumps suitable for liquid cooling applications are available from IDEX Health & Science (Oak Harbor, WA), Cole-Parmer (Vernon Hills, IL), KNF Neuberger (Trenton, NJ), and Iwaki America (Holliston, MA). Temperature controllers for precision thermal management may be obtained from Wavelength Electronics (Bozeman. MT), Thorlabs (Newton. NJ), Stanford Research Systems (Sunnyvale. CA), and Arroyo Instruments (San Luis Obispo, CA). Coolant reservoirs and associated fluid handling components are available from Swagelok (Solon, OH), Parker Hannifin (Cleveland, OH), and McMaster-Carr (Elmhurst, IL).

[0065] Monolithic Frame: As used herein, the term "monolithic frame" refers to a single, unitary structural component manufactured by metal additive manufacturing that integrates structural support, thermal management through integrated conformal cooling passages, and optical mounting features, wherein the monolithic frame is produced as a complete part in a single additive manufacturing build without requiring assembly of multiple discrete components. Suitable metal additive manufacturing services for producing monolithic frames may be obtained from commercial suppliers including Protolabs (Maple Plain, MN), Xometry (Derwood, MD), 3D Systems (Rock Hill, SC), EOS GmbH (Krailling, Germany), SLM Solutions (Lubeck, Germany), and GE Additive (Cincinnati, OH).AlSilOMg aluminum alloy powder suitable for selective laser melting of monolithic frames is available from AP&C (Saint-Eustache, Quebec. Canada). Carpenter Additive (Philadelphia, PA), and Hoganas AB (Hoganas, Sweden). Copper alloy powders for high thermal conductivity applications are available from GKN Powder Metallurgy (Bonn, Germany) and Sandvik Osprey (Neath, United Kingdom).

[0066] Mode Switching Without Mechanical Reconfiguration: As used herein, the term "mode switching without mechanical reconfiguration" refers to transitioning between a first detection mode and a second detection mode through software control by activating appropriate light sources and cameras based on the selected operational mode, wherein no physical movement of optical components, detectors, or structural elements is required to effect the transition between detection modes.

[0067] Teardrop-Shaped Cross-Sections: As used herein, the term "teardropshaped cross-sections" refers to a passage geometry wherein the channel cross-section incorporates a tapered or pointed upper surface that forms a self-supporting structure during metal additive manufacturing, enabling printing of enclosed internal passages without internal support structures that would otherwise be impossible to remove from the completedDocket No.: INST-01-PCTpart.

[0068] Sensitivity: As used herein, the term "sensitivity" when expressed in RU RMSD (resonance units root mean square deviation) refers to the noise floor of a surface plasmon resonance detection system, wherein lower RU RMSD values indicate higher sensitivity7and greater capability to detect small refractive index changes caused by molecular binding events, and wherein a sensitivity7of 0.03 RU RMSD enables detection of binding events producing response signals as small as approximately 0.1 RU.

[0069] SPR Detection and Imaging Systems. Surface plasmon resonance (SPR) biosensing has emerged as a technique for label-free detection and characterization of biomolecular interactions. The technique exploits the optical phenomenon occurring at metaldielectric interfaces where incident light at specific angles excites collective oscillations of free electrons in thin metal films, creating evanescent fields that extend into sample media. Changes in refractive index wi thin these evanescent fields caused by molecular binding events shift the resonance condition, enabling real-time monitoring of association and dissociation kinetics without requiring fluorescent or radioactive labels.

[0070] Multi-Modal Surface Plasmon Resonance Detector with Additively Manufactured Liquid-Cooled Frame for Measuring Molecular Interactions.

[0071] The present disclosure addresses the technical problem of providing a compact SPR instrument that integrates both high-sensitivity detection and imaging modes within a single device, with a liquid-cooled, additively manufactured frame achieving temperature stability7of ±0.003°C at the sensing surface, reduced acoustic signature compared to fan-cooled systems, and consolidated manufacturing complexity. The multi-modal surface plasmon resonance detector combines a detector frame manufactured by metal additive manufacturing with integrated conformal cooling passages, dual detection paths for surface plasmon resonance detection and surface plasmon resonance imaging detection, and an optional photochemistry path for ultraviolet activation of photoactive chemistries.

[0072] Surface plasmon resonance detection provides label-free analysis of biomolecular interactions by measuring changes in refractive index at a metal-dielectric interface. Surface plasmon resonance detection may be used for characterizing binding kinetics, including association rate constants, dissociation rate constants, and equilibrium dissociation constants. Surface plasmon resonance imaging detection provides array-based measurements that enable screening of multiple molecular interactions simultaneously. The multi-modal surface plasmon resonance detector integrates both detection modalities within aDocket No.: INST-01-PCTsingle device using orthogonal optical paths through a pyramidal prism geometry.

[0073] The detector frame may be manufactured from a thermally conductive metal alloy using metal additive manufacturing processes such as selective laser melting. Metal additive manufacturing enables formation of integrated conformal cooling passages within the detector frame that route temperature-controlled liquid around sensing zones and optical component mounting regions. The integrated conformal cooling passages may follow complex three-dimensional paths that would be impractical to produce using conventional machining methods. A liquid cooling system may circulate temperature-controlled liquid through the integrated conformal cooling passages to maintain thermal stability at a sensing surface.

[0074] The multi-modal surface plasmon resonance detector may include a prism providing a plurality of optical faces for a plurality of optical paths. A first detection path may be configured for surface plasmon resonance detection, and a second detection path may be configured for surface plasmon resonance imaging detection. In some cases, the first detection path and the second detection path are orthogonal to each other. The multi-modal surface plasmon resonance detector may further include a sensor chip interface configured to receive a sensor chip, and a microfluidic flow cell positioned above the sensor chip for delivery7of samples to a sensor surface.

[0075] In some cases, the multi-modal surface plasmon resonance detector includes a photochemistry path configured to deliver ultraviolet light to the sensor surface. The photochemistry path may enable activation of photoactivatable coupling chemistries, phototriggered release of captured molecules, or other photochemical processes during surface plasmon resonance monitoring. The photochemistry path may deliver ultraviolet light at 365 nm (e.g., in a range of 350 nm to 380 nm) for activating photoactivatable crosslinkers or other photoactive functional groups.

[0076] The detector frame may integrate structural support, thermal management, and optical mounting features in a single monolithic component. The integrated conformal cooling passages may provide heat transfer coefficients in a range of 2000 W / m2K to 5000 W / m2K, enabling precise temperature control at the sensing surface. In some cases, the liquid cooling system maintains a temperature stability of ±0.003°C at the sensing surface. The detector frame may include mounting features for optical components, including a prism mounting region, a light source mounting region, and a detector mounting region.

[0077] Referring to FIG. 5. a multi-modal SPR detector 500 for measuring molecular interactions is shown. The multi-modal SPR detector 500 comprises a detectorDocket No.: INST-01-PCTframe 502 manufactured by metal additive manufacturing. The detector frame 502 comprises integrated conformal cooling passages 504 formed within the detector frame 502. The detector frame 502 further comprises a prism interface 506 and optical mounting features 508. The detector frame 502 integrates structural support, thermal management through the integrated conformal cooling passages 504, and the optical mounting features 508 in a single component.

[0078] With continued reference to FIG. 5, the multi-modal SPR detector 500 comprises a prism 510 providing a plurality of optical faces for a plurality of optical paths. The prism 510 may be optically coupled to a first detection path 512, a second detection path 520, and a photochemistry path 528. The first detection path 512 is configured for surface plasmon resonance detection. The first detection path 512 comprises a light source 514, optical elements 516, and a detector 518. The second detection path 520 is configured for surface plasmon resonance imaging detection. The second detection path 520 comprises a light source 522, optical elements 524, and a camera 526. In some cases, the second detection path 520 is orthogonal to the first detection path 512.

[0079] The multi-modal SPR detector 500 further comprises a sensor chip interface 530 configured to receive a sensor chip. The sensor chip interface 530 may be positioned between a microfluidic flow cell 532 and the prism 510. The microfluidic flow cell 532 is positioned above the sensor chip interface 530 for delivery of samples to a sensing surface. A liquid cooling system 534 is configured to circulate liquid through the integrated conformal cooling passages 504. The integrated conformal cooling passages 504 are fluidically connected to the liquid cooling system 534, enabling circulation of temperature-controlled liquid through the detector frame 502 to maintain thermal stability at the sensing surface.

[0080] As further shown in FIG. 5, the photochemistry path 528 is configured to deliver ultraviolet light to the sensing surface. The photochemistry path 528 may enable activation of photoactivatable coupling chemistries or photo-triggered release of captured molecules during surface plasmon resonance monitoring. The detector frame 502 enables mode switching between the first detection path 512 and the second detection path 520 without mechanical reconfiguration.

[0081] The multi-modal SPR detector 500 may be configured for integration into robotic workcells for automated drug discovery platforms. In some cases, the multi-modal SPR detector 500 is configured for programmatic control via an application programming interface for autonomous experimental design and execution. The multi-modal SPR detector 500 may be configured for deployment in distributed instrument networks where individualDocket No.: INST-01-PCTsystems can operate independently or coordinate on larger programs. In some cases, the multi-modal SPR detector 500 is configured for benchtop clusters of 4-8 instruments operating simultaneously.

[0082] The first detection path 512 may be configured for surface plasmon resonance detection providing sensitivity' of 0.05 RU RMSD or less. The first detection path 512 comprises a light source 514, optical elements 516, and a detector 518. The light source 514 may comprise a near-infrared light source operating at a wavelength of 850 nm (e.g., in a range of 840 nm to 860 nm). In some cases, the light source 514 operates at alternative nearinfrared wavelengths in a range of 650 nm to 950 nm for surface plasmon resonance detection. The light source 514 may have a power in a range of 5 mW to 10 mW.

[0083] The light source 514 may be mounted directly to the liquid-cooled detector frame 502 with thermal interface material providing less than 0.5°C / W thermal resistance. The light source 514 may generate heat of 0.5 W to 2 W that is removed by the liquid cooling system 534. In some cases, the light source 514 achieves j unction temperature stability of ±0.01°C when mounted to the liquid-cooled detector frame 502. The thermal coupling between the light source 514 and the detector frame 502 enables heat removal from the light source 514 while maintaining stable operating temperature without separate thermal management.

[0084] The optical elements 516 in the first detection path 512 may comprise collimating optics, focusing optics, and polarizers configured for angular interrogation of a sensing surface. In some cases, the first detection path 512 is configured for angular interrogation at angles of 45° to 70° depending on prism geometry and metal film properties. The optical elements 516 may be mounted directly to the detector frame 502 through precision-machined mounting features.

[0085] The detector 518 in the first detection path 512 may comprise a line camera detector. In some cases, the detector 518 comprises a photodiode array as an alternative to a line camera. In some cases, the detector 518 comprises a two-dimensional camera with a region of interest restricted to a single line operating at 500 Hz to 1000 Hz frame rate. The detector 518 may operate at frame rates in a range of 100 Hz to 1000 Hz.

[0086] The first detection path 512 may be configured to provide a sensitivity of 0.05 RU RMSD or less. In some cases, the multi-modal SPR detector 500 achieves baseline noise of 0.035 RU RMSD and signal drift less than 0.5 RU / hour over 8-hour operation. The first detection path 512 may generate sensorgrams showing response units versus time with sensitivity sufficient to detect small molecules less than 100 Da binding to proteins.Docket No.: INST-01-PCT

[0087] The second detection path 520 comprises a light source 522, optical elements 524, and a camera 526. The second detection path 520 is configured for surface plasmon resonance imaging detection. The light source 522 may provide wide-field illumination for imaging the sensing surface. The optical elements 524 may comprise collimating optics, focusing optics, and polarizers configured for wide-field illumination of the sensing surface.

[0088] The camera 526 in the second detection path 520 may comprise a two-dimensional camera having at least 5 megapixels. In some cases, the camera 526 comprises a two-dimensional camera having at least 2 megapixels providing sufficient pixel density for array imaging with typical array spot sizes of 100 pm to 500 pm diameter. The camera 526 may comprise a CMOS camera. In some cases, the camera 526 comprises a CCD camera as an alternative to a CMOS camera.

[0089] The camera 526 may operate at frame rates from 1 Hz to 30 Hz for array imaging applications. In some cases, the camera 526 operates at frame rates up to 500 Hz when used as a surface plasmon resonance microscope for fluidic visualization. The camera 526 may generate heat of 2 W to 3 W that is removed by the liquid cooling system 534. In some cases, the liquid cooling system 534 maintains the camera 526 at an operating temperature of 30°C to 40°C for CMOS sensors to improve signal-to-noise ratio and dark current performance.

[0090] The second detection path 520 may be configured to provide a sensitivity of approximately 0.5 RU RMSD. The second detection path 520 may be configured to provide spatial resolution sufficient to resolve individual microarray spots with typical spot sizes of 100 pm to 500 pm diameter requiring 10 to 50 pixels per spot. In some cases, the second detection path 520 is configured to maintain thermal uniformity across larger sensing areas up to 20 mm x 40 mm for array applications.

[0091] Referring to FIG. 14, the multi-modal surface plasmon resonance detector comprises a prism providing a plurality of optical faces for a plurality of optical paths. The prism may comprise a pyramidal prism having four optical faces arranged around a central sensor platform. The pyramidal prism geometry enables independent, simultaneous interrogation of a shared sensing surface through orthogonal optical paths. As shown in FIG.14, a light source for SPR is positioned at a first optical face of the pyramidal prism, and a detector for SPR is positioned at an opposing optical face to receive reflected light from the sensing surface. A light source for SPR imaging is positioned at a third optical face of the pyramidal prism, and a detector for SPR imaging is positioned at a fourth optical face opposing the light source for SPR imaging.Docket No.: INST-01-PCT

[0092] With continued reference to FIG. 14, the plurality of optical paths comprise orthogonal optical paths. The second detection path is orthogonal to the first detection path. The orthogonal arrangement of the first detection path and the second detection path enables both detection modalities to address the shared sensing surface without optical interference between the two detection pathways. The first detection path, comprising the light source for SPR and the detector for SPR, forms a first diagonal pair across the pyramidal prism. The second detection path, comprising the light source for SPR imaging and the detector for SPR imaging, forms a second diagonal pair orthogonal to the first diagonal pair.

[0093] Referring to FIG. 15, the pyramidal prism structure is positioned at the intersection of multiple optical paths. A flow cell is positioned above the pyramidal prism at the sensor chip interface. The flow cell comprises fluidic ports for sample delivery to the sensing surface. A light source for excitation and detection via flow cell is positioned above the flow cell, providing a photochemistry path for delivering ultraviolet light to the sensing surface. The photochemistry path is oriented normal to the sensing surface to avoid internal reflection on the gold surface that would be encountered from imaging below. Light source excitation via prism components are arranged radially around the base of the pyramidal prism, representing the orthogonal optical paths for the first detection path and the second detection path.

[0094] The prism may be fabricated from BK7 glass. BK7 glass provides suitable optical properties for surface plasmon resonance detection in the near-infrared wavelength range. In some cases, the prism is fabricated from SF 10 glass. SF10 glass has a higher refractive index than BK7 glass and may be selected for applications requiring different SPR coupling angles or enhanced sensitivity. The prism may have an apex angle of 45° (e.g., in a range of 40° to 50°) for BK7 glass, with the apex angle adjusted based on metal film properties and desired sensitivity' range. In some cases, the apex angle is modified for SF10 glass or other optical glasses to achieve SPR coupling angles in a range of 45° to 70° depending on the metal film composition and thickness.

[0095] The prism may comprise a hemispherical prism as an alternative to the pyramidal prism geometry. The hemispherical prism geometry adds some distortion that would need to be corrected for in the optical path design or image processing. In some cases, the multi-modal surface plasmon resonance detector is adapted for grating-coupled SPR using diffraction gratings rather than prisms while maintaining integrated thermal control through the liquid-cooled detector frame.

[0096] Multiple SPR wavelengths may be employed concurrently in the detectionDocket No.: INST-01-PCTpaths for enhanced measurement capabilities. In some cases, the first detection path operates at a first wavelength and the second detection path operates at a second wavelength different from the first wavelength. The same camera may serve both the first detection path and second detection path functions by software-controlled region of interest switching. In some cases, a single two-dimensional camera is configured to capture data from both detection paths by switching between different regions of interest corresponding to each detection path.

[0097] The multi-modal surface plasmon resonance detector architecture may be adapted for other optical biosensing modalities beyond SPR including interferometry, ellipsometry, and fluorescence detection. The orthogonal optical path architecture enabled by the pyramidal prism geometry provides flexibility for integrating additional detection modalities while maintaining the integrated thermal control provided by the liquid-cooled detector frame.

[0098] The multi-modal surface plasmon resonance detector may further comprise a photochemistry path configured to deliver ultraviolet light at a wavelength of 365 nm (e.g., in a range of 350 nm to 380 nm) to a sensor chip interface. The photochemistry path may address a sensor surface from the top to avoid internal reflection on a gold surface as would be encountered from imaging below. In some cases, the photochemistry path delivers light through the bottom of a prism to provide direct illumination of the sensing surface without interfering with the SPR detection paths. The photochemistry path may be oriented normal to the sensing surface to enable uniform illumination across the sensor chip interface.

[0099] The photochemistry path may be used to activate diazirine groups at 350-365 nm for photoactivatable crosslinker chemistry7. Diazirine groups generate reactive carbene intermediates upon ultraviolet irradiation that form covalent bonds with nearby molecules. In some cases, the photochemistry path is used to activate benzophenone groups at 365 nm for photoactivatable crosslinker chemistry7. Benzophenone groups undergo hydrogen abstraction reactions upon ultraviolet irradiation to form covalent crosslinks with target molecules.

[0100] A light source in the photochemistry path may have a power in a range of 10 mW to 100 mW for photoactivatable crosslinker chemistry. The photochemistry path may include protective long-pass filters greater than 300 nm to protect proteins and biomolecules in bulk solution from photodamage while activating surface-bound photoreactive groups. The protective long-pass filters may transmit wavelengths above 300 nm while blocking shorter wavelength ultraviolet light that causes protein denaturation and nucleic acid damage.

[0101] The photochemistry path may7use alternative wavelengths including 405 nm, 470 nm, or other wavelengths in the UV -visible range depending on specific photoreactiveDocket No.: INST-01-PCTgroups employed. In some cases, the photochemistry path operates at 470 nm blue light for activating LOV2 domain-containing photoswitchable proteins. LOV2 domains undergo conformational changes upon blue light irradiation that can be coupled to binding domain function in engineered proteins.

[0102] The multi-modal surface plasmon resonance detector may be configured to operate with photo-releasable streptavidin capture reagents where ultraviolet light triggers conformational change to weaken biotin affinity for gentle surface regeneration. In some cases, the multi-modal surface plasmon resonance detector is configured to operate with OptoNanobodies as photoswitchable capture reagents where blue light at 470 nm triggers affinity switching for gentle regeneration. OptoNanobodies are engineered nanobodies with LOV2 photosensitive domains inserted into the protein framework that enable light-controlled binding affinity.

[0103] The photochemistry path may enable surface regeneration through phototriggered conformational changes in engineered capture proteins achieving greater than 100 regeneration cycles. Photo-triggered regeneration provides gentle, chemical-free surface regeneration that preserves protein activity and extends surface lifetime compared to harsh chemical regeneration conditions. In some cases, the multi-modal surface plasmon resonance detector is configured to characterize photoswitchable proteins by monitoring affinity switching in real-time during light activation. Real-time monitoring of photoswitching enables quantification of switching kinetics, fold-change in affinity between dark and lit states, and reversibility of the photoswitching process.

[0104] The sensor chip interface may be configured to receive a sensor chip at a top face of a prism. The sensor chip interface provides optical coupling between the sensor chip and the prism to enable surface plasmon resonance excitation at the metal-dielectric interface. The sensor chip interface may include a polymer-based optical matching interface for ensuring optical continuity' betw een the sensor chip and the prism. The polymer-based optical matching interface may comprise a silicone-based material having a refractive index matched to the prism and sensor chip substrate. In some cases, the sensor chip interface includes a refractive index matching fluid for ensuring optical continuity’ between the sensor chip and the prism. The refractive index matching fluid may be applied bet een the sensor chip and the prism to eliminate air gaps that w ould otherwise disrupt total internal reflection conditions.

[0105] The sensor chip may comprise a glass substrate with a refractive index of 1.52 (e.g., in a range of 1.50 to 1.54) coated with a 40 nm to 50 nm gold film. The gold filmDocket No.: INST-01-PCTthickness may be selected to optimize surface plasmon resonance coupling efficiency at the operating wavelength of the detection paths. In some cases, the sensor chip comprises a silver metal film coating as an alternative to gold. Silver metal films provide sharper surface plasmon resonance curves and higher sensitivity compared to gold films, but silver films are more susceptible to oxidation and may require protective coatings for stability7in aqueous environments.

[0106] The sensor chip may comprise a glass substrate with a refractive index of 1.52 (e.g., in a range of 1.50 to 1.54) coated with a 40 nm to 50 nm gold film. The gold film thickness may be selected to optimize surface plasmon resonance coupling efficiency at the operating wavelength of the detection paths. In some cases, the sensor chip comprises a silver metal film coating as an alternative to gold. Silver metal films provide sharper surface plasmon resonance curves and higher sensitivity compared to gold films, but silver films are more susceptible to oxidation and may require protective coatings for stability in aqueous environments.

[0107] A microfluidic flow cell may be positioned above the sensor chip for delivery of samples to a sensor surface. The microfluidic flow cell may operate at flow rates of 5 pL / min to 100 pL / min for a first detection path optimized for minimal sample consumption and rapid association / dissociation kinetics. The flow rate range for the first detection path enables efficient mass transport of analyte molecules to the sensor surface while minimizing sample volume requirements. In some cases, the microfluidic flow cell operates at flow rates of 1 pT / min to 10 pT / min for a second detection path suitable for array applications. The lower flow rate range for the second detection path provides uniform sample distribution across larger sensing areas while maintaining laminar flow conditions.

[0108] The multi-modal surface plasmon resonance detector may be configured for integration with nucleic acid programmable protein array technology to express proteins directly on the sensor surface. Nucleic acid programmable protein array technology enables high-throughput protein production and capture on microarray surfaces without requiring separate protein purification steps. The sensor chip interface may be configured to receive DNA microarray chips with positions containing plasmid DNA encoding proteins co-printed with anti-tag capture antibodies for in-situ protein expression. The plasmid DNA encodes proteins with affinity7tags that are captured by the co-printed anti-tag antibodies following cell-free expression.

[0109] The multi-modal surface plasmon resonance detector may be configured to incubate cell-free protein expression systems at 25°C for 1 hour to 2 hours for in-situ proteinDocket No.: INST-01-PCTexpression on DNA arrays. The liquid cooling system may maintain the sensor chip interface at the 25°C incubation temperature during the cell-free expression period. Following protein expression and capture, the multi-modal surface plasmon resonance detector may screen the expressed protein array against target molecules using the second detection path configured for surface plasmon resonance imaging detection.

[0110] The sensor chip interface may include a chip insertion mechanism that ensures reproducible optical coupling via the polymer-based optical matching interface for automated workflows. The chip insertion mechanism may provide consistent positioning of the sensor chip relative to the prism to maintain optical alignment across multiple chip exchanges. In some cases, the chip insertion mechanism comprises alignment features that register the sensor chip to the prism interface with positional repeatability suitable for automated experimental workflows.

[0111] The multi-modal surface plasmon resonance detector may be configured for manual or robotic sensor chip exchange when transitioning between experiments after fluidics retraction. The microfluidic flow cell may be retracted from the sensor chip to enable chip removal and replacement. In some cases, the multi-modal surface plasmon resonance detector is configured for integration with robotic sample handling systems that automate sensor chip exchange as part of high-throughput screening workflow s.

[0112] Referring to FIG. 7. a detector frame manufactured using metal additive manufacturing is shown in a side view. The detector frame comprises a monolithic frame that integrates structural support, thermal management, and optical mounting features in a single component. The detector frame includes a coolant inlet and a coolant outlet configured with barbed hose fittings for connection to flexible tubing. The coolant inlet and the coolant outlet are fluidically connected to integrated conformal cooling vias formed within the monolithic frame. The monolithic frame exhibits a complex geometric profile with multiple integrated features including mounting surfaces, structural supports, and internal conformal cooling channels.

[0113] The monolithic frame may be manufactured from a thermally conductive metal alloy. In some cases, the thermally conductive metal alloy comprises AlSilOMg aluminum alloy. AlSilOMg aluminum alloy provides athermal conductivity in a range of 130 W / m K to 180 W / m K. In some cases, the thermally conductive metal alloy comprises a copper alloy. Copper alloys provide a thermal conductivity in a range of 350 W / m K to 400 W / m K. In some cases, the thermally conductive metal alloy comprises titanium. In some cases, the thermally conductive metal alloy comprises stainless steel. The selection of theDocket No.: INST-01-PCTthermally conductive metal alloy may depend on application requirements including thermal performance, corrosion resistance, and mechanical properties.

[0114] With continued reference to FIG. 7, the monolithic frame may be manufactured by selective laser melting. Selective laser melting enables formation of the integrated conformal cooling passages during the metal additive manufacturing process. The monolithic frame may have dimensions of 120 mm x 80 mm x 60 mm (e.g., in a range of 100 mm to 140 mm x 60 mm to 100 mm x 40 mm to 80 mm). The monolithic frame may have a detector volume of 576 cm3. The monolithic frame may be manufactured with a print time in a range of 6 hours to 12 hours followed by 2 hours to 4 hours of post-processing for support removal and machining of mounting surfaces.

[0115] Referring to FIG. 8. a cutaway view of the detector frame reveals internal architecture including a temperature sensor positioned within the monolithic frame and coolant vias routed through the frame structure. The cutaway view shows a camera mounted to the detector frame and a prism positioned in a central region of the detector frame. The integrated conformal cooling vias are shown as channels routed through the frame material surrounding the prism interface area. Fluid connection ports with threaded fittings are visible at a lower portion of the assembly for connecting to an external liquid cooling system.

[0116] The monolithic frame consolidates functions of 15 to 25 precision-machined components of traditional SPR detector assemblies into a single part. Traditional SPR detector assemblies may include a base plate, multiple optical mounting blocks, a thermoelectric cooler housing, heat sinks, fan mounting brackets, thermal interface plates, a prism mount, a sensor chip holder, a light source mount, a detector mount, and various alignment and structural components. The monolithic frame manufactured by metal additive manufacturing replaces these discrete components with a single integrated structure.

[0117] Referring to FIG. 9, a cross-sectional view of the detector frame shows the internal cooling via geometry. The integrated conformal cooling vias are shown as a series of circular or rounded rectangular channels arranged within the detector body. The integrated conformal cooling vias are positioned to provide thermal regulation around sensing zones and optical component mounting regions. The cross-sectional view reveals mounting cavities and optical component housings shown as rectangular recesses within the frame structure.

[0118] With continued reference to FIG. 9, the monolithic frame may have a resonant frequency greater than 200 Hz for optical stability. The resonant frequency¬ specification ensures that mechanical vibrations do not couple to optical components during surface plasmon resonance measurements. The monolithic frame provides structural rigidityDocket No.: INST-01-PCTthrough the integrated design that combines structural support with thermal management and optical mounting features.

[0119] Referring to FIG. 10, a cutaway view of the detector frame reveals internal cooling architecture including prism cooling vias and structures designed to maximize heat transfer. The prism cooling vias are shown as a series of parallel elongated channels arranged in a horizontal pattern proximate to the prism interface region. The heat transfer enhancement structures appear as fin-like or ribbed internal geometries integrated within the cooling channel netw ork. The heat transfer enhancement structures increase surface area contact between circulating coolant and the thermally conductive metal frame material.

[0120] Referring to FIG. 11, a three-dimensional cutaway view shows a coolant flow path that routes temperature-controlled liquid throughout the monolithic frame. The coolant flow path is highlighted showing a serpentine routing pattern. A coolant inlet is positioned at a lower portion of the frame w here temperature-controlled liquid enters the detector body. A coolant outlet is positioned at an upper portion where the liquid exits after circulating through the internal passages. The cooling channels follow a conformal path that winds around thermal zones within the detector frame, passing through multiple levels and directions to provide uniform temperature distribution.

[0121] With continued reference to FIG. 11 , the integrated conformal cooling vias demonstrate three-dimensional complexity enabled by metal additive manufacturing. The integrated conformal cooling vias curve, branch, and traverse different depths within the monolithic frame structure. The conformal routing enables temperature-controlled liquid to be routed directly around a sensor chip footprint and optical support regions. The three-dimensional path of the integrated conformal cooling vias w ould be impractical to produce using conventional machining methods.

[0122] Referring to FIG. 12, a top-down cross-sectional view shows the internal cooling via geometry and design features. The integrated conformal cooling vias follow a serpentine or conformal routing pattern around a central rectangular region corresponding to a sensor chip and optical support area. A camera cooling via is identified in an upper left comer, demonstrating how the cooling channel network extends to provide thermal management for a detector mounting region. Teardrop-shaped vias are shown in a lower portion of the frame, where the teardrop geometry allows for printing without removable supports.

[0123] With continued reference to FIG. 12. the teardrop-shaped cross-sections represent a design-for-additive-manufacturing approach where the channel cross-sectionDocket No.: INST-01-PCTincorporates a tapered upper surface that can be printed without internal support structures. Internal support structures would otherwise be impossible to remove from enclosed internal passages. The cross-hatched regions indicate solid metal material of the frame, while white areas represent internal void spaces forming coolant flow passages.

[0124] Referring to FIG. 13, a top-dow n plan view' show s the internal cooling via network and structural features of the detector frame. The plan view depicts a rectangular frame structure with elongated oval-shaped recesses in left and right sections. A central section displays multiple circular openings of varying sizes arranged in a pattern. Larger circular holes are positioned at comers of the central region for mounting points or fastener locations. Smaller circular features distributed throughout the central area may represent cooling via entry and exit points or additional mounting provisions.

[0125] The mounting features for optical components may comprise precision-machined surfaces having a tolerance of ±0.01 mm. The precision-machined surfaces are achieved through post-processing of the monolithic frame following metal additive manufacturing. Post-processing may include machining of mounting surfaces to optical tolerances for positioning of optical components including a prism, light sources, and detectors. The combination of metal additive manufacturing for complex internal geometries and post-processing for precision mounting surfaces enables the monolithic frame to integrate structural support, thermal management, and optical mounting features in a single component.

[0126] The integrated conformal cooling passages may be formed during the metal additive manufacturing process. Formation of the integrated conformal cooling passages during metal additive manufacturing enables complex three-dimensional geometries that would be impractical to produce using conventional machining methods. The integrated conformal cooling passages may follow a serpentine channel pattern that routes temperature-controlled liquid through a continuous folded path within the detector frame. In some cases, the integrated conformal cooling passages follow a parallel channel pattern where multiple channels extend in parallel through the detector frame to provide uniform cooling across a sensing zone.

[0127] The integrated conformal cooling passages may have a diameter in a range of 2 mm to 4 mm. In some cases, the integrated conformal cooling passages have a diameter of 3 mm. The diameter range provides sufficient flow capacity for circulating temperature-controlled liquid while maintaining structural integrity of the detector frame. A liquid cooling system may be configured to circulate coolant at 200 mL / min flow rate through the integratedDocket No.: INST-01-PCTconformal cooling passages.

[0128] The integrated conformal cooling passages may comprise teardrop-shaped cross-sections configured for printing without removable supports. The teardrop-shaped cross-sections incorporate a tapered upper surface that enables the channel geometry to be printed without internal support structures that would otherwise be impossible to remove from enclosed internal passages. In some cases, the integrated conformal cooling passages comprise self-supporting geometries that eliminate the need for removable supports during metal additive manufacturing. The self-supporting geometries may include angled surfaces and gradual transitions that maintain structural stability during the layer-by-layer build process.

[0129] The integrated conformal cooling passages may be routed within 5 mm to 10 mm of a prism. In some cases, the integrated conformal cooling passages are spaced 6 mm to 8 mm from a prism interface to balance thermal uniformity with structural integrity. The spacing between the integrated conformal cooling passages and the prism interface provides thermal regulation of the sensing surface while maintaining sufficient material thickness for mechanical stability.

[0130] The integrated conformal cooling passages may be routed around a sensing zone and optical component mounting regions. The routing of the integrated conformal cooling passages may encompass a prism interface, optical mounting points, and a sample staging area. In some cases, the integrated conformal cooling passages are routed closer to higher heat-load regions including a light-emitting diode mounting region and a camera mounting region while maintaining uniform spacing around a temperature-sensitive sensing surface. The mounting features may comprise a prism mounting region, a light source mounting region, and a detector mounting region.

[0131] The integrated conformal cooling passages may comprise a camera cooling via routed proximate to a detector mounting region. The camera cooling via provides thermal management for a camera that may generate heat of 2 W to 3 W during operation. The integrated conformal cooling passages may be configured to provide unified thermal management for a light-emitting diode and a camera. Unified thermal management enables heat removal from multiple optical components through a single coolant loop, reducing system complexity' compared to separate thermal management solutions for each component.

[0132] The integrated conformal cooling passages may comprise heat transfer enhancement structures. The heat transfer enhancement structures may include fin-like or ribbed internal geometries that increase surface area contact between circulating coolant andDocket No.: INST-01-PCTthe thermally conductive metal frame material. The heat transfer enhancement structures improve thermal energy exchange efficiency between the coolant and the detector frame.

[0133] The integrated conformal cooling passages may follow a three-dimensional path traversing different depths within a monolithic frame. The three-dimensional path enables the integrated conformal cooling passages to simultaneously cool multiple regions at different depths within the detector frame while avoiding interference with optical mounting features, structural supports, and fluidic ports. The three-dimensional routing capability is enabled by metal additive manufacturing and would be impractical to achieve using conventional machining methods that are limited to straight drilled passages.

[0134] In some cases, the integrated conformal cooling passages comprise a first set of cooling passages and a second set of cooling passages configured for multi-zone temperature control. The first set of cooling passages may be connected to a first liquid circuit maintained at a first temperature, and the second set of cooling passages may be connected to a second liquid circuit maintained at a second temperature different from the first temperature. Multi-zone temperature control enables temperature-controlled sample staging areas to be maintained at different temperatures from the sensing region using independent cooling zones within the same printed structure.

[0135] A monolithic frame may be configured to maintain a temperature variation of less than 0.005°C between optical component mounting points. The temperature uniformity across optical component mounting points provides stable operating conditions for optical components including light sources, detectors, and cameras. The monolithic frame may comprise fluidic ports integrated into the frame structure for connection to an external liquid cooling system. The fluidic ports may include barbed hose fittings or quick-disconnect fittings for connection to flexible tubing or external chillers.

[0136] The monolithic frame may be configured to remove heat from electronic subsystems connected to a same coolant loop. Electronic subsystems may include signal conditioning electronics and power supplies that generate heat during operation. Connecting electronic subsystems to the same coolant loop as the detector frame consolidates heat removal into a single liquid cooling circuit, reducing total system complexity and component count compared to architectures requiring separate thermal management for each subsystem.

[0137] Referring to FIG. 6, a liquid cooling system 600 and a detector frame 610 for thermal management in an optical biosensing detector are shown. The liquid cooling system 600 includes a thermoelectric cooler module 602, a centrifugal pump 604, a coolant reservoir 606, and a temperature controller 608. The detector frame 610 includes a coolant inlet 612,Docket No.: INST-01-PCTconformal cooling passages 614. a coolant outlet 616, and a temperature sensor 618.

[0138] With continued reference to FIG. 6, the thermoelectric cooler module 602 is connected to the coolant reservoir 606 and regulates the temperature of coolant within the coolant reservoir 606. The thermoelectric cooler module 602 may have a capacity of 40 W (e.g., in a range of 30 W to 50 W) for regulating coolant temperature. The thermoelectric cooler module 602 is positioned external to the detector frame 610, enabling the thermoelectric cooler module 602 to be cooled independently without affecting thermal stability of the detector frame 610. The separation of the thermoelectric cooler module 602 from the detector frame 610 prevents heat generated at a hot side of the thermoelectric cooler module 602 from coupling to optical components mounted on the detector frame 610.

[0139] The centrifugal pump 604 is connected to the coolant reservoir 606 and circulates temperature-controlled liquid through the liquid cooling system 600. The centrifugal pump 604 delivers coolant to the coolant inlet 612 of the detector frame 610. The liquid cooling system 600 may be configured to circulate the temperature-controlled liquid at a flow rate in a range of 150 mL / min to 250 mL / min. In some cases, the centrifugal pump 604 circulates coolant at a flow rate of 200 mL / min through the conformal cooling passages 614. The separation of the centrifugal pump 604 from the detector frame 610 prevents vibration from the centrifugal pump 604 from coupling mechanically to optical components mounted on the detector frame 610.

[0140] As further shown in FIG. 6, the liquid cooling system 600 may circulate a water-glycol mixture with biocide to prevent algae growth as the temperature-controlled liquid. In some cases, the temperature-controlled liquid comprises a 50 / 50 water-glycol mixture with 0.05% biocide. The water-glycol mixture provides freeze protection and corrosion inhibition while maintaining thermal properties suitable for heat transfer in the conformal cooling vias 614.

[0141] The coolant flows from the coolant inlet 612 through the conformal cooling passages 614, which are routed within the detector frame 610 to provide thermal management around sensing zones and optical component mounting regions. The coolant exits through the coolant outlet 616 and returns to the coolant reservoir 606, completing a closed-loop circulation path. The closed-loop circulation of temperature-controlled liquid through the conformal cooling passages 614 enables the liquid cooling system 600 to maintain thermal stability- at a sensing surface of the detector frame 610.

[0142] With continued reference to FIG. 6, the temperature sensor 618 is positioned within the detector frame 610 proximate to a sensing zone. The temperature sensor 618 mayDocket No.: INST-01-PCTcomprise a thermistor embedded in the detector frame 610 near the sensing surface. The temperature sensor 618 provides real-time temperature feedback to the temperature controller 608. The temperature controller 608 receives the temperature feedback from the temperature sensor 618 and adjusts operation of the thermoelectric cooler module 602 to maintain precise temperature stability at the sensing surface.

[0143] The temperature controller 608 may implement a PID control loop for closed-loop temperature regulation. The closed-loop control arrangement enables the liquid cooling system 600 to compensate for ambient temperature variations and heat generated by optical components during operation. The combination of thermal mass of the detector frame 610, heat capacity of the temperature-controlled liquid, and precise temperature control provided by the thermoelectric cooler module 602 enables the liquid cooling system 600 to maintain stable operating conditions across a temperature range of 4°C to 40°C.

[0144] The liquid cooling system 600 may be configured to maintain a temperature stability' of ±0.003°C at the sensing surface. The temperature stability of ±0.003°C is suitable for high-sensitivity surface plasmon resonance measurements including detection of small molecules binding to proteins.

[0145] The liquid cooling system 600 may provide a heat transfer coefficient in a range of 2000 W / m2K to 5000 W / m2K in a turbulent flow regime within the conformal cooling passages 614. The heat transfer coefficient range of 2000 W / m2K to 5000 W / m2K exceeds heat transfer coefficients of 10-25 W / m2K for natural air convection and 50-250 W / m2K for forced air cooling used in traditional surface plasmon resonance detectors. The high heat transfer coefficient enables rapid thermal response and precise temperature control at the sensing surface.

[0146] The liquid cooling system 600 may enable temperature transitions from 25°C to 37°C in less than 10 minutes with minimal overshoot of less than 0.01°C. The rapid temperature transition capability enables the multi-modal surface plasmon resonance detector to transition between room temperature operation at 25°C and physiological temperature operation at 37°C for biological studies. The thermal mass of the circulating liquid provides buffering against rapid temperature fluctuations while the high heat transfer coefficient of liquid cooling enables active compensation for heat loads from optical components.

[0147] The detector frame 610 may comprise quick-disconnect fittings at the coolant inlet 612 and the coolant outlet 616 for connection to an external chiller. The quickdisconnect fittings enable users to substitute external recirculating chillers for extended temperature range without modifying the detector frame 610. The liquid cooling system 600Docket No.: INST-01-PCTmay be configured to interface with an external laboratory chiller for extended temperature range from -10°C to 70°C. In some cases, the liquid cooling system 600 maintains coolant temperature between 4°C and 40°C with stability better than ±0.1°C when connected to an external chiller.

[0148] The liquid cooling system 600 enables the detector frame 610 to produce an acoustic signature of 38 dB(A) to 40 dB(A) during operation. The acoustic signature of 38 dB(A) to 40 dB(A) represents a reduction of 15 dB to 20 dB compared to acoustic signatures of 54 dB(A) to 58 dB(A) for fan-cooled surface plasmon resonance systems with comparable thermal performance. The reduced acoustic signature addresses usability in laboratory environments where multiple instruments are clustered, as traditional surface plasmon resonance systems generate acoustic fatigue and interfere with communication in shared workspaces.

[0149] Referring to FIG. 1 , a prototype multi-modal detector assembly is shown in a three-dimensional CAD rendering. The prototype multi-modal detector assembly validates the independent beam path approach described herein, demonstrating how the pyramidal prism geometry enables orthogonal optical paths to address a shared sensing surface while maintaining a compact form factor suitable for benchtop deployment.

[0150] With continued reference to FIG. 16, the prototy pe multi-modal detector assembly comprises multiple integrated components arranged in a compact configuration. A cylindrical housing with a large circular aperture is visible on a left side of the assembly, corresponding to one of the optical paths for surface plasmon resonance detection. The cylindrical housing accommodates optical elements including collimating optics, focusing optics, and polarizers configured for angular interrogation of the sensing surface.

[0151] The prototype multi-modal detector assembly includes a central portion featuring a rectangular block structure with visible mounting holes. An electronic circuit board or sensor component is housed within a recessed cavity' on a front face of the rectangular block structure. The rectangular block structure provides structural support for the detector frame and optical components while maintaining thermal coupling to the liquid cooling system through the integrated conformal cooling passages.

[0152] As further shown in FIG. 16, an upper section of the prototype multi-modal detector assembly includes two rectangular block components with circular features that represent optical mounting points or adjustment mechanisms. The rectangular block components provide mounting locations for light sources and detectors associated with the first detection path and the second detection path. A linear rail or guide mechanism is visibleDocket No.: INST-01-PCTrunning horizontally across a top of the assembly, which may facilitate sensor chip positioning or flow cell alignment during operation.

[0153] The prototype multi-modal detector assembly exhibits evidence of precision machining with multiple threaded holes for fasteners distributed across various surfaces of the detector frame. The threaded holes enable secure mounting of optical components to the detector frame while maintaining positional stability' during surface plasmon resonance measurements. The combination of metal additive manufacturing for the detector frame and precision machining for mounting surfaces enables integration of structural support, thermal management, and optical mounting features in a single unified detector body.

[0154] The modular construction of the prototype multi-modal detector assembly allows for integration of both the first detection path configured for surface plasmon resonance detection providing sensitivity of 0.05 RU RMSD or less and the second detection path configured for surface plasmon resonance imaging detection within the single unified detector body. The pyramidal prism geometry at the center of the assembly provides the plurality of optical faces that enable the orthogonal optical paths to address the shared sensing surface without optical interference between the two detection pathways. The compact form factor of the prototype multi-modal detector assembly demonstrates suitability for benchtop deployment and integration into automation platforms where space constraints and acoustic requirements limit deployment of traditional surface plasmon resonance systems.

[0155] Referring to FIG. 1. a method for detecting molecular interactions using a multi-modal surface plasmon resonance detector is shown. The method begins with a step 100, where liquid is circulated through integrated conformal cooling passages in a detector frame to maintain thermal stability at a sensing surface. The detector frame may be additively- manufactured from a thermally conductive metal alloy. Circulating liquid may comprise circulating temperature-controlled liquid through the integrated conformal cooling passages. The temperature-controlled liquid may be circulated at a flow rate in a range of 150 mL / min to 250 mL / min to provide heat transfer coefficients in a range of 2000 W / m2K to 5000 W / m2K at the sensing surface.

[0156] With continued reference to FIG. 1, the method proceeds to a step 102, where the multi-modal surface plasmon resonance detector operates in a first detection mode using a first optical path for surface plasmon resonance detection. The first optical path may be configured for surface plasmon resonance detection providing sensitivity' of 0.05 RU RMSD or less. The first detection mode may provide a sensitivity of 0.05 RU RMSD or less for detecting molecular interactions including small molecules binding to proteins. The firstDocket No.: INST-01-PCToptical path may comprise a near-infrared light source, optical elements including collimating optics and polarizers, and a line camera detector configured for angular interrogation of the sensing surface.

[0157] Following the step 102, the method moves to a step 104, where the multimodal surface plasmon resonance detector operates in a second detection mode using a second optical path for surface plasmon resonance imaging detection. The second optical path may be orthogonal to the first optical path. The orthogonal arrangement of the first optical path and the second optical path enables both detection modalities to address a shared sensing surface without optical interference between the two detection pathways. The second detection mode may provide array-based measurements that enable screening of multiple molecular interactions simultaneously across a sensing area.

[0158] As further shown in FIG. 1, the method proceeds to a step 106, where switching occurs between the first detection mode and the second detection mode. Switching between the first detection mode and the second detection mode may comprise switching via software control without mechanical reconfiguration. Software-controlled mode switching enables users to transition between the first detection mode and the second detection mode by selecting the detection mode through a software interface rather than physically reconfiguring optical components or exchanging detector hardware. The softw are control may activate the appropriate light source and detector for the selected detection mode while deactivating components associated with the non-selected detection mode.

[0159] In some cases, the method comprises screening a plurality of molecular interactions in the second detection mode to identify hits. The second detection mode configured for surface plasmon resonance imaging detection enables simultaneous monitoring of binding responses across an array of molecular interactions. The array may comprise microarray spots having a diameter in a range of 100 pm to 500 pm, with each spot containing a different immobilized molecule for interaction analysis. Screening in the second detection mode may identify hits based on binding responses exceeding a threshold value indicative of molecular interaction.

[0160] Following screening in the second detection mode, the method may comprise validating identified hits in the first detection mode. The first detection mode configured for surface plasmon resonance detection providing sensitivity of 0.05 RU RMSD or less provides detailed kinetic characterization of the identified hits. Validation in the first detection mode may comprise extracting kinetic parameters including an association rate constant, a dissociation rate constant, and an equilibrium dissociation constant from bindingDocket No.: INST-01-PCTdata obtained for each identified hit. The software-controlled mode switching enables transition from screening in the second detection mode to validation in the first detection mode without requiring sensor chip exchange or mechanical reconfiguration of the multimodal surface plasmon resonance detector.

[0161] In some cases, the method comprises operating the first detection mode and the second detection mode concurrently. Concurrent operation of the first detection mode and the second detection mode may be enabled by the orthogonal arrangement of the first optical path and the second optical path, which prevents optical interference between the two detection pathways. Concurrent operation enables simultaneous acquisition of kinetic data with sensitivity of 0.05 RU RMSD or less from the first detection mode and spatial imaging data from the second detection mode during a single molecular interaction experiment.

[0162] Referring to FIG. 3, a method 300 for detecting molecular interactions using a multi-modal surface plasmon resonance detector with an optional photochemistry path is shown. The method 300 begins with a step 300, where temperature-controlled liquid is circulated through cooling passages in a detector frame to maintain thermal stability at a sensing surface. The temperature-controlled liquid may be circulated at a flow rate in a range of 150 mL / min to 250 mL / min through integrated conformal cooling passages formed wdthin the detector frame. Circulating the temperature-controlled liquid may comprise maintaining a temperature stability of ±0.003°C at the sensing surface for applications including fragmentbased drug discovery and kinase inhibitor characterization.

[0163] With continued reference to FIG. 3, the method 300 proceeds to a step 302, where a plurality of molecular interactions are screened in a second detection mode configured for surface plasmon resonance imaging. The second detection mode enables simultaneous monitoring of binding responses across an array of molecular interactions on the sensing surface. The multi-modal surface plasmon resonance detector may be configured for fragment-based drug discovery detecting weak binding interactions with equilibrium dissociation constants from 100 pM to 10 mM between small molecule fragments less than 300 Da and protein drug targets. Screening in the second detection mode may comprise monitoring binding responses across microarray spots having a diameter in a range of 100 pm to 500 pm.

[0164] Following the step 302, the method 300 moves to a step 304, where hits are identified from the screening results. Identify ing hits may comprise determining which molecular interactions exhibit binding responses exceeding a threshold value indicative of specific binding. The multi-modal surface plasmon resonance detector may be configured forDocket No.: INST-01-PCThigh-density kinase studies requiring 5,000 RU or more of immobilized protein for detecting fragments with equilibrium dissociation constants in a range of 1 pM to 100 pM. High protein density on the sensing surface generates sufficient signal magnitude for detection of weak binding interactions characteristic of fragment screening applications.

[0165] As further shown in FIG. 3, the method 300 reaches a step 306, where a determination is made whether to activate a photochemistry path. The determination at the step 306 may be based on the type of surface chemistry employed, the regeneration requirements of the assay, or the need for photoactivatable coupling during the molecular interaction analysis. In some cases, the determination at the step 306 is made automatically by software based on assay protocol parameters. In some cases, the determination at the step 306 is made by user selection through a software interface.

[0166] With continued reference to FIG. 3, if the photochemistry path is to be activated at the step 306, the method 300 proceeds along a first branch to a step 308A, where ultraviolet light is delivered to the sensing surface during surface plasmon resonance monitoring. Delivering ultraviolet light at the step 308A may comprise activating a photochemistry path configured to deliver ultraviolet light at approximately 365 nm to the sensing surface. The ultraviolet light may activate photoactivatable crosslinkers including diazirine groups or benzophenone groups for covalent coupling of molecules to the sensing surface. In some cases, the ultraviolet light triggers photo-release of captured molecules from photoswitchable capture reagents for gentle surface regeneration.

[0167] Monitoring photocoupling dynamics in real-time may be performed during ultraviolet activation at the step 308 A. Real-time monitoring of photocoupling dynamics comprises acquiring surface plasmon resonance data from the first detection mode while the photochemistry’ path delivers ultraviolet light to the sensing surface. The real-time monitoring enables observation of coupling efficiency, coupling kinetics, and completion of photochemical reactions at the sensing surface. In some cases, real-time monitoring of photocoupling dynamics enables quantification of switching kinetics and fold-change in affinity between dark and lit states of photoswitchable capture proteins.

[0168] From the step 308 A, the method 300 continues to a step 310A, where the identified hits are validated in a first detection mode configured for surface plasmon resonance detection providing sensitivity’ of 0.05 RU RMSD or less. Validation at the step 310A may comprise extracting kinetic parameters including an association rate constant, a dissociation rate constant, and an equilibrium dissociation constant from binding data obtained for each identified hit. The first detection mode provides sensitivity of 0.05 RUDocket No.: INST-01-PCTRMSD or less for detailed kinetic characterization of the identified hits following photochemistry’ activation.

[0169] As further show n in FIG. 3, if the photochemistry path is not to be activated at the step 306, the method 300 proceeds along a second branch to a step 308B, where the identified hits are validated in the first detection mode. Validation at the step 308B proceeds without photochemistry activation and comprises switching from the second detection mode to the first detection mode via software control without mechanical reconfiguration. From the step 308B, the method 300 continues to a step 310B, where the validation workflow ends. The method 300 enables complete screening-to-validation workflows within a single multimodal surface plasmon resonance detector, with optional integration of photochemistry' activation during the molecular interaction detection process.

[0170] Referring to FIG. 2, a method for manufacturing a detector frame for optical biosensing is shown. The method begins with a step 200, where a monolithic frame is fabricated using metal additive manufacturing. The monolithic frame may be fabricated from a thermally conductive metal alloy including AlSilOMg aluminum alloy, copper alloy, titanium, or stainless steel. Fabricating the monolithic frame may comprise selective laser melting, where a laser selectively fuses metal powder layer-by-layer to build the three-dimensional structure of the monolithic frame. Selective laser melting enables formation of complex internal geometries within the monolithic frame that would be impractical to produce using conventional machining methods.

[0171] With continued reference to FIG. 2, fabricating the monolithic frame at the step 200 may comprise a print time in a range of 6 hours to 12 hours. The print time depends on the size and complexity of the monolithic frame, the layer thickness selected for the selective laser melting process, and the density of internal features including cooling passages and mounting cavities. The monolithic frame consolidates functions of a plurality of precision-machined components into a single part, replacing 15 to 25 discrete components of traditional surface plasmon resonance detector assemblies w ith a single integrated structure.

[0172] Following the step 200. the method proceeds to a step 202, where integrated conformal cooling passages are formed within the monolithic frame. The integrated conformal cooling passages are formed during the metal additive manufacturing process at the step 200, enabling the cooling passages to follow^ complex three-dimensional paths that traverse different depths within the monolithic frame. Forming the integrated conformal cooling passages at the step 202 may comprise forming passages having a diameter in a range of 2 mm to 4 mm. The integrated conformal cooling passages may be routed within 5 mm toDocket No.: INST-01-PCT10 mm of a prism interface region to provide thermal regulation of a sensing surface while maintaining sufficient material thickness for mechanical stability.

[0173] As further shown in FIG. 2. forming the integrated conformal cooling passages at the step 202 may comprise forming teardrop-shaped via cross-sections configured for printing without removable supports. The teardrop-shaped cross-sections incorporate a tapered upper surface that enables the channel geometry to be printed without internal support structures. Internal support structures within enclosed internal passages would otherwise be impossible to remove following the metal additive manufacturing process. The teardrop-shaped via cross-sections represent a design-for-additive-manufacturing approach that enables formation of self-supporting geometries within the monolithic frame.

[0174] With continued reference to FIG. 2, forming the integrated conformal cooling passages at the step 202 may comprise forming a serpentine channel pattern or a parallel channel pattern depending on thermal management requirements. The serpentine channel pattern routes temperature-controlled liquid through a continuous folded path within the monolithic frame, while the parallel channel pattern provides multiple channels extending in parallel through the monolithic frame for uniform cooling across a sensing zone. Forming the integrated conformal cooling passages may further comprise forming heat transfer enhancement structures within the cooling passages, including fin-like or ribbed internal geometries that increase surface area contact between circulating coolant and the thermally conductive metal frame material.

[0175] Following the step 202, the method moves to a step 204, where mounting surfaces of the monolithic frame are post-processed. Post-processing the mounting surfaces at the step 204 may comprise machining mounting surfaces to a tolerance of ±0.01 mm for positioning of optical components. The precision-machined mounting surfaces provide optical tolerances suitable for mounting a prism, light sources, detectors, and cameras to the monolithic frame. Post-processing at the step 204 may comprise a post-processing time in a range of 2 hours to 4 hours for support removal and machining of mounting surfaces.

[0176] As further shown in FIG. 2, post-processing the mounting surfaces at the step 204 may comprise removing support structures from external surfaces of the monolithic frame. Support structures may be required during the selective laser melting process to support overhanging features and prevent warping during the layer-by-layer build process. The combination of metal additive manufacturing at the step 200 for complex internal geometries and post-processing at the step 204 for precision mounting surfaces enables the monolithic frame to integrate structural support, thermal management, and optical mountingDocket No.: INST-01-PCTfeatures in a single component.

[0177] The method may further comprise forming a coolant inlet port and a coolant outlet port integrated into the monolithic frame during the metal additive manufacturing at the step 200. The coolant inlet port and the coolant outlet port may be configured with barbed hose fittings or quick-disconnect fittings for connection to flexible tubing or external chillers. In some cases, the method further comprises forming a temperature sensor cavity within the monolithic frame proximate to a sensing zone for embedding a thermistor that provides realtime temperature feedback to a temperature controller.

[0178] The method may further comprise forming a prism mounting region, a light source mounting region, and a detector mounting region in the monolithic frame. The mounting regions may be formed during the metal additive manufacturing at the step 200 with near-net-shape geometry, followed by precision machining at the step 204 to achieve the ±0.01 mm tolerance for optical component positioning. In some cases, the method further comprises forming a camera cooling via routed proximate to the detector mounting region for thermal management of a camera that generates heat during operation.

[0179] The monolithic frame manufactured according to the method may be configured to achieve a resonant frequency greater than 200 Hz for optical stability . The resonant frequency specification ensures that mechanical vibrations do not couple to optical components during surface plasmon resonance measurements. The monolithic frame may be configured to provide unified thermal management for optical components and electronic subsystems through the integrated conformal cooling passages formed at the step 202.

[0180] Referring to FIG. 4, a comprehensive system workflow 400 for a multimodal surface plasmon resonance detector is shown. The workflow 400 integrates all system components for complete molecular interaction analysis from screening through validation. The workflow 400 begins with a step 400, where a detector frame with integrated conformal cooling passages routed proximate to a prism interface is provided. The detector frame may be manufactured by metal additive manufacturing from a thermally conductive metal alloy. The integrated conformal cooling passages may be formed during the metal additive manufacturing process, enabling complex three-dimensional geometries that route temperature-controlled liquid around sensing zones and optical component mounting regions. The detector frame integrates structural support, thermal management, and optical mounting features in a single monolithic component.

[0181] With continued reference to FIG. 4, the workflow 400 proceeds to a step 402, where temperature-controlled liquid is circulated through the cooling passages via aDocket No.: INST-01-PCTliquid cooling system. Circulating the temperature-controlled liquid at the step 402 may comprise circulating a water-glycol mixture at a flow rate in a range of 150 mL / min to 250 mL / min through the integrated conformal cooling passages. The liquid cooling system may comprise a thermoelectric cooler module, a centrifugal pump, a coolant reservoir, and a temperature controller configured to maintain temperature stability at a sensing surface. The temperature-controlled liquid may be maintained at a temperature stability of ±0.003°C at the sensing surface for molecular interaction analysis.

[0182] Following the step 402, the workflow 400 continues to a step 404, where a sensor chip is positioned at a sensor chip interface. Positioning the sensor chip at the step 404 may comprise inserting the sensor chip into the sensor chip interface such that the sensor chip is optically coupled to a prism through a polymer-based optical matching interface or a refractive index matching fluid. The sensor chip may comprise a glass substrate coated with a gold film and functionalized with surface chemistry for biomolecular immobilization. The sensor chip interface may include alignment features that register the sensor chip to the prism interface with positional repeatability suitable for automated experimental workflows.

[0183] As further shown in FIG. 4, the workflow 400 moves to a step 406, where samples are delivered to a sensor surface via a microfluidic flow cell. Delivering samples at the step 406 may comprise flowing analyte solutions over the sensor surface at flow rates of 5 pL / min to 100 pL / rnin for a first detection path or 1 pL / inin to 10 pL / min for a second detection path. The microfluidic flow cell may be positioned above the sensor chip for delivery of samples to the sensor surface. Sample delivery at the step 406 enables molecular interactions between analyte molecules in solution and ligand molecules immobilized on the sensor surface.

[0184] With continued reference to FIG. 4, the workflow 400 proceeds to a step 408, which is a decision point for selecting a detection mode. The selection at the step 408 may be made via software control based on assay protocol parameters or user selection through a softw are interface. The step 408 enables users to select between a first mode configured for high-sensitivity surface plasmon resonance detection and a second mode configured for surface plasmon resonance imaging detection. The selection at the step 408 determines which detection path is activated for the subsequent molecular interaction analysis.

[0185] If the first mode is selected at the step 408, the workflow- 400 proceeds along a first branch to a step 410A, where a first detection path is operated for surface plasmon resonance detection using a light source, optical elements, and a detector. Operating the firstDocket No.: INST-01-PCTdetection path at the step 410A may comprise activating a near-infrared light source operating at a wavelength of 850 nm (e.g.. in a range of 840 nm to 860 nm), directing light through collimating optics and polarizers to the sensing surface, and detecting reflected light with a line camera detector. The first detection path may provide a sensitivity of 0.05 RU RMSD or less for detailed kinetic characterization of molecular interactions.

[0186] If the first mode is selected at the step 408, the workflow 400 proceeds along a first branch to a step 410A, where a first detection path is operated for surface plasmon resonance detection using a light source, optical elements, and a detector. Operating the first detection path at the step 410A may comprise activating a near-infrared light source operating at a wavelength of 850 nm (e.g., in a range of 840 nm to 860 nm), directing light through collimating optics and polarizers to the sensing surface, and detecting reflected light with a line camera detector. The first detection path may provide a sensitivity’ of 0.05 RU RMSD or less for detailed kinetic characterization of molecular interactions.

[0187] As further shown in FIG. 4, following the step 410A, the workflow 400 proceeds to a step 412A, where modes are switched without mechanical reconfiguration. Switching modes at the step 412A may comprise transitioning from the first detection path to the second detection path or from the second detection path to the first detection path via software control. The software control activates the appropriate light source and detector for the selected detection mode while deactivating components associated with the non-selected detection mode. Mode switching at the step 412A enables users to transition between high-sensitivity kinetic measurements and high-throughput array screening within a single experimental workflow without requiring sensor chip exchange or physical reconfiguration of optical components.

[0188] Following the step 410B, the workflow 400 proceeds to a step 412B, where a photochemistry path is activated to deliver ultraviolet light to the sensor surface. Activating the photochemistry path at the step 412B may comprise delivering ultraviolet light at a wavelength of 365 nm (e.g., in a range of 350 nm to 380 nm) to the sensor surface for activating photoactivatable crosslinkers or triggering photo-release of captured molecules from photoswitchable capture reagents. The photochemistry path may be activated during surface plasmon resonance monitoring to enable real-time observation of photochemical reactions at the sensor surface. In some cases, the photochemistry path delivers blue light at 470 nm for activating LOV2 domain-containing photoswitchable proteins.

[0189] The workflow 400 enables complete molecular interaction analysis from screening through validation within a single multi-modal surface plasmon resonance detector.Docket No.: INST-01-PCTThe workflow 400 may begin with screening a plurality of molecular interactions in the second detection mode at the step 41 OB to identify hits from an array of immobilized molecules. Following hit identification, the workflow 400 may proceed to the step 412A to switch modes without mechanical reconfiguration, enabling transition to the first detection mode at the step 410A for detailed kinetic validation with sensitivity7of 0.05 RU RMSD or less of the identified hits. The integration of the step 412B for photochemistry activation enables gentle surface regeneration through photo-triggered release of captured molecules, extending surface lifetime for screening campaigns that test multiple compounds per surface.

[0190] EXAMPLES

[0191] Aspects of the present teachings may be further understood in light of the following examples, which should not be construed as limiting the scope of the present teachings in any way.

[0192] Example 1: Dual-Mode Detector Validation with Carbonic Anhydrase II Small Molecule Binding

[0193] A prototype multi-modal surface plasmon resonance detector was constructed according to the present disclosure. The detector frame was manufactured from AlSilOMg aluminum alloy using selective laser melting with integrated conformal cooling passages having a diameter of 3 mm routed within 6-8 mm of the prism interface. A pyramidal BK7 prism was mounted at the prism interface. The first detection path comprised an 850 nm near-infrared LED light source operating at 5 mW, polarizing optics, and a line camera detector with 2048 pixels operating at 500 Hz frame rate. The second detection path comprised the same 850 nm illumination wavelength with wide-field optics and a 5-megapixel CMOS camera for imaging mode operation.

[0194] The liquid cooling system circulated a 50 / 50 water-glycol mixture with 0.05% biocide at 200 mL / min through the integrated conformal cooling passages. A thermoelectric cooler module with 40 W capacity regulated the coolant temperature. A thermistor embedded in the detector frame proximate to the sensing surface provided realtime temperature feedback to a PID temperature controller. The system maintained temperature stability7of ±0.003°C at the sensing surface during continuous 8-hour operation at a 25°C setpoint.

[0195] For validation of the first detection path, the classical carbonic anhydrase II (CAII) small molecule interaction system was employed. CAII is a well-characterized modelDocket No.: INST-01-PCTsystem for surface plasmon resonance validation, with published binding kinetics available for comparison (Navratilova, I., et al., "Fragment Screening by Surface Plasmon Resonance." ACS Med. Chem. Lett. 2010, 1, 44-48). A CM5 sensor chip functionalized with carboxymethyl dextran was prepared by amine coupling of bovine carbonic anhydrase II (Sigma- Aldrich, catalog number C2522) to achieve an immobilization level of 5,000 RU (e.g., in a range of 4,000 RU to 6,000 RU). The amine coupling procedure followed established protocols using 0.4 M l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 0.1 M N-hydroxysuccinimide (NHS) activation for 7 minutes, followed by injection of CAII at 50 pg / mL in 10 mM sodium acetate buffer pH 5.0 for 10 minutes, and deactivation with 1 M ethanolamine-HCl pH 8.5 for 7 minutes.

[0196] Two sulfonamide inhibitors were tested: 4-carboxybenzenesulfonamide (molecular weight 201 Da) and furosemide (molecular weight 331 Da). Analyte solutions were prepared in HBS-EP running buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.005% v / v Surfactant P20) at concentrations ranging from 41.2 pM to 3.33 mM in a 2-fold dilution series. Multi-cycle kinetics were performed with 60-second association phases, 120-second dissociation phases, and regeneration with 50 mM NaOH for 30 seconds between cycles. Flow rate was maintained at 30 pL / min.

[0197] The first detection path achieved baseline noise of 0.035 RU RMSD and signal drift less than 0.5 RU / hour over 8-hour operation. Sensorgrams were fitted to a 1: 1 Langmuir binding model using standard kinetic analysis software. For 4-carboxybenzenesulfonamide, the measured equilibrium dissociation constant (KD) was 1.1 pM, with association rate constant (ka) of 2.3 x 104 M-ls-1 and dissociation rate constant (kd) of 0.025 s-1. For furosemide, the measured KD was 0.8 pM, with ka of 3.1 x 104 M-ls-1 and kd of 0.024 s-1. These values are consistent with published literature values for these interactions (Myszka, D.G., "Improving Biosensor Analysis," J. Mol. Recognit. 1999, 12, 279-284).

[0198] For validation of the second detection path, the imaging mode was used during fluidics development to visualize fluid flow patterns across the sensor surface. The system operated as a surface plasmon resonance microscope, providing real-time images of the flow cell where grayscale intensity was proportional to local refractive index. The imaging mode captured complete spatial mapping of the flow cell at 30 Hz frame rate, showing fluid distribution and confirming proper hy drodynamic addressing. Real-time visualization of sample injection, diffusion, and wash steps enabled rapid optimization of injection protocols. Direct observation of artifacts including air bubbles (appearing as lightDocket No.: INST-01-PCTregions) and sample aggregates (creating localized high-intensity spots) allowed immediate identification and correction of experimental issues.

[0199] The detector frame achieved a detector volume of 576 cm3, compared to 3,800-4,500 cm3 for traditional TEC -fan assemblies. Acoustic measurements recorded 38 dB(A) at 1 m distance during operation, compared to 54-58 dB(A) for fan-cooled systems with comparable thermal performance, representing a 15-20 dB noise reduction.

[0200] Example 2: Integrated Thermal Management for Detector and Light Source

[0201] A detector frame was manufactured according to the present disclosure with the integrated conformal cooling passages configured to provide unified thermal management for both the sensing surface and the LED light source. The detector frame was manufactured from AlSilOMg aluminum alloy using selective laser melting. The integrated conformal cooling passages were routed to pass within 5 mm of both the prism interface and the LED mounting region.

[0202] The 850 nm LED light source (Thorlabs LED850L, 5 mW typical output) was mounted directly to the liquid-cooled detector frame using thermal interface material (Bergquist Gap Pad 5000S35) providing thermal resistance less than 0.5°C / W. The LED generated approximately 2.5 W of heat during operation. Without active cooling. LED junction temperature would rise by 15-20°C above ambient, causing wavelength drift of 0.3-0.5 nm per °C and intensity variations that manifest as apparent baseline drift in surface plasmon resonance measurements.

[0203] The liquid cooling system circulated temperature-controlled coolant (50 / 50 water-glycol mixture) at 200 mL / min through the integrated conformal cooling passages. Coolant entered the detector frame at 22°C and exited at approximately 26°C under continuous operation, after which it passed through a radiator and was returned to the thermoelectric cooler assembly for recooling.

[0204] LED junction temperature stability was monitored via forward voltage measurement, which provides a linear relationship to junction temperature with a coefficient of approximately -2 mV / °C for GaAlAs LEDs. The integrated thermal management achieved LED junction temperature stability of ±0.01°C during continuous operation, eliminating wavelength drift as a source of measurement noise.

[0205] The unified thermal management approach reduced total system component count by 8 parts compared to traditional architectures requiring separate LED heat sink.Docket No.: INST-01-PCTthermal interface plates, fan assembly, and mounting brackets. The system footprint was reduced because the thermoelectric cooler and radiator were located in a separate compartment, enabling a thinner and more compact detector assembly. Temperature variation between the LED mounting point and the sensing surface was less than 0.005°C during continuous operation.

[0206] Example 3: High-Affinity Antibody Interaction with Extended Dissociation Phase

[0207] The multi-modal surface plasmon resonance detector was used to characterize a high-affinity antibody-antigen interaction requiring extended dissociation phase monitoring. High-affinity interactions with dissociation rate constants (kd) less than 10'4s’1require dissociation phases of 3,600 seconds or longer to accurately determine the off-rate, placing stringent demands on thermal stability and baseline drift performance (Karlsson, R., et al., "Analyzing a Kinetic Titration Series Using Affinity Biosensors," Anal. Biochem.2006, 349, 136-147).

[0208] A CM5 sensor chip was prepared with anti-human IgG Fc capture antibody (Cytiva, catalog number BR- 1008-39) immobilized by amine coupling to achieve 10,000 RU (e.g., in a range of 8,000 RU to 12,000 RU) capture level. A therapeutic antibody candidate (human IgGl, molecular weight 150 kDa) was captured at 200 RU (e.g., in a range of 150 RU to 250 RU). The target antigen (recombinant human protein, molecular weight 25 kDa) was injected at concentrations ranging from 0.1 nM to 10 nM in HBS-EP running buffer.

[0209] The liquid cooling system maintained temperature stability of ±0.003°C at the sensing surface throughout the extended measurement period. Association phases were 300 seconds, and dissociation phases were 3,600 seconds (60 minutes) to enable accurate determination of the slow off-rate. Regeneration was performed with 10 mM glycine-HCl pH 2.0 for 60 seconds between cycles.

[0210] The first detection path provided baseline noise of 0.03 RU RMSD and signal drift less than 0.3 RU / hour over the 8-hour experimental period. The extended dissociation phase data showed clean exponential decay without baseline drift artifacts. Kinetic analysis yielded kaof 2.1 x 106M'1, kd of 4.2 x 10'5s'1, and KD of 20 pM. The thermal stability provided by the liquid-cooled detector frame was essential for accurate determination of the slow off-rate, as temperature-induced refractive index drift would otherwise obscure the dissociation kinetics.Docket No.: INST-01-PCT

[0211] Example 4: Automated Antibody Screening-to-Validation Workflow (Prophetic)

[0212] This prophetic example demonstrates an integrated workflow where initial screening in the second detection mode identifies high-affinity antibodies, followed by automated validation in the first detection mode, all within a single device without user intervention or sample transfer.

[0213] A multi-modal surface plasmon resonance detector is configured according to the present disclosure with both the first detection path and the second detection path operational. The liquid cooling system maintains temperature stability of ±0.003°C at the sensing surface. An autosampler is loaded with a 96-well plate containing antibody variant library at 50 p.g / mL in PBS, target antigen at 10 pg / mL, regeneration solution (10 mM glycine-HCl pH 2.0), and running buffer (HBS-EP).

[0214] For Phase 1 (Array Screening in Second Detection Mode), a pre-fabricated antibody microarray chip with all 96 antibody variants immobilized in defined array positions is loaded into the sensor chip interface. The second detection path is activated for surface plasmon resonance imaging mode operation. A baseline image is captured showing all 96 array spots. Target antigen is injected across the array at 5 pL / min flow rate. The second detection path captures time-lapse images at 10 Hz frame rate showing binding events at each spot. Software analyzes binding response at each array position, calculating relative response units for each antibody variant. The array is regenerated with 10 mM glycine-HCl and the process is repeated with antigen variants or orthologues to evaluate specificity. The system automatically ranks antibodies by binding signal strength and specificity profile, identifying top 8 candidates for detailed characterization. Phase 1 is completed in 2-3 hours including specificity testing.

[0215] For Phase 2 (Automated Transition), the system completes the imaging protocol and ejects the antibody array chip. An Fc capture chip (Protein A / G surface) is loaded into the sensor chip interface. The system switches to the first detection mode via software control without mechanical reconfiguration.

[0216] For Phase 3 (Kinetics Validation with Sensitivity of 0.05 RU RMSD or Less in First Detection Mode), for each of the 8 identified hit antibodies, the system retrieves antibody from the original autosampler position and captures it on the Fc surface at 200 RU (e.g., in a range of 150 RU to 250 RU) immobilization level. A multi-cycle kinetics protocol is executed: inject target antigen at 5 concentrations (2-fold dilution series, 100 nM to 6.25Docket No.: INST-01-PCTnM), monitor association for 180 seconds, monitor dissociation for 300 seconds, regenerate with glycine-HCl, and repeat for next concentration. The system fits data to a 1 : 1 Langmuir binding model, extracting ka, kj. and KD for each antibody-antigen pair. Phase 3 is completed in 4-5 hours.

[0217] Expected results include complete binding profile for all 96 antibodies obtained in 2-3 hours during imaging mode screening, identification of 8 antibodies with strong target binding and acceptable specificity7, and kinetics data with sensitivity of 0.05 RU RMSD or less for 8 hits obtained in 4-5 hours. Typical results include KD values ranging from 0.1 nM to 10 nM for successful antibodies, with kaof 105- 106M^s'1and kd of 10"3- 1 O’4s'1. Total time for complete screening and validation is 6-8 hours, fully automated after initial setup. Traditional workflow using separate imaging device for screening, sample transfer, and kinetics on separate surface plasmon resonance device would require 2-3 days with significant manual intervention.

[0218] Example 5: Photochemistry-Enabled Surface Coupling (Prophetic)

[0219] This prophetic example demonstrates real-time surface plasmon resonance monitoring of photoactivatable coupling chemistry7, eliminating the need for NHS / EDC reagents and enabling precise temporal control of surface immobilization.

[0220] A sensor chip is prepared with a gold surface functionalized with a carboxyl-terminated self-assembled monolayer that has been pre-coupled with benzophenone-PEG4-amine linker. Benzophenone groups are stable and non-reactive until ultraviolet activation (Dorman, G., et al., "The Application of Photoreactive Compounds in Drug Discovery," Trends Biotechnol. 2016, 34, 291-303). The multi-modal surface plasmon resonance detector is configured with the photochemistry path comprising a 365 nm ultraviolet LED (Thorlabs M365LP1, 10-100 mW adjustable power) positioned to deliver light normal to the sensor surface through the flow cell.

[0221] For Phase 1 (Protein Injection and Ultraviolet Activation), the sensor chip is loaded into the detector. The first detection path is operated to monitor surface events in realtime. Streptavidin solution at 20 pg / mL in 10 mM sodium acetate buffer pH 4.5 (below the isoelectric point of streptavidin, pl 5.0) is injected. The first detection path shows initial response of approximately 2,000 RU as protein electrostatically pre-concentrates on the surface. While protein solution continues flowing, the photochemistry path is activated: the 365 nm ultraviolet LED is pulsed at 50 mW for 30 seconds. During ultraviolet exposure,Docket No.: INST-01-PCTbenzophenone groups are photoactivated to form reactive biradical intermediates that insert into nearby C-H bonds in protein molecules, forming covalent crosslinks between surface and protein. The first detection path monitors the coupling process in real-time. Protein flow is stopped and the surface is washed with buffer. The first detection path shows stable baseline with covalently coupled protein remaining on surface with minimal dissociation.

[0222] For Phase 2 (Coupling Efficiency Validation), ethanolamine at 1 M pH 8.5 is injected to block any remaining reactive groups. Elution solution (50 mM NaOH) is injected to remove any non-covalently bound protein. Comparison to traditional NHS / EDC coupling shows photoactivated coupling achieves equivalent or better stability with greater than 95% retention. Functional activity is tested by injecting biotinylated protein; streptavidin-biotin based capture confirms that immobilized streptavidin retains activity.

[0223] Expected results include final immobilization level of at least 2,000 RU streptavidin, comparable to NHS / EDC coupling but achieved with simpler reagents and precise temporal control. Coupling is complete in 5 minutes (protein injection plus ultraviolet pulse plus wash) versus 20-30 minutes for NHS / EDC protocol. Ultraviolet pulse duration and intensity provide precise control over coupling density with variation less than 5% between runs, superior to NHS / EDC where reagent age affects coupling efficiency.

[0224] Example 6: Fragment-Based Drug Discovery Screening (Prophetic)

[0225] This prophetic example demonstrates the use of the multi-modal surface plasmon resonance detector for fragment-based drug discovery, where high protein density’ is required to detect weak binding interactions characteristic of small molecule fragments.

[0226] Fragment-based drug discovery requires detection of weak binding interactions with KD values from 100 pM to 10 mM between small molecule fragments (molecular weight less than 300 Da) and protein drug targets (Erlanson, D.A., et al., "Twenty Years On: The Impact of Fragments on Drug Discovery," Nat. Rev. Drug Discov. 2016, 15, 605-619). Small molecules produce weak surface plasmon resonance signals proportional to their molecular weight; a 400 Da compound binding to a 50 kDa protein produces only approximately 8 RU per 1,000 RU immobilized protein. Detecting fragments with KD values of 1-100 pM requires 5,000 RU or more of immobilized protein to achieve measurable signals of 40-400 RU.

[0227] A CM5 sensor chip is prepared with high-density' immobilization of EGFR kinase domain (molecular weight approximately 50 kDa). Amine coupling is performed using extended activation (10 minutes with EDC / NHS) and high protein concentration (200 pg / mLDocket No.: INST-01-PCTin 10 mM sodium acetate pH 5.0) to achieve immobilization level of 5,000-6,000 RU. The liquid cooling system maintains temperature stability of ±0.003°C at the sensing surface, which is adequate for fragment screening applications where signal magnitudes of 40-400 RU are well above the noise floor.

[0228] A fragment library of 500 compounds (molecular weight 150-300 Da, solubility greater than 1 mM in 5% DMSO) is screened using single-concentration injections at 500 pM in HBS-EP buffer with 5% DMSO. Association phases are 60 seconds and dissociation phases are 60 seconds. Flow rate is 50 pL / min to minimize mass transport limitations. Reference subtraction using an unmodified reference surface corrects for bulk refractive index effects from DMSO.

[0229] Primary screening in the first detection mode identifies fragments showing binding responses greater than 5 RU (approximately 3 times the noise floor) as potential hits. Hit rate of 2-5% is typical for fragment libraries against validated drug targets. Confirmed hits are characterized with dose-response curves using 8-point concentration series from 15.6 pM to 2 mM. Kinetic analysis extracts KD values for ranking hits by affinity.

[0230] Reference compound gefitinib (molecular weight 446 Da, literature KD approximately 10 nM for EGFR) is included as a positive control to validate surface activity. The multi-modal surface plasmon resonance detector has demonstrated the ability to measure binding kinetics with 95 Da molecules, showcasing no limitations on molecular weight with the prototype design.

[0231] Example 7: Integration with Nucleic Acid Programmable Protein Array Technology (Prophetic)

[0232] This prophetic example demonstrates integration of the multi-modal surface plasmon resonance detector with nucleic acid programmable protein array (NAPPA) technology to express proteins directly on the sensor surface, screen them in the second detection mode, and characterize hits in the first detection mode.

[0233] NAPPA is an established technology for creating protein microarrays by printing DNA constructs that encode proteins of interest (Ramachandran, N., et al., "SelfAssembling Protein Microarrays," Science 2004, 305, 86-90). The DNA is co-printed with capture reagents (anti-tag antibodies). When supplied with cell-free protein expression reagents, the DNA is transcribed and translated in situ, producing proteins that are immediately captured at their array positions.

[0234] A DNA microarray chip with 384 positions is prepared. Each positionDocket No.: INST-01-PCTcontains plasmid DNA encoding a different antibody variant from a computationally designed library, co-printed with anti-His6 capture antibody. The chip is loaded into the multi-modal surface plasmon resonance detector.

[0235] For Phase 1 (In-Situ Protein Expression), cell-free expression mix (wheat germ extract with translation components, Promega TNT SP6 High-Yield Wheat Germ system) is flowed over the array. The liquid cooling system maintains the sensor chip interface at 25°C during the 1-2 hour incubation period. During expression, DNA is transcribed and translated, and newly synthesized proteins are captured by anti-His6 antibodies at each array position. Expression reagents are washed away with HBS-EP buffer.

[0236] For Phase 2 (Array Screening in Second Detection Mode), the second detection path is activated for surface plasmon resonance imaging mode. A baseline image is captured showing expression levels at all 384 positions based on surface plasmon resonance signal from captured proteins. Expression levels vary from 50-500 RU captured protein per spot. Target antigen at 10 pg / mL is injected. The second detection mode monitors binding across the entire array simultaneously at 10 Hz frame rate. Software identifies array positions showing strong antigen binding (response greater than 10 RU above baseline). The array is regenerated and tested with off-target antigens to confirm specificity.

[0237] For Phase 3 (Hit Validation), the system provides DNA sequences of hit antibodies to the user. Following synthesis and purification of hit antibodies by commercial vendors (2-3 week turnaround), purified proteins are loaded into the autosampler. The first detection mode is activated for kinetics characterization with sensitivity of 0.05 RU RMSD or less with multi-concentration injections to determine ka, kd, and KD values.

[0238] Expected results include 384 antibody variants expressed on array in 2 hours with expression levels of 50-500 RU per spot. Complete binding screen of 384 variants is obtained in 1-2 hours, identifying 10-20 strong binders with good specificity. Following synthesis and purification of hit antibodies, the first detection mode characterization provides KD values for the hits, identify ing 2-3 antibodies with sub-nanomolar affinity suitable for therapeutic development.

[0239] Example 8: Photo-Releasable Capture Reagents for Gentle Surface Regeneration (Prophetic)

[0240] This prophetic example demonstrates the use of engineered photoswitchable capture reagents with the photochemistry path for gentle, chemical-free surface regenerationDocketNo.: INST-01-PCTin high-density kinase studies.

[0241] Standard regeneration conditions (glycine-HCl, DMSO) degrade kinase activity within 20-50 cycles, limiting the number of compounds that can be screened per surface. Photo-releasable capture reagents enable gentle regeneration through light-triggered conformational changes that weaken binding affinity7without harsh chemical treatment (Gil, A. A., et al., "Optogenetic Control of Protein Binding Using Light-Switchable Nanobodies." Nat. Commun. 2020. 11. 4044).

[0242] A CM5 sensor chip is prepared with high-density immobilization of engineered photo-releasable streptavidin (Photo-SA) at 10,000 RU (e.g., in a range of 8,000 RU to 12,000 RU). Photo-SA is a computationally designed streptavidin variant with an AsLOV2 photosensitive domain inserted near the biotin-binding pocket. In the dark state, the LOV2 Ja-helix is docked and the binding pocket maintains wild-type geometry with KD of 10'15M (e.g., in a range of 10’14M to 10'16M) for biotin. Upon 365 nm ultraviolet illumination, the LOV2 Ja-helix undocks, causing allosteric distortion of the binding pocket and weakening biotin affinity to KD of 10'9M (e.g., in a range of 10'8M to IO'10M) (1,000-fold weaker). Dark recovery occurs within 60 seconds as the LOV2 domain re-docks.

[0243] Biotinylated EGFR kinase domain is captured on the Photo-SA surface at 5,000-6,000 RU. Small molecule kinase inhibitors are screened using the first detection mode. Following each screening cycle, the photochemistry path is activated: the 365 nm ultraviolet LED is pulsed at 50 mW for 30 seconds. The first detection mode monitors the photo-release process in real-time, showing signal decrease as the kinase dissociates from the weakened Photo-SA binding site. Buffer wash removes released kinase. Dark recovery for 60 seconds restores Photo-SA binding capacity7. Fresh biotinylated kinase is captured for the next screening cycle.

[0244] Expected results include maintenance of 5,000 RU or greater kinase capture level across more than 100 regeneration cycles, compared to 20-50 cycles with traditional harsh regeneration. Real-time monitoring of photo-release kinetics enables optimization of ultraviolet pulse duration and intensity7. The gentle regeneration preserves kinase activity7throughout extended screening campaigns testing 100 or more compounds per surface.

[0245] The photochemistry path enables characterization of the Photo-SA switching performance by monitoring affinity changes in real-time during light activation. Dark state KD of approximately 10’15M (tight capture) transitions to lit state KD of approximately 10'9M (1, 000-fold weaker) upon 365 nm illumination. Switch kinetics occur on the seconds-to-Docket No.: INST-01-PCTminutes timescale. Reversibility is demonstrated across more than 10 cycles. This characterization capability is essential as computational protein design produces new photoswitchable proteins that require experimental validation.

[0246] Other Embodiments

[0247] The detailed description set forth above is provided to aid those skilled in the art in practicing the present invention. However, the invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed because these embodiments are intended as illustration of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description which do not depart from the spirit or scope of the present inventive discover}'. Such modifications are also intended to fall within the scope of the appended claims.

[0248] References Cited

[0249] All publications, patents, patent applications and other references cited in this application are incorporated herein by reference in their entirety' for all purposes to the same extent as if each individual publication, patent, patent application or other reference was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Citation of a reference herein shall not be construed as an admission that such is prior art to the present invention.

Claims

Docket No.: INST-01-PCTCLAIMSWhat is claimed is:

1. A multi-modal surface plasmon resonance detector for measuring molecular interactions, the detector comprising:a detector frame manufactured by metal additive manufacturing, the detector frame comprising integrated conformal cooling passages;a prism providing a plurality of optical faces for a plurality of optical paths;a first detection path configured for surface plasmon resonance detection;a second detection path configured for surface plasmon resonance imaging;a sensor chip interface configured to receive a sensor chip; anda liquid cooling system configured to circulate liquid through the integrated conformal cooling passages.

2. The multi-modal surface plasmon resonance detector according to claim 1, wherein the detector frame is manufactured from a thermally conductive metal alloy.

3. The multi-modal surface plasmon resonance detector according to claim 1 or 2, wherein the thermally conductive metal alloy comprises AlSilOMg aluminum alloy.

4. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 3, wherein the thermally conductive metal alloy comprises a copper alloy.

5. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 4, wherein the thermally conductive metal alloy comprises titanium.

6. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 5, wherein the thermally conductive metal alloy comprises stainless steel.

7. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 6, wherein the integrated conformal cooling passages are formed during the metal additive manufacturing.

8. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 7, wherein the integrated conformal cooling passages have a diameter in a range of 2 mm to 4 mm.

9. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 8, wherein the integrated conformal cooling passages follow a serpentine channel pattern.

10. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 9, wherein the integrated conformal cooling passages follow a parallel channel pattern.Docket No.: INST-01-PCT11. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 10, wherein the integrated conformal cooling passages are routed within 5 mm to 10 mm of the prism.

12. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 11, wherein the integrated conformal cooling passages comprise teardrop-shaped cross-sections having a self-supporting geometry with an apex angle of less than 45 degrees.

13. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 12, wherein the plurality of optical paths comprise orthogonal optical paths.

14. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 13, wherein the second detection path is orthogonal to the first detection path.

15. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 14, wherein the first detection path is configured for surface plasmon resonance detection having a sensitivity of 0.05 RU RMSD or less.

16. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 15, wherein the sensor chip interface is configured to receive the sensor chip at a top face of the prism.

17. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 16, wherein the liquid cooling system is configured to circulate temperature-controlled liquid through the integrated conformal cooling passages.

18. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 17, wherein the liquid cooling system is configured to maintain a temperature stabi 1 i ty of ±0.003°C at a sensing surface.

19. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 18, wherein the liquid cooling system is configured to operate over a temperature range of 4°C to 40°C.

20. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 19, wherein the liquid cooling system is configured to circulate the temperature-controlled liquid at a flow rate in a range of 150 mL / min to 250 mL / min.

21. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 20, wherein the liquid cooling system provides a heat transfer coefficient in a range of 2000 W / m2K to 5000 W / m2K.

22. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 21, wherein the prism comprises a pyramidal prism.

23. The multi-modal surface plasmon resonance detector according to any one ofDocket No.: INST-01-PCTclaims 1 to 22, wherein the prism is fabricated from BK7 glass.

24. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 23, wherein the prism is fabricated from SF10 glass.

25. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 24, wherein the first detection path comprises a near-infrared light source operating at a wavelength in a range of 840 nm to 860 nm.

26. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 25, wherein the first detection path comprises a light source having a power in a range of 5 mW to 10 mW.

27. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 26, wherein the first detection path comprises a line camera detector.

28. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 27, wherein the second detection path comprises a two-dimensional camera having at least 5 megapixels.

29. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 28, wherein the first detection path is configured to provide a sensitivity of 0.05 RU RMSD or less.

30. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 29, wherein the second detection path is configured to provide a sensitivity of 0.5 RU RMSD.

31. The multi-modal surface plasmon resonance detector according to any one of claims 1 to 30, further comprising a photochemistry path configured to deliver ultraviolet light at a wavelength in a range of 350 nm to 380 nm to the sensor chip interface.

32. A liquid-cooled detector frame for optical biosensing, the detector frame comprising:a monolithic frame manufactured by metal additive manufacturing;integrated conformal cooling passages formed within the monolithic frame;a coolant inlet and a coolant outlet fluidically connected to the integrated conformal cooling passages; andmounting features for optical components.

33. The liquid-cooled detector frame according to claim 32, wherein the monolithic frame is manufactured from a thermally conductive metal alloy.

34. The liquid-cooled detector frame according to claim 32 or 33, wherein the thermally conductive metal alloy comprises AlSilOMg aluminum alloy having athermalDocket No.: INST-01-PCTconductivity in a range of 130 W / m K to 180 W / m K.

35. The liquid-cooled detector frame according to any one of claims 32 to 34. wherein the thermally conductive metal alloy comprises a copper alloy having a thermal conductivity in a range of 350 W / m K to 400 W / m K.

36. The liquid-cooled detector frame according to any one of claims 32 to 35, wherein the integrated conformal cooling passages are formed during the metal additive manufacturing.

37. The liquid-cooled detector frame according to any one of claims 32 to 36, wherein the integrated conformal cooling passages are routed around a sensing zone and optical component mounting regions.

38. The liquid-cooled detector frame according to any one of claims 32 to 37. wherein the mounting features comprise a prism mounting region, a light source mounting region, and a detector mounting region.

39. The liquid-cooled detector frame according to any one of claims 32 to 38, wherein the integrated conformal cooling passages have a diameter in a range of 2 mm to 4 mm.

40. The liquid-cooled detector frame according to any one of claims 32 to 39, wherein the integrated conformal cooling passages comprise teardrop-shaped cross-sections having a self-supporting geometry with an apex angle of less than 45 degrees.

41. The liquid-cooled detector frame according to any one of claims 32 to 40. wherein the integrated conformal cooling passages are spaced 5 mm to 10 mm from a prism interface.

42. The liquid-cooled detector frame according to any one of claims 32 to 41, further comprising a temperature sensor embedded in the monolithic frame proximate to the sensing zone.

43. The liquid-cooled detector frame according to any one of claims 32 to 42, wherein the monolithic frame is configured to interface with a thermoelectric cooler module positioned external to the monolithic frame.

44. The liquid-cooled detector frame according to any one of claims 32 to 43, wherein the mounting features comprise precision-machined surfaces having a tolerance of ±0.01 mm.

45. The liquid-cooled detector frame according to any one of claims 32 to 44, wherein the monolithic frame has a resonant frequency greater than 200 Hz.

46. The liquid-cooled detector frame according to any one of claims 32 to 45,Docket No.: INST-01-PCTwherein the monolithic frame has a detector volume of 576 cm3.

47. The liquid-cooled detector frame according to any one of claims 32 to 46. wherein the integrated conformal cooling passages are configured to provide unified thermal management for a light-emitting diode and a camera.

48. The liquid-cooled detector frame according to any one of claims 32 to 47, wherein the integrated conformal cooling passages comprise a camera cooling passage routed proximate to the detector mounting region.

49. The liquid-cooled detector frame according to any one of claims 32 to 48, wherein the integrated conformal cooling passages comprise heat transfer enhancement structures.

50. The liquid-cooled detector frame according to any one of claims 32 to 49. wherein the monolithic frame is configured to produce an acoustic signature of 38 dB(A) to 40 dB(A) during operation.

51. The liquid-cooled detector frame according to any one of claims 32 to 50, further comprising quick-disconnect fittings at the coolant inlet and the coolant outlet for connection to an external chiller.

52. The liquid-cooled detector frame according to any one of claims 32 to 51, wherein the integrated conformal cooling passages comprise a first set of cooling passages and a second set of cooling passages configured for multi-zone temperature control.

53. The liquid-cooled detector frame according to any one of claims 32 to 52. wherein the monolithic frame is manufactured by selective laser melting.

54. The liquid-cooled detector frame according to any one of claims 32 to 53, wherein the integrated conformal cooling passages comprise self-supporting geometries having tapered upper surfaces configured for printing without removable supports.

55. The liquid-cooled detector frame according to any one of claims 32 to 54, wherein the monolithic frame integrates structural support, thermal management, and optical mounting features in a single component.

56. The liquid-cooled detector frame according to any one of claims 32 to 55. wherein the monolithic frame is configured to maintain a temperature variation of less than 0.005°C between optical component mounting points.

57. The liquid-cooled detector frame according to any one of claims 32 to 56, wherein the integrated conformal cooling passages are routed around a prism interface, optical mounting points, and a sample staging area.

58. The liquid-cooled detector frame according to any one of claims 32 to 57,Docket No.: INST-01-PCTwherein the monolithic frame comprises fluidic ports integrated into the frame structure.

59. The liquid-cooled detector frame according to any one of claims 32 to 58. wherein the integrated conformal cooling passages follow a three-dimensional path traversing different depths within the monolithic frame.

60. The liquid-cooled detector frame according to any one of claims 32 to 59, wherein the monolithic frame is configured to remove heat from electronic subsystems connected to a same coolant loop.

61. A method for detecting molecular interactions using a multi-modal surface plasmon resonance detector, the method comprising:circulating liquid through integrated conformal cooling passages in a detector frame to maintain thermal stability at a sensing surface;operating in a first detection mode using a first optical path for surface plasmon resonance detection;operating in a second detection mode using a second optical path for surface plasmon resonance imaging detection; andswitching between the first detection mode and the second detection mode.

62. The method according to claim 62, wherein the detector frame is additively manufactured.

63. The method according to claim 62 or 63, wherein circulating liquid comprises circulating temperature-controlled liquid through the integrated conformal cooling passages.

64. The method according to any one of claims 61 to 63, wherein the first optical path is configured for surface plasmon resonance detection providing sensitivity of 0.05 RU RMSD or less.

65. The method according to any one of claims 62 to 65, wherein the second optical path is orthogonal to the first optical path.

66. The method according to any one of claims 62 to 66, wherein switching between the first detection mode and the second detection mode comprises switching via software control without mechanical reconfiguration.

67. The method according to any one of claims 62 to 67, further comprising screening a plurality of molecular interactions in the second detection mode to identify hits.

68. The method according to any one of claims 62 to 68, further comprising validating identified hits in the first detection mode following screening in the second detection mode.

69. The method according to any one of claims 62 to 69, further comprisingDocket No.: INST-01-PCToperating the first detection mode and the second detection mode concurrently.

70. The method according to any one of claims 62 to 70, further comprising activating a photochemistry path to deliver ultraviolet light to the sensing surface during surface plasmon resonance monitoring.

71. The method according to any one of claims 62 to 71, wherein the photochemistry path delivers ultraviolet light at a wavelength in a range of 350 nm to 380 nm for activating photoactivatable crosslinkers.

72. The method according to any one of claims 62 to 72, further comprising monitoring photocoupling dynamics in real-time using the first detection mode during ultraviolet activation.

73. The method according to any one of claims 61 to 72, wherein circulating the temperature-controlled liquid comprises maintaining a temperature stability of ±0.003°C at the sensing surface.

74. The method according to any one of claims 62 to 74, wherein circulating the temperature-controlled liquid comprises transitioning from 25°C to 37°C in less than 10 minutes.

75. The method according to any one of claims 62 to 75, further comprising screening an array of molecular interactions in the second detection mode, wherein the array comprises microarray spots having a diameter in a range of 100 pm to 500 pm.

76. The method according to any one of claims 62 to 76, further comprising extracting kinetic parameters comprising an association rate constant, a dissociation rate constant, and an equilibrium dissociation constant from data obtained in the first detection mode.

77. The method according to any one of claims 62 to 77, further comprising characterizing antibody-antigen binding interactions.

78. The method according to any one of claims 62 to 78, further comprising performing epitope binning using the second detection mode.

79. The method according to any one of claims 62 to 79, further comprising integrating with nucleic acid programmable protein array technology to express proteins directly on the sensing surface.

80. The method according to any one of claims 62 to 80, further comprising activating photoactivatable coupling chemistry' using the photochemistry path while monitoring coupling efficiency using the first detection mode.

81. The method according to any one of claims 62 to 81, wherein the firstDocket No.: INST-01-PCTdetection mode provides a sensitivity of 0.05 RU RMSD or less.

82. The method according to any one of claims 62 to 82, wherein the second detection mode provides a sensitivity of approximately 0.5 RU RMSD.

83. The method according to any one of claims 62 to 83, further comprising visualizing fluid flow patterns across the sensing surface using the second detection mode.

84. The method according to any one of claims 62 to 84, further comprising detecting small molecules having a molecular weight of less than 500 Da binding to proteins.

85. The method according to any one of claims 62 to 85, further comprising performing fragment-based drug discovery screening.

86. The method according to any one of claims 62 to 86, further comprising regenerating the sensing surface using photo-triggered release of captured molecules.

87. The method according to any one of claims 62 to 87, further comprising monitoring binding kinetics at a plurality of array spots simultaneously using the second detection mode.

88. The method according to any one of claims 62 to 88, further comprising operating the first detection mode at a frame rate in a range of 100 Hz to 1000 Hz.

89. The method according to any one of claims 62 to 89, further comprising operating the second detection mode at a frame rate in a range of 1 Hz to 500 Hz.

90. The method according to any one of claims 62 to 90, further comprising maintaining the sensing surface at a temperature of 25°C or 37°C for biological studies.

91. A method for manufacturing a detector frame for optical biosensing, the method comprising:fabricating a monolithic frame using metal additive manufacturing;forming integrated conformal cooling passages within the monolithic frame; and post-processing mounting surfaces of the monolithic frame.

92. The method according to claim 92, wherein fabricating the monolithic frame comprises fabricating from a thermally conductive metal alloy.

93. The method according to claim 92 or 93. wherein forming the integrated conformal cooling passages comprises forming during the metal additive manufacturing.

94. The method according to any one of claims 92 to 94, wherein post-processing the mounting surfaces comprises post-processing to optical tolerances.

95. The method according to any one of claims 92 to 95, wherein fabricating the monolithic frame comprises selective laser melting.

96. The method according to any one of claims 92 to 96, wherein the thermallyDocket No.: INST-01-PCTconductive metal alloy comprises AlSilOMg aluminum alloy.

97. The method according to any one of claims 92 to 97, wherein the thermally conductive metal alloy comprises a copper alloy.

98. The method according to any one of claims 92 to 98, wherein forming the integrated conformal cooling passages comprises forming teardrop-shaped passage crosssections configured for printing without removable supports.

99. The method according to any one of claims 92 to 99, wherein forming the integrated conformal cooling passages comprises forming self-supporting geometries having tapered upper surfaces configured for printing without removable supports.

100. The method according to any one of claims 92 to 100, wherein fabricating the monolithic frame comprises a print time in a range of 6 hours to 12 hours.

101. The method according to any one of claims 92 to 101, wherein postprocessing the mounting surfaces comprises a post-processing time in a range of 2 hours to 4 hours.

102. The method according to any one of claims 92 to 102, wherein postprocessing the mounting surfaces comprises machining mounting surfaces to a tolerance of ±0.01 mm.

103. The method according to any one of claims 92 to 103, wherein forming the integrated conformal cooling passages comprises forming passages having a diameter in a range of 2 mm to 4 mm.

104. The method according to any one of claims 92 to 104, wherein forming the integrated conformal cooling passages comprises routing the passages within 5 mm to 10 mm of a prism interface region.

105. The method according to any one of claims 92 to 105, wherein forming the integrated conformal cooling passages comprises forming a serpentine channel pattern.

106. The method according to any one of claims 92 to 106, wherein forming the integrated conformal cooling passages comprises forming a parallel channel pattern.

107. The method according to any one of claims 92 to 107, further comprising integrating structural support features, thermal management features, and optical mounting features in the monolithic frame as a single component.

108. The method according to any one of claims 92 to 108, further comprising forming heat transfer enhancement structures within the integrated conformal cooling passages.

109. The method according to any one of claims 92 to 109, further comprisingDocket No.: INST-01-PCTforming a coolant inlet port and a coolant outlet port integrated into the monolithic frame.

110. The method according to any one of claims 92 to 110, further comprising forming a temperature sensor cavity within the monolithic frame proximate to a sensing zone.

111. The method according to any one of claims 92 to 111, wherein forming the integrated conformal cooling passages comprises forming passages that traverse different depths within the monolithic frame.

112. The method according to any one of claims 92 to 112, further comprising forming a camera cooling passage routed proximate to a detector mounting region.

113. The method according to any one of claims 92 to 113, further comprising forming a prism mounting region, a light source mounting region, and a detector mounting region in the monolithic frame.

114. The method according to any one of claims 92 to 114, wherein the monolithic frame consolidates functions of a plurality of precision-machined components into a single part.

115. The method according to any one of claims 92 to 115, further comprising removing support structures from the integrated conformal cooling passages during postprocessing.

116. The method according to any one of claims 92 to 116, wherein the monolithic frame is configured to achieve a resonant frequency greater than 200 Hz for optical stability.

117. The method according to any one of claims 92 to 117. further comprising forming a first set of cooling passages and a second set of cooling passages for multi-zone temperature control.

118. The method according to any one of claims 92 to 118, wherein the monolithic frame is configured to provide unified thermal management for optical components and electronic subsystems.

119. A multi-modal surface plasmon resonance detector for measuring molecular interactions, the detector comprising:a detector frame manufactured by metal additive manufacturing, the detector frame comprising integrated conformal cooling passages routed proximate to a prism interface and around a sensing zone;a prism providing a plurality7of optical faces for a plurality7of optical paths;a first detection path configured for surface plasmon resonance detection, the first detection path comprising a light source, optical elements, and a detector;a second detection path configured for surface plasmon resonance imaging detection,Docket No.: INST-01-PCTthe second detection path comprising a light source, optical elements, and a camera; a photochemistry path configured to deliver ultraviolet light to a sensor surface; a sensor chip interface configured to receive a sensor chip;a liquid cooling system configured to circulate temperature-controlled liquid through the integrated conformal cooling passages; anda microfluidic flow cell positioned above the sensor chip for delivery of samples to the sensor surface;wherein the detector frame integrates structural support, thermal management, and optical mounting features; andwherein the first detection path and the second detection path are configured for mode switching without mechanical reconfiguration.

120. A multi-modal surface plasmon resonance detector for measuring molecular interactions, the detector comprising:a prism providing a plurality of optical faces for a plurality of optical paths; a first detection path configured for surface plasmon resonance detection, the first detection path comprising a first light source, first optical elements, and a first detector;a second detection path configured for surface plasmon resonance imaging detection, the second detection path comprising a second light source, second optical elements, and a two-dimensional camera;wherein the first detection path and the second detection path are configured to interrogate a common sensing surface through different optical faces of the prism; and wherein the first detection path and the second detection path are configured for mode switching without mechanical reconfiguration.

121. A multi-modal surface plasmon resonance detector for measuring molecular interactions, the detector comprising:a pyramidal prism having a plurality of optical faces arranged to enable a plurality of orthogonal optical paths addressing a common sensing surface;a first detection path configured for surface plasmon resonance detection providing sensitivity of 0.05 RU RMSD or less, the first detection path comprising a light source and a detector arranged along a first diagonal pair of optical faces of the pyramidal prism;Docket No.: INST-01-PCTa second detection path configured for surface plasmon resonance imaging detection, the second detection path comprising a light source and a two-dimensional camera arranged along a second diagonal pair of optical faces of the pyramidal prism orthogonal to the first diagonal pair; anda sensor chip interface configured to receive a sensor chip at a top face of the pyramidal prism;wherein the orthogonal arrangement of the first detection path and the second detection path enables simultaneous or sequential interrogation of the common sensing surface without optical interference.

122. A multi-modal surface plasmon resonance detector for measuring molecular interactions, the detector comprising:a prism providing a plurality of optical faces for a plurality of optical paths; a first detection path configured for surface plasmon resonance detection; a second detection path configured for surface plasmon resonance imaging detection, the second detection path being orthogonal to the first detection path; a photochemistry path configured to deliver light at a wavelength suitable for activating photoactivatable functional groups to a sensing surface; anda sensor chip interface configured to receive a sensor chip;wherein the first detection path, the second detection path, and the photochemistry path are configured for independent or concurrent operation.

123. A method for detecting molecular interactions using a multi-modal surface plasmon resonance detector, the method comprising:operating in a first detection mode using a first optical path through a prism for surface plasmon resonance detection;operating in a second detection mode using a second optical path through the prism for surface plasmon resonance imaging detection, wherein the second optical path is different from the first optical path;switching between the first detection mode and the second detection mode via software control without mechanical reconfiguration of optical components; and wherein the first detection mode and the second detection mode interrogate aDocket No.: INST-01-PCTcommon sensing surface.

124. A method for detecting molecular interactions, the method comprising: screening a plurality of molecular interactions in a surface plasmon resonance imaging mode using a first optical path through a prism to identify hits;switching to a surface plasmon resonance detection mode using a second optical path through the prism without mechanical reconfiguration; and validating identified hits in the surface plasmon resonance detection mode with sensitivity of 0.05 RU RMSD or less;wherein the screening and validating are performed using a common sensing surface within a single device.

125. A method for detecting molecular interactions, the method comprising: monitoring a sensing surface using surface plasmon resonance detection through a first optical path of a prism;activating a photochemistry path to deliver ultraviolet light at a wavelength in a range of 350 nm to 380 nm to the sensing surface while continuing to monitor the sensing surface; anddetecting photocoupling dynamics in real-time using the surface plasmon resonance detection;wherein the photochemistry path is configured to activate photoactivatable crosslinkers on the sensing surface without requiring chemical activation reagents.

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