Systems and methods for in VIVO cytometry analysis

Abstract

This disclosure provides a method for identifying and counting unlabeled (native) blood cells and analytes in whole blood. In particular, this disclosure relates to devices used within the body equipped with microfluidic cytometry optical sensors and detectors that recognize analytes or histological architecture from within a patient to provide clinically actionable data related to the subject's health.

Description

[0001] Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0002] SYSTEMS AND METHODS FOR IN VIVO CYTOMETRY ANALYSIS

[0003] Cross-Reference to Related Applications

[0004] This application claims priority to, and the benefit of, U.S. Provisional Application No. 63 / 694,769, filed September 13, 2024, the content of which is incorporated by reference herein in its entirety.

[0005] Field of the Invention

[0006] This disclosure relates to intravascular, implantable medical devices or portions of the device, implanted short or long term, having one or more optical sensors as part of a miniaturized in vivo cytometer for identifying and taking a measurement of the blood cell count within a given time segment.

[0007] Background

[0008] Blood cells can be differentiated from the background blood matrix by their specific light scattering properties, absorbance, and reflection. The types of blood cells include red blood cells (RBCs), white blood cells (WBCs), and platelets. WBCs are further differentiated by their biochemical composition, size, and function: Polymorphonuclear white blood cells (PMNs) include neutrophils, basophils, and eosinophils, while mononuclear cells include monocytes and lymphocytes. Because of their unique shape, size, and biochemical structure, their light scattering profiles are distinguishable. RBCs contain hemoglobin, which absorbs violet light to a high degree, whereas WBCs have a lower absorbance of violet light and higher absorbance in the blue spectrum.

[0009] In addition to blood cell counts, in vitro methods for analyzing blood analytes such as proteins, circulating nucleic acids, metabolites, hormones, glucose, and others, have used technologies that are now standard of care. These technologies include biochemical assays using colorimetric or fluorescently labelled detection systems.

[0010] Summary

[0011] The present invention relates to short or long-term implantable medical devices having one or more optical sensors as part of a miniaturized in vivo cytometer for identifying and counting blood cell types intravascularly. The present invention recognizes the limitations of currently utilized methods for blood analysis, i.e., current ex vivo systems, to provide an important alternative to conventional blood draws. Furthermore, the systems and methods of the Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION invention may include the analysis of other variables from sensors separate from or integrated into the device. In particular, the invention addresses the limitations of in vitro analysis and monitoring of a patient’s physiological parameters in both inpatient and outpatient settings. It provides systems and methods for identifying and counting native blood cells and analytes in vivo.

[0012] Most blood analysis is conducted in vitro, requiring a blood draw. However, optical biosensors are now being applied to analyte detection in fields such as healthcare, biomedical research, and environmental monitoring. Miniaturization of these biosensors enables their use in implantable devices, including continuous glucose monitors.

[0013] This invention describes systems and methods of utilizing optical sensors within implantable medical devices to measure blood cells and other blood analytes. In particular, this disclosure describes systems and methods for identifying and counting blood cells, blood analytes, and measuring blood flow properties in vivo, utilizing a microfluidic cytometer embedded in an intravascular access device. Flow cytometry has been used to identify human blood cell subsets for over 50 years. These instruments, including more recent microfluidic versions that have helped miniaturize them for blood analyses, are used routinely to process blood samples from venous blood draws. High-speed flow cytometry techniques enable the realtime classification of cell types using machine-learning algorithms. The invention utilizes these techniques with a novel, in vivo cytometer embedded in an intravascular catheter. Accordingly, the invention provides for immediate blood cell analysis within defined time segments, which may be used, for example, by clinicians to determine if patients need to be transported to clinical facilities or emergency rooms.

[0014] As disclosed herein, the invention provides systems and methods for using flow cytometry techniques in a miniaturized device that is small enough to be placed in vivo. The device, also referred to herein as an optical probe assembly, is embedded in an intravascular catheter. The device may be placed using the catheter to position the miniaturized device within, for example, the vena cava of the human heart, thereby allowing numbers and types of blood cells to be measured in vivo without the need for venous blood draws. In vivo analysis provides for remote monitoring, for example, in cancer patients, other patients with catheters, as well as in pediatric newborns, for whom the amount of blood needed for blood draws might compromise the health of the patient. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0015] In particular embodiments, light scatter results may be used to identify unlabeled human blood cells or other blood analytes using light at blue (roughly 480 nm) and / or blue and violet (roughly 405 nm) light-emitting diodes.

[0016] In one embodiment, light scatter pulse width time of flight (LSPWTOF) is used in conjunction with high-angle light scatter using micro blue and violet LEDs to correlate LSPWTOF to cell size and composition, similar to Forward Scatter FSC used in flow cytometry. LSPWTOF may be used to extract cell size information at any scattering angle, allowing scattering based on cell internal structures to be used for the classification of blood cell types in 3-part and 5-part blood differentials. Two Micro LED light sources operating at wavelengths of approximately 405 nm and 480 nm may be used for the exciting light and may be placed within a port housing component or an intravascular catheter. The 405 nm light is strongly absorbed by hemoglobin in RBCs and also helps distinguish platelets in a 5-part blood differential. The LEDs may be pulsed to help the signal processing electronics know when and where to look for the cell signals.

[0017] Specifically designed microfluidic channels are utilized to minimize turbulent blood flow and enable measurements of individual blood cell types in vivo. These channels may be successively branched with decreasing cross-sections to increase cell-cell inter-distances for improved single-cell measurements and more reasonable cell count rates. This design also provides a chamber with specified and predictable dimensions, which can yield an indirect, yet useful, measurement of overall blood flow, a measurement of the blood cell count, based on realtime measurements within given time segments and back-calculations.

[0018] In a second embodiment, the blue or violet light is sent from the port to the catheter via optical fibers. The beams exiting the fibers intersect within the microchannel or blood flow interface through optically transparent windows which also create light source beam widths of the appropriate size as well as defining the angular ranges of back scattered light from the blood cells and collected by similar fibers taking the light back to scatter sensors in the port which also contains digital signal processing circuitry to convert these signals to digital single cell data that can be furthered processed using machine learning algorithms.

[0019] In a third embodiment, to identify white blood cells without using micro channels, the underlying measurement method may involve sending light (blue or violet) to an interface between whole blood and an optical surface (polymer or glass, flat or otherwise), while Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION collecting the light scattered from the blood cells at that interface. The central angle between the source and collection optical paths at the interface may be, ideally, between 0 and 90 degrees. With this arrangement, there is a higher magnitude of collected scattered signal from polymorphonuclear leukocytes (PMNs), also known as granulocytes, compared to all other blood cell types. With this difference in the collected scattered signal as the blood flows by, the signal from the PMNs may be quantified over time.

[0020] Devices of the invention may include any intravascular access device, such as a portcatheter or other catheters, including peripherally inserted central catheters (PICCs) or other central venous catheters (CVCs), in which a “port” can be externally secured to the outside of a patient's body. With this invention, devices can also be placed in the peripheral vasculature to count different types of blood cells.

[0021] The sensor data collected in the patient population described herein, may also include, for example, information related to one or more of red blood cells, white blood cells, platelets, proteins, electrolytes, circulating tumor cells, other types of cells or cluster of cells that may be present, microbes, nucleic acids, hemodynamics, including blood flow rate and velocity and cardiac output of the subject, or histological architecture of organs. In some embodiments, sensor data may include identifiable chemicals or drugs for use in clinical pain management applications.

[0022] Aspects of the invention provide implantable devices for in vivo analysis. The implantable devices include a micro cytometer comprising one or more sensor systems configured to sense one or more of light scatter, light absorption, light reflection, light dispersion, light diffraction, and light interference.

[0023] In some embodiments, the implantable device further includes at least one microfluidic chamber configured to allow blood from a vein or artery to flow into the chamber. In some embodiments, the chamber comprises one or more branches configured to suppress turbulent flow and provide a known and measurable blood volume, allowing for the acquisition of, a measurement of the blood cell count, based on real-time measurements within given time segments and back-calculations. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0024] In some embodiments, the implantable device further comprises an optical interface positioned to allow blood from a vein or artery to flow near the interface for optical measurement of the blood.

[0025] In some embodiments, the device comprises a light scatter sensor system comprising at least one light-emitting diode placed at appropriate angles, emitting a plurality of distinct wavelengths of light between 300 and 1000 nanometers.

[0026] In some embodiments, the implantable device further comprises at least one light collecting optic placed at appropriate angles with the ability to capture light scatter, and / or light absorption and / or light reflection and / or light dispersion, and / or light diffraction and / or light interference sensor systems to measure the pulse height and pulse width time-of-flight from individual blood cell signals from the photodetectors.

[0027] In some embodiments, the device comprises a catheter comprising of a cannula, wherein the catheter comprises one or more of a Peripherally Inserted Central Catheter (PICC), a Centrally Inserted Central Catheter (CICC), a tunneled catheter, a central venous catheter (CVC), and an arterial line (ART), and wherein the microfluidic optical sensor system is attached to the intravascular portion of said cannula. In some embodiments, the catheter is fitted for pediatric and neonatal patients, wherein the catheter is implanted to enable infusion of a pharmaceutical, withdraw blood, or administer liquid nutrition. In some embodiments, the device is a catheter fitted for animals, with or without a harness to protect the device port, to enable infusion of a pharmaceutical and / or removal of fluid.

[0028] In some embodiments, the device is operably associated with a machine learning system.

[0029] In some embodiments, the device further comprises a digitally processed multi-angle scattered light from individual cells as they pass near one or more optical sensors.

[0030] In some embodiments, the device further comprises a communication module operable to provide data to a computing device that is external to the subject. In some embodiments, the communication module is operable to provide data to a health care provider or animal care provider.

[0031] In some embodiments, the implantable device is configured to remain in place for at least two days. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10

[0032] PCT APPLICATION

[0033] In some embodiments, the device is protected by one or more of a specialized harness, collar, wristband, and chest band configured to protect the microfluidic optical sensor system when implanted in a subject or animal.

[0034] Aspects of the invention provide methods for monitoring patient health. The methods include the steps of implanting a device into a subject, the device comprising a microfluidic optical sensor system configured to sense an analyte in the subject; and sensing the analyte via the sensor system to thereby assess a health status of the subject.

[0035] In some embodiments of the methods, the analyte comprises a blood cell, a circulating tumor cell, a protein, a microbe, an organic compound, a chemical, a chemical composition, a drug, or a nucleic acid. In some embodiments of the methods, the device is further configured to measure one or more vital signs and parameters comprising one or more of blood pressure, body temperature, cardiac function through heart rate, hemodynamics blood flow rate, velocity and cardiac output, ECG, oxygen level, and electrolyte concentration from the skin. In some embodiments of the methods, the method further comprises sensing, with the microfluidic optical sensor system, one or more of light dispersion, light scattering, light diffraction, or light interference, light reflection and / or light absorption. In some embodiments of the methods, the microfluidic optical sensor system comprises one or more photodetectors, wherein light scattering and / or light diffraction, and / or light interference and / or light reflection and / or light absorption sensed by the one or more photodetectors is useful to assess one or more of size, granularity, nuclear size, shape, organic chemical composition, or cytoplasmic density, of the blood cells. In some embodiments of the methods, the microfluidic sensor system comprises light emitting and light detecting components, wherein the light emitting components comprises one or more of, light-emitting diode of distinct wavelengths of light between 300 and 1000 nanometers, wherein the emitted light may or may not be polarized to aid in differentiating cell types, wherein the emitted light which may or may not be pulsed to aid in signal processing timing, wherein the light detecting components analyze a plurality of distinct light scattering profiles indicative of one or more properties of the blood cells.

[0036] In some embodiments of the methods, the sensor system is configured to begin sensing and to collect data immediately upon implanting the device.

[0037] In some embodiments of the methods, the device further comprises one or more LED excitation light sources and one or more photodetector sensors. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10

[0038] PCT APPLICATION

[0039] In some embodiments of the methods, the device further comprises a catheter comprising a port subcutaneously implanted or held in place external to the body and connected to a reservoir for receiving or injecting fluid by a needle.

[0040] In some embodiments of the methods, the device, when implanted, extends into at least one of a superior vena cava, a right atrium, a peripheral vein or artery, or a central vein or artery.

[0041] In some embodiments of the methods, the device is configured to assess the health status of the subject remotely.

[0042] Aspects of the invention provide methods for collecting research or clinical data. The methods include the steps of receiving, via a device, data based on light scatter properties and other signals sensed by a microfluidic optical sensor system implanted in a subject.

[0043] In some embodiments of the methods, the data is provided from a remote location by a wireless data network.

[0044] In some embodiments of the methods, the data comprises information related to one or more of red blood cells, white blood cells, platelets, circulating tumor cells, microbes, organic compounds, chemicals, chemical composition, drugs, nucleic acids.

[0045] In some embodiments of the methods, the data comprises information on one or more vital signs and parameters, including blood pressure, body temperature, cardiac function through heart rate, hemodynamics, blood flow rate, velocity, and cardiac output, ECG, oxygen level, and electrolyte concentration from the skin.

[0046] In some embodiments of the methods, the data comprises light scatter and / or light absorption and / or light reflection and / or light dispersion, and / or light diffraction and / or light interference data of cells or analytes circulating in the bloodstream of the subject.

[0047] In some embodiments of the methods, the data collected by sensors associated with the device are transmitted to a remote server via local transmission protocols, such as Bluetooth or similar protocols, and then through a secure router to storage on Cloud servers that can be accessed remotely by trained medical professionals.

[0048] In some embodiments of the methods, the method further comprises analyzing the data to generate an assessment of the subject’s health. In some embodiments of the methods, the analyzed data is shown to the recipients as a report. In some embodiments of the methods, the method further comprises providing an alert to a physician based on the assessment of the subject’s health. In some embodiments of the methods, analyzing involves correlating the data Attorney Docket No.: NXGE-003 / 01WG 36543 / 10

[0049] PCT APPLICATION from the subject with other pre-determined parameters associated with a physiological condition. In some embodiments of the methods, the physiological condition comprises one of a genetic disease, an autoimmune disease, a neurologic disease, a metabolic disease, or a chemotherapy- related condition. In some embodiments of the methods, the parameters are established based on input from a health care professional. In some embodiments of the methods, the assessment is used to identify the subject as needing a treatment. In some embodiments of the methods, the method further includes providing the assessment to a healthcare professional and / or the subject. In some embodiments of the methods, wherein the assessment is indicative of a change in the subject’s health.

[0050] In some embodiments of the methods, the method further includes de-identifying or blinding the subject from recipients. In some embodiments of the methods, the subject is a clinical trial participant or de-identified relevant data from other sources. In some embodiments of the methods, the method further comprises providing access control to unblind the data.

[0051] In some embodiments of the methods, the method further comprises providing data auditing and / or tracing capabilities for received data. In some embodiments, the subject is a patient undergoing treatment, an animal, a pediatric patient, or a clinical trial subject. In some embodiments of the methods, the animal comprises one of a pet, a non-human primate, a research animal, a horse, or a cow. In some cases, a harness may be necessary to protect the port from damage due to the movement of the animal.

[0052] Brief Description of the Drawings

[0053] FIG. 1 depicts an implantable venous access port with remote physiological monitoring capabilities, according to one embodiment of the invention.

[0054] FIG. 2 illustrates an implantable device with an optical probe assembly according to one embodiment of the invention.

[0055] FIG. 3 depicts an optical probe assembly in a catheter, according to one embodiment of the invention. FIG. 4 illustrates an in vivo micro flow cytometer configuration of an optical probe assembly according to one embodiment of the systems of the invention. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0056] FIG. 5 illustrates a probe assembly with an in vivo flow cytometer connected to the outside of the catheter according to one embodiment of the invention.

[0057] FIG. 6A illustrates the Hagen-Poiseuille equation used to calculate the flow through the micro-channel device, thereby obtaining the flow rate, flow velocity, and cell separation required to measure the light scatter from cell types and other analytes.

[0058] FIG. 6B illustrates an example of a micro-channel design according to one embodiment of the invention.

[0059] FIG. 7 illustrates a typical implantation of a chemo-port implanted in a pediatric subject.

[0060] FIG. 8A illustrates a tunneled catheter implanted in a neonate, according to one embodiment of the systems of the invention.

[0061] FIG. Figure 8B illustrates a peripherally inserted central catheter (PICC) line, according to one embodiment of the systems of the invention, implanted in a pediatric subject.

[0062] FIG. 9 illustrates examples of implantable intravenous devices with which the systems of the invention may be integrated.

[0063] FIG. 10 illustrates an example of an implantable system for use in animals, according to one embodiment of the invention.

[0064] FIGS. HA through 1 ID depict examples of device harnesses for animals that come in various animal sizes.

[0065] FIG. 12 illustrates a method for remote monitoring of a patient.

[0066] FIG. 13 illustrates a method for remote monitoring of a pediatric subject, according to one embodiment of the invention.

[0067] FIG. 14 illustrates a method for remote monitoring of animal health, according to one embodiment of the invention.

[0068] FIG. 15 illustrates a method for remote monitoring of clinical trial subjects, according to one embodiment of the invention.

[0069] Detailed Description

[0070] This disclosure provides systems and methods for monitoring remote in vivo physiological parameters. In particular, the systems and methods of the invention provide optical probe assemblies integrated with an intravascular device for identifying and quantifying blood cell types intravascularly, determining and quantifying blood analytes, and measuring Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION blood flow properties. The systems and methods of the invention provide implantable devices with one or more optical sensors as part of a miniaturized in vivo cytometer. The systems and methods of the invention address the drawbacks of current ex vivo systems by providing an alternative to conventional blood draws that allows for remote monitoring of patient physiological properties.

[0071] Patients with chronic illnesses, e.g., cancer, make frequent visits to healthcare facilities. The purpose of these visits may be to receive routine treatments and / or undergo health assessments that provide a snapshot of the patient’s health. However, for many patients, visiting a healthcare facility is unduly difficult. Some patients, for example, must commute long distances to reach the nearest healthcare facility. Depending on the severity of a patient’s condition, commuting may require personal assistance, which is not always available. Thus, many patients may not be able to keep scheduled appointments and / or are not monitored in between treatments. As such, physicians are unable to determine whether a treatment is effective or if it is causing harmful side effects, such as organ damage. As such, complications associated with treatment may go undetected.

[0072] The systems and methods address these challenges by providing a microfluidic cytometer embedded in an intravascular access device positioned within the vena cava vessel of a heart. Thus, remote monitoring of patients, for example, cancer patients and / or newborns, for whom the amount of blood needed for blood draws might compromise the health of the patient. Accordingly, systems and methods of the invention are useful for collecting data from one or more analytes in a natural, unperturbed state, the collection of which may not otherwise be possible. As such, the systems of the invention provide in vivo data collection, assessment, and monitoring of patient health, while also offering improved access for remote patients compared to conventional laboratory procedures.

[0073] Systems for in vivo analysis

[0074] Most blood analysis is conducted in vitro, requiring a blood draw. The systems and methods of the invention provide microfluidic devices, for example, with a cytometer embedded within the device. The device is configured for intravascular access for in vivo analysis of physiological parameters.

[0075] Aspects of the invention provide systems for in vivo blood analysis. The systems include an implantable device comprising an optical probe assembly, alternatively referred to herein as a Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION sensor system, which consists of a micro cytometer comprising one or more sensor systems configured to sense one or more of light scatter, light absorption, light reflection, light dispersion, light diffraction, and light interference.

[0076] The systems provide an optical probe assembly suitable for flow cytometry that may be integrated with a catheter, e.g., a port-catheter, such as those described in International PCT Application No. PCT / US2022 / 024635, filed April 13, 2022, incorporated herein by reference in its entirety.

[0077] FIG. 1 illustrates an implantable venous access port catheter with remote physiological monitoring capabilities with which the systems of the invention may be integrated, according to one embodiment of the invention.

[0078] The port-catheter may be dimensioned for complete implantation within a subject’s body, e g., placed under the skin of a subject. The port-catheter may include a reservoir with a selfsealing septum to provide a point of entry to a subject’s central venous system for periodic delivery of treatments, for example, chemotherapy agents.

[0079] FIG. 2 illustrates an implantable device 100 with an optical probe assembly 101.

[0080] The device 100 may include, among other things, a light source 103, e.g., optical fiber, lightemitting diode, a spectrometer 105, a sensor / probe assembly 101, a power supply 107, a computer 109 comprising memory to control various components and / or record measurements, and a communications module 111 to transmit data obtained from the optical probe assembly 101 to a computing device. The computing device may be external to the subject.

[0081] The components may be encased within one housing 113. The housing 113 may be made of a biocompatible metal (e.g., titanium), plastic, polymer (e.g., polyether ether-ketone), or some combination thereof. The material may be selected for its biologically inert properties, which allow the device to be implanted for at least one week and preferably longer, such as at least one month or at least two months, without eliciting an adverse reaction.

[0082] Preferably, the device 100 is dimensioned for surgical insertion under the skin of a subject. The housing 113 may be inserted, for example, in an upper chest region, or an arm, of the subject. After insertion, the housing 113 may appear as a small bump under the skin. The device 100, once inserted, preferably requires no special maintenance.

[0083] The device 100 may further include a cannula 115 that is surgically inserted into a blood vessel (e.g., the jugular vein or artery, or the subclavian vein or artery). Ideally, the cannula 115 Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION may terminate in the superior vena cava or the right atrium. As illustrated, the optical probe assembly 101 may be disposed at a distal portion of the cannula 115.

[0084] In preferred embodiments, devices of the invention are further equipped with at least one additional component operable to sense autofluorescence. Accordingly, in some embodiments, a distal portion of the cannula may include a sensor assembly including a plurality of photosensors. The plurality of photosensors may be arranged in an array format and configured to measure autofluorescence emitted by a circulating analyte in combination with absorbance and / or reflectance of light at one or more specific wavelengths.

[0085] The device 100 may include one or more fiber optic conduits that relay, for example, a two-dimensional array of instantaneous light intensities to a two-dimensional photosensor array. The fiber optic image conduit may be disposed within the cannula 115. The two-dimensional photosensor array may comprise one or more of a complementary metal-oxide-semi conductor (CMOS), a charge-coupled device (CCD), or photodiode arrays of adequate resolution, so that each pixel of the fiber optic image conduit is represented by at least one pixel of the photometric sensor array. Preferably, the two-dimensional photometric sensor array is capable of acquiring light intensity information at a high sampling rate, for example, such as more than 10 frames per second. The high sampling rates enable the detection and analysis of one or more analytes in rapidly moving blood.

[0086] The device 100 may be constructed of discrete optical and optoelectronic components or integrated into an optical and optoelectronic construct, such as, for example, a micro-electro- mechanical system or a photonic integrated circuits-based sensor.

[0087] The device may include a reservoir 117 covered by a self-sealing septum for receiving fluids (e.g., chemotherapy agents).

[0088] The device may include one or more power modules 107. The power module 107 may include several components, including a power manager, a battery, and a charging circuit. The power manager may be configured to manage and maintain the power supply provided by the battery for the various components of the device, including the distal sensor assembly.

[0089] FIG. 3 depicts an optical probe assembly 101 in a catheter, according to one embodiment of the invention.

[0090] The optical probe assembly may include a flow cytometer. Thus, flow cytometry may be used to measure intrinsic physical and / or chemical properties of cells. The chemical and / or Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION physical properties measured may be, in non-limiting examples, cell counting to determine a number of cells in a sample, cell sorting to, for example, separate and identify cells based on specific characteristics / properties, biomarker detection to identify proteins or other molecules on or inside the cells, and size and granularity analysis.

[0091] The optical components may include, in non-limiting examples, a focused light source, such as one or more lasers, optical filters (e.g., dichroic or bandpass filters), and one or more detectors (e.g., photodiodes and / or photomultiplier tubes). The optical components may also include associated lenses and mirrors configured to focus and direct light, thereby optimizing signal collection and resolution.

[0092] The optical probe assembly may have any diameter suitable for surgical insertion into a blood vessel and to contain the detection and microfluidic channel elements. As illustrated in FIG. 3, the optical probe may have an outer diameter of 3 mm to 5 mm. The optical probe assembly may include a source path and a detection path. The optical assembly may include a Gradient-Index (GRIN) lens. The GRIN lens may be configured to couple light between the fibers of the fiber optic system and the sensors and / or detectors. As is understood by a person skilled in the art, GRIN relates to a branch of optics covering optical effects produced by a gradient of the refractive index of a material. The refractive index of a GRIN lens varies gradually from the center to the edge, rather than being uniform throughout as in a conventional lens. Such gradual variation can be used to produce lenses with flat surfaces or lenses that do not have the aberrations typical of traditional spherical lenses. Gradient-index lenses may have a refraction gradient that is spherical, axial, or radial. Accordingly, light entering a GRIN lens follows a sinusoidal path due to continuous bending, allowing for precise control of the beam shape and direction. The GRIN lens may be utilized to shape the laser beam and / or for optical biosensing for light delivery and collection.

[0093] The optical probe assembly may include one or more light-blocking features, one or more prisms, and an interrogation area.

[0094] The flow cytometry may be label-free. Flow cytometry may utilize light scattering, such as forward scattering or side scatter, to determine cell size and / or assess internal complexity / granularity. For example, flow cytometry may include measuring light scattered in the direction of the laser, i.e., forward scatter (FSC), and / or side scatter (SSC), i.e., light scattered at a 90-degree angle. The flow cytometer may be configured to measure autofluorescence. The Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION flow cytometer may be configured to measure changes in electrical impedance. The flow cytometer may be configured to measure Raman scattering.

[0095] In particular embodiments, light scatter results may be used to identify unlabeled human blood cells or other blood analytes using light at blue (roughly 480 nm) and / or blue and violet (approximately 405 nm) light-emitting diodes.

[0096] In one embodiment, light scatter pulse width time of flight (LSPWTOF) is used in conjunction with high-angle light scatter using micro blue and violet LEDs to correlate LSPWTOF to cell size and composition, similar to Forward Scatter FSC used in flow cytometry. Light scatter pulse width, also referred to as “time of flight” (TOF), is a parameter that provides information about the longitudinal size of a cell particle as it passes through the light source. LSPWTOF may be used to extract cell size information at any scattering angle, allowing scattering based on cell internal structures to be used for the classification of blood cell types in 3-part and 5-part blood differentials.

[0097] In some embodiments, two micro LED light sources at wavelengths of approximately 405 nm and 480 nm may be used for the exciting light. It may be placed within a port housing component or an intravascular catheter. The 405 nm light is strongly absorbed by hemoglobin in RBCs and also helps distinguish platelets in a 5-part blood differential. The LEDs may be pulsed to help the signal processing electronics know precisely when and where to look for the cell signals.

[0098] The light source may be, for example, a laser, a superluminescent diode, a light-emitting diode, or a wavelength-tunable light source. The light may be delivered through the cannula 115 via a separate light guide or fiber optic cable, or it may be coupled with an imaging fiber optic bundle using a beam splitter. In preferred embodiments, all optical components are constructed from high-quality optical-grade materials and include antireflection coatings, as needed, to enhance the optical efficiency of the system and minimize stray light dispersion and reflection.

[0099] A single multimode fiber may be used for both light delivery and collection.

[0100] As disclosed herein, in some embodiments, the systems include an optical probe assembly comprising at least one microfluidic chamber configured to receive blood from a vein or artery. Thus, in some embodiments, the system may further comprise an optical interface positioned to allow blood from a vein or artery to flow near the interface for optical measurement of the blood. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0101] FIG. 4 illustrates an in vivo micro flow cytometer configuration of an optical probe assembly according to one embodiment of the systems of the invention. The in vivo micro flow cytometer may include a microfluidic channel configured to spread the spacing between single cells, as well as to suppress turbulent and pulsatile flow, in preparation for single-cell measurement.

[0102] As disclosed in more detail herein, in some embodiments, specifically designed microfluidic channels are utilized to minimize the turbulent flow of blood and to enable the measurement of individual blood cell types in vivo. These channels may be successively branched with decreasing cross-sections to increase cell-cell inter-distances for improved singlecell measurements and more reasonable cell count rates. This design also provides a microfluidic chamber with specified and predictable dimensions, which can yield an indirect, yet useful, measurement of overall blood flow, a measurement of the blood cell count, based on realtime measurements within given time segments and back-calculations.

[0103] As illustrated in FIG. 4, the optical interface can function directly in the blood with or without a micro channel that guides the blood over the optical interface. The channel may be configured to spread the spacings between the cells such that a single cell is analyzed at a time and / or change the speed of blood cells near the optical interface. The channel may be configured to suppress turbulent and pulsatile flow at an inlet of the microfluidic channel and / or through the channel for improved single-cell analysis. As disclosed herein, the probe may include one or more light sources. For example, as illustrated in FIG. 3, the optical probe may include a blue source and a violet source delivered by one or more separate optical fibers. One or more detection channels may sense manipulation of the delivered light. For example, the flow cytometer / optical probe may include one detection channel pair at a 65-degree angle from the microfluidic channel. Alternatively, the flow cytometer / optical probe may have two detection channel pairs at 20 degrees and 65 degrees. The optical fibers and apertures may be embedded in the microfluidic channel structure. The microfluidic channel may include absorbing surfaces to minimize stray light and to define the beam angle. As disclosed in more detail herein, the data collected via the cytometer may be related to one or more analytes and provide clinically useful information. The information may relate to one or more of red blood cells, white blood cells, platelets, circulating tumor cells, microbes, chemicals, drugs, nucleic acids, and / or hemodynamics, and cardiac function, including blood flow rate and velocity and cardiac output, Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION oxygen level, heart rate, body temperature, blood pressure, and ECG measurements of the patient.

[0104] FIG. 5 illustrates a probe assembly with an in vivo flow cytometer connected to the outside of the catheter according to one embodiment of the invention.

[0105] In some embodiments, the system may include a light scatter sensor system comprising at least one light-emitting diode placed at appropriate angles, emitting a plurality of distinct wavelengths of light between 300 and 1000 nanometers.

[0106] As illustrated in FIG. 5, the optical probe assembly may include a sheath that surrounds the catheter, the flow cytometer, and additional associated devices. As disclosed herein, the flow cytometer may include a microfluidic channel for whole blood to enter and exit. The whole blood may be, for example, whole venous blood. As the blood enters the microfluidic channel, the channel design allows for the analysis of a single cell. The flow cytometer may include separate pulsed blue and violet LED light sources. The scattered light may be detected as high- angle scatter violet (VSC) and / or blue scatter (BSC). The flow cytometer may include separate detectors for high-angle violet scatter and blue scatter.

[0107] In some embodiments, the blue or violet light may be sent from the port to the catheter via optical fibers. The beams exiting the fibers may intersect within the microchannel or blood flow interface through optically transparent windows, which also create light source beam widths of the appropriate size, as well as defining the angular ranges of back-scattered light from the blood cells and collected by similar fibers, taking the light back to scatter sensors in the port. The port may also contain digital signal processing circuitry to convert these signals to digital single-cell data that can be further processed using machine learning algorithms.

[0108] In some embodiments, the system may further include at least one light collecting optic placed at appropriate angles with the ability to capture light scatter, and / or light absorption and / or light reflection and / or light dispersion, and / or light diffraction and / or light interference sensor systems to measure the pulse height and pulse width time-of-flight from individual blood cell signals from the photodetectors.

[0109] In some embodiments, to identify white blood cells, the underlying measurement method involves sending light (blue or violet) to an interface between whole blood and an optical surface (polymer or glass, flat or otherwise), while collecting the light scattered from the blood cells at that interface. The central angle between the source and collection optical paths at the interface Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION ideally ranges from 0 to 90 degrees. With this arrangement, there is a higher magnitude of collected scattered signal from the polymorphonuclear leukocytes (PMNs) as compared to all other blood cell types. With this difference in the collected scattered signal as the blood flows by, the signal from the PMNs may be quantified over time.

[0110] As disclosed in more detail herein, in some embodiments, the system includes a catheter comprising a cannula, wherein the catheter comprises one or more of a Peripherally Inserted Central Catheter (PICC), a Centrally Inserted Central Catheter (CICC), a tunneled catheter, a central venous catheter (CVC), and an arterial line (ART), and wherein the microfluidic optical sensor system is attached to the intravascular portion of said cannula. In some embodiments, the catheter is designed for pediatric and neonatal patients, allowing for the implantation to enable infusion of a pharmaceutical, withdrawal of blood, or administration of liquid nutrition. In some embodiments of the system, the catheter is designed for use in animals to enable the infusion of a pharmaceutical and / or the removal of fluid.

[0111] The catheter may include one or more lumens for delivering a treatment to the blood vessel. As illustrated in the example in FIG. 5, the catheter includes a lumen attached to an external port and configured to deliver a chemotherapy drug into the venous system. Treatment of certain medical conditions requires frequent intravascular access. For example, cancer treatment often involves frequent access to a subject’s central venous system to deliver chemotherapy agents. Unfortunately, repeated needle insertions into a subject's blood vessels can lead to narrowing or collapse of the vessels. To avoid these unwanted side effects, implantable port-catheters may be used to provide long-term direct access to a subject’s central venous system. Accordingly, the systems of the invention may include a chemotherapy access port that incorporates a plurality of sensors, which are integrated with, operably or communicatively coupled to, and / or otherwise connected to the chemotherapy access port. The sensors may be part of a sensor assembly, which may be embodied as a system-on-a-chip, such as, for example, a field-programmable gate array, an application-specific integrated circuit, and / or another programmable hardware device.

[0112] The systems of the invention may include a sensor interface module configured to communicate with various physiological sensors that may be integrated into the device. The sensor interface module may be configured to communicate one or more physiological indicators to a computing device, such as a central server via a data network, a healthcare professional’s Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION device, or a local computing device. The sensor interface may include a power bus that is integrated with the sensor connection, supplying power to the sensor micro-assembly, as well as a data bus included in the sensor connection for communicating data between the sensor microassembly and the sensor interface. In some embodiments, this interface may be configured to use the Inter-Integrated Circuit (“I2C”) protocol, a half-duplex bidirectional two-wire bus system for serial communication between different devices, or the Serial Port Interface (SPI) bus protocol, which is a higher-speed bidirectional communication bus between integrated circuits.

[0113] Other physiological sensors may be integrated into systems of the invention, for example, at the tip, or near the tip, of the cannula, or into the optical probe assembly. For example, in some embodiments, a fiber optic-based pressure sensor or a fiber optic-based temperature sensor may be incorporated into the cannula. In some embodiments, a multimodal fiber having a Fiber Bragg grating may be etched within the cannula. The Fiber Bragg grating may comprise a short segment of optical fiber that reflects particular wavelengths of light and transmits all others. In some embodiments, this is achieved by creating a periodic variation in the refractive index of the fiber core, which generates a wavelength-specific dielectric mirror. Accordingly, a fiber Bragg grating may be used as an inline optical filter to block certain undesired wavelengths or may be used as a wavelength-specific reflector. The Fiber Bragg grating may be coupled to other reactive elements, for example, to translate instantaneous changes in pressure or temperature into a spectral pattern that corresponds to such changes in pressure, temperature, or both simultaneously.

[0114] FIG. 6A illustrates the Hagen-Poiseuille equation used to calculate the flow through the micro-channel device, thereby obtaining the flow rate, flow velocity, and cell separation required to measure the light scatter from cell types and other analytes. The Hagen-Poiseuille equation is relevant to the fluid dynamics of the microfluidic channel(s) that guide the cells through the interrogation point. Precise control of fluid flow is essential. The systems of the invention enable single-cell passage through the beam (hydrodynamic focusing), maintain a stable flow rate for consistent signal detection, and prevent turbulence that disrupts measurements. Thus, the microfluidic channel(s) of the optical probe assembly are designed to optimize sample throughput and signal resolution, while also balancing flow rate with detection sensitivity.

[0115] FIG. 6B illustrates an example of a micro-channel design according to one embodiment of the invention. The microchannel design provides a flow rate, flow velocity, and cell Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION separation for improved single or few-cell analysis that allows for accurate light scatter measurement, and therefore measurement of cell type and other analyte ratios. As illustrated in FIG. 6B, the microchannel design is based on the Hagen-Poiseuille equation.

[0116] As disclosed herein, in some embodiments, the system may include, or be operably associated with, a machine learning system. In some embodiments, the system may further include digitally processed, via the machine learning system, multi-angle scattered light from individual cells as they pass near one or more optical sensors. In some embodiments, the system may further include a communication module that is operable to provide data to a computing device external to the subject. In some embodiments, the communication module may be operable to provide data to a health care provider or animal care provider.

[0117] Utilization of an implanted port-catheter device, as provided by this disclosure, for in vivo assessments can enable active measurements at clinically-appropriate interval reporting time points (e.g., hourly, daily, weekly, etc.) as determined by a healthcare professional for early detection of blood count aberrations, such as, anemia, neutropenia, leukocytosis or thrombocytopenia, which can indicate increased risk for infection, inflammation, or bleeding. Furthermore, port-catheter devices of this disclosure also provide for blood flow rate and velocity assessments, which can enable longitudinal measurements at reporting intervals that are clinically appropriate for the early detection of heart aberrations, for example, as measured by cardiac output parameters.

[0118] The systems of the invention may be integrated with an implantable venous access device (IV AD). Systems of the invention may include any intravascular access device, including a portcatheter, or other catheters, including peripherally inserted central catheter (PICC) or other central venous catheters (CVC) in which a “port” can be externally secured to the outside body of a patient. With this invention, devices can also be placed in the peripheral vasculature to count different types of blood cells. IVADs, either centrally or peripherally implanted, are used to assist treatment in children with a variety of diseases. These diseases include neoplasms, hemophilia, long-term supplement needs, and metabolic / endocrine diseases, as examples.

[0119] In the area of neoplasms, approximately 85% of pediatric cancer patients have a longterm chemo-port catheter in use to facilitate drug infusions and blood sampling. Totally implantable port-catheters are preferred in children with solid and hematological malignancies because of decreased pain related to injections, the rate of infection, and the ability to maintain Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION patency for the long term. Despite widespread use, complications in central venous access devices have been reported to be as high as 40%. Pediatric patients receiving conventional chemotherapy for cancer have an increased risk of infections, and represent acute life-threatening events in these immunocompromised patients. Pediatric patients with infections are normally immediately hospitalized and treated with IV antibiotics. Symptom monitoring in pediatric patients through use of patient-reported outcomes (PROs) is uncommon due to difficulty obtaining accurate and consistent information from children experiencing complications.

[0120] The systems and methods of the invention are useful for remotely and passively monitoring pediatric patients for complications by measuring physiologic functions through an implantable port-catheter or other implanted intravenous access devices equipped with flow cytometry technology.

[0121] Thus, the invention provides systems comprising an implantable device (e.g., a port) with remote monitoring capabilities for assessing the health of a patient with an implanted chemo-port or another implanted intravenous access device (IVAD). For example, as disclosed herein, the systems of the invention may include a port with an optical probe assembly capable of remotely measuring symptoms of physiological distress (e.g., fever, heart rate aberrations, blood cell count fluctuations, blood flow velocity, and blood flow rate) via the spectral sensors. In some embodiments, the ports may be used to deliver chemotherapy treatments and monitor patient health throughout the course of chemotherapy. In some embodiments, IVADs may be utilized in non-cancer disease states, such as endocrine, cardiovascular, and autoimmune diseases, and can also be equipped with optical sensors to monitor patient health status.

[0122] The optical probe assembly of the current systems are configured to measure light scattered by one or more analytes present in the body to allow for earlier detection, identification, and quantification of cells indicative of a health status. This is useful for, among other things, early detection of complications associated with implantable ports, such as infections or thrombosis, and to evaluate patients prior to scheduled chemotherapy treatments. Early detection is especially critical for pediatric patients who may not have the ability to describe symptoms of complications, and frequently are hospitalized over the course of cancer treatment or other types of treatment associated with chronic or acute diseases in children.

[0123] The type of IVADs utilized may depend on the access site and include the following: Peripheral venous, midline access, peripheral inserted central catheter (PICC), non-tunneled Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION central catheter, tunneled central venous access (Hickman’s / Broviac or similar), implantable port access, intra-osseous access, arterial access, umbilical (arterial and venous). IVADs can be used to infuse cancer therapy and adjuvant therapies, provide liquid nutrition, and infuse antibiotics and antifungal medicines in the case of infection.

[0124] FIG. 7 illustrates a typical implantation of a chemo-port implanted in a pediatric subject. Catheter sizes may depend on the type and age of the pediatric patient, with ranges of 20-26 G and 1-7 French. Dwell time can also vary depending on the type of IV AD, which can range from short-term (days) to intermediate-term (up to 6 months) or long-term (6 months or longer). As the length of the catheter is typically sized to the individual, systems of the invention may be embodied in one of the following manners: a) the probe assembly, microfluidic channel, and / or the flow cytometer, may be molded into the catheter such that the catheter may be cut to size without damage to the sensng components, b) the system may have various pre-sized catheter lengths, or c) the system may have a dual or triple lumen catheter where the optical probe assembly will be threaded through the secondary lumen to desired length allowing for shortening and attachment at the port. The implant time may vary depending on the treatment, which can range from days (short-term), up to 6 months (intermediate-term) or 6 months or longer (longterm).

[0125] The systems of the invention may be equipped with wireless communication components that provide for remote transmission of clinically actionable data to one or more locations, such as one or more treatment facilities. Systems of the invention, for use in pediatric patients, may further include any of the features described more generally above.

[0126] As disclosed herein, in some embodiments, the assembly of these components may be designed into the “port” of a chemo-port, which is implanted subcutaneously in the subject. In some embodiments, the port may not be implanted subcutaneously, but rather attached to the end of one of the catheter lumens accessible outside the body, and held into place with surgical tape, arm band, chest band, or similar.

[0127] The system may include an optical probe assembly, such as a flow cytometer with a microfluidic channel, placed directly in the bloodstream via a catheter for measuring various analytes as they flow or circulate through the bloodstream. For example, the system / device may be placed in the superior vena cava, and blood may be analyzed as it flows through this vein. Measurements are recorded from the blood, and data are preferably transmitted, e.g., via a Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION communication module, to one or more computing devices external to the subject. For example, the optical probe assembly may be configured to detect and measure various circulating elements in the bloodstream, such as red blood cells, platelets, white blood cells, and / or other circulating components, such as, for example, DNA, proteins, and cancer cells.

[0128] The port of the device may be equipped with components as described herein, and may be placed under the skin near a large vein in the upper chest.

[0129] The data collected in the patient population described herein, may also include, for example, information related to one or more of red blood cells, white blood cells, platelets, proteins, electrolytes, circulating tumor cells, other types of cells or cluster of cells that may be present, microbes, nucleic acids, hemodynamics, including blood flow rate and velocity and cardiac output, oxygen level, heart rate, body temperature, blood pressure, and ECG measurements of the patient.

[0130] The invention may be used to address a long-standing problem in the healthcare industry, which is that throughout patient treatment, the clinical health status of a patient (including pediatric patients) is largely unknown on a cellular level.

[0131] Additionally, systems of the invention are useful to address problems associated with the rising volume of emergency department (ED) visits for pediatric cancer treatment-related toxicities when new or worsening symptoms emerge in between clinic visits. Such problems are well documented. For example, one study in pediatric cancer patients showed that the two most common diagnoses in the ED are fever and fever with neutropenia. Of the patients studied, 44% were admitted to the hospital, and those with febrile neutropenia were admitted at a rate of 82%. Furthermore, recognizing these complications is more difficult and delayed in pediatric patients due to diminished communication ability.

[0132] FIG. 8A illustrates a tunneled catheter implanted in a neonate, according to one embodiment of the systems of the invention. A tunneled catheter is a type of central venous catheter (CVC) with which the systems of the invention may be integrated. In some embodiments, the tunneled central venous catheter may be inserted into a central vein. The system may include an external device port with a lumen, which may be equipped with components such as those described herein. The port may be attached to one end of the doubleaccess catheter. In one embodiment, the loose device port may be taped to the skin of the Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION pediatric subject with surgical tape. The catheter may also have clamps that provide access to or to close off access to the catheter.

[0133] FIG. 8B illustrates a peripherally inserted central catheter (PICC) line, according to one embodiment of the systems of the invention implanted in a pediatric subject. The catheter may be inserted into the vein of the arm. The system may include an external device port with a lumen. The external device may be equipped with the components described herein and may be attached to one end of the double-access catheter. In one embodiment, the loose device port is taped to the skin of the pediatric subject with surgical tape. The catheter may also have clamps that provide access to or close off access to the catheter.

[0134] FIG. 9 illustrates examples of implantable intravenous devices with which the systems of the invention may be integrated. The systems may be used in patients undergoing various treatments for diseases. In some embodiments, the component of the systems containing microelectronics and other components may be external to the body and held in place with, for example, surgical tape, straps, or harnesses. Externally positioned components may be preferable for shorter-term implantation of a catheter that does not require a port for infusing or withdrawing blood. As disclosed herein, a peripherally inserted central catheter (PICC) central venous catheter may also be used to administer therapy. A catheter is inserted into the vein of the arm 1503 until it reaches the heart. PICC lines may be used for short-term (weeks) or longer- term (months) use.

[0135] In some embodiments, the device has an external port with a single or double lumen and is attached to one end of the double-access catheter. In some embodiments, a tunneled venous catheter is inserted into a central vein, the jugular vein, or the femoral vein. In each type of central vein catheter, an external device port with a lumen may be equipped with the components described herein.

[0136] FIG. 10 illustrates an example of an implantable system for use in animals, according to one embodiment of the invention. The system 1001 is illustrated from both external and internal views. The optical probe assembly 1002 is placed directly in a bloodstream via a catheter for measuring various analytes in the blood as it flows or circulates in the bloodstream, for example, through the central vein 1003. Measurements may be recorded from the blood and preferably transmitted, e.g., via a communication module, to one or more computing devices external to the subject. For example, the sensors may be configured to detect and measure various circulating Attorney Docket No.: NXGE-003 / 01WG 36543 / 10

[0137] PCT APPLICATION elements in the bloodstream such as red blood cells, platelets, white blood cells, and / or other circulating components such as, for example, DNA, proteins, and cancer cells. The port 1004 of the device, which may be equipped with components as described herein. The port may be placed under the skin near a large vein in the upper chest. In one embodiment, the port is external and taped to the skin of the animal with surgical tape, and / or protected and enclosed in a harness or collar designed for the device.

[0138] FIGS. 11A through 1 ID depict examples of device harnesses 1100 for animals that come in various animal sizes. The harness 1100 has extra protection 1101 to protect the ports and catheters, as well as the incision on the animal. FIG. HA illustrates an example animal harness designed to secure any portion of the device that extends outside the body, according to one embodiment. FIG. 1 IB illustrates an example of a small animal wearing a harness according to one embodiment of the invention. FIG. 11C illustrates a dog wearing a harness to house the device, according to one embodiment of the invention. FIG. 1 ID illustrates an example harness designed for larger animals. FIGs. 11A and 11C depict the protective pocket 1102 inside the harness, which allows any portion of the device that extends outside the body to be safely tucked in when not in use, and provides easy access to the implanted septum for the delivery and / or removal of fluids. The harness 1100 may have adjustable leg straps or pre-sized openings 1103 for common-sized animals. The harness may have a Velcro, or similar, closure 1104 at the back of the harness. FIG. 1 IB illustrates a harness 1100 according to one embodiment of the invention. FIG. 1 ID depicts a close-up of the opening of the protective pocket 1 102 for easy access to the port and / or catheter openings. The protective pocket may be inside the harness and configured to house any portion of the device that extends outside the body.

[0139] As disclosed herein, the system of the invention may be configured to remain in place for at least two days. In some embodiments, the system may include one or more specialized components, such as a harness, collar, wristband, or chest band, configured to protect the microfluidic optical sensor system when implanted in a subject or animal.

[0140] Methods for in vivo analysis

[0141] As disclosed herein, the invention provides methods for identifying and counting blood cells, blood analytes, and measuring blood flow properties, in vivo, utilizing a microfluidic cytometer embedded in an intravascular access device. The implantable system, referred to herein interchangeably as a device, sensor system, and optical probe assembly, provides remote Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION physiological monitoring capabilities. Thus, the invention provides methods for remotely monitoring a subject and for providing clinically actionable data related to the subject's health.

[0142] Aspects of the invention provide methods for monitoring patient health. The methods include the steps of providing a system for in vivo analysis as disclosed herein; implanting a device into a subject, the device comprising a microfluidic optical sensor system configured to sense an analyte in the subject; and sensing the analyte via the sensor system to thereby assess a health status of the subject.

[0143] FIG. 12 illustrates a method 1200 for remote monitoring of a patient. The system, as described herein, includes an optical probe assembly with a micro cytometer configured to sense an analyte in the subject. The optical probe assembly is implanted in the patient 1202. The method further includes measuring 1204 of an analyte by the sensor to generate data useful for assessing the subject's health status. In further embodiments, the method 1200 includes transmitting 1206 the physiological condition indicators / parameters to a local computing device, which may also be referred to as a local data collection system. In some embodiments, the method 1200 may include transmitting 1208 the physiological condition indicators / parameters to a central server.

[0144] Assessing a health status may involve aggregating and analyzing the data 1210 generated by the sensing step. For example, analyzing 1210 may involve correlating signature profiles with signature profiles taken from a subject with a known health status. The methods of the invention can involve determining 1212 that the physiological condition indicators / parameters are within a predefined / expected range. Then the method 1200 continues to measure 1204 one or more physiological condition indicators / parameters using one or more sensors integrated with the IV AD device.

[0145] If data is in range, the system may generate a report 1214 at intervals determined by the healthcare provider, and send the report 1218 to a treatment facility or the patient’s health care provider. The report may facilitate interactions between the patient and his / her healthcare provider 1220. In certain embodiments, the remote monitoring system may be configured to send patient health alerts 1222 to one or more healthcare providers, such as the subject’s treating physician, when physiologic parameters deviate from the set thresholds for a designated period of time. Alert messages may be generated by the platform and sent to the healthcare providers via wireless communication links (e.g., email or text message), and facilitating action to be taken Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION by the healthcare provider 1224, such as ordering lab tests, blood transfusions, or growth factors, and the method ends.

[0146] In some embodiments, data from the apparatus, system, and methods disclosed herein may provide data that, upon proper authorization, may be entered into a patient's electronic medical records (“EMR”). The apparatus, system, and methods described in the various embodiments above provide several important improvements over existing methods for monitoring the physiological conditions of patients with IVADs. For example, among the beneficial aspects of the apparatus, system, and methods described in the present disclosure and previous disclosures are the ability to monitor physiological functions in an integrated and realtime fashion, and to detect earlier evidence of physiological deviations from expected levels. This saves time for the patient, physician, and laboratory, as well as reducing overall costs to the health care system.

[0147] Methods of the invention may be useful to monitor changes in the health status of chronically ill patients. For example, methods of the invention may be useful for identifying when a chronically ill patient requires treatment and determining the urgency of the treatment. For example, such as a growth factor treatment or a steroid treatment, in conjunction with chemotherapy.

[0148] Regular monitoring of physiological parameters is important in chronically ill patients, particularly cancer patients who are receiving cytotoxic or immunomodulating therapies and who are potentially immunosuppressed. Additionally, monitoring a patient’s physiological responses (e g., body temperature, heart rate and variability, signs of infection, sleep, oxygen levels, glucose, and cortisol, etc.) before and after therapy infusion would be a desirable method to determine if the patient is benefiting from the current treatment strategy. Current methods for monitoring a patient’s physiological parameters primarily focus on the use of external measurement devices such as thermometers (oral, rectal, axillary, ear, or temporal), electrocardiograms (ECG), blood pressure cuffs, and laboratory-based blood analysis. The device may alert physicians if laboratory -based blood analysis is necessary prior to next scheduled treatment.

[0149] As disclosed herein, the systems include an implantable device comprising an optical probe assembly that includes a micro cytometer comprising one or more sensor systems Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION configured to sense one or more of light scatter, light absorption, light reflection, light dispersion, light diffraction, and light interference.

[0150] The systems provide an optical probe assembly suitable for flow cytometry that may be integrated with a catheter, e.g., a port-catheter. The port-catheter may be dimensioned for complete implantation within a subject’s body, e.g., placed under the skin of a subject. The portcatheter may include a reservoir with a self-sealing septum to provide a point of entry to a subject’s central venous system for periodic delivery of treatments, for example, chemotherapy agents. The catheter may include components such as a light source, e.g., optical fiber, lightemitting diode, a spectrometer, a sensor / probe assembly, a power supply, a computer comprising memory to control various components and / or record measurements, and a communications module configured to transmit data obtained from the optical probe assembly to a computing device. The computing device may be external to the subject.

[0151] The components may be encased within one housing. The components may be internal to the device and / or external to the patient. The housing may be made of a biocompatible metal (e.g., titanium), plastic, or polymer (e.g., polyether ether-ketone) or some combination thereof. The material may be selected for its biologically inert properties, which allow the device to be implanted for at least one week and preferably longer, such as at least one month or at least two months, without eliciting an adverse reaction.

[0152] Preferably, the implantable portion of the device is dimensioned for surgical insertion under the skin of a subject. The housing may be inserted, for example, in an upper chest region or in an arm of the subject. After insertion, the housing may appear as a small bump under the skin. The device, once inserted, preferably requires no special maintenance.

[0153] The device may further include a cannula that may be surgically inserted into a blood vessel (e.g., into the jugular vein or artery, or subclavian vein or artery). Ideally, the cannula may terminate in the superior vena cava or the right atrium. As illustrated, the optical probe assembly 101 may be disposed at a distal portion of the cannula.

[0154] In preferred embodiments, devices of the invention are further equipped with at least one additional component operable to sense autofluorescence. Accordingly, in some embodiments, a distal portion of the cannula may include a sensor assembly including a plurality Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION of photosensors. The plurality of photosensors may be arranged in an array format and configured to measure autofluorescence emitted by a circulating analyte in combination with absorbance and / or reflectance of light at one or more specific wavelengths.

[0155] The device may include one or more fiber optic conduits that relay, for example, a two- dimensional array of instantaneous light intensities to a two-dimensional photosensor array. The fiber optic image conduit may be disposed within the cannula. The two-dimensional photosensor array may comprise one or more of a complementary metal-oxide-semiconductor, a charge- coupled device, or photodiode arrays of adequate resolution so that each pixel of the fiber optic image conduit is represented by at least one pixel of the photometric sensor array. Preferably, the two-dimensional photometric sensor array is capable of acquiring light intensity information at a high sampling rate, for example, such as more than 10 frames per second. The high sampling rates allows for one or more analytes in fast moving blood to be readily detected and analyzed.

[0156] The device may be constructed of discrete optical and optoelectronic components or integrated into an optical and optoelectronic construct, such as, for example, a micro-electro- mechanical system or a photonic integrated circuits-based sensor.

[0157] The device may include a reservoir 1 covered by a self-sealing septum for receiving fluids (e.g., chemotherapy agents).

[0158] The device may include one or more power modules. The power module 107 may include several components, including a power manager, a battery, and a charging circuit. The power manager may be configured to manage and maintain the power supply that the battery supplies for the various components of the device including the distal sensor assembly.

[0159] As described herein, the optical probe assembly of the device / system may include a flow cytometer. Thus, flow cytometry may be used to measure intrinsic physical and / or chemical properties of cells. The chemical and / or physical properties measured may be, in non-limiting examples, a measurement of the blood cell count to determine a number of cells in a sample, cell sorting to, for example, separate and identify cells based on specific characteristics / properties, biomarker detection to identify proteins or other molecules on or inside the cells, size and granularity analysis.

[0160] The optical components may include, in non-limiting examples, a focused light source such as one or more lasers, optical filters, for example, dichroic or bandpass filters, and one or more detectors, such as a photodiode and / or photomultiplier tubes. The optical components may Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION also include associated lenses and mirrors configured to focus and direct light and optimize signal collection and resolution.

[0161] The optical probe assembly may have any diameter suitable for surgical insertion into a blood vessel and to contain the detection and microfluidic channel elements. In non-limiting examples, the optical probe assembly may have an outer diameter of 3 mm to 5 mm. The optical probe assembly may include a source path and a detection path. The optical assembly may include a Gradient-Index (GRIN) lens. The GRIN lens may be configured to couple light between the fibers of the fiber optic system and the sensors and / or detectors. As is understood by a person skilled in the art, GRIN relates to a branch of optics covering optical effects produced by a gradient of the refractive index of a material. The refractive index of a GRIN lens varies gradually from the center to the edge, rather than being uniform throughout as in a conventional lens. Such gradual variation can be used to produce lenses with flat surfaces, or lenses that do not have the aberrations typical of traditional spherical lenses. Gradient-index lenses may have a refraction gradient that is spherical, axial, or radial. Accordingly, light entering a GRIN lens follows a sinusoidal path due to the continuous bending, allowing precise control of beam shape and direction. The GRIN lens may be utilized to shape the laser beam and / or for optical biosensing for light delivery and collection.

[0162] The optical probe assembly may include one or more light blocking features, one or more prisms, and an area of interrogation.

[0163] The flow cytometry may be label-free flow cytometry. The flow cytometry may utilize light scattering, such as forward scattering or side scatter to determine cell size and / or determine internal complexity / granularity. For example, the flow cytometry may include measuring light scattered in the direction of the laser, i.e., forward scatter (FSC), and or sided scatter (SSC), i.e., light scattered at a 90-degree angle. The flow cytometer may be configured to measure autofluorescence. The flow cytometer may be configured to measure changes in electrical impedance. The flow cytometer may be configured to measure Raman scattering.

[0164] In particular embodiments of the methods, light scatter results may be used to identify unlabeled human blood cells or other blood analytes using light at blue (roughly 480 nm) and / or blue and violet (roughly 405 nm) light emitting diodes.

[0165] In one embodiment of the methods, light scatter pulse width time of flight (LSPWTOF) is used in conjunction with high angle light scatter using micro blue and violet LEDs to correlate Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0166] LSPWTOF to cell size and composition, similar to Forward Scatter FSC used in flow cytometry. Light scatter pulse width, also referred to as “time of flight” (TOF) is a parameter that provides information about the longitudinal size of a cell particle as it passes through the light source. LSPWTOF may be used to extract cell size information at any scattering angle and allows scattering based on cell internal structures to be used for classification of blood cell types in 3- part and 5-part blood differentials.

[0167] In some embodiments of the methods, two micro LED light sources at wavelengths of approximately 405 nm and 480 nm may be used for the exciting light and may be placed within a port housing component or an intravascular catheter. The 405 nm light is strongly absorbed by hemoglobin in RBCs and also helps distinguish platelets in a 5-part blood differential. The LEDs may be pulsed to help the signal processing electronics know exactly when and where to look for the cell signals.

[0168] The light source may be, for example, a laser, a superluminescent diode, a light-emitting diode, or a wavelength-tunable light source. The light may be delivered through the cannula via a separate light guide, fiber optic cable, or may be coupled with an imaging fiber optic bundle via a beam splitter. In preferred embodiments of the methods, all optical components are constructed from high-quality optical-grade materials and include antireflection coatings, as needed, to enhance the optical efficiency of the system and minimize stray light dispersion and reflection.

[0169] A single multimode fiber may be used for both light delivery and collection.

[0170] As disclosed herein, in some embodiments of the methods, the systems include an optical probe assembly comprising at least one microfluidic chamber configured to receive blood from a vein or artery. Thus, in some embodiments, the system may further comprise an optical interface positioned to allow blood from a vein or artery to flow near the interface for optical measurement of the blood.

[0171] In some embodiments of the methods, the in vivo micro flow cytometer may include a microfluidic channel configured to spread the spacings between single cells, as well as to suppress turbulent and pulsatile flow, in preparation for single-cell measurement.

[0172] As disclosed in more detail herein, in some embodiments of the methods, specifically designed microfluidic channels are utilized to minimize the turbulent flow of blood and to enable the measurement of individual blood cell types in vivo. These channels may be Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION successively branched with decreasing cross-sections to increase cell-cell interdistances for improved single-cell measurements and more reasonable cell count rates. This design also provides a chamber with exact and predictable dimensions, which can yield an indirect, yet useful, measurement of overall blood flow, a measurement of the blood cell count, based on realtime measurements within given time segments and back-calculations.

[0173] In some embodiments of the methods, the microfluidic chamber may include one or more branches configured to suppress turbulent flow and provide a known and measurable blood volume, allowing for the acquisition of a measurement of the blood cell count information.

[0174] In some embodiments of the methods, the flow cytometer may include a microfluidic channel configured for blood to flow through. The channel may be configured to spread the spacings between the cells such that a single or fewer cells are analyzed at a time and / or to change the speed of the blood cells. The channel may be configured to suppress turbulent and pulsatile flow at an inlet of the microfluidic channel and / or through the channel for improved single or few-cell analysis. As disclosed herein, the probe may include one or more light sources. For example, the optical probe assembly may include a blue source and a violet source delivered by one or more separate optical fibers. One or more detection channels may sense the light returned to the detector(s). For example, the flow cytometer / optical probe may include one detection channel pair at a 35-degree angle from the microfluidic channel surface normal. Alternatively, the flow cytometer / optical probe may have two detection channel pairs, one at 20 degrees and 35 degrees to the surface normal. The optical fibers and apertures may be embedded in the microfluidic channel structure. The microfluidic channel may include surfaces to minimize stray light and to define the beam angle. As disclosed in more detail herein, the data collected via the cytometer may be related to one or more analytes and provide clinically useful information. The information may relate to one or more of red blood cells, white blood cells, platelets, circulating tumor cells, microbes, chemicals, drugs, nucleic acids, and / or hemodynamics, and cardiac function, including blood flow rate and velocity and cardiac output, oxygen level, heart rate, body temperature, blood pressure, and ECG measurements of the patient.

[0175] In some embodiments of the methods, the system may include a light scatter sensor system comprising at least one light-emitting diode placed at appropriate angles, emitting a plurality of distinct wavelengths of light between 300 and 1000 nanometers. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0176] The optical probe assembly may include a sheath that surrounds the catheter and the flow cytometer, and additional associated devices. As disclosed herein, the flow cytometer may include a microfluidic channel for whole blood to enter and exit. The whole blood may be, for example, whole venous blood. As the blood enters the microfluidic channel, the channel design allows for the analysis of a single cell. The flow cytometer may include separate pulsed blue and violet LED light sources. The scattered light may be detected as high-angle scatter violet (VSC) and / or blue scatter (BSC). The flow cytometer may include separate detectors for high-angle violet scatter and blue scatter.

[0177] In some embodiments of the methods, the blue or violet light may be sent from the port to the catheter via optical fibers. The beams exiting the fibers may intersect within the microchannel through optically transparent windows, which also create light source beam widths of the appropriate size, as well as defining the angular ranges of back-scattered light from the blood cells and collected by similar fibers, taking the light back to scatter sensors in the port. The port may also contain digital signal processing circuitry to convert these signals to digital single-cell data that can be further processed using machine learning algorithms.

[0178] In some embodiments of the methods, the system may further include at least one light collecting optic placed at appropriate angles with the ability to capture light scatter, and / or light absorption and / or light reflection and / or light dispersion, and / or light diffraction and / or light interference sensor systems to measure the pulse height and pulse width time-of-flight from individual blood cell signals from the photodetectors.

[0179] In some embodiments of the methods, to identify white blood cells, the underlying measurement method may be to send light (blue or violet) to an interface between whole blood and an optical surface (polymer or glass, flat or otherwise) while collecting the light that is scattered from the blood cells at that interface. The central angle between the source and collection optical paths at the interface may be, ideally, between 0 and 90 degrees. With this arrangement, there is a higher magnitude of collected scattered signal from the polymorphonuclear leukocytes (PMNs) as compared to all other blood cell types. With this difference in the collected scattered signal as the blood flows by, the signal from the PMNs may be quantified over time.

[0180] As disclosed in more detail herein, in some embodiments of the methods, the system includes a catheter comprising of a cannula, wherein the catheter comprises one or more of a Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0181] Peripherally Inserted Central Catheter (PICC), a Centrally Inserted Central Catheter (CICC), a tunneled catheter, a central venous catheter (CVC), and an arterial line (ART), and wherein the microfluidic optical sensor system is attached to the intravascular portion of said cannula. In some embodiments, the catheter is fitted for pediatric and neonatal patients, wherein the catheter is implanted to enable infusion of a pharmaceutical or withdraw blood or to administer liquid nutrition. In some embodiments of the system, the catheter is designed for use in animals to enable the infusion of a pharmaceutical and / or the removal of fluid.

[0182] The catheter may include one or more lumens for delivering a treatment to the blood vessel. The catheter may include a lumen attached to an external port and configured to deliver a chemotherapy drug into the venous system. Treatment of certain medical conditions requires frequent intravascular access. For example, cancer treatment often involves frequent access to a subject’s central venous system to deliver chemotherapy agents. Unfortunately, repeated needle insertions into the blood vessels of a subject can lead to narrowing or collapse of the blood vessels. To avoid these unwanted side effects, implantable port-catheters may be used to provide long-term direct access to a subject’s central venous system. Accordingly, the systems of the invention may include a chemotherapy access port that includes a plurality of sensors integrated with, operably or communicatively coupled to, and / or otherwise connected to the chemotherapy access port. The sensors may be part of a sensor assembly, which may be embodied as a system on a chip, such as, for example, a field-programmable gate array, an application-specific integrated circuit, and / or another programmable hardware device.

[0183] The systems of the invention may include a sensor interface module that is configured to communicate with various physiological sensors that may be integrated into the device. The sensor interface module may be configured to communicate one or more physiological indicators to a computing device, such as a central server via a data network, a healthcare professional’s device, or a local computing device. The sensor interface may include a power bus that is included with the sensor connection that supplies power to the sensor micro-assembly, as well as a data bus included in the sensor connection for communicating data between the sensor microassembly and the sensor interface. In some embodiments of the methods, this interface may be configured to use the inter-integrated circuit (“I2C”) protocol, which is a half-duplex bidirectional two-wire bus system for serial communication between different devices, or a Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0184] Serial Port Interface (SPI) bus protocol, which is a higher speed bidirectional communication bus between integrated circuits.

[0185] Other physiological sensors may be integrated into systems of the invention, for example at the tip, or near the tip, of the cannula, or into the optical probe assembly. For example, in some embodiments, a fiber optic-based pressure sensor or a fiber optic-based temperature sensor may be incorporated into the cannula. In some embodiments, a multimodal fiber having a Fiber Bragg grating may be etched within the cannula. The Fiber Bragg grating may comprise a short segment of optical fiber that reflects particular wavelengths of light and transmits all others. In some embodiments, this is achieved by creating a periodic variation in the refractive index of the fiber core, which generates a wavelength-specific dielectric mirror. Accordingly, a fiber Bragg grating may be used as an inline optical filter to block certain undesired wavelengths or may be used as a wavelength-specific reflector. The Fiber Bragg grating may be coupled to other reactive elements, for example, to translate instantaneous changes in pressure or temperature to a spectral pattern that corresponds to such a change in pressure, temperature, or both simultaneously.

[0186] In some embodiments of the methods, the optical probe assembly may include one light source and no microfluidic channels. Thus, light scatter may be measured using only one LED, for example. The flow cytometry takes place via blood flowing near an optical interface positioned on the optical probe assembly. The input light for is directed to the blood flowing near the optical interface, and a detector then measures the scattered (output) light.

[0187] The methods of the invention provide for ensuring single-cell passage through the beam (hydrodynamic focusing), maintaining a stable flow rate for consistent signal detection, and avoiding measurement-disrupting turbulence. Thus, the microfluidic channel(s) of the optical probe assembly are designed to optimize sample throughput and signal resolution, as well as to balance flow rate with detection sensitivity.

[0188] The microchannel design provides a flow rate, flow velocity, and cell separation for improved single or few-cell analysis that allows for accurate light scatter measurement, and therefore measurement of cell type and other analyte ratios. As disclosed herein, the microchannel design may be based on the Hagen-Poiseuille equation.

[0189] As disclosed herein, in some embodiments of the methods, the system may include, or be operably associated with, a machine learning system. In some embodiments, the system may Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION further include digitally processed, via the machine learning system, multi-angle scattered light from individual cells as they pass near one or more optical sensors. In some embodiments, the system may further include a communication module operable to provide data to a computing device that is external to the subject. In some embodiments of the methods, the communication module may be operable to provide data to a health care provider or animal care provider.

[0190] Utilization of an implanted port-catheter device, as provided by this disclosure, for in vivo assessments can enable active measurements at clinically-appropriate interval reporting time points (e.g., hourly, daily, weekly, etc.) as determined by a healthcare professional for early detection of blood count aberrations, such as, anemia, neutropenia, leukocytosis or thrombocytopenia, which can indicate increased risk for infection, inflammation, or bleeding. Furthermore, the port-catheter devices of this disclosure also provide for blood flow rate and velocity assessments, which can enable longitudinal measurements at reporting intervals that are clinically appropriate for early detection of heart aberrations, for example, as measured by cardiac output parameters.

[0191] As disclosed herein, the systems of the invention may be integrated with an implantable venous access device (IVADs). Systems of the invention may include any intravascular access device, including a port-catheter, or other catheters, including peripherally inserted central catheter (PICC) or other central venous catheters (CVC) in which a “port” can be externally secured to the outside body of a patient. With this invention, devices can also be placed in the peripheral vasculature to count different types of blood cells. IVADs, either centrally or peripherally implanted, are used to assist treatment in children with a variety of diseases. These diseases include neoplasms, hemophilia, long-term supplement needs, and metabolic / endocrine diseases, as examples.

[0192] The methods of the invention are useful for remotely and passively monitoring pediatric patients for complications by measuring physiologic functions through an implantable portcatheter or other implanted intravenous access devices equipped with flow cytometry technology.

[0193] FIG. 13 illustrates a method 1300 for remote monitoring of a pediatric subject, according to one embodiment of the invention. The methods include providing a system comprising a device / optical probe assembly as disclosed herein, and implanting 1302 the device. The methods may further comprise the steps of sensing / measuring 1304, via the device / optical probe Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION assetnbly / microfluidic sensor system, one or more physiological parameters; transmitting 1306 data to a local data collection system; transmitting 1308 data to a central server; aggregating and analyzing 1310 the data; determining 1312 if the physiological parameter data is within a defined range; and based on the results of the analysis, continue to monitor, i.e., sense and measure physiological parameters or send 1314 one or more alerts to a healthcare provide. The one or more alerts sent to the healthcare provider may then trigger action 1316 by the healthcare provider.

[0194] The method may further include generating 1318 a report of the measured / detected physiological parameters, and sending 1320 the report to a managing physician, e.g., a healthcare provider. Further, the methods may include facilitating 1322 instructions between the patient’s guardian and the care provider.

[0195] As disclosed herein, the type of IVADs utilized may depend on the access site and include the following: Peripheral venous, midline access, peripheral inserted central catheter (PICC), non-tunneled central catheter, tunneled central venous access (Hickman’ s / Broviac or similar), implantable port access, intra-osseous access, arterial access, umbilical (arterial and venous). IVADs can be used to infuse cancer therapy and adjuvant therapies, provide liquid nutrition, and infuse antibiotics and antifungal medicines in the case of infection.

[0196] Catheter sizes may depend on the type and age of the pediatric patient, with ranges of 20- 26 G and 1-7 French. Dwell time can also vary depending on the type of IV AD, which can range from days (short-term), up to 6 months (intermediate-term), or 6 months or longer (longterm). As the length of the catheter is typically sized to the individual, systems of the invention may be embodied in one of the following manners: a) the probe assembly, microfluidic channel, and / or the flow cytometer, may be molded into the catheter such that the catheter may be cut to size without damage to the sensing components, b) the system may have various pre-sized catheter lengths, or c) the system may have a dual or triple lumen catheter where the optical probe assembly will be threaded through the secondary lumen to desired length allowing for shortening and attachment at the port.

[0197] The methods of the invention may be equipped with wireless communication components that provide for remote transmission of clinically actionable data to one or more locations, such as one or more treatment facilities. As disclosed herein, in some embodiments of the methods, the assembly of these components may be designed into the “port” of a chemo-port, which is Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION implanted subcutaneously in the subject. Tn some embodiments, the port may not be implanted subcutaneously, but rather attached to the end of one of the catheter lumens accessible outside the body, and held into place with surgical tape, arm band, chest band or similar.

[0198] The system may include an optical probe assembly, e.g., a flow cytometer with a microfluidic channel, placed directly in the bloodstream via a catheter for measuring various analytes in the blood as it flows or circulates in the bloodstream. For example, the system / device may be placed in the superior vena cava, and blood may be analyzed as it flows through the superior vena cava. Measurements are recorded from the blood and data preferably transmitted, e.g., via a communication module, to one or more computing devices external to the subject. For example, the optical probe assembly may be configured to detect and measure various circulating elements in the bloodstream, such as red blood cells, platelets, white blood cells, and / or other circulating components, such as, for example, DNA, proteins, and cancer cells.

[0199] The port of the device may be equipped with components as described herein, and may be placed under the skin near a large vein in the upper chest.

[0200] The data collected in the patient population described herein, may also include, for example, information related to one or more of red blood cells, white blood cells, platelets, proteins, electrolytes, circulating tumor cells, other types of cells or cluster of cells that may be present, microbes, nucleic acids, hemodynamics, including blood flow rate and velocity and cardiac output, oxygen level, heart rate, body temperature, blood pressure, and ECG measurements of the patient.

[0201] In some embodiments, the device has an external port with a single or double lumen and is attached to one end of the double-access catheter. In some embodiments, a tunneled venous catheter is inserted into a central vein, the jugular vein, or the femoral vein. In each type of central vein catheter, an external device port with a lumen may be equipped with the components described herein.

[0202] As disclosed herein, in some embodiments of the methods, the analyte detected and analyzed comprises a blood cell, a circulating tumor cell, a protein, a microbe, an organic compound, a chemical, a chemical composition, a drug, or a nucleic acid. Further, in some embodiments of the methods, the device may be further configured to measure one or Attorney Docket No.: NXGE-003 / 01WG 36543 / 10

[0203] PCT APPLICATION more vital signs and parameters comprising one or more of blood pressure, body temperature, cardiac function through heart rate, hemodynamics, blood flow rate, velocity, and cardiac output, ECG, oxygen level, and electrolyte concentration from the skin.

[0204] In some embodiments of the methods, the methods may include sensing, with the microfluidic optical sensor system, i.e., optical probe assembly, one or more of light dispersion, light scattering, light diffraction, or light interference, light reflection, and / or light absorption. In some embodiments of the methods, the microfluidic optical sensor system may include one or more photodetectors, wherein light scattering and / or light diffraction, and / or light interference and / or light reflection and / or light absorption sensed by the one or more photodetectors is useful to assess one or more of size, granularity, nuclear size, shape, organic chemical composition, or cytoplasmic density, of the blood cells. In some embodiments of the methods, the microfluidic sensor system may include light emitting and light detecting components, wherein the light emitting components comprises one or more of, light-emitting diode of distinct wavelengths of light between 300 and 1000 nanometers, wherein the emitted light may or may not be polarized to aid in differentiating cell types, wherein the emitted light which may or may not be pulsed to aid in signal processing timing, wherein the light detecting components analyze a plurality of distinct light scattering profdes indicative of one or more properties of the blood cells.

[0205] In some embodiments of the methods, the sensor system may be configured to begin sensing and to collect data immediately upon implanting the device.

[0206] In some embodiments of the methods, the device may further include one or more LED excitation light sources and one or more photodetector sensors.

[0207] In some embodiments of the methods, the device may further include a catheter comprising a port that is subcutaneously implanted or held in place externally to the body and connected to a reservoir for receiving or injecting fluid via a needle.

[0208] In some embodiments of the methods, the device, i.e., the optical probe assembly, when implanted, may extend into at least one of the following: the superior vena cava, the right atrium, a peripheral vein or artery, or a central vein or artery.

[0209] In some embodiments of the methods, the device may be configured to remotely assess the subject's health status.

[0210] Aspects of the invention provide methods for remote monitoring of animal health. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0211] FIG. 14 illustrates a method 1400 for remote monitoring of animal health, according to one embodiment of the invention. The methods include providing a system comprising a device / optical probe assembly as disclosed herein, and implanting 1402 the device. The methods may further comprise the steps of sensing / measuring 1404, via the device / optical probe assembly / microfluidic sensor system, one or more physiological parameters; transmitting 1406 data to a local data collection system; transmitting 1408 data to a central server; aggregating and analyzing 1410 the data and de-identifying animal information if required; determining 1412 if the physiological parameter data is within a defined range; and based on the results of the analysis, continue to monitor, i.e., sense and measure physiological parameters or send 1414 one or more alerts to an animal care team. The one or more alerts sent to the animal care team may then trigger action 1416 by an appropriate caregiver, e.g., a veterinarian, lead investigator.

[0212] The method may further include generating 1418 a report of the measured / detected physiological parameters, and sending 1420 the report to the animal care team. Further, the methods may include facilitating 14322 instructions between the appropriate animal care team members.

[0213] Aspects of the invention provide methods for remote monitoring of clinical trial subjects.

[0214] FIG. 15 illustrates a method 1500 for remote monitoring of clinical trial subjects, according to one embodiment of the invention. The methods include providing a system comprising a device / optical probe assembly as disclosed herein, and implanting 1502 the device. The methods may further comprise the steps of sensing / measuring 1504, via the device / optical probe assembly / microfluidic sensor system, one or more physiological parameters; transmitting 1506 data to a local data collection system; transmitting 1508 data to a central server; aggregating and analyzing 1510 the data and de-identifying subject information if required; determining 1512 if the physiological parameter data is within a defined range; and based on the results of the analysis, continue to monitor, i.e., sense and measure physiological parameters or send 1514 one or more alerts to principal investigator, physician, nurse, or clinician. The one or more alerts sent to the principal investigator, physician, nurse, or clinician may then trigger action 1516 by an appropriate physician, nurse, or clinician.

[0215] The method may further include generating 1518 a report of the measured / detected physiological parameters, and sending 1520 the report to the animal care team. Further, the Attorney Docket No.: NXGE-003 / 01WG 36543 / 10

[0216] PCT APPLICATION methods may include facilitating 1422 appropriate communication between the subject and a principal investigator, physician, clinician, or nurse.

[0217] Aspects of the invention provide methods for collecting research or clinical data. The methods include receiving, via a system / device as disclosed herein, data based on light scatter properties and other signals sensed by a microfluidic optical sensor system, i.e., optical probe assembly / device implanted in a subject.

[0218] In some embodiments of the methods, the data may be provided from a remote location by a wireless data network. As disclosed herein, in some embodiments of the methods, the data may include information related to one or more of the following: red blood cells, white blood cells, platelets, circulating tumor cells, microbes, organic compounds, chemicals, chemical composition, drugs, and nucleic acids. In some embodiments of the methods, the data may include information on one or more vital signs and parameters, including blood pressure, body temperature, cardiac function through heart rate, hemodynamics, blood flow rate, velocity, and cardiac output, ECG, oxygen level, and electrolyte concentration from the skin. In some embodiments of the methods, the data may include light scatter and / or light absorption and / or light reflection and / or light dispersion, and / or light diffraction and / or light interference data of cells or analytes circulating in the blood stream of the subject. In some embodiments of the methods, the data may be collected by sensors associated with the device. In some embodiments of the methods, the data may be transmitted to a remote server via local transmission protocols, such as Bluetooth or similar protocols, and then through a secure router to storage on Cloud servers that can be accessed remotely by trained medical professionals.

[0219] In some embodiments of the methods, the method may further include analyzing the data to generate an assessment of the subject’s health. In some embodiments of the methods, the analyzed data may be shown to the recipients as a report.

[0220] In some embodiments of the methods, the method may further include providing an alert to a physician based on the assessment of the subject’s health.

[0221] In some embodiments of the methods, analyzing the data may include correlating the data from the subject with other pre-determined parameters associated with a physiological condition. In some embodiments of the methods, the physiological condition may include one or more of a genetic disease, an autoimmune disease, a neurologic disease, a metabolic disease, or a chemotherapy-related condition. In some embodiments of the methods, the parameter may be Attorney Docket No.: NXGE-003 / 01WG 36543 / 10

[0222] PCT APPLICATION established based on input from a health care professional. In some embodiments of the methods, the assessment may be used to identify the subject as needing a treatment. In some embodiments of the methods, the method may further include providing the assessment to a health care professional and / or the subject. In some embodiments of the methods, the assessment may be indicative of a change in the subject’s health.

[0223] In some embodiments of the methods, the method may further include de-identifying or blinding the subject from recipients. For example, in some embodiments, the subject may be a clinical trial participant or de-identified relevant data from other sources. In some embodiments, the method includes methods for providing access control to unblind the data. In some embodiments, the methods provide for data auditing and / or tracing capabilities for received data.

[0224] As disclosed herein, in some embodiments of the methods, the subject may be a patient undergoing treatment, an animal, a pediatric patient, or a clinical trial subject. In some embodiments of the methods, the animal may be one of a pet, a non-human primate, a research animal, a horse, or a cow.

[0225] As disclosed herein, it is an insight of the invention that one or more analytes (e.g., cells, nucleic acids, proteins, etc.) exhibit characteristic light absorption and scattering properties that are unique to clinically relevant information (e.g., size, cellular content, molecular structure) of the analytes. Devices and methods of the invention take advantage of these unique properties to characterize one or more analytes in a subject and, based on the characterizations, assess the health status of a subject. The health assessments are useful for identifying early signs of infection, changes in heart function, blood cell counts, oxygen levels, body temperature, and protein levels (i.e., hemoglobin), as well as monitoring for cancer recurrence and / or detecting device leakage or failure. As disclosed herein, the systems and methods of the invention may employ algorithms, such as machine learning algorithms, to differentiate and assess blood cell subtypes. For example, after appropriate training using blood cell subsets, logistic regression machine learning algorithms can result in automated classification of the cell types for a five-part blood differential including probabilities of correct classifications. Such logistic regression algorithms can be embedded as firmware in the electronics for near real-time classification of blood cell types flowing through the in-vivo cytometer device. Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION

[0226] The systems and methods of the invention may include a control architecture configured to control the components of the system. In some embodiments, the systems may include a console, i.e., a control unit configured to be operably associated with the device and to exchange data therewith, the console comprising a hardware processor coupled to non-transitory, computer-readable memory containing instructions executable by the processor. The instructions cause the console to receive data associated with physiological parameters, process, and combine the received data to generate a report and / or alert.

[0227] The system may generally include one or more processors (e.g., a central processing unit (CPU), a graphics processing unit (GPU), or both) and storage, such as main memory, static memory, or a combination of both, which communicate with each other via a bus or the like. The memory, according to embodiments of the invention, can include a machine-readable medium on which may be stored one or more sets of instructions (e.g., software) embodying any one or more of the methodologies or functions described herein. The software may also reside, completely or at least partially, within the main memory and / or within the processor during execution thereof by the computer system, the main memory and the processor also constituting machine-readable media. The software may further be transmitted or received over a network via the network interface device.

[0228] As used in any embodiment herein, the term “module” may refer to software, firmware, and / or circuitry configured to perform any of the aforementioned operations. Software may be embodied as a software package, code, instructions, instruction sets, and / or data recorded on a non-transitory computer-readable storage medium. Firmware may be embodied as code, instructions, or instruction sets and / or data that are hard-coded (e.g., nonvolatile) in memory devices. “Circuitry”, as used in any embodiment herein, may comprise, for example, singly or in any combination, hardwired circuitry, programmable circuitry such as computer processors comprising one or more individual instruction processing cores, state machine circuitry, and / or firmware that stores instructions executed by programmable circuitry. The modules may, collectively or individually, be embodied as circuitry that forms part of a larger system, such as an integrated circuit (IC), system-on-chip (SoC), desktop computer, laptop computer, tablet computer, server, or smartphone.

[0229] Any of the operations described herein may be implemented in a system that includes one or more storage mediums having stored thereon, individually or in combination, instructions that Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION when executed by one or more processors perform the methods. Here, the processor may include, for example, a server CPU, a mobile device CPU, and / or other programmable circuitry.

[0230] Also, it is intended that operations described herein may be distributed across a plurality of physical devices, such as processing structures at more than one different physical location. The storage medium may include any type of tangible medium, for example, any type of disk including hard disks, floppy disks, optical disks, compact disk read-only memories (CD-ROMs), compact disk rewritables (CD-RWs), and magneto-optical disks, semiconductor devices such as read-only memories (ROMs), random access memories (RAMs) such as dynamic and static RAMs, erasable programmable read-only memories (EPROMs), electrically erasable programmable read-only memories (EEPROMs), flash memories, Solid State Disks (SSDs), magnetic or optical cards, or any type of media suitable for storing electronic instructions. Other embodiments may be implemented as software modules executed by a programmable control device. The storage medium may be non-transitory.

[0231] As described herein, various embodiments may be implemented using hardware elements, software elements, or any combination thereof. Examples of hardware elements may include processors, microprocessors, circuits, circuit elements (e.g., transistors, resistors, capacitors, inductors, and so forth), integrated circuits, application specific integrated circuits (ASIC), programmable logic devices (PLD), digital signal processors (DSP), field programmable gate array (FPGA), logic gates, registers, semiconductor device, chips, microchips, chip sets, and so forth.

[0232] Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0233] The term "non-transitory" is to be understood to remove only propagating transitory signals per se from the claim scope and does not relinquish rights to all standard computer- readable media that are not only propagating transitory signals per se. Stated another way, the meaning of the term "non-transitory computer-readable medium" and "non-transitory computer- readable storage medium" should be construed to exclude only those types of transitory Attorney Docket No.: NXGE-003 / 01WG 36543 / 10 PCT APPLICATION computer-readable media which were found in In Re Nuijten to fall outside the scope of patentable subject matter under 35 U.S.C. § 101.

[0234] The terms and expressions which have been employed herein are used as terms of description and not of limitation. There is no intention, in the use of such terms and expressions, of excluding any equivalents of the features shown and described (or portions thereof). It is recognized that various modifications are possible within the scope of the claims. Accordingly, the claims are intended to cover all such equivalents.

[0235] Incorporation by Reference

[0236] References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, and web content, have been made throughout this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes.

[0237] Equivalents

[0238] Various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including references to the scientific and patent literature cited herein. The subject matter herein contains important information, exemplification and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.

Claims

Attorney Docket No.: NXGE-003 / 01WG 36543 / 10PCT APPLICATIONClaims1. An implantable device comprising of a microfluidic cytometer with one or more sensor systems configured to sense one or more of light scatter, light intensity, light absorption, light reflection, light dispersion, light diffraction, and light interference.

2. The implantable device of claim 1, further comprising at least one microfluidic chamber configured for blood from a vein or artery to flow into and / or through the microfluidic chamber for improved single or few-cell analysis.

3. The implantable device of claim 2, wherein the chamber comprises one or more branches configured to suppress turbulent flow, alter blood speed, and provide a known and measurable blood volume so that a measurement of the blood cell count information can be obtained.

4. The implantable device of claim 1, comprising of an optical interface without a micro channel positioned for blood to flow near the interface for optical measurement of the blood.

5. The implantable device of claim 1, wherein the device comprises a light scatter, reflection, absorption, and transmission sensor system comprising at least one light source placed at appropriate angles, emitting a plurality of distinct wavelengths of light between 300 and 1000 nanometers.

6. The implantable device of claim 1 further comprising at least one light collecting optic placed at appropriate angles with the ability to capture light scatter, and / or light absorption and / or light reflection and / or light dispersion, and / or light diffraction and / or light interference sensor systems to measure the pulse height, pulse width time-of-flight and collected intensity of light from individual blood cell signals from the photodetectors.

7. The implantable device of claim 1, wherein the device comprises a catheter comprising of a cannula, wherein the catheter comprises one or more of a Peripherally Inserted Central Catheter (PICC), a Centrally Inserted Central Catheter (CICC), a tunneled catheter, a central venousAttorney Docket No.: NXGE-003 / 01WG 36543 / 10PCT APPLICATION catheter (CVC), and an arterial line (ART), and wherein the microfluidic optical sensor system is attached to the intravascular portion of said cannula.

8. The implantable device of claim 7, wherein the catheter is fitted for pediatric and neonatal patients, wherein the catheter is implanted to enable infusion of a pharmaceutical, withdraw blood, or administer liquid nutrition.

9. The implantable device of claim 7, wherein the device is a catheter fitted for animals to enable infusion of a pharmaceutical and / or removal of fluid.

10. The implantable device of claim 1, wherein the device is operably associated with a machine learning system.

11. The implantable device of claim 1, further comprising analog and / or digitally processed multi-angle scattered light from individual cells as they pass near one or more optical sensors.

12. The implantable device of claim 1, further comprising a communication module operable to provide data to a computing device that is external to the subject.

13. The implantable device of claim 12, wherein the communication module is operable to provide data to a health care provider or animal care provider.

14. The implantable device of claim 1, configured to remain in place for at least two days.

15. The implantable device of claim 1, wherein the device is protected by one or more of a specialized harness, collar, wristband, and chest band configured to protect the microfluidic optical sensor system when implanted in a subject or animal.

16. A method to monitor patient health, the method comprising:Implanting a device into a subject, the device comprising a microfluidic optical sensor system configured to sense an analyte in the subject;Attorney Docket No.: NXGE-003 / 01WG 36543 / 10PCT APPLICATION sensing the analyte via the sensor system to thereby assess the health status of the subject.

17. The method of claim 16, wherein the analyte comprises a blood cell, a circulating tumor cell, a protein, a microbe, an organic compound, a chemical, a chemical composition, a drug, or a nucleic acid.

18. The method of claim 16, wherein the device is further configured to measure one or more vital signs and parameters comprising one or more of blood pressure, body temperature, cardiac function through heart rate, hemodynamics, blood flow rate, velocity, and cardiac output, ECG, blood oxygenation level, and electrolyte concentration from the skin.

19. The method of claim 16, further comprising sensing, with the microfluidic optical sensor system, one or more of light dispersion, light scattering, light intensity, light diffraction, or light interference, light reflection, and / or light absorption.

20. The method of claim 19, wherein the microfluidic optical sensor system comprises one or more photodetectors, wherein light scattering, and / or light intensity, and / or light diffraction, and / or light interference and / or light reflection and / or light absorption sensed by the one or more photodetectors is useful to assess one or more of size, granularity, nuclear size, shape, organic chemical composition, or cytoplasmic density, of the blood cells.

21. The method of claim 19, wherein the microfluidic sensor system comprises light emitting and light detecting components, wherein the light emitting components comprises one or more of, light-emitting diode of distinct wavelengths of light between 300 and 1000 nanometers, wherein the emitted light may or may not be polarized to aid in differentiating cell types, wherein the emitted light which may or may not be pulsed to aid in signal processing timing, wherein the light detecting components analyze a plurality of distinct light scattering profiles indicative of one or more properties of the blood cells.

22. The method of claim 16, wherein the sensor system is configured to begin sensing and to collect data immediately upon implanting the device.Attorney Docket No.: NXGE-003 / 01WG 36543 / 10PCT APPLICATION23. The method of claim 16, wherein the device further comprises one or more LED, laser, and / or a Super Luminescent Diode (SLD) excitation light source and one or more photodetector sensors.

24. The method of claim 16, wherein the device further comprises a catheter comprising a port subcutaneously implanted or held in place external to the body and connected to a reservoir for receiving or injecting fluid by a needle.

25. The method of claim 16, wherein the device, when implanted, extends into at least one of a superior vena cava, a right atrium, a peripheral vein or artery, or a central vein or artery.

26. The method of claim 16, wherein the device is configured to assess the health status of the subject remotely.

27. A method for collecting research or clinical data, the method comprising:Receiving, via a device, data based on light scatter properties and other signals sensed by a microfluidic optical sensor system implanted in a subject.

28. The method of claim 27, wherein the data is provided from a remote location by a wireless data network.

29. The method of claim 27, wherein the data comprises information related to one or more of red blood cells, white blood cells, platelets, circulating tumor cells, microbes, organic compounds, chemicals, chemical composition, drugs, nucleic acids.

30. The method of claim 27, wherein the data comprises information on one or more vital signs and parameters, including blood pressure, body temperature, cardiac function through heart rate, hemodynamics, blood flow rate, velocity, and cardiac output, ECG, oxygen level, and electrolyte concentration from the skin.Attorney Docket No.: NXGE-003 / 01WG 36543 / 10PCT APPLICATION31 . The method of claim 27, wherein the data comprises light scatter, and / or light intensity, and / or light absorption, and / or light reflection, and / or light dispersion, and / or light diffraction, and / or light interference data of cells or analytes circulating in the blood stream of the subject.

32. The method of claim 27, wherein the data collected by sensors associated with the device are transmitted to a remote server through local transmission protocols such as Bluetooth or similar protocols and then through a secure router to storage on Cloud servers that can be accessed remotely by trained medical professionals.

33. The method of claim 27, further comprising analyzing the data to generate an assessment of the subject’s health.

34. The method of claim 33, in which the analyzed data is shown to the recipients as a report.

35. The method of claim 33, further comprising providing an alert to a physician based on the assessment of the subject’s health.

36. The method of claims 33, wherein analyzing involves correlating the data from the subject with other pre-determined parameters associated with a physiological condition.

37. The method of claim 36, wherein the physiological condition comprises one of a genetic disease, an autoimmune disease, a neurologic disease, a metabolic disease, or a chemotherapy- related condition.

38. The method of claim 36, wherein the parameters are established based on input of a health care professional.

39. The method of claim 33, wherein the assessment is used to identify the subject as needing a treatment.Attorney Docket No.: NXGE-003 / 01WG 36543 / 10PCT APPLICATION40. The method of claim 33, further comprising providing said assessment to a health care professional and / or the subject.

41. The method of claim 33, wherein the assessment is indicative of a change in the subject’s health.

42. The method of claim 33, further comprising de-identifying or blinding the subject from recipients.

43. The method of claim 42, wherein the subject is a clinical trial participant or de-identified relevant data from other sources.

44. The method of claim 42, further comprising providing access control to unblind the data.

45. The method of claim 27, further comprising providing data auditing and / or tracing capabilities for received data.

46. The method of claim 27, wherein the subject is a patient undergoing treatment, an animal, a pediatric patient, or a clinical trial subject.

47. The method of claim 46, wherein the animal comprises one of a pet, a non-human primate, a research animal, a horse, or a cow.

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