WO2022133307A1 - Device and methods for sample analysis - Google Patents

Abstract

Methods, apparatuses, and computer program products for analyzing samples for detecting one or more analytes. For example, certain embodiments may relate to diagnostic tests performed on human subjects for a variety of applications related to health, medical, and wellness testing. Other embodiments may relate to testing in various applications. Further embodiments may relate to processing environmental samples for detection of various analytes of biological or non- biological origin. Additional embodiments may relate to an adaptor or analyzer device compatible with another device for analyzing various analytes of biological or non-biological origin.

Description

DEVICE AND METHODS FOR SAMPLE ANALYSIS

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0001] This invention was made with government support in part by grants 1R43 AH 18180-01 Al, 2R44AI118180-02, and 5R44AI118180-03 awarded by the National Institutes of Health (NIH) and contract 75N93019C00024 awarded by NIH. The government has certain rights in the invention.

CROSS-REFERENCES TO RELATED APPLICATIONS

[0002] This applications claims the benefit of priority to U.S. Provisional Application No. 63/128,046, filed December 19, 2020, and U.S. Provisional Application. No. 63/141,907, filed January 26, 2021, the contents of which are hereby incorporated in the entirety and for all purposes.

FIELD

[0003] Some embodiments may generally relate to diagnostic testing, analysis, and monitoring. For example, certain embodiments may relate to diagnostic tests performed on human subjects for a variety of applications related to health, medical, and wellness testing. Other embodiments may relate to testing in various applications. Further embodiments may relate to processing environmental samples for detection of various analytes of biological or non-biological origin. Additional embodiments may relate to an adaptor or analyzer device compatible with another device for analyzing various analytes of biological or non-biological origin.

BACKGROUND

[0004] Conventional lab-based in vitro diagnostics (IVDs) are tests that are designed to be carried out in a laboratory that contains the essential equipment and supplies needed for sample preparation, running the test or assay, and analyzing the results. For lab-based tests, the sample may be collected from the patient offsite and sent to the lab for analysis. The slow turnaround time to results for lab-based tests has inspired the development of point-of-care (POC) testing technologies that allow testing near the patient for more immediately actionable results. [0005] The field of IVDs enable sensitive and specific detection of a variety of analytes in POC, low-resource, and at-home settings without reliance on sophisticated instrumentation or medical laboratories. Test formats such as the lateral flow assay (LFA) may be used in POC testing and have been adapted for various use applications.

[0006] POC tests that have workflows that may be reasonably straightforward for a trained medical practitioner are often too complex for lay users, and, thus, are not feasible for general consumer at-home self-testing, due to the potential for inaccurate results from user error and variability. Inter-operator variability is an even greater concern for lay users, where it is highly likely that an appreciable percentage of the users may perform the sample preparation step incorrectly. This inter-operator variability is a major concern for developers and manufacturers of medical devices, and for products that are subject to IVD regulations this inter-operator variability and related issues encountered when people use the product must be evaluated and analyzed in well-defined human factors studies.

[0007] Thus, it may be desirable to provide a rapid POC testing/analytic device that has high confidence that the human factor studies and verification and validation studies will be successfully completed, that reduces risk to the user, and is simple to use. Additionally, as the trend of personalized medicine and at-home diagnostics gains traction and interest by the general public and the healthcare and medical industry, there is a need for sample analysis methods and devices that are affordable and can be easily used by a lay person in his or her home, yet still deliver laboratory quality.

SUMMARY

[0008] Certain embodiments may be directed to an active adapter configured to secure/attach and align a processing device with a sampling device including an analyte and at least one reporter or label. In some embodiments, the reporter exhibits luminescence, and preferably persistent luminescence. The active adapter may include a power source, a circuit board connected to the power source, at least one excitation source connected to the circuit board, and a port configured to receive the sampling device. The processing device may be communicatively and wirelessly paired with the circuit board and configured to execute an analysis procedure to analyze the analyte, wherein said analysis procedure may include at least one luminescence imaging cycle including excitation of the reporter followed by termination of the excitation, and then capture of at least one image of the reporter, e.g. bound to the analyte.

[0009] In some embodiments, the processing device may be configured to execute the analysis procedure to analyze the analyte at a plurality of time points. In other embodiments, the analysis procedure may be performed at predefined time intervals, [e.g. at least every 30/60/90 seconds, at least every 2, 3, 4, 5, 6, 7, 8, 9, 10 minutes.

[00010] In some embodiments, the analysis procedure may include a plurality of luminescence imaging cycles, each of which may include an excitation of the reporter followed by capturing an image of the luminescence, and the captured luminescence images are averaged together to improve the signal -to-noise ratio. In other embodiments, a plurality of luminescence images may be captured during each luminescence imaging cycle, and the captured luminescence images are averaged together to improve the signal-to-noise ratio. According to further embodiments, the processing device may communicate with the circuit board in the active adapter to turn on and off the excitation source, and the processing device may be configured to capture one ore more images of the luminescence within tens to hundreds of milliseconds after the excitation source turns off.

[00011] In certain embodiments, the active adapter may include a mechanical switch, the active adapter may be turned on when the sampling device is inserted into the port initiating the switch, wherein the processing device may be turned on separately from the active adapter, and wherein the active adapter may be configured to be paired with the processing device after the active adapter and the processing device are turned on. In other embodiments, pairing of the active adapter and the processing device may occur automatically when the sampling device is inserted into the port. In further embodiments, after the active adapter and the processing device are paired, the processing device may be configured to capture continuous video of a portion of the sampling device.

[0010] In certain embodiments, the active adapter may further include an enclosure base comprising a first side and a second side opposite the first side. The power source may be integrally connected on the first side of the enclosure base and the circuit board may be attached to the second side of the enclosure base. Further, a top section cover may include a lens, the top section cover integrally connected to the second side of the enclosure base. In some embodiments, the active adapter may also include a middle section cover disposed on the second side of the enclosure base and configured to support the processing device, and an enclosure cover configured to secure the processing device to the sample analysis device. According to certain embodiments, the processing device may be an imaging device, a portable mobile communication device, or a smartphone.

[0011] Certain embodiments may be directed to sample analysis device. The sample analysis device may include the active adapter according to any of the embodiments described above, and a processing device attached to and communicatively paired with the active adapter. The processing device and the active adapter may be synchronized, and the synchronization may be repeated periodically. According to certain embodiments, the active adapter and the processing device may each include a respective clock source that are configured to oscillate at a predetermined frequency. According to other embodiments, the clock source of the active adapter and the processing device may be a crystal oscillator, a micro-electromechanical system oscillator, or a quartz crystal oscillator.

[0012] In some embodiments, during the synchronization, the processing device may be configured to perform a low latency time reading of a counter value of the clock of the active adapter, repeat the low latency time reading of the counter value a plurality of times, and take a minimum time reading of the plurality of time readings. In other embodiments, the processing device may be configured to store ticks of the processing device and ticks of the active adapter in two vectors of equal length. In certain embodiments, the processing device may be configured to generate a third vector based on the two vectors, and to calculate a clock offset between the clock source of the processing device and the clock source of the active adapter. According to certain embodiments, the processing device may be configured to establish a consistent time base between the processing device and the active adapter based on the clock offset, and the processing device may be configured to send commands to the active adapter with one or more time-stamps in active adapter tick units based on the clock offset. According to some embodiments, the processing device may be configured to control, based on the clock offset, the active adapter to trigger an excitation for a defined period of time, after which the excitation may be turned off and the processing device may be configured to capture a time-gated image of the reporter, including e.g. any reporter//analyte complex. [0013] Certain embodiments may be directed to a sample analysis kit for analyzing an extracted analyte, including the sample analysis device according to any of the embodiments described above and the sampling device. The sampling device may be adapted and configured to provide a biological analyte into the active adapter. According to certain embodiments, the sampling device may include a lateral flow assay cartridge comprising a test strip, a result window, and a sample well. According to other embodiments, the result window may be aligned with the lens of a camera of the processing device. According to further embodiments, the test strip may include an absorbent pad, a conjugate pad, a filtration pad, a sample pad, and a membrane section comprising a control line and a test line. Further, the membrane section may be aligned with the result window.

[0014] In some embodiments the lateral flow assay cartridge contains a reporter or label for detecting the analyte. The reporter may be a nanoparticle ranging in size from 10 nm to 1000 nm, or a particle in the micron size range ranging from 1 micron to 10 microns. The reporter may exhibit luminescence, persistent luminescence, phosphorescence, long-lived phosphorescence, or fluorescence. The reporter may comprise an inorganic phosphor capable of emitting photons with a wavelength suitable for detection by the optics in the device. In some embodiments the reporter may comprise a particulate label such as a virus, virus fragment, or biological assembly, macromolecule, or other biological particulate conjugate to a phosphorescent molecule capable of time-gate phosphorescence or time-resolved fluorescence detection. In some embodiments the label is an inorganic phosphor that emits persistent luminescence such as strontium aluminate doped with europium and dysprosium. In some embodiments the inorganic phosphor is a material with similar luminescence properties to strontium aluminate.

[0015] Certain embodiments may be directed to a method for analyzing an analyte with the sample analysis device according any of the embodiments described above. The method may include inserting a sampling device into the active adapter and executing, via the processing device, an analysis procedure. The analysis procedure may include pairing the processing device with the active adapter in response to inserting the sampling device into the sample analysis device. The analysis procedure may also include performing a time synchronization procedure to synchronize the processing device with the active adapter in response to inserting the sampling device into the active adapter. The analysis procedure may further include capturing a first brightfield image of the test strip. In addition, the analysis procedure may include determining a position of the test line and the control line by scanning through the brightfield image. Further, the analysis procedure may include adding the analyte to the sample well of the sampling device. The analysis procedure may also include capturing, via the processing device, a plurality of luminescence images of a detection zone located within the result window during each luminescence imaging cycle and in response to each corresponding excitation of the reporter. The analysis procedure may further include analyzing the plurality of luminescence images to calculate test line and control line intensities. In addition, the analysis procedure may include determining a test result based on the intensities of the test line and control line intensities.

[0016] In certain embodiments, the first brightfield image may be captured by utilizing a white light emitting diode. In other embodiments, the test result may be positive if a test line signal is above a predefined threshold, and the test result may be negative if a test line signal is below a predefined threshold. In some embodiments, the test result may be valid if a control line signal is above a predefined threshold, and the test result may be invalid if a control line signal is below a predefined threshold. According to certain embodiments, the plurality of images comprises at least twenty images. According to other embodiments, the processing device may continuously image the detection zone of the strip after the sampling device is inserted into the active adapter in real time / in brightfield mode, and begin the first luminescence imaging cycle when the fluid front flows past a predefined region in the detection zone. According to further embodiments, the predefined time interval may be between one and thirty minutes, preferably at least every 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes, or fractions thereof.

[0017] According to certain embodiments, the processing device may communicate with the active adapter during the time intervals to control a light emitting diode of the active adapter for each time interval. According to other embodiments, the light emitting diode may be controlled to turn on and off. According to further embodiments, when the light emitting diode is turned on, the light emitting diode may be configured to excite reporters, e.g., phosphors on the test strip.

[0018] In certain embodiments, when the light emitting diode is turned off, the processing device may be triggered to capture the plurality of luminescence images. In other embodiments, the method may also include averaging intensity values of each of the plurality of images at the control line and the test line. In some embodiments, the analysis procedure may further include selecting and collapsing, at a time interval, a 2-dimensional image to a 1 -dimensional vector to create a luminescence intensity profile, and applying a global background correction to the luminescence profile. According to certain embodiments, the global background correction may include fitting a polynomial to one or more regions of the luminescence intensity profile that fall outside of test line and the control line. According to other embodiments, the analysis procedure may further include refining a local background correction at the test line. According to further embodiments, refining the local background correction at the test line may include estimating positions for test line peak bounds by analyzing dye bounds calculated from the brightfield image, and by analyzing the global background corrected luminescence profile.

BRIEF DESCRIPTION OF THE DRAWINGS

[0019] For proper understanding of example embodiments, reference should be made to the accompanying drawings, wherein:

[0020] FIG. 1 illustrates an exploded view of an analyzer device, according to certain embodiments.

[0021] FIG. 2 illustrates an assembled analyzer device, according to certain embodiments.

[0022] FIG. 3(a) illustrates a side view of an enclosure base, according to certain embodiments.

[0023] FIG. 3(b) illustrates a top end view of the enclosure base, according to certain embodiments.

[0024] FIG. 3(c) illustrates a bottom end view of the enclosure base, according to certain embodiments.

[0025] FIG. 3(d) illustrates another side view of the enclosure base, according to certain embodiments.

[0026] FIG. 3(e) illustrates a bottom view of a bottom portion of the enclosure base, according to certain embodiments.

[0027] FIG. 3(f) illustrates a top view of a top portion of the enclosure base, according to certain embodiments. [0028] FIG. 3(g) illustrates an isometric view of the enclosure base, according to certain embodiments.

[0029] FIG. 4(a) illustrates a top view of a battery door, according to certain embodiments.

[0030] FIG. 4(b) illustrates a bottom view of the battery door, according to certain embodiments.

[0031] FIG. 4(c) illustrates an isometric view of the battery door, according to certain embodiments.

[0032] FIG. 4(d) illustrates a rear view of the battery door, according to certain embodiments.

[0033] FIG. 4(e) illustrates a front view of the batter door, according to certain embodiments.

[0034] FIG. 4(f) illustrates a side view of the battery door, according to certain embodiments.

[0035] FIG. 5(a) illustrates a top view of a printed circuit board assembly (PCBA) bracket, according to certain embodiments.

[0036] FIG. 5(b) illustrates a bottom view of the PCBA bracket, according to certain embodiments.

[0037] FIG. 5(c) illustrates a back upright view of the PCBA bracket, according to certain embodiments.

[0038] FIG. 5(d) illustrates a bottom view of the PCBA bracket, according to certain embodiments.

[0039] FIG. 5(e) illustrates an isometric view of the PCBA bracket, according to certain embodiments.

[0040] FIG. 5(f) illustrates a side view of the PCBA bracket, according to certain embodiments.

[0041] FIG. 5(g) illustrates another side view of the PCBA bracket, according to certain embodiments.

[0042] FIG. 6(a) illustrates a side view of a bottom securing piece, according to certain embodiments.

[0043] FIG. 6(b) illustrates a side interior of the bottom securing piece, according to certain embodiments.

[0044] FIG. 6(c) illustrates a top view of the bottom securing piece, according to certain embodiments. [0045] FIG. 6(d) illustrates a bottom side view of the bottom securing piece, according to certain embodiments.

[0046] FIG. 6(e) illustrates another side view of the bottom securing piece, according to certain embodiments.

[0047] FIG. 6(f) illustrates a bottom view of the bottom securing piece, according to certain embodiments.

[0048] FIG. 6(g) illustrates an isometric view of the bottom securing piece, according to certain embodiments.

[0049] FIG. 7(a) illustrates a right side view of a middle section cover, according to certain embodiments.

[0050] FIG. 7(b) illustrates a top view of the middle section cover, according to certain embodiments.

[0051] FIG. 7(c) illustrates a left side view of the middle section cover, according to certain embodiments.

[0052] FIG. 7(d) illustrates a bottom view of the middle section cover, according to certain embodiments.

[0053] FIG. 7(e) illustrates top side view of the middle section cover, according to certain embodiments.

[0054] FIG. 7(f) illustrates a bottom end side view of the middle section cover, according to certain embodiments.

[0055] FIG. 7(g) illustrates an isometric view of the middle section cover, according to certain embodiments.

[0056] FIG. 8(a) illustrates a right side view of a top section cover, according to certain embodiments.

[0057] FIG. 8(b) illustrates a top view of the top section cover, according to certain embodiments.

[0058] FIG. 8(c) illustrates a left side view of the top section cover, according to certain embodiments.

[0059] FIG. 8(d) illustrates a bottom view of the top section cover, according to certain embodiments. [0060] FIG. 8(e) illustrates a top end view of the top section cover, according to certain embodiments.

[0061] FIG. 8(f) illustrates an interior view of the top section cover, according to certain embodiments.

[0062] FIG. 8(g) illustrates an isometric view of the top section cover, according to certain embodiments.

[0063] FIG. 9 illustrates a signal processing pipeline output, according to certain embodiments.

[0064] FIG. 10 illustrates a test strip and components, according to certain embodiments.

[0065] FIG. 11 illustrates an assembled test cartridge, according to certain embodiments.

[0066] FIG. 12(a) illustrates an analyzer device with the test cartridge inserted, according to certain embodiments.

[0067] FIG. 12(b) illustrates a PCBA, according to certain embodiments.

[0068] FIG. 13 illustrates a method of assembling the analyzer device, according to certain embodiments.

[0069] FIG. 14(a) illustrates an apparatus, according to certain embodiments.

[0070] FIG. 14(b) illustrates another apparatus, according to certain embodiments.

[0071] FIG. 15 illustrates a reusable analyzer and kit, according to certain embodiments.

DETAILED DESCRIPTION

Introduction:

[0072] POC tests may range in complexity and ease of use with some tests requiring access to certain kinds of equipment found in a conventional medical lab, while other POC tests are capable of being administered by a healthcare professional in low-resource or field-use settings. A subset of POC tests are sufficiently simple such that they can be carried out by a layperson for convenient-at-home self-testing. The field of IVDs has seen a wide number of innovations in assay formats and detection methods that enable sensitive and specific detection of a variety of analytes in POC, low-resource, and at-home settings without reliance on sophisticated instrumentation or medical laboratories. [0073] The purpose of the sample analysis step may involve sample preparation, which may in turn vary depending on the nature of the test, but generally may be used to convert the sample into a form that is more compatible with the format or chemistry of the assay or to make the analyte more available for detection. Sample preparation may involve chemical or physical breakdown, removal, separation, or processing of the sample material into components that are more easily detected by the assay, and may introduce chemical species that enhance sensitivity or specificity, reduce interference, reduce the coefficient of variation, enhance quantitation, or generally improve the assay accuracy, precision, or performance. In some applications it may be essential to dilute the original sample into a buffer or reagent solution, at a controlled volume and dilution factor, to improve assay consistency and performance by decreasing the concentration of interfering components in the sample. In some applications, sample preparation steps may include access to equipment used in medical laboratories, such as pipettes to measure out quantities of various chemical reagents, vortexers, vortex mixers, agitators, or shakers to mix the sample and reagents, sonication equipment or sonicators to aid in extraction of the analyte or for lysis, centrifuges or other tools for separation of plasma or cells from biological fluids such as blood, and a variety of other tools for mixing, reagent dispensing, separation, heating or other chemical, mechanical, or physical processes.

[0074] With regard to the design of sample analysis devices and methods, an often- overlooked area of importance is hedonomics, which examines the pleasure or satisfaction the user experiences while engaging with the device. Thus, even if the basic ergonomic considerations are properly accounted for, and a functional sample analysis process and accompanying assay can be run properly by an untrained lay user, there remains the possibility that the device and sample analysis process or other steps needed to run the test are so complex that many users would be unsatisfied with the experience and would not want to use the device in the future.

[0075] As the trend of personalized medicine and at-home diagnostics gains traction and interest by the general public and the healthcare and medical industry, there is a need for sample analysis methods and devices that are affordable and can be easily used by a lay person in his or her home, yet still deliver laboratory quality performance. Faced with the challenge of sample analysis in low-resource or OTC settings, the available options are either cheap, but inaccurate, highly variable, and potentially hazardous to lay users, or precise and safe but complex and prohibitively expensive. The methods and devices of certain embodiments described herein address the unmet need of sample analytical tools that greatly simplify the workflow of sample analysis, such that both a lay user and trained professional using the same devices and methods would be able to achieve comparable performance and consistency of sample preparation of a variety of sample types for analysis in assays or analytical procedures. The methods and devices of certain embodiments described herein may be particularly advantageous in processing or analysis of samples such as saliva, blood, urine, feces, sputum, and others. Further, the methods and devices of certain embodiments described herein may have broad applications outside of human medical testing and diagnostics, such as veterinary testing, environmental monitoring, contamination detection, or preparing any arbitrary sample type for analysis by an analytical technique. In addition, the methods and devices of certain embodiments described herein may also create new opportunities in mail-order or mail-in diagnostics, wherein a user collects and processes a sample at home, and sends or transports the sample to a laboratory for analysis.

[0076] It will be readily understood that the components of certain example embodiments, as generally described and illustrated in the figures herein, may be arranged and designed in a wide variety of different configurations. The following is a detailed description of some example embodiments of methods and apparatuses for sample preparation.

[0077] The features, structures, or characteristics of example embodiments described throughout this specification may be combined in any suitable manner in one or more example embodiments. For example, the usage of the phrases “certain embodiments,” “an example embodiment,” “some embodiments,” or other similar language, throughout this specification refers to the fact that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment. Thus, appearances of the phrases “in certain embodiments,” “an example embodiment,” “in some embodiments,” “in other embodiments,” or other similar language, throughout this specification do not necessarily all refer to the same group of embodiments, and the described features, structures, or characteristics may be combined in any suitable manner in one or more example embodiments. In addition, as discussed herein, a sample preparation device may be referred to as the sample prep device, the preparation pod, the prep pod, the sample preparation pod, the sample prep pod, or simply the device. [0078] Where a numerical value is specified herein as qualified by the term “about”, it is understood that the disclosed value is intended to include without limitation both the specific value specified and a range of values of ±10%.

[0079] One having ordinary skill in the art will readily understand that the invention as discussed above may be practiced with steps in a different order, and/or with hardware elements in configurations which are different than those which are disclosed. Therefore, although the invention has been described based upon these example embodiments, it would be apparent to those of skill in the art that certain modifications, variations, and alternative constructions would be apparent, while remaining within the spirit and scope of example embodiments.

[0080] Further, it is to be understood that both the foregoing general description and the following detailed description are illustrative and explanatory, and are not restrictive of the subject matter, as claimed. In this application, the use of the singular includes the plural, the word “a” or “an” means “at least one”, and the use of “or” means “and/or”, unless specifically stated otherwise. Furthermore, the use of the term “including”, as well as other forms, such as “includes” and “included”, is not limiting. Also, terms such as “element” or “component” may encompass both elements or components comprising one unit and elements or components that comprise more than one unit unless specifically stated otherwise. Moreover, parameters disclosed herein (e.g., volume, temperature, time, concentrations, length, etc.) may be approximate.

Devices and Methods:

[0081] As described herein, certain embodiments enable simple analysis of analytes from biological or non-biological samples. The device may include a mechanism to enable analysis of a dispensed extracted liquid sample on a secondary device such as an assay or analyte detection device. In some embodiments the sample analysis device may have features that enable it to analyze an extracted liquid sample on a secondary device such as an assay or analyte detection device. In other embodiments, the sample analysis device may include features that enable it to mate with a lateral flow test cartridge or cassette. According to certain embodiments, the device may be used for on-site sample analysis.

[0082] FIG. 1 illustrates an exploded view of an analyzer device 100, according to certain embodiments. As illustrated in FIG. 1, the analyzer device 100 may include an enclosure base 105 that is configured to receive various attachable components. For instance, in certain embodiments, a battery holder 110 may be assembled onto one side of the enclosure base 105 via two screws 115. According to certain embodiments, the battery holder 110 may be configured to hold one or more batteries to serve as a power source for the analyzer device 100. Once assembled onto the enclosure base 105, the battery holder 110 may be covered by a battery door 120, which may also be attached to the enclosure base 105. In certain embodiments, the enclosure base 105 may include a printed circuit board assembly (PCBA) bracket 125 configurable to receive a PCBA 130. As illustrated in FIG. 1, the PCBA bracket 125 may be attached to the enclosure base 105 via two screws 135 into corresponding through-holes of the PCBA bracket 125. Further, the PCBA 130 may define two through-holes 185, which may be configured to receive the two screws 135 threaded through the PCBA bracket.

[0083] In certain embodiments, the analyzer device 100 may also include a top section cover 140. The top section cover 140 may be attached to another side of the enclosure base 105, opposite the side that the battery holder 110 is attached. According to certain embodiments, a lens bracket 145 may be placed in an opening 150 defined by the top section cover 140. In certain embodiments, the lens bracket 145 may be configured to hold a lens. According to other embodiments, the lens bracket 145 may include pegs (not shown) that may be visible and face outward, away from a bottom surface 155 of the top section cover 140. In certain embodiments, the lens that is placed within the lens bracket 145 may have a curved surface that also faces away from the bottom surface 155 of the top section cover 140. As illustrated in FIG. 1, the top section cover 140 may be attached to the enclosure base 105 via screws 160, which can be threaded into corresponding through-holes of the enclosure base 105 and the top section cover 140.

[0084] According to certain embodiments, the analyzer device 100 may also include a middle section cover 165. As illustrated in FIG. 1, the middle section cover 165 may be attached to the enclosure base 105 via screws 170, which may be threaded into corresponding through-holes of the enclosure base 105 and the middle section cover 165. In other embodiments, the analyzer device 100 may include a bottom securing piece or enclosure cover 180. As described herein, the bottom securing piece and enclosure cover may be used interchangeably. The bottom securing piece 180 may be attached to the enclosure base 105 via a screw 185, which may be threaded into corresponding through-holes of the enclosure base 105 and the bottom securing piece 180. According to certain embodiments, the top section cover 140 and the bottom securing piece 180 may be integrally connected to opposite ends of the middle section cover 165.

[0085] In some embodiments, the analyzer device 100 may include a temperature sensor (not shown), which may measure the temperature of the environment in which the analyzer device 100 is being operated. In certain embodiments, if the temperature of the operating environment is not within a predefined acceptable range, the analyzer device may be configured to transmit a warning or similar type message to a processing device communicatively connected (e.g., wirelessly) to the analyzer device, indicating to a user or operator that accuracy of the results produced by the analyzer device and processing device may be compromised. In other embodiments, if the measured temperature is not within an acceptable predefined operating range, the analyzer device may be configured to not operate until the temperature is within the acceptable operating range. According to other embodiments, phosphor luminescence emission may depend on temperature, and it may be possible to leverage the temperature data to normalize luminescence signal(s) and improve accuracy of quantization of results.

[0086] FIG. 2 illustrates an assembled analyzer device 200, according to certain embodiments. As illustrated in FIG. 2, the analyzer device 200 may be configured to receive a portable electronic device (not shown). According to certain embodiments, the portable electronic device may include a smartphone, tablet, wireless terminal, or the like. As illustrated in FIG. 2, the portable electronic device may be mounted and secured on the assembled analyzer device 200. For example, the portable electronic device may be secured onto the assembled analyzer device 200 by the bottom securing piece 205 and the top section 210. Once secured, there may be no gaps that appear between portions of the assembled analyzer device 200 and the portable electronic device, and it may be determined that the portable electronic device is properly aligned in the assembled analyzer device 200.

[0087] According to other embodiments, the analyzer device 200 may include a cartridge port 215 located on the enclosure base 220. In certain embodiments, the cartridge port 215 may be configured to receive a lateral flow assay cartridge (not shown). In other embodiments, the portable electronic device may be used for signal acquisition and readout or analysis of the test results of the lateral flow assay cartridge. According to certain embodiments, the rear camera on the portable electronic device, which may be aligned with the lens on the analyzer device 200, may be used to capture images of a result window of the lateral flow assay cartridge to analyze the signal by image processing and image analysis. In certain embodiments, a software application or app on the portable electronic device may continuously capture video or images of the lateral flow assay cartridge result window or analyte detection zone to determine if a sample (e.g., biological sample, non-biological sample, or liquid, etc.) has been added to the cartridge for automated timing of the assay duration and automated timing of when to initiate signal acquisition. [0088] FIGs. 3(a) - 3(g) illustrate different views of the enclosure base 300, according to certain embodiments. For instance, FIG. 3(a) illustrates a side view of the enclosure base 300. As illustrated in FIG. 3(a), the enclosure base 300 may include a bottom face 310 and a top face 315. As discussed above, the top face 315 of the enclosure base 300 may be configured to receive a portable electronic device. Further, as illustrated in FIG. 3(a), the enclosure base 300 may include a cartridge port 305. The cartridge port 305 may be located at a side surface 320 of the enclosure base 300.

[0089] FIG. 3(b) illustrates a top end view of the enclosure base 300, and FIG. 3(c) illustrates a bottom end view of the enclosure base 300. In both FIGs. 3(b) and 3(c), the cartridge port 305 may be disposed on a side portion of the enclosure base 300. Additionally, FIG. 3(d) illustrates another side view of the enclosure base 300. As illustrated in FIG. 3(d), the bottom face 310 may be facing upwards, and the top face 315 may be facing downwards. Further, FIG. 3(d) illustrates another side surface 325 of the enclosure base 300, which is opposite the side surface 320 illustrated in FIG. 3(a).

[0090] FIG. 3(e) illustrates a bottom view of a bottom portion of the enclosure base 300. As illustrated in FIG. 3(e), the enclosure base 300 may include a backside plate 330, and battery holder 335. The backside plate 330 may include a plurality of through-holes 340 that may be configured to receive corresponding screws to attach the top section and middle section cover of the analyzer device to the enclosure base 300. FIG. 3(e) also illustrates additional through-holes 345 that may be configured to receive corresponding screws to attach the middle section cover to the enclosure base 300. FIG. 3(e) further illustrates that the enclosure base 300 may include a further through-hole 350, which may be configured to receive a corresponding screw to attach the bottom securing piece to the enclosure base 300. [0091] FIG. 3(f) illustrates a top view of a top portion of the enclosure base 300, and FIG. 3(g) illustrates an isometric view of the enclosure base 300. As illustrated in FIGs. 3(f) and 3(g), the enclosure base 300 may include through-holes 340, which may be configured to receive corresponding screws to attach the top section of the analyzer device to the enclosure base 300. The enclosure base 300 may also include through -holes 360, which may be configured to receive corresponding screws to attach the middle section cover to the enclosure base 300. In addition, the enclosure base 300 may include a through-hole 350, which may be configured to receive a corresponding screw to attach the bottom securing piece to the enclosure base 300. Further, FIGs. 3(f) and 3(g) illustrate that the enclosure base 300 may include through-holes 365, which may be configured to receive corresponding screws to attach the battery holder to the enclosure base. According to certain embodiments, the enclosure base 300 may define an opening 370 through which the PCBA and corresponding wires may be passed through, enabling the PCBA to be attached to the top portion of the enclosure base 300. In addition, the enclosure base 300 may include through-holes 355, which may be configured to receive corresponding screws to attach the PCBA bracket to the enclosure base 300.

[0092] FIGs. 4(a) - 4(f) illustrate different views of the battery door 400, according to certain embodiments. For instance, FIG. 4(a) illustrates a top view of the battery door 400, and FIG. 4(b) illustrates a bottom view of the battery door 400. In addition, FIG. 4(c) illustrates an isometric view of the battery door 400, and FIG. 4(d) illustrates a rear view of the battery door 400. Further, FIG. 4(e) illustrates a front view of the battery door 400, and FIG. 4(f) illustrates a side view of the battery door 400.

[0093] As illustrated in FIGs. 4(a) - 4(f), the battery door may 400 include a flange 405, and two attachment pieces 410. According to certain embodiments, the flange 405 and attachment pieces 410 may be configured to secure the battery door 400 to the enclosure base. As illustrated in FIG. 4(d), the back surface of the flange 405 may include ribbed portions 415 extending along the length of the flange 405. According to other embodiments, the back surface of the battery door 400 may also include ribbed portions 420 that extend in a vertical and horizontal direction across the back surface of the battery door 400, as illustrated in FIG. 4(b). In some embodiments, the ribbed portions 420 may provide rigidity and strength to the battery door 400. [0094] FIGs. 5(a) - 5(g) illustrate different views of the PCBA bracket 500, according to certain embodiments. FIG. 5(a) illustrates a top view of the PCBA bracket 500, and FIG. 5(b) illustrates a bottom view of the PCBA bracket 500. Further, FIG. 5(c) illustrates a back upright view of the PCBA bracket 500, and FIG. 5(d) illustrates a bottom view of the PCBA bracket 500. In addition, FIG. 5(e) illustrates an isometric view of the PCBA bracket 500. Further, FIG. 5(f) illustrates a side view of the PCBA bracket 500, and FIG. 5(g) illustrates another side view of the PCBA bracket 500.

[0095] As illustrated in FIGs. 5(a) - 5(g), the PCBA bracket may include a crossbar 505 connecting screw mounts 515 on opposite ends of the crossbar 505. In certain embodiments, the screw mounts 515 may define respective through-holes 520, which may be configured to receive corresponding screws to attach the PCBA bracket 500 to the enclosure base. The PCBA bracket 500 may also include a pair of hooks 510 located at respective end pieces 525 of the PCBA bracket 500. According to certain embodiments, the hooks 510 may be configured to grasp and secure the PCBA onto the enclosure base, and prevent the PCBA from moving in the enclosure base.

[0096] FIGs. 6(a) - 6(g) illustrate different views of the bottom securing piece or enclosure cover 600, according to certain embodiments. For instance, FIG. 6(a) illustrates a side view of the bottom securing piece 600, and FIG. 6(b) illustrates a side interior of the bottom securing piece 600. In addition, FIG. 6(c) illustrates a top view of the bottom securing piece 600, and FIG. 6(d) illustrates a bottom side view of the bottom securing piece 600. Further, FIG. 6(e) illustrates another side view of the bottom securing piece 600, FIG. 6(f) illustrates a bottom view of the bottom securing piece 600, and FIG. 6(g) illustrates an isometric view of the bottom securing piece 600.

[0097] As illustrated in FIGs. 6(a) and 6(e), the bottom securing piece may include a bottom side surface 605 and a top side surface 610. The bottom securing piece 600 may also include a central opening 615, and two side openings 620. In addition, the bottom securing piece 600 may include a through-hole 625, which may be configured to receive a corresponding screw to attach the bottom securing piece to the enclosure base.

[0098] FIGs. 7(a) - 7(g) illustrate different views of the middle section cover 700, according to certain embodiments. For instance, FIG. 7(a) illustrates a right side view of the middle section cover 700, FIG. 7(b) illustrates a top view of the middle section cover 700, and FIG. 7(c) illustrates a left side view of the middle section cover 700. In addition, FIG. 7(d) illustrates a bottom view of the middle section cover 700, and FIG. 7(e) illustrates a top end side view of the middle section cover 700. Further, FIG. 7(f) illustrates a bottom end side view of the middle section cover 700, and FIG. 7(g) illustrates an isometric view of the middle section cover 700.

[0099] As illustrated in FIGs. 7(a) - 7(f), the middle section cover 700 may include a bottom surface 705, and a top surface 710. According to certain embodiments, the middle section cover 700 may include a plurality through-holes 715, which may be configured to receive corresponding screws to attach the middle section cover 700 to the enclosure base. According to other embodiments, the middle section cover may include a first notch 720, and a second notch 725. As illustrated in FIGs. 7(a) - 7(g), the first notch 720 and the second notch 725 may be different sizes. In certain embodiments, the first notch 720 and the second notch 725 may be configured to provide access to functionalities of the portable electronic device when attached to the analyzer device.

[00100] FIGs. 8(a) - 8(g) illustrate different views of the top section cover 800, according to certain embodiments. For instance, FIG. 8(a) illustrates a right side view of the top section cover 800, and FIG. 8(b) illustrates a top view of the top section cover 800. Further, FIG. 8(c) illustrates a left side view of the top section cover 800, and FIG. 8(d) illustrates a bottom view of the top section cover 800. In addition, FIG. 8(e) illustrates a top end view of the top section cover 800, and FIG. 8(f) illustrates an interior view of the top section cover 800. Furthermore, FIG. 8(g) illustrates an isometric view of the top section cover 800.

[0100] As illustrated in FIGs. 8(a) - 8(g), the top section cover 800 may include an interior hole 820 configured to receive a lens bracket 825 and a lens 830 in the lens bracket. The top section cover 800 may also include through-holes 815, which may be configured to receive corresponding screws to fix the top section cover 800 to the enclosure base. According to other embodiments, the top section cover 800 may include one or more pegs (not shown).

[0101] FIG. 9 illustrates a signal processing pipeline output, according to certain embodiments. As previously noted, the analyzer device may be configured to receive a portable electronic device such as, for example, a smartphone. In certain embodiments, the analyzer device may use the smartphone to capture images of luminescence from reporters and/or labless that have been designed to detect specific targets including, for example, viruses, bacteria, proteins, molecules, or other analytes. According to certain embodiments, this may be accomplished by using a lateral flow test format (e.g., lateral flow test cartridge) comprising appropriate reporters, e.g. phosphors. According to other embodiments, the analyzer device may be incorporated into a sample analysis kit for analyzing an extracted biological analyte along with the test cartridge (i.e., biological sampling device). In certain embodiments, the test cartridge may be configured to provide a biological analyte into the analyzer device. In other embodiments, the test cartridge may include a lateral flow assay cartridge including a test strip, result window, and sample well. According to certain embodiments, the result window may be aligned with a camera of the smartphone and the lens of the analyzer device.

[0102] According to certain embodiments, the combination of the portable electronic device and the analyzer device may be used to perform image processing of a test sample contained in the test cartridge. For example in certain embodiments, a user may be provided with the analyzer device that may include a portable electronic device such as a smartphone. The smartphone may be fitted with an adapter of the analyzer device, which enables the user to run a lateral flow test with the combination of the analyzer device and the smartphone. To run the test, the user may open a software application that has been pre-installed in the smartphone, and insert a test cartridge into the analyzer device.

[0103] In certain embodiments, the analyzer device may use the smartphone camera to capture a brightfield image of the test strip located inside the test cartridge. According to certain embodiments, the test strip may include a detection zone that has a control line (CL) and a test line (TL). In certain embodiments, the positions of the TL and CL may be determined by scanning through the brightfield image. Further, in other embodiments, the CL and TL may be striped with a dye that allows these regions to be detected in the brightfield image. For example, the edges of the CL and TL may be determined by an algorithm and defined as the dye bounds.

[0104] According to certain embodiments, once the positions of the TL and CL have been registered, fluid-front tracking and automatic analysis may be initiated. In this procedure, the user may add a sample to the sample well of the test cartridge. In certain embodiments, the sample may be added before the test cartridge is inserted into the analyzer device, or while the test cartridge is inserted in the analyzer device. The analyzer device may continuously image the detection zone of the test strip in real-time in a brightfield mode. While this is occurring, a fluid-front tracking algorithm of the software application may look for the liquid sample flowing across the detection zone. When the liquid flows past a certain predefined region in the detection zone, the analyzer device, via the smartphone, may begin to capture luminescence images.

[0105] In certain embodiments, the luminescence images may be captured at various timepoints after the sample has been added. For example, the analyzer device may capture luminescence images of the detection zone using the smartphone at some predefined time interval. According to certain embodiments, a set of luminescence images may be captured every 2.5 minutes, and the total assay time may be set to 30 minutes (i.e., timepoints at 0 min, 2.5 min, 5 min, . . . , 30 min). In some embodiments, the analyzer device may be communicatively paired with the circuit board and configured to execute an analysis procedure to analyze the biological analyte at a plurality of time points and at predefined time intervals (e.g., at least every 30/60/90 seconds, at least every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 minutes, or fractions thereof). In certain embodiments, about 20 images may be captured in each 2.5 min imaging cycle, and in other embodiments, multiple images of the luminescence may be captured within tens to hundreds of milliseconds after the excitation source turns off. That is, the analysis procedure may include a plurality of luminescence imaging cycles, each of which may include an excitation of the biological analyte. In other embodiments, during each luminescence imaging cycle, the smartphone application may communicate with a circuit board inside the analyzer device to turn on and off a light emitting diode (LED) used for excitation of the phosphors on the test strip. In some embodiments, the analyzer device may use the LED of the smartphone, and in other embodiments, the analyzer device may use an LED built into the analyzer device. In certain embodiments, the wavelength of the LED may be about 400 to 450 nm. In other embodiments, the LED wavelength may be near ultraviolet wavelength range of about 100 to 400 nm.

[0106] According to certain embodiments, the phosphors may exhibit a persistent luminescence phenomenon (i.e., glow-in-the-dark), and the particles may continue to glow after the excitation LED is turned off. When the LED is switched off, the smartphone camera may be triggered to capture an image of the luminescence within tens to hundreds of milliseconds after the LED turns off. In other embodiments, the phosphors may be re-excited with the LED and imaged for multiple cycles, and the captured luminescence images may be averaged together to improve the signal-to-noise ratio. [0107] In certain example embodiments, the captured luminescence images may be analyzed to calculate TL and CL intensities. For example, a global background correction may be applied. According to certain embodiments, the luminescence image from a particular timepoint (e.g., 30 min), may be selected and collapsed from a 2-dimensional image to a 1 -dimensional vector called the luminescence intensity profile. In certain embodiments, the dye bounds (determined previously from the brightfield image) may be overlaid on the luminescence profile, and a global background correction may be applied to the luminescence profile. In other embodiments, the background may be calculated by fitting a polynomial to the regions of the luminescence intensity profile that fall outside of the dye bounds. Further, the background corrected luminescence profile may be calculated by subtracting the raw data (curve 915) from the background fit (curve 920).

[0108] According to certain embodiments, a refined local background correction may be applied at the TL. For example, an algorithm may be run to revise the estimated positions for the TL peak bounds by analyzing the dye bounds (calculated from the brightfield image), and the global background corrected luminescence profile. The revised TL peak bounds (lines 925) may be located near the positions of the dye bounds (lines 910), but may be shifted slightly to the left or right depending on noise in the luminescence intensity profile. The average pixel intensity within the refined bounds may be calculated to give a single numerical value for the TL signal. In other embodiments, a similar algorithm may be run to calculate the signal for the CL.

[0109] In certain embodiments, it may be possible to determine whether the result is positive, negative, or invalid. For example, in some embodiments, a test may be called positive if the TL signal falls above some predefined threshold, and the test may be called negative if the TL signal is less than or equal to the threshold. According to some embodiments, the CL may be invoked to determine if the test is invalid or not. For example, in certain embodiments, if the control line is not visible, then there may be an indication that potential issue exists. In certain embodiments, the CL signal may not help determine if the result is positive or negative. Rather, the CL signal may help reduce inaccurate results that may occur due to a defective test strip or user error.

[0110] As illustrated in FIG. 9, at 900, an enhanced luminescence image (detection zone region) may be provided. In particular, the image 900 illustrates a 2-dimensional image of the detection zone of the lateral flow test strip. As can be seen in FIG. 9, there are two lines near the middle of the image with the CL on the left and the TL on the right. [0111] As illustrated in FIG. 9, an enhanced luminescence image of the TL 905 may be provided. This feature illustrates a 2-dimensional image of the test line region of the lateral flow strip. Further, FIG. 9 illustrates dye bounds 910. According to certain embodiments, the dye bounds may be calculated by analyzing the brightfield image of the strip, which may be captured before the sample is added to the strip. In certain embodiments, the dye bounds may be incorporated into the dispensing solution when the strips are manufactured. In other embodiments, the dye may improve quantitation accuracy as it may allow the analyzer device to more precisely register the exact positions of the CL and TL.

[0112] FIG. 9 also illustrates a luminescence intensity profile 915. As illustrated in FIG. 9, the luminescence intensity profile 915 may be a 1 -dimensional vector that is calculated from the 2-dimensional luminescence image. In certain embodiments, the luminescence intensity profile may have two main peaks; one peak at the CL position, and one peak at the TL position. According to certain embodiments, these two peaks may be used to determine whether the test result is negative, positive, or invalid.

[0113] As further illustrated in FIG. 9, a global background correction (polynomial fit) 920 may be applied. For example, in certain embodiments, the luminescence intensity profile may be background-corrected by a polynomial to estimate the background signal. According to certain embodiments, the background correction may be used to better refine the exact positions of the TL and CL peaks for improved signal quantitation. FIG. 9 also illustrates refined peak bounds 925. As illustrated in FIG. 9, the refined peak bounds 925 for the TL peak may be determined by analyzing the background-corrected luminescence intensity profile. In addition, FIG. 9 illustrates a luminescence intensity profile 930. In certain embodiments, the luminescence intensity profile 930 illustrated in FIG. 9 is a zoomed in version of the intensity profile, but focused on the TL region, whereas the luminescence intensity profile 915 shows the entire luminescence profile.

[0114] FIG. 9 also illustrates a local background correction 935 at a TL region (polynomial fit). According to certain embodiments, a second background correction may be applied by looking at the luminescence intensity profile in the TL region. By leaving out the other regions of the detection zone, a more accurate background correction may be applied to the TL peak for improved signal quantitation. FIG. 9 further illustrates final calculated TL and CL values 940, where the average intensity value for the CL and TL peaks are calculated and displayed. According to certain embodiments, these numbers 940 may be proportional to how bright the signal is. For instance, in certain embodiments, if a lot of the detected sample is present, a brighter line will be visible (e.g., brighter test line).

[0115] FIG. 10 illustrates a test strip and components, according to certain embodiments. In particular, FIG. 10 illustrates a test strip in the test cartridge. As illustrated in FIG. 10, the test cartridge includes a cartridge top 1000, and the cartridge top 1000 includes a result window (detection zone) 1005, and a sample well 1010 through which a sample may be added to the test strip. The test cartridge also includes a cartridge bottom 1015, on which a lateral flow strip is placed. The lateral flow strip may include a control line 1020 and a test line 1025. The lateral flow strip may also include an absorbent pad 1030, membrane 1035, conjugate pad 1040, filtration pad 1045, and sample pad 1050. According to certain embodiments, the cartridge top 1000 may be assembled onto the cartridge bottom 1015 such that the result window 1005 overlaps the membrane 1035. The overlap may allow the control line 1020 and the test line 1025 to be visible through the result window 1005. In addition, the sample well 1010 may be aligned with the sample pad 1050 when the cartridge top 1000 and the cartridge bottom 1015 are assembled together. As such, a sample added through the sample well 1010 may initially be added onto the sample pad 1050.

[0116] FIG. 11 illustrates an assembled test cartridge 1105, according to certain embodiments. As illustrated in FIG. 11, the assembled test cartridge 1105 may include a result window 1100 (e.g., detection zone). As described above, when the test cartridge is assembled, the result window 1100 may overlap and align with the membrane 1035. The alignment enables the control line and the test line to be visible through the result window. In addition, FIG. 11 illustrates that once the test cartridge is assembled, the sample well overlaps and is aligned with the sample pad 1050.

[0117] According to certain embodiments, the test cartridge 1105 may be packaged in a container such as, for example a pouch, which may include a quick response (QR) code printed on the container. The container may also include a lot number. In certain embodiments, a camera on the processing device may be configured to scan the QR code, after which the processing device may know the lot number for the cartridge and link the cartridge information to a database with information on how to reach a certain threshold for that particular lot. According to certain embodiments, the threshold may be set for a specific lot that indicates if it is a positive or a negative result depending on luminescence intensity. In other words, according to certain embodiments, it may be possible to correlate phosphors with analytics of the analyzer device and processing device, and make adjustments tailored to the phosphors of the test cartridge.

[0118] FIG. 12(a) illustrates the analyzer device with the test cartridge inserted, according to certain embodiments. According to certain embodiments, the sample may be added into the test cartridge after the test cartridge has been inserted into the analyzer device. As illustrated in FIG. 12, a smartphone device that is fitted with the analyzer device. In addition, after the test cartridge is inserted, the detection zone of the test cartridge may be aligned with the camera of the smartphone. According to certain example embodiments, the smartphone may be connected to the analyzer device via a wired or wireless communication. For instance, in some embodiments, the smartphone may connect to the analyzer device by way of a wireless connection such as via Bluetooth® low energy (BLE), near-field communication (NFC), or the like. In certain embodiments, the smartphone may wirelessly connect to the PCBA that may include a BLE module. Further, in some embodiments, insertion of the test cartridge into the analyzer device may trigger the analyzer device and/or the smartphone to automatically turn on, and pairing of the analyzer device and the smartphone may occur automatically when the test cartridge is inserted into the port of the analyzer device. In other embodiments, the analyzer device may include a manual switch that enables the user to turn on or off the analyzer device. In further embodiments, after the cartridge is inserted into the analyzer device, the pre-installed software application in the smartphone may automatically recognize that the cartridge has been inserted, and may automatically pair with the smartphone. According to other embodiments, after the smartphone is paired with the test cartridge, the software application of the smartphone may prompt the user to collect the sample.

[0119] FIG. 12(b) illustrates a PCBA, according to certain embodiments. As illustrated in FIG. 12(b), the PCBA may include BLE module configured to execute wireless communication with a processing device. The PCBA may also include one or more excitation LEDs, which may have a power output of about 405 nm. Further, the PCBA may include one or more white LEDs, and a cartridge detection switch. According to certain embodiments, the cartridge detection switch may be configured to detect when a test cartridge is inserted into the active adapter/analyzer device, and configured to power on the PCBA in response to detecting that a test cartridge has been inserted.

[0120] FIG. 13 illustrates a method of assembling the analyzer device, according to certain embodiments. The method may include, at 1300, soldering wires of the battery holder to the PCBA. The method may also include, at 1302, placing batteries in the battery holder. The method may further include, at 1304, programming the PCBA, and at 1306, removing the batteries from the battery holder. At 1308, the method may include verifying that the assembly area is clear of materials and documentation from previous assembly builds, and any other materials and documentation not associated with the current assembly build.

[0121] Further, at 1310, the method may include routing the PCBA through the rectangular opening of the enclosure base. At 1312, the method may include assembling the battery holder to the enclosure base with two screws. In certain embodiments, the screws may be M2.5 x 6 mm screws. The method may also include at 1314, turning over the enclosure base, and at 1316, routing the battery holder wires around the bosses in the enclosure base. Further, the method may include, at 1318, placing the PCBA onto the mounting bosses in the enclosure base, and at 1320, placing the PCBA bracket onto the PCBA, and assembling the parts together with two screws. During the assembling process, the PCBA bracket holding the PCBA may be fixed to the enclosure base via the two screws. According to certain embodiments, the two screws may be #4 x 5/8 screws.

[0122] In certain embodiments, the method may further include, at 1322, placing the lens bracket into the interior hole of the top section or enclosure cover. In some embodiments, when placing the lens bracket into the interior hole, the method may include ensuring that the pegs are visible and face outward (away from a surface on which the top section is placed). The method may also include, at 1324, placing the lens into the lens bracket. At this step, it may be desirable to ensure that the curved surface of the lens faces outward (away from the surface on which the top section is placed). In addition, the method may include, at 1326, assembling the lens bracket, lens, and top section together by heat staking the pegs or using ultraviolet (UV) glue. In this step, it may be desirable to ensure that the pegs melt onto the lens bracket and lens, and that the parts are secure or that there is enough UV glue.

[0123] Additionally, at 1328, the method may include placing the top section onto the enclosure base. Further, at 1330, the method may include placing the middle section cover onto the enclosure base. At 1332, the method may include sliding a portable electronic device into the bottom section of the middle section cover. The method may also include, at 1334, sliding the portable electronic device into the middle and top sections of the enclosure cover, and ensure that all parts fit over the enclosure base. Further, at 1336, the method may include turning over the assembled parts, and at 1338, assembling the parts together using seven screws. In certain embodiments, the seven screws may include #4 x 3/8 screws. At 1340, the method may include placing the batteries in the battery holder, and at 1342, the method may include attaching the battery door to the enclosure base. Further, at 1344, the method may include visually inspecting the assembled unit, and confirm alignment of housings and no gaps.

[0124] FIG. 14(a) illustrates an apparatus 10 according to certain embodiments. In certain example embodiments, apparatus 10 may be a user equipment (UE), mobile equipment (ME), mobile station, mobile device, stationary device, loT device, or other device. It should be noted that one of ordinary skill in the art would understand that apparatus 10 may include components or features not shown in FIG. 14(a).

[0125] In some embodiments, apparatus 10 may include one or more processors, one or more computer-readable storage medium (for example, memory, storage, or the like), one or more radio access components (for example, a modem, a transceiver, or the like), and/or a user interface. In some example embodiments, apparatus 10 may be configured to operate using one or more radio access technologies, such as GSM, LTE, LTE-A, NR, 5G, WLAN, WiFi, NB-IoT, Bluetooth®, NFC, MulteFire, and/or any other radio access technologies. It should be noted that one of ordinary skill in the art would understand that apparatus 10 may include components or features not shown in FIG. 14(a).

[0126] As illustrated in the example of FIG. 14(a), apparatus 10 may include or be coupled to a processor 12 for processing information and executing instructions or operations. Processor 12 may be any type of general or specific purpose processor. In fact, processor 12 may include one or more of general-purpose computers, special purpose computers, microprocessors, digital signal processors (DSPs), field-programmable gate arrays (FPGAs), application-specific integrated circuits (ASICs), and processors based on a multi-core processor architecture, as examples. While a single processor 12 is shown in FIG. 14(a), multiple processors may be utilized according to other example embodiments. For example, it should be understood that, in certain example embodiments, apparatus 10 may include two or more processors that may form a multiprocessor system (e.g., in this case processor 12 may represent a multiprocessor) that may support multiprocessing. According to certain example embodiments, the multiprocessor system may be tightly coupled or loosely coupled (e.g., to form a computer cluster).

[0127] Processor 12 may perform functions associated with the operation of apparatus 10 including, any of the processes described and illustrated herein.

[0128] Apparatus 10 may further include or be coupled to a memory 14 (internal or external), which may be coupled to processor 12, for storing information and instructions that may be executed by processor 12. Memory 14 may be one or more memories and of any type suitable to the local application environment, and may be implemented using any suitable volatile or nonvolatile data storage technology such as a semiconductor-based memory device, a magnetic memory device and system, an optical memory device and system, fixed memory, and/or removable memory. For example, memory 14 can be comprised of any combination of random access memory (RAM), read only memory (ROM), static storage such as a magnetic or optical disk, hard disk drive (HDD), or any other type of non-transitory machine or computer readable media. The instructions stored in memory 14 may include program instructions or computer program code that, when executed by processor 12, enable the apparatus 10 to perform tasks as described herein.

[0129] In certain embodiments, apparatus 10 may further include or be coupled to (internal or external) a drive or port that is configured to accept and read an external computer readable storage medium, such as an optical disc, USB drive, flash drive, or any other storage medium. For example, the external computer readable storage medium may store a computer program or software for execution by processor 12 and/or apparatus 10 to perform any of the methods described herein.

[0130] In some embodiments, apparatus 10 may also include or be coupled to one or more antennas 15 for receiving a downlink signal and for transmitting via an uplink from apparatus 10. Apparatus 10 may further include a transceiver 18 configured to transmit and receive information. The transceiver 18 may also include a radio interface (e.g., a modem) coupled to the antenna 15. The radio interface may correspond to a plurality of radio access technologies including one or more of GSM, LTE, LTE-A, 5G, NR, WLAN, NB-IoT, Bluetooth®, BT-LE, NFC, RFID, UWB, and the like. The radio interface may include other components, such as filters, converters (for example, digital-to-analog converters and the like), symbol demappers, signal shaping components, an Inverse Fast Fourier Transform (IFFT) module, and the like, to process symbols, such as OFDMA symbols, carried by a downlink or an uplink.

[0131] For instance, transceiver 18 may be configured to modulate information on to a carrier waveform for transmission by the antenna(s) 15 and demodulate information received via the antenna(s) 15 for further processing by other elements of apparatus 10. In other example embodiments, transceiver 18 may be capable of transmitting and receiving signals or data directly. Additionally or alternatively, in some example embodiments, apparatus 10 may include an input and/or output device (VO device). In certain example embodiments, apparatus 10 may further include a user interface, such as a graphical user interface or touchscreen.

[0132] In certain embodiments, memory 14 stores software modules that provide functionality when executed by processor 12. The modules may include, for example, an operating system that provides operating system functionality for apparatus 10. The memory may also store one or more functional modules, such as an application or program, to provide additional functionality for apparatus 10. The components of apparatus 10 may be implemented in hardware, or as any suitable combination of hardware and software. According to certain embodiments, apparatus 10 may optionally be configured to communicate with apparatus 20 via a wireless or wired communications link 70.

[0133] FIG. 14(b) illustrates an apparatus 20 according to certain embodiments. In certain example embodiments, the apparatus 20 may be an analyzer device, a PCBA, or a combination of the analyzer device and the PCBA. It should be noted that one of ordinary skill in the art would understand that apparatus 20 may include components or features not shown FIG. 14(b).

[0134] As illustrated in the example of FIG. 14(b), apparatus 20 may include a processor 22 for processing information and executing instructions or operations. Processor 22 may be any type of general or specific purpose processor. For example, processor 22 may include one or more of general-purpose computers, special purpose computers, microprocessors, digital signal processors (DSPs), field-programmable gate arrays (FPGAs), application-specific integrated circuits (ASICs), and processors based on a multi-core processor architecture, as examples. While a single processor 22 is shown in FIG. 14(b), multiple processors may be utilized according to other example embodiments. For example, it should be understood that, in certain example embodiments, apparatus 20 may include two or more processors that may form a multiprocessor system (e.g., in this case processor 22 may represent a multiprocessor) that may support multiprocessing. In certain example embodiments, the multiprocessor system may be tightly coupled or loosely coupled (e.g., to form a computer cluster).

[0135] According to certain embodiments, processor 22 may perform functions associated with the operation of apparatus 20, which may include, for example, any of the processes described and/or illustrated herein.

[0136] Apparatus 20 may further include or be coupled to a memory 24 (internal or external), which may be coupled to processor 22, for storing information and instructions that may be executed by processor 22. Memory 24 may be one or more memories and of any type suitable to the local application environment, and may be implemented using any suitable volatile or nonvolatile data storage technology such as a semiconductor-based memory device, a magnetic memory device and system, an optical memory device and system, fixed memory, and/or removable memory. For example, memory 24 can be comprised of any combination of random access memory (RAM), read only memory (ROM), static storage such as a magnetic or optical disk, hard disk drive (HDD), or any other type of non-transitory machine or computer readable media. The instructions stored in memory 24 may include program instructions or computer program code that, when executed by processor 22, enable the apparatus 20 to perform tasks as described herein.

[0137] In certain embodiments, apparatus 20 may further include or be coupled to (internal or external) a drive or port that is configured to accept and read an external computer readable storage medium, such as an optical disc, USB drive, flash drive, or any other storage medium. For example, the external computer readable storage medium may store a computer program or software for execution by processor 22 and/or apparatus 20 to perform the methods described herein.

[0138] In certain embodiments, apparatus 20 may also include or be coupled to one or more antennas 25 for transmitting and receiving signals and/or data to and from apparatus 20. Apparatus 20 may further include or be coupled to a transceiver 28 configured to transmit and receive information. The transceiver 28 may include, for example, a plurality of radio interfaces that may be coupled to the antenna(s) 25. For example, the radio interface may correspond to a plurality of radio access technologies including one or more of WLAN or Bluetooth®, and the like.

[0139] In certain embodiments, memory 24 may store software modules that provide functionality when executed by processor 22. The modules may include, for example, an operating system that provides operating system functionality for apparatus 20. The memory may also store one or more functional modules, such as an application or program, to provide additional functionality for apparatus 20. The components of apparatus 20 may be implemented in hardware, or as any suitable combination of hardware and software.

[0140] FIG. 15 illustrates a reusable analyzer and kit, according to certain embodiments. As described herein, certain embodiments may utilize a smartphone (e.g., processing device) to capture images of the luminescence from nanoparticles that detect specific targets such as viruses, bacteria, proteins, molecules, or other analytes. As illustrated in FIG. 15, certain embodiments may include a reusable analyzer 1500 (e.g., active adapter) and kit (e.g., the combination of 1505 - 1525) of single-use consumable components. The consumables may include a nasal swab 1515, an extraction tube 1520 for removing viral antigens from the swab 1515, a dropper cap 1525, and a lateral flow cartridge 1510. According to certain embodiments, the extraction tube 1520 may include a viral lysis buffer and may include a “shipping cap” that contains the buffer within the tube until a user is ready to run a sample. In certain embodiments, a user may dip the swab 1515 into the extraction tube 1520 and lysis buffer for about one minute, after which the user may remove and discard the swab 1515, then attach a dropper cap 1525 onto the tube 1520 for dispensing of the sample.

[0141] The analyzer according to certain embodiments may include a smartphone 1505 and an active adapter 1500. The active adapter may include components such as a plastic housing designed with a form factor that allows it to mate with a particular smartphone model, a power source (e.g., AA batteries) for the electronics within the active adapter, a switch that turns the active adapter on when a user inserts a cartridge into the sample port of the active adapter, a lens that magnifies the result window of the cartridge to allow the phone camera to focus on the test strip within the cartridge, and a circuit board that connects with the phone wirelessly. According to certain embodiments, the board may include a white LED that illuminates the result window for brightfield image acquisition (i.e., a brightfield LED) and a 405 nm near-UV LED used for excitation of the phosphors (i.e., an excitation LED).

[0142] According to certain embodiments, it may be possible to enable time-gated luminescence imaging, wherein the smartphone controls the circuit board to switch on the near- UV LED for excitation of the phosphors for a defined period of time, after which the LED may be turned off and almost immediately the smartphone camera captures a time-gated image of the luminescence from the phosphors. That is, certain embodiments may establish a consistent time base between the smartphone and the circuit board via wireless communication between the devices (i.e., clock synchronization).

[0143] In certain embodiments, the smartphone (i.e., host device) and the circuit board (i.e., the peripheral) within the active adapter may be synchronized. According to certain embodiments, for time synchronization between the host (smartphone) and the peripheral (circuit board) to work, both devices may include a clock source. The smartphone may include a built-in clock. For the peripheral to have a clock that can be synchronized with the host’s clock, the circuit board may incorporate a component that can function as a clock and establish a time base within the peripheral. In certain embodiments, this clock may be something that oscillates at a known frequency with high accuracy and low or negligible drift over time. A variety of devices may fulfill this oscillation requirement to establish a time base within the peripheral such as, for example, a crystal oscillator, a MEMS oscillator, a quartz crystal oscillator, a temperature controlled temperature oscillator, or another device. Certain embodiments may use a quartz crystal oscillator as it may be inexpensive and sufficient enough to limit any drift.

[0144] According to certain embodiments, quartz crystal oscillators may be used as watch crystals, and oscillate at a frequency of 32.768 kHz. This is a power of 2 (2¹⁵ = 32,768), which make sit easy to drive a binary counter, since it may be possible to obtain a precise 1 second period with a 15-stage binary counter. In certain embodiments, the frequency of the oscillatory in the host or the peripheral may vary. In other embodiments, the oscillators may have frequencies of 20 kHz, 100kHz, or others. According to certain embodiments, the host and peripheral may use oscillators that have the same frequency, but in other embodiments, the frequencies may differ, and it may be possible to convert from one oscillator’s time base to the other. [0145] In certain embodiments, the peripheral device may include a counter that may be incremented at a known 32.768 kHz rate. The counter may initially be set at 200,000 tickets (device ticks or peripheral ticks) or some arbitrary number. The counter value on the peripheral may increase at a known rate based on the frequency (e.g., 32.768 kHz). Likewise, the host device may have a counter that is being incremented at some other known frequency, and may initially be set at some arbitrary number of ticks. Thus, there may be two counters, but the clock offset between the two counters may be unknown. To determine the clock offset, certain embodiments may provide a method that is analogous to a network time protocol (NTP) (i.e.., an NTP analogous protocol), where a phone or computer may set its time from the internet. In certain embodiments, the host (e.g., smartphone) and peripheral (e.g., circuit board) may implement the NTP protocol.

[0146] In addition to NTP, certain embodiments may provide an alternative where the host may perform a low latency read over BLE of the counter value on the peripheral, and repeat that read command multiple times (e.g., 10 times), and take the minimum time reading. Multiple time readings are taken because wireless communications may be delayed due to interference or other reasons causing transmission retries that may result in delayed delivery. According to certain embodiments, by reading the counter value multiple times and taking the minimum clock offset observed, it may be possible to obtain 1-2 clock offset readings that are larger than they should be because the packet transmitted via BLE was delayed. However, it may be known that when the host obtains a reading of the peripheral counter value from the host perspective, the true counter value on the peripheral device at the time the host receives that communication, is greater than or equal to the number that was transmitted by the peripheral.

[0147] In certain embodiments, the information on host ticks and peripheral ticks may be stored (e.g., on the smartphone) in two vectors of equal length: a vector of peripheral ticks received by the host and an equal length vector of host ticks at the time the peripheral tick value was received. These values of host and peripheral ticks may be written to the vectors in lock-step. According to certain embodiments, a third vector may be generated to calculate the clock offset between the host and peripheral clocks, which may done by subtracting the host ticks from the peripheral ticks. The offset may be taken as the minimum value in the third vector.

[0148] According to certain embodiments, the clock offset may serve as a link to convert host times into device times (e.g., host ticks into device ticks). The synchronization may be repeated on a regular basis (e.g., once per minute) to preserve accuracy of the offset since drift between the time bases can increase inaccuracy of the offset. For example, if one clock is running slightly slower or faster than the other, there may be some clock drift. In certain embodiments, this drift may be computed in milliseconds per seconds. Once the number of host ticks to peripheral ticks offset (clock offset) is obtained, a consistent time base between the two devices may be established. This may be made possible by having the ability to send commands from the host to the peripheral device with time-stamps in the peripheral device tick units, and knowing when these events are going to occur, such as board trigger events. For instance, according to certain embodiments, an illumination command may be sent from the host (e.g., smartphone) to the peripheral (e.g., circuit board) with 2 times, tO and tl, where tO is the time at which to turn on the LED for excitation and tl being the time to turn off the LED. The difference between tl and tO may be the desired excitation time, and the command may be constructed such that tO is some time in the future that accounts for latency. According to certain embodiments, the host (e.g., smartphone) may manage the clock conversion and bookkeeping, and instruct the peripheral on what to do in peripheral tick units.

[0149] With the above time synchronization, it may be possible to establish a consistent time base between the host and peripheral devices so that commands may be sent from the host (e.g., smartphone) to the peripheral device (e.g., circuit board), and know when those commands will be executed. In certain embodiments, these commands may involve executing some action on the circuit board such as, for example, turning on or off the LEDs for illumination or excitation. According to certain embodiments, both the host and the peripheral may include clocks that oscillate at a known frequency. The host may include a built-in oscillator, whereas the circuit board may include a quartz crystal oscillator. However, in other embodiments, any arbitrary oscillator with a known frequency may be used. As noted above, the host and peripheral may communicate wirelessly over Bluetooth®, exchanging information on current clock ticks to allow the host to establish a time base. However, in other embodiments, BLE, NFC, or the like may be used instead of Bluetooth®. According to further embodiments, the time synchronization may be repeated periodically to keep the clocks synchronized to mitigate effects of clock drift.

*** [0150] One having ordinary skill in the art will readily understand that the invention as discussed above may be practiced with steps in a different order, and/or with hardware elements in configurations which are different than those which are disclosed. Therefore, although the invention has been described based upon these preferred embodiments, it would be apparent to those of skill in the art that certain modifications, variations, and alternative constructions would be apparent, while remaining within the spirit and scope of the invention.

Claims

WE CLAIM:

1. An active adapter configured to secure and align a processing device with a sampling device comprising an analyte and at least one reporter, wherein the active adapter comprises a power source, a circuit board connected to the power source, at least one excitation source connected to the circuit board, and a port configured to receive the sampling device, and wherein the processing device is communicatively and wirelessly paired with the circuit board and configured to execute an analysis procedure to analyze the analyte, wherein said analysis procedure comprises at least one luminescence imaging cycle comprising excitation followed by termination of excitation, and then capture of at least one image of the reporter and/or analyte.

2. The active adapter according to claim 1, wherein the processing device is configured to execute the analysis procedure to analyze the analyte at a plurality of time points.

3. The active adapter according to claim 2, wherein the analysis procedure is performed at predefined time intervals, [e.g. at least every 30/60/90 seconds, at least every 2, 3, 4, 5, 6, 7, 8, 9, 10 minutes.

4. The active adapter according to claim 1, wherein each of the luminescence imaging cycles comprises a predetermined time period during which excitation and image capturing is halted.

5. The active adapter according to claim 1, comprising a plurality of luminescence imaging cycles, each of which comprises excitation of the reporter followed by capturing at least one image of the luminescence, wherein the captured luminescence images are averaged together to improve the signal -to-noise ratio.

6. The active adapter according to claim 1 or 5, wherein a plurality of luminescence images are captured during each luminescence imaging cycle, and the captured luminescence images are averaged together to improve the signal-to-noise ratio.

36

7. The active adapter according to any preceding claim, wherein the active adapter comprises a mechanical switch, wherein the active adapter is turned on when the sampling device is inserted into the port initiating the switch, and is configured to be paired with the processing device when the processing device is turned on.

8. The active adapter according to any preceding claim, wherein pairing of the active adapter and the processing device occurs automatically when the sampling device is inserted into the port.

9. The active adapter according to claim 8, wherein after the active adapter and the processing device are paired, the processing device is configured to capture continuous video of a portion of the sampling device.

10. The active adapter according to any preceding claim, further comprising an enclosure base comprising a first side and a second side opposite the first side, wherein the power source is integrally connected on the first side of the enclosure base and the circuit board is attached to the second side of the enclosure base; and a top section cover comprising a lens, the top section cover integrally connected to the second side of the enclosure base.

11. The active adapter according to claim 10, further comprising a middle section cover disposed on the second side of the enclosure base and configured to support the processing device; and an enclosure cover configured to secure the processing device to the sample analysis device.

12. The active adapter according to any preceding claim, wherein the processing device is an imaging device, a portable mobile communication device, or a smartphone.

13. A sample analysis device comprising the active adapter according to any of claims 1-12 and a processing device attached to and communicatively paired with the active adapter, wherein the processing device and the active adapter are synchronized, and wherein the synchronization is repeated periodically.

14. The sample analysis device according to claim 13, wherein the active adapter and the

37 processing device each comprise a respective clock source that are configured to oscillate at a predetermined frequency.

15. The sample analysis device according to claim 14, wherein the clock source of the active adapter and the processing device is a crystal oscillator, a micro-electromechanical system oscillator, or a quartz crystal oscillator.

16. The sample analysis device according to any of claims 13-15, wherein during the synchronization, the processing device is configured to perform a low latency time reading of a counter value of the clock of the active adapter, repeat the low latency time reading of the counter value a plurality of times, and take a minimum time reading of the plurality of time readings.

17. The sample analysis device according to claim 16, wherein the processing device is configured to store ticks of the processing device and ticks of the active adapter in two vectors of equal length.

18. The sample analysis device according to claim 17, wherein the processing device is configured to generate a third vector based on the two vectors, and to calculate a clock offset between the clock source of the processing device and the clock source of the active adapter.

19. The sample analysis device according to claim 18, wherein the processing device is configured to establish a consistent time base between the processing device and the active adapter based on the clock offset, and wherein the processing device is configured to send commands to the active adapter with one or more time-stamps in active adapter tick units based on the clock offset.

20. The sample analysis device according to claim 18, wherein the processing device is configured to control, based on the clock offset, the active adapter to trigger an excitation for a defined period of time, after which the excitation is turned off and the processing device is configured to capture a time-gated image of the reporter and/or analyte.

21. A sample analysis kit for analyzing an extracted analyte, comprising the sample analysis device according to any of claims 13-19 and the sampling device, wherein the sampling device is adapted and configured to provide a biological analyte into the active adapter.

22. The sample analysis kit according to claim 21, wherein the sampling device comprises a lateral flow assay cartridge comprising a test strip, a result window, and a sample well.

23. The sample analysis kit according to claim 22, wherein the result window is aligned with the lens of a camera of the processing device.

24. The sample analysis kit according to claim 23, wherein the test strip comprises an absorbent pad, a conjugate pad, a filtration pad, a sample pad, and a membrane section comprising a control line and a test line, and wherein the membrane section is aligned with the result window.

25. A method for analyzing an analyte with the sample analysis device according to any of claims 13-20, comprising: inserting a sampling device into the active adapter and executing, via the processing device, an analysis procedure, wherein the analysis procedure comprises: pairing the processing device with the active adapter in response to inserting the sampling device into the sample analysis device; performing a time synchronization procedure to synchronize the processing device with the active adapter in response to inserting the sampling device into the active adapter; capturing a first brightfield image of the test strip; determining a position of the test line and the control line by scanning through the brightfield image; adding the analyte to the sample well of the sampling device; capturing, via the processing device, a plurality of luminescence images of a detection zone located within the result window during one or more luminescence imaging cycle(s); analyzing the plurality of luminescence images to calculate test line and control line intensities; and determining a test result based on the intensities of the test line and control line intensities.

26. The method according to claim 25, wherein the first brightfield image is captured by utilizing a white light emitting diode.

27. The method according to claim 25, wherein the test result is positive if a test line signal is above a predefined threshold, and the test result is negative if a test line signal is below a predefined threshold.

28. The method according to claim 25, wherein the test result is valid if a control line signal is above a predefined threshold, and the test result is invalid if a control line signal is below a predefined threshold.

29. The method according to any one of claims 25-28, wherein the plurality of images comprises at least twenty images.

30. The method according to any of claims 25-29, wherein the processing device continuously images the detection zone of the strip after the sampling device is inserted into the active adapter in brightfield mode, and begins the first luminescence imaging cycle when the fluid front flows past a predefined region in the detection zone.

31. The method according to any of claims 25-30, wherein the predefined time interval is between one and thirty minutes, preferably at least every 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes, or fractions thereof.

32. The method according to claim 31, wherein the processing device communicates with the active adapter during the time intervals to control a light emitting diode of the active adapter for each time interval.

33. The method according to claim 32, wherein the light emitting diode is controlled to turn on and off.

34. The method according to claim 33, wherein when the light emitting diode is turned on, the light emitting diode is configured to excite phosphors on the test strip.

35. The method according to claim 33, wherein when the light emitting diode is turned off, the processing device is triggered to capture at least one image.

36. The method according to any one of claims 25-35, further comprising averaging intensity values of each of the plurality of images at the control line and the test line.

37. The method according to any one of claims 25-36, wherein the analysis procedure further comprises: selecting and collapsing, at a time interval, a 2-dimensional image to a 1 -dimensional vector to create a luminescence intensity profile; and applying a global background correction to the luminescence profile, wherein the global background correction comprises fitting a polynomial to one or more regions of the luminescence intensity profile that fall outside of test line and the control line.

38. The method according to any one of claims 25-36, wherein the analysis procedure further comprises refining a local background correction at the test line.

39. The method according to claim 38, wherein refining the local background correction at the test line comprises estimating positions for test line peak bounds by analyzing dye bounds calculated from the brightfield image, and by analyzing the global background corrected luminescence profile.

41