Devices, systems, and methods for measuring solution characteristic of sample using multi-layered active sensor

Abstract

To provide devices, systems and methods for measuring a solution characteristic of a sample using a multi-layered active sensor.SOLUTION: Various devices, systems, and methods for measuring a solution characteristic of a sample comprising microorganisms are disclosed. In one embodiment, a sensor device is disclosed comprising a sample chamber having a chamber lateral wall surrounding a chamber cavity configured to receive the sample, a reference sensor comprising a transfer component to transfer the sample to a reference electrode material, and an active sensor made of a substrate covered in part by an active electrode layer. The active sensor can be coupled to at least part of the chamber lateral wall at a window opening defined along the chamber lateral wall. The active sensor can be positioned such that the active electrode layer faces the chamber cavity to allow the sample within the chamber cavity to be in fluid contact with the active electrode layer through at least part of the chamber lateral wall surrounding the window opening.SELECTED DRAWING: Figure 1A

Description

[Technical Field]

[0001] The present disclosure relates generally to diagnostic devices for measuring solution properties of a sample. More specifically, the present disclosure relates to devices, systems, and methods for measuring solution properties of a sample using multi-layered active sensors. [Background technology]

[0002] Infections caused by anti-infective drug-resistant bacteria and microorganisms are a major challenge in healthcare settings such as hospitals and nursing homes. For example, in infections, chemicals released into the bloodstream by infectious pathogens can trigger dangerous systemic inflammatory and vasoactive responses, leading to a potentially life-threatening complication known as sepsis, which can lead to fever, low blood pressure, and even death. When faced with such infections, clinicians are encouraged to use anti-infective drugs judiciously, preferably only as needed to mitigate the infection.

[0003] However, the most common current practice is to administer broad-spectrum anti-infective drugs, often multiple drugs, to confirm the adequacy of treatment until the organism is identified and its drug susceptibility is tested. As a result, multi-drug resistant infections tend to emerge. Ideally, the sensitivity of an infectious agent would be detected as soon as its presence is confirmed. To determine the susceptibility of such an infectious agent to anti-infective drugs, samples containing the infectious agent must be quantified, which requires analyzing the sample for the presence or absence of microbial growth.

[0004] Existing biosensors used to analyze infectious agents and pathogens in biological and other sample types often include an active sensing element and a reference sensing element in fluid contact or communication with the sample of interest. Current in vitro diagnostic measurement devices, particularly those for detecting oxidation-reduction potential (ORP) and pH in biological and fluid samples, are often not designed for both high performance and low cost. Furthermore, because it is important to prevent cross-contamination of patient samples, the sensing elements of diagnostic measurement systems are preferably disposable consumables. Therefore, disposable sensing elements place significant emphasis on cost and manufacturability.

[0005] Conventional biosensors often use expensive glass or silicon substrates, which increase costs and require multiple manufacturing steps. Furthermore, the active sensing elements of such biosensors can malfunction if a biological or other fluid sample inadvertently contacts conductive portions of the active sensing element that are not intended to come into contact with the sample. Summary of the Invention [Problem to be solved by the invention]

[0006] Therefore, there is a need for a solution that overcomes the above challenges and limitations. It is desirable that such a solution be disposable and cost-effective to manufacture. However, such a solution should also be capable of accurate measurements and be made from highly biocompatible materials. [Means for solving the problem]

[0007] Disclosed are devices, systems, and methods for measuring solution properties (e.g., ORP or pH) of a sample containing microorganisms. In one embodiment, a sensor device for measuring solution properties of a sample is disclosed. The sensor device can comprise a sample container comprising a sample chamber. The sample chamber comprises a chamber sidewall surrounding a chamber cavity configured to receive the sample. The sensor device can also include a reference sensor comprising a reference electrode material and a wick in fluid communication with the sample chamber. At least a portion of the sample can be wicked by the wick toward the reference electrode material.

[0008] It may also be configured with an active sensor formed from a portion of a conductive substrate covered with an active electrode layer. The active sensor may be coupled to at least a portion of the chamber sidewall at a window opening formed along the chamber sidewall. In some embodiments, no portion of the active sensor extends into the chamber cavity. The active electrode layer may face the chamber cavity such that a sample in the chamber cavity can be in fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening. The reference sensor and the active sensor may be electrically connected to a reader device, and solution properties of the sample may be determined based on the potential difference measured between the active sensor and the reference sensor.

[0009] The solution property measured can be redox potential. The solution property measured can also be pH. The active electrode layer can be comprised of at least one of a platinum oxide layer (formed on a platinum layer) and a tantalum oxide layer. The conductive substrate can be comprised of stainless steel. The active electrode layer can have an active electrode layer thickness of approximately 50 nm to 500 nm. The active sensor can further include an adhesion layer between the conductive substrate and the active electrode layer. The adhesion layer can have a thickness of 5 nm to 50 nm. The ratio of the thickness of the adhesion layer to the thickness of the active electrode layer can be approximately 1:10 to 1:20. The adhesion layer can be a chromium layer, a gold layer, or a nickel layer.

[0010] The active layer can have an active electrode layer thickness of about 400 nm. The active sensor can be insert molded into the chamber sidewall while the sample container is formed by injection molding. The active sensor can be press molded into the chamber sidewall after the sample container is formed by injection molding. The chamber sidewall can be configured with a recess surrounding the window opening. The recess can be formed along an outer side surface of the chamber sidewall. The active sensor can be adhered to at least a portion of the chamber sidewall within the recess via an adhesive.

[0011] The active sensor may be comprised of an active electrode side, a conductive substrate side opposite the active electrode side, and a side surface. The side surface may be covered by at least one of a chamber sidewall and an adhesive to prevent contact of the side surface with the sample. The sample chamber may be formed in part from at least one of polyoxymethylene, polyamide, polyethylene, acrylonitrile-butadiene-styrene, polycarbonate, and polypropylene. The reference electrode material may be a cured conductive ink deposited or otherwise applied to the proximal end of a wick.

[0012] In some embodiments, the active sensor can be formed of a non-conductive printed circuit (PCB) substrate partially covered by an electrode layer. The active electrode layer can be electrically coupled to conductive contacts of the PCB substrate by conductive vias extending through the PCB substrate. The PCB substrate can be a flexible PCB substrate.

[0013] The PCB substrate can be formed in part from polyimide, FR-4 composite, copper, etc. The solution property measured can be redox potential. The active electrode layer can be a platinum layer or a gold layer. The active electrode layer can have an active electrode layer thickness of at least 50 nm. In certain embodiments, the active electrode layer can have an active electrode layer thickness of at least 400 nm.

[0014] In some embodiments, the active sensor can be formed from a non-conductive polymer substrate containing through-holes. One side of the polymer substrate and one end of the through-holes can be covered with a conductive layer. The active electrode layer can be electrically coupled to the conductive layer via a conductive coating covering the lateral sides of the through-holes.

[0015] The active electrode layer may be a platinum layer or a gold layer. The active electrode layer may have an active electrode layer thickness of at least 50 nm. In certain embodiments, the active electrode layer may have an active electrode layer thickness of at least 400 nm.

[0016] The active sensor can be a rectangular component having a width dimension between about 100 μm and 6.0 mm and a length dimension between about 100 μm and 6.0 mm. The diameter of the through-hole can be between about 10 μm and 100 μm. The conductive coating covering the lateral sides of the through-hole can be a coating such as platinum or gold.

[0017] In some embodiments, the active sensor can be formed of a conductive dowel partially covered by an active electrode layer. The active sensor can be coupled to at least a portion of the chamber sidewall at a window opening formed along the chamber sidewall. The portion of the conductive dowel covered by the active electrode layer can extend into the chamber cavity so that a sample in the chamber cavity can be in fluid contact with the active electrode layer. The end of the conductive dowel that does not extend into the chamber cavity can extend outside the chamber sidewall. The conductive dowel can be formed in part from stainless steel and can be shaped as a cylinder with substantially rounded edges.

[0018] Also disclosed is a method for measuring solution properties of a sample. The method may include cleaning a conductive substrate with an acid and base treatment, depositing an adhesive layer on one side of the conductive substrate, and depositing an active electrode layer on the adhesive layer. The method may further include singulating the conductive substrate covered with the adhesive layer and the active electrode layer to obtain active sensors sized to cover a window opening formed along a chamber sidewall of a sample chamber. The method may further include bonding the active sensor to at least a portion of the chamber sidewall such that no portion of the active sensor extends into a chamber cavity within the sample chamber and the active electrode layer faces the chamber cavity, such that any sample within the chamber cavity is in fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening.

[0019] The method can further include treating the conductive substrate with ammonium hydroxide, isopropyl alcohol, or acetone after treating the conductive substrate with nitric acid. The method can further include laser cutting, metal shearing, hot wire cutting, die cutting, stamping, or sawing the conductive substrate.

[0020] The method may further include applying a bead of adhesive to a portion of the chamber sidewall within a recess formed along the chamber sidewall surrounding the window opening. The method may further include pressing the active sensor onto the bead of adhesive within the recess and allowing the adhesive to cure.

[0021] The method can further include insert molding the active sensor into the chamber sidewall during injection molding to form the sample chamber. The method can further include locally melting a portion of the chamber sidewall surrounding the window opening, pressing the active sensor into the melted portion of the chamber sidewall, and cooling the melted portion of the chamber sidewall to affix the active sensor to the chamber sidewall.

[0022] The method can further include depositing the active electrode material comprising the active electrode layer until the active electrode layer has a thickness of at least 50 nm. In some embodiments, the method can include depositing the active electrode material comprising the active electrode layer until the active electrode layer has a thickness of at least 400 nm. In some embodiments, the active electrode material can be platinum when the solution property being measured is the oxidation-reduction potential (ORP) of the sample. The active electrode material can be deposited using sputter deposition, evaporative deposition, electrodeposition, ink screen printing, or the like.

[0023] The method can further include depositing an adhesive material that constitutes the adhesive layer using sputter deposition. The adhesive layer can be deposited in a vacuum chamber, and the active electrode layer can be deposited subsequently to the adhesive layer in the same vacuum chamber.

[0024] When the solution property to be measured is the pH of the sample, the active electrode material can be a metal oxide, hi some embodiments, the metal oxide can be platinum oxide, which can cover a platinum layer deposited on the adhesion layer.

[0025] In some embodiments, a method for manufacturing a sensor device for measuring solution properties of a sample can include providing a non-conductive printed circuit (PCB) substrate and depositing an active electrode layer on one side of the PCB substrate. The active electrode layer can be electrically coupled to conductive contacts of the PCB substrate by conductive vias extending through the PCB substrate. The method can further include singulating the PCB substrate covered with the active electrode layer to obtain active sensors sized to cover window openings formed along chamber sidewalls of a sample chamber. The active sensors can include at least one conductive via extending through the PCB substrate. The method can further include bonding the active sensors to at least a portion of the chamber sidewalls such that no portion of the active sensor extends into a chamber cavity within the sample chamber, the active electrode layer faces the chamber cavity, and any sample within the chamber cavity is in fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening.

[0026] The method can further include depositing an active electrode material comprising the active electrode layer using sputter deposition, evaporative deposition, and electrodeposition. The active electrode material can be deposited until the active electrode layer has a thickness of at least 50 nm. In certain embodiments, the active electrode material can be deposited until the active electrode layer has a thickness of at least 400 nm. When the solution property being measured is the oxidation-reduction potential (ORP) of the sample, the active electrode material can be platinum or gold. The conductive contacts can be formed in part of gold.

[0027] In some embodiments, another method for manufacturing a sensor device can include providing a non-conductive polymer substrate including a plurality of through-holes and depositing a conductive layer on one side of the polymer substrate. The method can further include depositing an active electrode layer on the other side of the polymer substrate. One end of each through-hole can be covered with the active electrode layer, and the other end of each through-hole can be covered with the conductive layer. After the depositing step, the active electrode layer can be electrically coupled to the conductive layer via a conductive coating covering the lateral side of the through-hole. The method can further include singulating the polymer substrate covered with the active electrode layer and the conductive layer to obtain active sensors sized to cover window openings formed along the sidewalls of the sample chamber. The active sensors can include an active electrode layer and at least one through-hole covered with the conductive layer.

[0028] The method can further include bonding the active sensor to at least a portion of the chamber sidewall such that no portion of the active sensor extends into the chamber cavity within the sample chamber, the active electrode layer faces the chamber cavity, and any sample within the chamber cavity is in fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening. Depositing a conductive layer can include depositing a conductive material on the other side of the polymeric substrate. In some embodiments, the conductive material can be gold. [Brief explanation of the drawings]

[0029] [Figure 1A] FIG. 1A is a front view of a sensor device for measuring solution properties of a sample according to one embodiment. [Figure 1B] FIG. 1B is a side cross-sectional view showing a portion of the sensor device. [Figure 1C] FIG. 1C is an enlarged perspective view showing the active sensor of the sensor device adhered to the side wall of the chamber of the sensor device. [Figure 1D] FIG. 1D is a cross-sectional view showing a sample-filled sensor device. [Figure 2]FIG. 2 illustrates an active sensor of a sensor device according to one embodiment, insert molded into the sidewall of the chamber of the sensor device. [Figure 3A] 3A is a black and white top plan view showing the side of an active sensor covered with an active electrode layer, which in this image is molded into a portion of the chamber sidewall. [Figure 3B] Figure 3B is a black and white image showing the opposite side of the active sensor shown in Figure 3A, which in this image is molded into a portion of the chamber sidewall. [Figure 4A] FIG. 4A is a perspective view illustrating an active sensor according to one embodiment. [Figure 4B] FIG. 4B is a side view of an active sensor according to one embodiment used to measure ORP. [Figure 4C] FIG. 4C is a side view of another example active sensor used to measure pH. [Figure 4D] FIG. 4D is a side view of another example active sensor used to measure pH. [Figure 5A] FIG. 5A is a side view of another example active sensor formed using a PCB substrate. [Figure 5B] FIG. 5B shows a single PCB substrate covered with an active electrode layer that can be singulated into multiple individual active sensors. [Figure 6A] FIG. 6A is a black and white image showing an active sensor consisting of three individual active electrodes. [Figure 6B] FIG. 6B is a black and white image showing a close-up of the contact side of the active sensor shown in FIG. 6A. [Figure 6C] FIG. 6C is a black and white image showing a single PCB substrate that can be singulated into multiple individual active sensors. [Figure 7] FIG. 7 shows yet another example of an active sensor formed by covering a non-conductive polymer substrate containing through-holes with an active electrode layer and a conductive contact layer. [Figure 8A]8A and 8B are side cross-sectional views showing two different embodiments of an active sensor. [Figure 8B] 8A and 8B are side cross-sectional views showing two different embodiments of an active sensor. [Figure 9] FIG. 9 illustrates that a large non-conductive plastic sheet or a large PCB board can be singulated into multiple active sensors. [Figure 10] FIG. 10 is a side view of an alternative sensor device comprising an active sensor formed from a conductive dowel. [Figure 11] FIG. 11 is a graph showing the time course of the oxidation-reduction potential (ORP) of three samples containing E. coli, measured using three types of sensors. [Figure 12] FIG. 12 is a graph showing the change in pH over time measured using the sensor device disclosed herein for four samples containing different starting concentrations of E. coli. [Figure 13A] FIG. 13A is a perspective view of a reader device configured to receive a sensor device and measure solution properties of a sample within the sensor device. [Figure 13B] FIG. 13B is a partial cross-sectional view showing a reader device with a sample-loaded sensor device positioned within the reader device. [Figure 13C] FIG. 13C is a perspective view of a portion of the reader device with the reader housing removed. [Figure 13D] FIG. 13D is an enlarged view showing how the gas nozzle of the reader device is connected to the bottom of the sensor device to aerate the sample within the sensor device. [Figure 14] FIG. 14 illustrates a method for fabricating a sensor device for measuring solution properties of a sample according to one embodiment. [Figure 15] FIG. 15 illustrates a method for fabricating an alternative sensor device for measuring solution properties of a sample. [Figure 16] FIG. 16 illustrates a method for fabricating yet another example sensor device for measuring solution properties of a sample. DETAILED DESCRIPTION OF THE INVENTION

[0030] Aspects of the devices, systems, and methods described herein are best understood from the detailed description when read in conjunction with the accompanying drawings. It is emphasized that, according to common practice, the various elements of the drawings are not to scale. Dimensions of certain elements have been expanded or reduced for clarity, and not every element may be shown or labeled in every drawing. The drawings are provided for illustrative purposes only and are not intended to define or limit the scope of the claims to those shown.

[0031] 1A-1D illustrate an embodiment of a sensor device 100 for measuring a solution property of a sample. In some embodiments, the solution property measured may be the oxidation-reduction potential (ORP) of the sample. In other embodiments, the solution property measured may be the pH of the sample.

[0032] In some embodiments, the sample may be obtained from a patient or subject, hi other embodiments, the sample may be a biological sample, an environmental sample, or a food sample.

[0033] When the sample is obtained from a patient or subject, the sample may include at least one of a bodily fluid of the patient or subject and a swab obtained from the patient or subject.

[0034] In some embodiments, the patient or subject may be a human patient or subject, while in other embodiments, the patient or subject may be a non-human animal patient or subject.

[0035] In some embodiments, the bodily fluid may consist of blood, urine, serum, plasma, saliva, sputum, semen, breast milk, joint fluid, spinal fluid such as cerebrospinal fluid, wound material, mucus, stool-associated fluid, vaginal secretions, synovial fluid, pleural fluid, peritoneal fluid, pericardial fluid, amniotic fluid, or a combination thereof.

[0036] In these and other embodiments, the swabs obtained from the patient or subject may consist of wound swabs, rectal swabs, vaginal swabs, resuspended instances of the swabs described above, or combinations thereof.

[0037] In all such embodiments, the sample may contain multiple microorganisms or infectious agents. The devices, systems, and methods disclosed herein can be used to evaluate a sample for microbial growth or lack thereof as part of a microbial quantification procedure or an antibiotic susceptibility testing (AST) procedure.

[0038] In certain embodiments, the sample can include or refer to a bacterial culture derived from at least one of a sample obtained from a patient or subject, a biological sample, an environmental sample, and a food sample. For example, the sample can include or refer to a bacterial culture or resuspended bacterial culture derived from a bodily fluid or swab obtained from a patient or subject. As a more specific example, the sample can include a bacterial culture or resuspended bacterial culture derived from a bodily fluid or swab obtained from a patient or subject that has tested positive for microbial growth.

[0039] More specifically, the sample can include a bacterial culture derived from blood obtained from a patient or subject that tests positive for microbial growth. In some embodiments, the sample is or can indicate a positive blood culture. For purposes of this disclosure, a positive blood culture can be a bacterial culture derived from blood drawn from a patient or subject that tests positive for bacterial growth. For example, a patient may exhibit symptoms of sepsis (high fever, chills, etc.), and blood (e.g., 5 mL to 10 mL) may be drawn from the patient and transferred to a commercially available blood culture container or vessel containing bacterial growth medium (e.g., 30 mL to 40 mL of growth medium). The blood culture container or vessel may then be incubated at 35°C plus or minus 2°C to allow bacteria to grow. If the patient's blood is contaminated with bacteria, the bacteria will multiply within the container or vessel. A blood culture system or device can then be used to monitor bacterial growth (e.g., by monitoring bacterial CO2 production within the container or vessel), and the system or device can determine the sample as "positive" for bacterial growth if a critical CO2 threshold is met. Depending on the type and growth rate of the pathogen, blood cultures may become positive between 7 hours and 3 days. Such "positive blood cultures" may be used for further downstream testing, such as using any of the devices, systems, and methods disclosed herein.

[0040] In additional embodiments, the sample can include an environmental sample obtained from a stream, river, lake, ocean, contamination site, quarantine area, emergency area, or combinations thereof, hi other embodiments, the sample can include a food sample obtained from a food preparation facility, a diner, a waste facility, or combinations thereof.

[0041] In some embodiments, an aqueous growth medium can be added to the sample before it is introduced into the sample container 104 of the sensor device 100. In other embodiments, an aqueous growth medium can be added to the sample once the sample is injected, delivered, poured, or otherwise introduced into the sample container 104.

[0042] In one embodiment, the aqueous growth medium can be Mueller-Hinton Broth (MHG) supplemented with glucose, while in other embodiments, the aqueous growth medium can be a solution containing Bactotryptone, tryptic soybean digest, yeast extract, beef extract, cation-adjusted Mueller-Hinton Broth (CAMHB), starch, acid hydrolysate of casein, calcium chloride, magnesium chloride, sodium chloride, blood or hemolyzed horse serum (LHB), CAMHB-LHB, glucose or other carbohydrates, or a combination thereof.

[0043] Microorganisms or infectious agents that can be evaluated using the devices, methods, and systems disclosed herein can be any metabolically active unicellular or multicellular organism, including bacteria and fungi. In certain embodiments, the microorganism or infectious agent is Acinetobacter, Acetobacter, Actinomyces, Aerococcus, Aeromonas, Agrobacterium, Anaplasma, Azorhizobium, Azotobacter, Bacillus, Bacteriodes, Bartonella, Bordesella, Borrelia, Brucella, Burkholderia, Campylobacter, Chlamydia, Chlamydophila, Citrobacter, Clostridium, Corynebacterium, Coxiella, Ehrlichia, Enterobacter, Enterococcus, Escherichia, Francisella, Fusobacterium, Gardnerella, Haemophilus, Helicobacter, Klebsiella, Lactobacillus, Legionella, Listeria, Methioninae ... The microorganism or infectious agent may be a bacterium, including, but not limited to, Tanobacterium, Microbacterium, Micrococcus, Morganella, Moraxella, Mycobacterium, Mycoplasma, Neisseria, Pandrea, Pasteurella, Peptostreptococcus, Porphyromonas, Prevotella, Proteus, Providencia, Pseudomonas, Ralstonia, Raoultella, Rhizobium, Rickettsia, Localimaea, Rothia, Salmonella, Serratia, Shewanella, Shigella, Spirillum, Staphylococcus, Streptomyces, Treponema, Vibrio, Wolbachia, Yersinia, or a combination thereof. In other embodiments, the microorganism or infectious agent may be one or more fungi selected from the genera Candida or Cryptococcus, or molds.

[0044] Other specific bacteria that can be evaluated using the methods and systems disclosed herein include Staphylococcus aureus, Staphylococcus lugdunensis, coagulase-negative Staphylococcus species (including, but not limited to, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus aureus, Staphylococcus capitis, and undifferentiated), Enterococcus faecalis, Enterococcus faecium (Enterococcus faecium and other Enterococcus species, including, but not limited to, undifferentiated, excluding Enterococcus faecalis), Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus species (Streptococcus mitis, Streptococcus pyogenes, Streptococcus gallolyticus, Streptococcus agalactiae, Streptococcus pneumoniae, and undifferentiated). The bacterial strains may include, but are not limited to, Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella spp. (including but not limited to Klebsiella pneumoniae, Klebsiella oxytoca, undifferentiated), Escherichia coli, Enterobacter spp. (including but not limited to Enterobacter cloacae, Enterobacter aerogenes, undifferentiated), Proteus spp. (including but not limited to Proteus mirabilis, Proteus vulgaris, undifferentiated), Citrobacter spp. (including but not limited to Citrobacter freundii, Citrobacter koseri, undifferentiated), Serratia marcescens, Candida albicans, Candida glabrata, and Candida tropicalis.

[0045] Other more specific bacteria that may be evaluated include, among others, Acinetobacter baumannii, Actinobacillus spp., Actinomyces spp. (including but not limited to Actinomyces israelii, and Actinomyces naeslundii), Aeromonas spp. (including but not limited to Aeromonas hydrophila, Aeromonas veronii biovar sobria (Aeromonas sobria), and Aeromonas caviae), Anaplasma phagocytophilum, Alcaligenes psilocydans, and Actinomyces spp. Bacterial pathogens include Bacillus actinomycetemcomitans, Bacillus spp. (including but not limited to Bacillus anthracis, Bacillus cereus, Bacillus subtilis, Bacillus thuringiensis, and Bacillus stearothermophilus), Bacteroides spp. (including but not limited to Bacteroides fragilis), Bartonella spp. (including but not limited to Bartonella bacilliformis and Bartonella henselae), Bifidobacterium spp., Bordetella spp. (including but not limited to Bordetella pertussis, Bordetella parapertussis, and and Bordetella bronchiseptica), Borrelia spp. (including but not limited to Borrelia recurrentis and Borrelia burgdorferi), Brucella spp. (including but not limited to Brucella abortus, Brucella canis, Brucella melintensis, and Brucella suis), Burkholderia spp. (including but not limited to Burkholderia pseudomallei and Burkholderia cepacia), Campylobacter spp. (including but not limited to Campylobacter pneumoniae, Campylobacter spp. ... Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter fetus), Capnocytophaga spp., Cardiobacterium hominis, Chlamydia trachomatis, Chlamydophila pneumoniae, Chlamydophila psittaci, Citrobacter spp., Coxiella burnetii, Corynebacterium spp. (including but not limited to Corynebacterium diphtheriae, Corynebacterium jacqueum, and Corynebacterium spp.),Clostridium spp. (including, but not limited to, Clostridium perfringens, Clostridium difficile, Clostridium botulinum, and Clostridium tetani), Eikenella colobodens, Enterobacter spp. (Enterobacter aerogenes, Enterobacter agglomerans, Enterobacter cloacae), and Escherichia coli (including, but not limited to, opportunistic E. coli, which also includes enterotoxigenic E. coli, enteroinvasive E. coli, enterohemorrhagic E. coli, enterocollagenous E. coli, and urinary tract E. coli) including, but not limited to, pathogenic Escherichia coli), Enterococcus spp. (including, but not limited to, Enterococcus faecalis and Enterococcus faecium), Ehrlichia spp. (including, but not limited to, Ehrlichia chaffeensia and Ehrlichia canis), Erysipelothrix rhusiopathiae, Eubacterium spp., Francisella tularensis, Fusobacterium nucleatum, Gardnerella vaginalis, Gemella morvirorum, Haemophilus spp. (Haemophilus influenzae), Helicobacter spp. (including but not limited to Helicobacter pylori, Helicobacter cinerea, and Helicobacter fenneri), Kingella kingii, Klebsiella spp. (including but not limited to Klebsiella pneumoniae, Klebsiella granulomatis, and Klebsiella oxytoca) (including, but not limited to, Lactobacillus spp., Listeria monocytogenes, Leptospira interrogans, Legionella pneumophila, Leptospira interrogans, Peptostreptococcus spp., Moraxella catarrhalis, Morganella spp., Mobiluncus spp., Micrococcus spp., Mycobacterium spp. (including, but not limited to, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium intracellulare, Mycobacterium avium, Mycobacterium bovis, and Mycobacterium marinum)(including but not limited to Mycoplasma pneumoniae, Mycoplasma hominis, and Mycoplasma genitalium), Nocardia (including but not limited to Nocardia asteroides, Nocardia ciliasigeorgica, and Nocardia brasiliensis), Neisseria (including but not limited to Neisseria gonorrhoeae and Neisseria meningitidis), Pasteurella mulhera Tosida, Plesiomonas shigelloides, Prevotella spp., Porphyromonas spp., Prevotella melaninogenica, Proteus spp. (including but not limited to Proteus vulgaris and Proteus mirabilis), Providencia spp. (including but not limited to Providencia alcalifaciens, Providencia rettgeri, and Providencia stuartii), Pseudomonas aeruginosa, Propionibacterium acnes, Rhodococcus equi, Rickettsia spp. Rickettsia rickettsii, Rickettsia akari, and Rickettsia prowazekii, Orientia tsutsugamushi (formerly Rickettsia tsutsugamushi), and Rickettsia typhi), Rhodococcus spp., Stenotrophomonas maltophilia, Salmonella spp. (including but not limited to Salmonella enterica, Salmonella typhi, Salmonella paratyphi, Salmonella enteritidis, Salmonella cholerae suis, and Salmonella typhimurium) ), Serratia spp. (including, but not limited to, Serratia marcesans and Serratia liquifaciens), Shigella spp. (including, but not limited to, Shigella dysenteriae, Shigella flexneri, Shigella boydii, and Shigella sonnei), Staphylococcus spp. (including, but not limited to, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hemolyticus, and Staphylococcus saprophyticus), Streptococcus spp. (e.g., Streptococcus pneumoniae (e.g., chloramphenicol-resistant serotype 4 Streptococcus pneumoniae, spectinomycin-resistant serotype 6B Streptococcus pneumoniae, streptomycin-resistant serotype 9V Streptococcus pneumoniae, erythromycin-resistant serotype 14 Streptococcus pneumoniae,Optocin-resistant serotype 14 Streptococcus pneumoniae, rifampicin-resistant serotype 18C Streptococcus pneumoniae, tetracycline-resistant serotype 19F Streptococcus pneumoniae, penicillin-resistant serotype 19F Streptococcus pneumoniae, and trimethoprim-resistant serotype 23F Streptococcus pneumoniae, chloramphenicol-resistant serotype 4 Streptococcus pneumoniae, spectinomycin-resistant serotype 6B Streptococcus pneumoniae, streptomycin-resistant serotype 9V Streptococcus pneumoniae, optocin-resistant serotype 14 Streptococcus pneumoniae, rifampicin-resistant serotype 18C Streptococcus pneumoniae, penicillin-resistant serotype 23F Streptococcus pneumoniae, and chloramphenicol-resistant serotype 4 Streptococcus pneumoniae. Streptococcus pneumoniae (either methicillin-resistant serotype 19F or trimethoprim-resistant serotype 23F), Streptococcus agalactiae, Streptococcus mutans, Streptococcus pyogenes, group A streptococcus, Streptococcus pyogenes, group B streptococcus, Streptococcus agalactiae, group C streptococcus, Streptococcus anginosus, Streptococcus equimiris, group D streptococcus, Streptococcus bovis, group F streptococcus, Streptococcus anginosus, and group G streptococcus (including but not limited to), Spirillum minus, Streptobacillus moniliformi, Treponema spp. (including but not limited to Treponema calatheum, Treponema petenue, Treponema pallidum, and Treponema endemicum), Tropherima whipperi, Ureaplasma urealyticum, Veillonella spp., Vibrio spp. (Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio parahaemolyticus Vibrio species may include, but are not limited to, Vibrio vulnificus, Vibrio alginolyticus, Vibrio mimicus, Vibrio holisae, Vibrio fluvialis, Vibrio metschnikowii, Vibrio damsela, and Vibrio furnicee, Yersinia species (including but not limited to Yersinia enterocolitica, Yersinia pestis, and Yersinia pseudotuberculosis), and Xanthomonas maltophilia.

[0046] Additionally, other microorganisms or infectious agents that can be evaluated using the methods and systems disclosed herein include Candida species (including, but not limited to, Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, and Candida krusei), Aspergillus species (including, but not limited to, Aspergillus fumigatos, Aspergillus flavus, and Aspergillus clavatus), Cryptococcus species (including, but not limited to, Cryptococcus neoformans, Cryptococcus thuringiensis, Cryptococcus thuringiensis), and the like. Fungi or molds may include, but are not limited to, fungi such as Cryptococcus gattii, Cryptococcus laurentii, and Cryptococcus albidus, Fusarium species (including but not limited to Fusarium oxporum, Fusarium solani, Fusarium verticillioides, and Fusarium proliferatum), Rhizopus oryzae, Penicillium marneffei, Coccidiodes immitis, and Blastomyces dermatitidis.

[0047] 1A is a front view illustrating a sensor device 100 according to one embodiment for measuring solution properties of a sample. The sensor device 100 can include a sample container 104 including a sample chamber 108, a reference sensor 122 formed as part of a container cap 116 (see, e.g., FIG. 1D), and an active sensor 106 coupled to at least a portion of the sample chamber 108. The container cap 116 can be removably or detachably coupled or secured to the sample container 104 (e.g., threaded or pressed onto the top of the sample container 104).

[0048] The sample vessel 104 can be formed in part from an inert or non-conductive material. In some embodiments, the sample vessel 104 can be formed in part from a polymeric material, a ceramic material, or glass, or a combination thereof. As a more specific example, the sample vessel 104 can be formed in part from polyvinyl chloride (PVC), poly(methyl methacrylate) (PMMA), polydimethylsiloxane (PDMS), or a combination thereof.

[0049] 1B is a side cross-sectional view of a portion of the sensor device 100. FIG. 1B shows that the sample chamber 108 can be comprised of a chamber sidewall 112 that encloses a chamber cavity 109 configured to receive a sample. The active sensor 106 can be affixed, glued, or otherwise coupled to the chamber sidewall 112 of the sample container 104. In other embodiments not shown, the active sensor 106 can be coupled to or positioned along the bottom of the sample container 104.

[0050] The active sensor 106 may be coupled to at least a portion of the chamber sidewall 112 at a window opening 114 formed along the chamber sidewall 112. The chamber sidewall 112 may include a recess 134 surrounding the window opening 114. The recess 134 may be formed along an outer side of the chamber sidewall 112.

[0051] With regard to the placement of the active sensor 106, the active sensor 106 can be configured such that no portion of the active sensor 106 extends into the chamber cavity 109, as shown in FIG. 1C.

[0052] As described in more detail below, the active sensor 106 can be formed of a conductive substrate partially covered by an active electrode layer 132. The active electrode layer 132 of the active sensor 106 can face the chamber cavity 109 such that a sample in the chamber cavity 109 is in fluid contact with the active electrode layer 132 through at least a portion of the chamber sidewall 112 surrounding the window opening 114.

[0053] 1C is a close-up perspective view showing an active sensor 106 adhered to a chamber sidewall 112. In the embodiment shown in FIG. 1C, the active sensor 106 is adhered to a recess 134 in the chamber sidewall 112. At least a portion of the active electrode layer 132 of the active sensor 106 can cover a window opening 114 formed along the chamber sidewall 112 such that this portion of the active electrode layer 132 covering the window opening 114 is positioned in fluid communication with the chamber cavity 109 of the sample chamber 108. When the sample chamber 108 is filled with sample, the sample can be in fluid contact with the portion of the active electrode layer 132 covering the window opening 114.

[0054] 1C also shows that the active sensor 106 can be covered on its sides by adhesive 138. Because the active sensor 106 can be composed of multiple layers, the adhesive 138 can protect certain layers of the active sensor 106 from unwanted contact with the fluid sample. The adhesive 138 can act as a barrier to prevent the fluid sample from contacting the lateral sides 136 of the active sensor 106. In other embodiments not shown but contemplated by the present disclosure, the recess 134 in the chamber sidewall 112 can be sized so that the active sensor 106 fits tightly within the recess 134, with the walls of the recess 134 adjacent to or bonded to the lateral sides 136 of the active sensor 106. This can ensure that only the exposed portions of the active electrode layer 132 contact the fluid sample, thereby enabling more accurate measurement of the solution properties of the fluid sample.

[0055] To adhere the active sensor 106 to the sample chamber 108, a bead of adhesive 138 can be applied to the inner ledge 140 and / or side edge 142 of the recess 134, after which the active sensor 106 can be pressed into the recess 134 with an end effector of a pick-and-place machine. The active sensor 106 can be pressed or otherwise urged into the recess 134 until the exterior-facing surface of the active sensor 106 is flush with the exterior surface of the chamber sidewall 112.

[0056] The adhesive 138 can then be cured to secure the active sensor 106 in place. In some embodiments, the adhesive 138 can be a medical-grade, UV-curable adhesive. For example, the adhesive 138 can be Dymax® 1405M-T-UR-SC adhesive (curable using LED light with a wavelength of approximately 405 nm). In other embodiments, the adhesive 138 can be any low-outgassing medical-grade adhesive.

[0057] As described above, the active sensor 106 can be formed of a conductive substrate partially covered by an active electrode layer 132. The active sensor 106 can be positioned such that the active electrode layer 132 faces the chamber cavity 109, allowing a sample within the chamber cavity 109 to come into fluid contact with the active electrode layer 132 through at least a portion of the chamber sidewall 112 surrounding the window opening 114. In this embodiment, the active sensor 106 (including the active electrode layer 132) is positioned radially outward from the inward-facing or cavity-facing side of the chamber sidewall 112, and the lateral sides 136 of the active sensor 106 are not exposed to the fluid sample.

[0058] In some embodiments, the solution property being measured or monitored may be the pH of the sample. When the solution property being measured or monitored is pH, the active electrode layer 132 may be a pH-sensitive material. For example, the pH-sensitive material may be or include any one or combination of silicon dioxide (SiO), aluminum oxide (AlO), titanium dioxide (TiO), tantalum oxide / tantalum pentoxide (TaO), hafnium dioxide (HfO), iridium dioxide (IrO), ruthenium dioxide (RuO), and zirconium dioxide (ZrO).

[0059] In these and other embodiments, the solution property being measured or monitored can be the oxidation-reduction potential (ORP) of the sample. When the solution property being measured or monitored is the ORP of the sample, the active electrode layer 132 can be a redox-sensitive material. For example, the redox-sensitive material can be or consist of platinum (Pt), gold (Au), a redox-sensitive metal oxide, or a combination thereof. More specifically, the redox-sensitive material can be or include silicon dioxide (SiO), aluminum oxide (AlO), titanium dioxide (TiO), tantalum pentoxide (TaO), hafnium dioxide (HfO), iridium dioxide (IrO), ruthenium dioxide (RuO), or zirconium dioxide (ZrO). Formation of the active sensor 106 is described in more detail below.

[0060] Although not shown, it is contemplated by the present disclosure that the sensor device 100 may be designed to simultaneously measure both the pH and ORP of a sample. For example, the sample chamber 108 of the sensor device 100 may include multiple window openings 114 formed along the chamber sidewall 112 of the sample chamber 108. Each of these window openings 114 may then be covered by a different active sensor 106 (e.g., one window opening 114 may be covered by an active sensor 106 having an active electrode layer 132 formed from a redox-sensitive material, and another window opening 114 may be covered by an active sensor 106 having an active electrode layer 132 formed from a pH-sensitive material).

[0061] The sensor device 100 can have a device height. In some embodiments, the device height can be between about 20.0 mm and about 50.0 mm. In some embodiments, the device height can be between about 25.0 mm and about 35.0 mm. For example, the device height can be about 31.3 mm.

[0062] 1D illustrates that the reference sensor 122 can be formed as part of the vessel cap 116. The reference sensor 122 can include a reference conduit 118 that includes a reference conduit cavity 120 (see, e.g., FIG. 1B). The reference conduit cavity 120 can have first and second openings at opposite ends of the reference conduit cavity 120. The reference conduit 118 can be an elongate channel or passage configured to extend into the chamber cavity 109 of the sample chamber 108.

[0063] The reference sensor 122 may further include a reference electrode material 149 and a wick element 150 in fluid communication with the chamber cavity 109. The reference conduit cavity 120 may house the wick element 150. At least a portion of the sample may be wicked by the wick element 150 toward the reference electrode material 149.

[0064] The reference conduit 118 can be tapered such that the volume of the reference conduit cavity 120 tapers or narrows from the reference conduit proximal end 126 to the reference conduit distal end 128 (see, for example, FIG. 1B ). The shape of the core element 150 can match or correspond to the shape of the reference conduit cavity 120. The shape of the core element 150 can be configured to taper or narrow from the core proximal end 152 to the core distal end 154.

[0065] The core element 150 can extend through the length of the reference conduit cavity 120. In some embodiments, the core element 150 can fill or occupy all of the space within the reference conduit cavity 120. In other embodiments, the core element 150 can partially fill or occupy the space within the reference conduit cavity 120.

[0066] At least a portion of the wick element 150 can be in fluid communication with the chamber cavity 109 of the sample chamber 108 such that when the sample chamber 108 is filled with sample, at least a portion of the sample in the sample chamber 108 is wicked, absorbed, or otherwise drawn by at least a portion of the wick distal end 154 toward the wick proximal end 152. The wick element 150 can be formed of a polymeric material that wicks the fluid sample toward the reference electrode material 149 by capillary action.

[0067] In some embodiments, at least a portion of the wick distal end 154 can extend beyond the second opening of the reference conduit such that the wick distal end 154 protrudes or extends into the chamber cavity 109 of the sample chamber 108. In these embodiments, the wick distal end 154 can extend or protrude into the sample when the sample chamber 108 is filled with the sample.

[0068] In other embodiments, the wick distal end 154 is positioned proximal to or above the second opening of the reference conduit such that the wick distal end 154 does not protrude or extend into the chamber cavity 109 of the sample chamber 108. In these embodiments, the wick distal end 154 can still be in fluid communication with the sample chamber 108, and the fluid sample can still reach or contact the wick distal end 154 by being drawn into the reference conduit 118 by capillary action or by rocking the sample container 104.

[0069] As described above, core element 150 can be formed in part from a porous material. Core element 150 can be formed in part from a material with pores measuring 15 μm to about 150 μm (e.g., about 50 μm). In some embodiments, core element 150 can be formed in part from a polymeric material. As a more specific example, core element 150 can be formed in part from a porous polymeric material with pores measuring 15 μm to about 150 μm. In one embodiment, core element 150 can be formed in part from high-density polyethylene (HDPE). For example, core element 150 can be formed in part from HDPE with pores measuring about 50 μm. In other embodiments, core element 150 can be formed in part from natural fibers. For example, core element 150 can be formed in part from cellulose fibers, pulp, paper, cotton, or a combination thereof.

[0070] Additionally, core element 150 can be treated with a surfactant such that at least one surface of core element 150 is coated with the surfactant. In some embodiments, core element 150 can be saturated with surfactant or immersed in a solution containing surfactant before being introduced into reference conduit cavity 120. The surfactant can be configured to increase the hydrophilicity of core element 150 (i.e., to make a substantially hydrophobic surface of core element 150 more hydrophilic). In some embodiments, the surfactant can be a fluorosurfactant. In other embodiments, the surfactant can be a non-ionic surfactant, such as one or more poloxamers. As a more specific example, the surfactant can be Pluronic® F-68.

[0071] In one embodiment, the reference conduit 118 is generally conical or frustoconical in shape and has a reference conduit cavity 120 that is also generally conical or frustoconical in shape. In other embodiments, the reference conduit 118 can be substantially shaped as an elongated pyramid with a polygonal base. For example, the reference conduit 118 can be substantially shaped as an elongated triangular, square, or pentagonal pyramid. In additional embodiments, the reference conduit 118 can be substantially shaped as a cylinder with a generally cylindrical reference conduit cavity 120. In these embodiments, the reference conduit 118 can have a tapered reference conduit distal end 128 (see, for example, FIG. 1B ).

[0072] As shown in FIG. 1D , at least a portion of the wick element 150 can be in fluid contact with the sample in the sample chamber 108. At least a portion of the sample can be wicked by the wick element 150 toward the wick proximal end 152. A reference electrode material 149 can be disposed at the wick proximal end 152. FIG. 1D also illustrates that at least a portion of the active electrode layer 132 can be in fluid contact with the sample in the sample chamber 108. As the wick element 150 attracts or wicks the sample, the sample reaches the reference electrode material 149, and charge carriers in the sample can establish an electrical connection between the reference electrode material 149 of the reference sensor 122 and the active electrode layer 132 of the active sensor 106. When both the reference sensor 122 and the active sensor 106 are electrically coupled to a reader device 190, the reader device 190 can be used to measure solution properties of the sample.

[0073] The solution properties of the sample can be determined based on the potential difference measured between the active sensor 106 and the reference sensor 122 when the reference sensor 122 and the active sensor 106 are electrically coupled to the reader device 190. For example, the reference sensor 122 can provide a stable half-cell potential compared to the active sensor 106 when both the reference electrode material 149 and the active electrode layer 132 are in fluid contact with the sample in the sample chamber 108.

[0074] In some embodiments, the reference electrode material 149 can be a conductive ink that is painted or dispensed onto the wick proximal end 152. The conductive ink that is painted or dispensed onto the wick proximal end 152 can be hardened by curing. More specifically, the conductive ink can be a silver-silver chloride (Ag-AgCl) ink.

[0075] At least a portion of the reference electrode material 149 can be bonded to the core element 150. For example, the reference electrode material 149 can be a hardened mass disposed at the core proximal end 152. In certain embodiments, the reference electrode material 149 can be centrally disposed in the container cap 116. In some embodiments, at least a portion of the reference electrode material 149 can protrude or extend beyond the container cap 116.

[0076] One advantage of the wick element 150 disclosed herein is that the wick element 150 can wick up the sample, allowing the sample to advance by capillary action through the pores of the wick element 150 toward the reference electrode material 149. For example, a liquid sample can be wicked up into the wick proximal end 152, which is in fluid contact with the reference electrode material 149. If the reference electrode material 149 is formed of a material such as silver-silver chloride (Ag-AgCl), the wick element 150 can act as a barrier or obstacle to silver ions (Ag) that would otherwise freely diffuse into the sample in the sample chamber 108. Such silver ions can be detrimental to the growth of microorganisms and infectious agents in the sample, or otherwise affect their function. The wick element 150 can act as a barrier or obstacle to the silver ions by slowing or stopping their diffusion into the sample. A wick element 150 having the dimensions and shape disclosed herein is effective in slowing or stopping the diffusion of such harmful ions.

[0077] When the reference sensor 122 is implemented as a container cap 116, the container cap 116 can have dimensions defined by a cap width (or diameter) and a cap height. In some embodiments, the cap width can be between about 10.0 mm and about 20.0 mm. For example, the cap width can be about 15.7 mm. In some embodiments, the cap height can be between about 5.0 mm and about 20.0 mm. For example, the cap height can be about 10.5 mm. When the container cap 116 is fastened, affixed, or otherwise coupled to the sample container 104, the sensor device 100 can have a device height measured from the bottom of the sample container 104 to the cap top 130 of the container cap 116.

[0078] The core element 150 can have a core height as measured from the core proximal end 142 to the core distal end. In some embodiments, the core height can be from about 10.0 mm to about 20.0 mm. More specifically, the core height can be from about 14.0 mm to about 15.0 mm. For example, the core height can be about 14.8 mm.

[0079] 1D , the reference electrode material 149 can be positioned or disposed at least partially within a central divot, depression, or recessed area of ​​the container cap 116 above the wick element 150. In the case where the reference sensor 122 is a cured conductive ink or solution (e.g., Ag-AgCl ink), the divot, depression, or recessed area can serve as a receiving space for the liquid ink or solution to be cured.

[0080] In some embodiments, the reference electrode material 149 can have a reference electrode height and a reference electrode width. The reference electrode height can be between about 0.2 mm and 1.0 mm. For example, the reference electrode height can be about 0.4 mm. The reference electrode width can be between about 2.0 mm and about 5.0 mm. For example, the reference electrode width can be about 3.0 mm. One advantage of the reference sensor 122 disclosed herein is that the reference sensor 122 can function as a stable reference electrode or provide a stable reference potential for up to 10 hours of testing or operation.

[0081] 1D also shows that the sensor device 100 can include an aeration port 160 formed along the bottom side of the sample chamber 108. In other embodiments not shown, the aeration port 160 can be formed along the chamber sidewall 112 of the sample chamber 108.

[0082] The aeration port 160 may be covered by a first air permeable membrane. The aeration port 160 and the first air permeable membrane may be configured to allow gas 162 to enter the sample chamber 108.

[0083] In some embodiments, the gas 162 can be ambient air (e.g., the air in a laboratory, clinical environment, or testing facility). In other embodiments, the gas 162 consists of a combination of pressurized oxygen, carbon dioxide, nitrogen, and argon. Aerating the sample can accelerate the growth of microbial populations within the sample by providing an oxygen-rich environment within the sample chamber 108.

[0084] In an alternative embodiment not shown, an aeration port 160 can be formed along the cap top 130 of the container cap 116 and gas 162 can be pumped from the top of the sample container 104 into the sample chamber 108.

[0085] The gas 162 (e.g., ambient air) can be pumped into the sample chamber 108 by a micropump or other pump-type device integrally incorporated within the reader device 190. The gas 162 (e.g., ambient air) can be pumped or directed into the sample chamber 108 through the aeration port 160 and the first air-permeable membrane at a constant flow rate of about 1.0 to 10.0 mL / min. In other embodiments, the gas 162 (e.g., ambient air) can be pumped or directed into the sample chamber 108 through the aeration port 160 and the first air-permeable membrane at a specific duty cycle or interval.

[0086] In certain embodiments, the second air permeable membrane can cover at least a portion of the underside of the container cap 116. The second air permeable membrane can allow any gas 162 pumped or otherwise directed into the sample chamber 108 to exit the sample chamber 108 while preventing any liquid within the sample chamber 108 from spilling out of the sample container 104.

[0087] In some embodiments, the first and second air permeable membranes can be formed of the same material. The first and second air permeable membranes can be formed of a hydrophobic air permeable film or thin sheet. For example, both the first and second air permeable membranes can be formed of or include polytetrafluoroethylene (PTFE).

[0088] 1D , the container cap 116 can be removably or detachably coupled or secured to the sample container 104 by being screwed onto a proximal portion of the sample container 104 via a threaded connection 164. When the container cap 116 (which functions as part of the reference sensor 122) is secured or coupled to the sample container 104 by the threaded connection 164, air can enter the sample chamber 108 through the aeration port 160 through the first air permeable membrane, thereby forming an airflow path 166. The air then exits the sample chamber 108 through a second air permeable membrane and an air gap 168 formed between the threads of the container cap 116 and the threads of the sample container 104.

[0089] The container cap 116 can be partially formed of a transparent or translucent material, or a transparent or non-conductive material. In other embodiments, the container cap 116 can be partially formed of a translucent or transparent material. For example, at least a portion of the wick element 150 is visible through the side of the container cap 116. This allows a user or operator of the sensor device 100 to observe the drawing of the fluid sample from the wick distal end 154 to the wick proximal end 152 when the container cap 116 is secured to the sample container 104, ensuring that at least a portion of the sample reaches the reference electrode material 149 at the wick proximal end 152. In some embodiments, the container cap 116 can be partially formed of a transparent or translucent polymeric material, glass, or a combination thereof.

[0090] In some embodiments, the sample container 104, the container cap 116, or a combination thereof can be formed in part from an inert polymeric material. For example, the sample container 104, the container cap 116, or a combination thereof can be formed in part from at least one of polyoxymethylene, polyamide, polyethylene, acrylonitrile-butadiene-styrene, polycarbonate, polypropylene, or copolymers or composites thereof. In other embodiments, the sample container 104, the container cap 116, or a combination thereof can be formed in part from a glass material, such as borosilicate glass, or a ceramic material.

[0091] 2 illustrates that when the sample container 104 is formed from a polymeric material, the active sensor 106 can be insert molded into a portion of the chamber sidewall 112. For example, the active sensor 106 can be insert molded into the chamber sidewall 112 while the sample container 104 is being formed by injection molding.

[0092] When the active sensor 106 is insert molded into a portion of the chamber sidewall 112 of the sample chamber 108, the active sensor 106 can have its lateral sides 136 sealed by the polymer material used to form the chamber sidewall 112.

[0093] In the embodiment shown in FIG. 2, the active sensor 106 can be insert molded such that the active electrode layer 132 faces the chamber cavity 109, allowing a sample within the chamber cavity 109 to come into fluid contact with the active electrode layer 132 through at least a portion of the chamber sidewall 112 surrounding the window opening 114.

[0094] 3A and 3B are black and white images showing an active sensor 106 insert molded into a polymeric material representative of the material used to construct the chamber sidewalls 112 of the sample chamber 108 (see, for example, FIGS. 1A-1D). In some embodiments, the sample chamber 108 can be formed in part from an inert polymeric material such as polyoxymethylene, polyamide, polyethylene, acrylonitrile-butadiene-styrene, polycarbonate, or polypropylene.

[0095] 3A is a top plan view showing the side of the active sensor 106 covered by the active electrode layer 132. As mentioned above, the active sensor 106 can be insert molded such that the active electrode layer 132 faces the chamber cavity 109 so that a sample in the chamber cavity 109 (see, for example, FIG. 1D ) can be in fluid contact with the exposed area of ​​the active electrode layer 132.

[0096] 3B is a top view showing the side of the active sensor 106 opposite the active electrode layer 132. The side of the active sensor 106 shown in FIG. 3B can be used to contact the conductive connections of a reader device 190 (see, for example, FIGS. 14 and 15). As described in more detail below, this side of the active sensor 106 can be referred to as the conductive layer.

[0097] 3A and 3B, the lateral sides 136 of the active sensor 106 can be sealed with a polymeric material to ensure that only the exposed portions of the active electrode layer 132 are in contact with the fluid sample in the sample chamber 108, thereby enabling more accurate measurement of the solution properties of the fluid sample.

[0098] Although not shown, it is contemplated by the present disclosure that the active sensor 106 can be affixed or otherwise coupled to the chamber sidewall 112 by locally melting (e.g., by ultrasonic welding) a portion of the chamber sidewall 112 surrounding the window opening 114 (see, e.g., FIGS. 1B-1D for the location of the window opening 114) and pressing the active sensor 106 against the melted portion on the chamber sidewall 112. Once the melted portion of the chamber sidewall 112 has cooled, the active sensor 106 is then affixed or coupled to the chamber sidewall 112.

[0099] 4A is a perspective view of an active sensor 106 according to one embodiment, with two of its lateral sides 136 visible. As shown in FIG. 4A, the active sensor 106 can be shaped as a generally flattened or truncated rectangular prism. In other embodiments, the active sensor 106 can be substantially disc-shaped or shaped as a flattened or truncated polygonal prism (e.g., a flattened or truncated pentagonal or hexagonal prism).

[0100] 4A also shows that when the active sensor 106 is shaped as a generally rectangular prism, the active sensor 106 can have a sensor length dimension 400, a sensor width dimension 402, and a sensor height dimension 404. In some embodiments, the sensor length dimension 400 can be between about 100 μm and 6.0 mm, the sensor width dimension 402 can be between about 100 μm and 6.0 mm, and the sensor height dimension 404 can be between about 10 μm and 0.70 mm. For example, when the active sensor 106 is shaped as a generally rectangular prism, the active sensor 106 can have a sensor length dimension 400 of about 6.0 mm, a sensor width dimension 402 of about 6.0 mm, and a sensor height dimension 404 of about 0.61 mm.

[0101] 4B is a side view illustrating an active sensor 106 according to one embodiment used to measure the oxidation-reduction potential (ORP) of a sample. In this embodiment, the active sensor 106 can have an active electrode layer 132 formed from a precious metal. For example, the active electrode layer 132 can be made of platinum, gold, or a combination or composite thereof.

[0102] The active electrode layer 132 may be adhered to one side of a conductive substrate 406 via an adhesive layer 408. The conductive substrate 406 may be formed of a conductive material such as stainless steel (SS). For example, the conductive substrate 406 may be SS316. In other embodiments, the conductive substrate 406 may be formed of aluminum, copper, or any combination or composite of aluminum, copper, or stainless steel.

[0103] In some embodiments, the adhesion layer 408 may be a thin layer of chromium (Cr). Alternatively, the adhesion layer 408 may be a thin layer of gold, nickel, titanium, or tantalum. The adhesion layer 408 may be disposed between the conductive substrate 406 and the active electrode layer 132.

[0104] In an alternative embodiment, the active electrode layer 132 can be deposited directly onto one side of the conductive substrate 406 without the adhesive layer 408 .

[0105] The active electrode layer 132 may have an active electrode layer thickness 410 of about 50 nm to 500 nm (e.g., about 400 nm). The adhesion layer 408 may have an adhesion layer thickness 412 of about 5 nm to 50 nm (e.g., about 20 nm). The ratio of the adhesion layer thickness 412 to the active electrode layer thickness 410 may be about 1:10 to 1:20.

[0106] The conductive substrate 406 can have a substrate layer thickness 414. The substrate layer thickness can be between about 10 μm and 0.70 mm (e.g., about 0.61 mm).

[0107] 4C shows a side view of another example active sensor 106 used to measure the pH of a sample. In this embodiment, the active sensor 106 can have an active electrode layer 132 made of a pH-sensitive material. The pH-sensitive material can be deposited as a layer directly on a conductive substrate 406 or via an adhesion layer 408.

[0108] For example, the active electrode layer 132 can be formed from a metal oxide. For example, the active electrode layer 132 can be formed from tantalum pentoxide (TaO). In other embodiments, the active electrode layer 132 can be formed from silicon dioxide (SiO), silicon nitride (SiN), aluminum oxide (AlO), titanium dioxide (TiO), hafnium dioxide (HfO), iridium dioxide (IrO), ruthenium dioxide (RuO), zirconium dioxide (ZrO), or combinations or composites thereof.

[0109] The conductive substrate 406 may be formed of a conductive material such as stainless steel (SS). For example, the conductive material may be SS316. In other embodiments, the conductive substrate 406 may be formed of aluminum, copper, or any combination or composite of aluminum, copper, or stainless steel.

[0110] In some embodiments, the adhesion layer 408 may be a thin layer of chromium (Cr). Alternatively, the adhesion layer 408 may be a thin layer of gold, nickel, titanium, or tantalum. The adhesion layer 408 may be disposed between the conductive substrate 406 and the active electrode layer 132.

[0111] As mentioned above, the active electrode layer 132 can be deposited directly onto the conductive substrate 406 without the adhesion layer 408 .

[0112] The active electrode layer 132 can have an active electrode layer thickness 410 of about 50 nm to 500 nm (e.g., about 400 nm). The adhesive layer 408 can have an adhesive layer thickness 412 of about 5 nm to 50 nm (e.g., about 20 nm). The ratio of the adhesive layer thickness 412 to the active electrode layer thickness 410 can be about 1:10 to 1:20.

[0113] The conductive substrate 406 can have a substrate layer thickness 414. The substrate layer thickness can be between about 10 μm and 0.70 mm (e.g., about 0.61 mm).

[0114] 4D is a side view of another example active sensor 106 used to measure the pH of a sample. In this embodiment, a surface modification technique can be used to modify the platinum layer 416. For example, oxygen plasma treatment can be used to oxidize the platinum layer 416 to form a platinum oxide / platinum dioxide (PtO) layer 418. The platinum oxide layer 418 thus formed reacts to hydrogen ions and can be used as a pH-sensitive layer. In this manner, the platinum oxide layer 418 can function as the active electrode layer 132.

[0115] The platinum layer 416 may be adhered to the conductive substrate 406 via an adhesive layer 408. The conductive substrate 406 may be formed of a conductive material such as stainless steel (SS). For example, the conductive substrate may be SS316. In other embodiments, the conductive substrate 406 may be formed of aluminum, copper, or any combination or composite of aluminum, copper, or stainless steel.

[0116] In some embodiments, the adhesion layer 408 may be a thin layer of chromium (Cr). Alternatively, the adhesion layer 408 may be a thin layer of gold, nickel, titanium, or tantalum. The adhesion layer 408 may be disposed between the conductive substrate 406 and the active electrode layer 132.

[0117] In an alternative embodiment, the platinum layer 416 can be deposited directly onto one side of the conductive substrate 406 without the adhesion layer 408 .

[0118] The platinum layer 416 can have a layer thickness of about 50 nm to 500 nm (e.g., about 400 nm). The adhesion layer 408 can have an adhesion layer thickness 412 of about 5 nm to 50 nm (e.g., about 20 nm).

[0119] The conductive substrate 406 can have a substrate layer thickness 414. The substrate layer thickness can be between about 10 μm and 0.70 mm (e.g., about 0.61 mm).

[0120] The platinum oxide layer 418 may have an oxide layer thickness 420. The oxide layer thickness 420 may be between about 10 nm and 100 nm.

[0121] As mentioned above, the deposited layer can be selected to achieve a desired sensitivity or specificity for a particular analyte. Other surface modification techniques, such as biofunctionalization with self-assembled monolayers (SAMs), antibodies, binding antibody fragments, binding aptamers, binding DNA, and plasma treatment, can also be employed to alter the surface properties of the deposited layer and thereby tune specificity and sensitivity.

[0122] 5A is a side view of yet another example active sensor 106. This embodiment of the active sensor 106 takes advantage of the scale and efficiency of printed circuit board (PCB) manufacturing techniques.

[0123] The active sensor 106 may be formed of a non-conductive PCB substrate 500 partially covered by the active electrode layer 132. In some embodiments, the non-conductive PCB substrate 500 may be formed of polyimide. In other embodiments, the non-conductive PCB substrate 500 may be formed of a glass-reinforced epoxy laminate material, such as FR-4 composite. In particular embodiments, the PCB substrate 500 may be a flexible PCB material.

[0124] In some embodiments, the active electrode layer 132 can be formed of a precious metal. For example, the active electrode layer 132 can be formed of platinum (see, e.g., FIGS. 5A, 5B, and 6A-6C), gold (see, e.g., FIGS. 6A-6C), or a combination or composite thereof. The platinum or gold can be deposited on the PCB substrate 500 by electrodeposition or sputtering.

[0125] The active electrode layer 132 can have an active electrode layer thickness of at least 50 nm. In certain embodiments, the active electrode layer 132 can have an active electrode layer thickness of at least 400 nm. When the active electrode layer 132 is formed from platinum, the active sensor 106 can be used to measure or monitor the ORP of a sample.

[0126] In an alternative embodiment, a platinum layer deposited on the non-conductive PCB substrate 500 can be modified using a surface modification technique to make the platinum layer pH-sensitive (see, for example, FIG. 4D). For example, an oxygen plasma treatment can be used to oxidize the platinum layer to form a platinum oxide (PtO) layer. The platinum oxide layer thus formed responds to hydrogen ions and can be used as a pH-sensitive layer. In this embodiment, the active sensor 106 can be used to measure or monitor the pH of a sample.

[0127] The PCB substrate 500 may have conductive contacts or a conductive contact layer 502 patterned on the side of the substrate opposite the active electrode layer 132. In some embodiments, the conductive contact layer 502 may be a gold layer. In other embodiments, the conductive contact layer 502 may be formed of another type of conductive metal, such as platinum, nickel, copper, or an alloy or composite thereof.

[0128] 5A, the active electrode layer 132 may be electrically coupled to the conductive contact or conductive contact layer 502 by one or more conductive vias 504. In one embodiment, the conductive vias 504 may be formed in part from copper or a copper alloy. In other embodiments, the conductive vias 504 may be formed from another type of conductive metal, such as gold.

[0129] In some embodiments, each active sensor 106 can have at least one conductive via 504 located in the center of the sensor package. In other embodiments, the conductive vias 504 can be located near the periphery or edge of the sensor package.

[0130] The conductive vias 504 can be formed by electroplating, vapor deposition, or a combination thereof. Additionally, standard PCB etching processes can be used to form additional features or patterns on the PCB substrate 500.

[0131] 5B illustrates a single PCB substrate covered with an active electrode layer 132 (e.g., platinum) that can be singulated into multiple individual active sensors 106. For example, a single PCB substrate can be singulated to produce 400-500 active sensors 106.

[0132] 6A is a black-and-white image showing an active sensor 106 consisting of three individual active electrodes, including a gold (Au) active electrode 600A, a first platinum (Pt) active electrode 600B, and a second platinum active electrode 600C. The active sensor 106 can be fabricated using the PCB fabrication techniques described above. The only difference is that different active electrode materials (e.g., Au and Pt) are electroplated or vapor-deposited onto the same non-conductive PCB substrate 500. For example, one section or strip of the non-conductive PCB substrate 500 can be covered with a first active electrode material or layer (e.g., Au), and another section or strip of the same non-conductive PCB substrate 500 can be covered with a second active electrode material or layer (e.g., Pt).

[0133] As shown in Figure 6A, the active sensor 106 may be bonded (e.g., glued or insert molded) to a portion of the chamber sidewall 112 of the sample chamber 108. The sample container 104 shown in Figure 6A is formed as a transparent container, allowing the active electrode side of the active sensor 106 to be viewed through the transparent wall of the sample chamber 108. When the sample chamber 108 is filled with a fluid sample (not shown in Figure 6A), the fluid sample can contact the gold active electrode 600A, the first platinum active electrode 600B, and the second platinum active electrode 600C through window openings 114 formed along the chamber sidewall 112.

[0134] One advantage of an active sensor 106 consisting of multiple active electrodes is that each electrode can report a unique potential relative to the same reference electrode or sensor (e.g., reference sensor 122). Additionally, the active electrodes can be formed of different materials so that different solution properties of a sample (e.g., ORP and pH) can be measured or monitored simultaneously.

[0135] Although not shown, it is contemplated by the present disclosure that multiple active electrodes arranged in an active electrode array (e.g., electrode array 96) to form an active sensor 106 may be integrally formed in a single sensor device 100 to simultaneously measure multiple solution properties of a sample. The multiple active electrodes may be patterned on a non-conductive PCB substrate 500 using techniques common in the PCB industry, including selective etching, photoresist layers, shadow masking, or combinations thereof.

[0136] Figure 6B is a black-and-white image showing a close-up of the contact side of the active sensor 106 shown in Figure 6A. For example, as shown in Figure 6B, each of the active electrodes can have a unique conductive contact strip 602 or segment deposited or plated on the opposite side of the non-conductive PCB substrate 500. For example, the conductive contact strips 602 or segments can be formed of gold. The conductive contact strips 602 or segments can be segmented instances of the conductive contact layer 502 (see, e.g., Figure 5A). The active electrodes can be electrically coupled to the conductive contacts by conductive vias (not shown in Figure 6B) extending through the non-conductive PCB substrate 500.

[0137] FIG. 6C is a black-and-white image showing a single PCB substrate that can be singulated into individual active sensors 106 (see, e.g., FIGS. 6A and 6B). As described above, a non-conductive PCB substrate can be covered on one side with an active electrode layer 132 using electroplating or sputter deposition. The other side of the PCB substrate can be partially covered with a conductive contact layer 502 (also via electroplating or deposition techniques). The active electrode layer 132 can be electrically coupled to the conductive contact layer 502 by conductive vias 504 extending through the non-conductive PCB substrate. As described above, a single PCB substrate processed in this manner can be singulated to produce 400 to 500 active sensors 106.

[0138] FIG. 7 shows yet another example of an active sensor 106 formed by covering a non-conductive polymer substrate 700 defining through-holes 702 with an active electrode layer 132 and a conductive contact layer 502 .

[0139] The non-conductive polymer substrate 700 can be a substrate formed from any type of injection molded plastic, such as polyamide, polycarbonate, polyoxymethylene, polystyrene, acrylonitrile-butadiene-styrene, polypropylene, polyethylene, or copolymers or blends thereof.

[0140] In some embodiments, the active electrode layer 132 is a precious metal layer. For example, the active electrode layer 132 can be a platinum layer, a gold layer, or a combination or composite thereof. The platinum or gold layer can be deposited or otherwise applied to the conductive polymer substrate 700 via sputter deposition (e.g., physical vapor deposition (PVD) sputter deposition), evaporative deposition, or electrodeposition. In some embodiments, the platinum or gold layer can be printed using screen printing.

[0141] The active electrode layer 132 can have an active electrode layer thickness of at least 50 nm. In certain embodiments, the active electrode layer 132 can have an active electrode layer thickness of at least 400 nm. When the active electrode layer 132 is formed from platinum or gold, the active sensor 106 can be used to measure or monitor the ORP of a sample.

[0142] In an alternative embodiment, the platinum layer deposited on the non-conductive polymer substrate 700 can be modified using a surface modification technique to make the platinum layer pH-sensitive (see, for example, FIG. 4D). For example, an oxygen plasma treatment can be used to oxidize the platinum layer to form a platinum oxide (PtO) layer. The platinum oxide layer thus formed responds to hydrogen ions and can be used as a pH-sensitive layer. In this embodiment, the active sensor 106 can be used to measure or monitor the pH of a sample.

[0143] In some embodiments, the conductive contact layer 502 may be a gold layer, while in other embodiments, the conductive contact layer 502 may be formed of another type of conductive metal, such as platinum, nickel, copper, or alloys or composites thereof.

[0144] The through-holes 702 can have a diameter of about 10 μm to 100 μm. In some embodiments, the active sensor 106 can have a width dimension of about 100 μm to 6.0 mm and a length dimension of about 100 μm to 6.0 mm. For example, the active sensor 106 can have a width dimension of about 100 μm and a length dimension of about 100 μm.

[0145] 8A and 8B are side cross-sectional views showing two different embodiments of the active sensor 106. In both embodiments, the ends of the through-holes 702 are covered by the active electrode layer 132 and the conductive contact layer 502. As shown in FIGS. 8A and 8B, a conductive coating can cover the lateral sides of the through-holes 702.

[0146] In the embodiment shown in Figure 8A, the conductive coating is made of the same material as the active electrode layer 132. In the embodiment shown in Figure 8B, the conductive coating is made of the same material as the conductive contact layer 502. Whether the sides of the through-holes 702 are covered with the active electrode material or the conductive coating material can be determined by which layer is deposited first on the non-conductive polymer substrate 700.

[0147] As a more specific example, if the active electrode layer 132 is a platinum layer and the platinum layer is first deposited on the non-conductive polymer substrate 700, the conductive coating covering the lateral sides of the through-holes may be a platinum coating. Alternatively, if the conductive contact layer 502 is a gold layer and the gold layer is first deposited on the non-conductive polymer substrate 700, the conductive coating covering the lateral sides of the through-holes may be a gold coating.

[0148] In some embodiments (e.g., as shown in FIGS. 8A and 8B ), the entire through-hole 702 need not be filled, as long as the lateral sides of the through-hole 702 are covered with the conductive coating. The conductive coating can serve as an electrical connection or conductive path between the two sides of the active sensor 106. Alternatively, at least a portion of the through-hole 702 may be filled with the conductive coating.

[0149] In some embodiments, the non-conductive polymer substrate 700 can start as a sheet of plastic with an array of small through-holes 702 formed throughout the sheet of plastic. The plastic sheet can then be first covered with either the active electrode layer 132 or the conductive contact layer 502. At least one of the lateral sides and ends of the through-holes 702 can then be coated with the material used to initially cover the plastic sheet. The other side of the plastic sheet, including the remaining open ends of the through-holes 702, can then be covered with either the conductive contact layer 502 or the active electrode layer 132, depending on which layer was applied first. Once both sides of the plastic sheet are covered, the plastic sheet can be singulated to produce individual active sensors 106. Active sensors 106 produced in this manner can be formed with dimensions as small as 100 μm × 100 μm (W × L).

[0150] 9 illustrates that a large sheet of non-conductive plastic or a large PCB can be processed using the methods disclosed herein (coated with an active electrode layer, an adhesive layer, a conductive layer, or a combination thereof) and then singulated into a plurality of active sensors 106. In some embodiments, the large sheet of non-conductive plastic or a large PCB can be singulated using sawing, laser cutting, metal shearing, hot wire cutting, die cutting, stamping, or a combination thereof.

[0151] 10 is a side view of another example sensor apparatus 100 including an active sensor 106 formed from a conductive dowel 1000. In some embodiments, the conductive dowel 1000 may be a stainless steel dowel. The conductive dowel 1000 may be partially covered by an active electrode layer 132 that extends into the chamber cavity 109. The portion of the conductive dowel 1000 that is covered by the active electrode layer 132 may extend into the chamber cavity 109 such that a sample in the chamber cavity 109 can be in fluid contact with the active electrode layer 132.

[0152] The conductive dowel 1000 may be coupled to at least a portion of the chamber sidewall 112 at a window opening formed along the chamber sidewall 112. The opposite end of the conductive dowel 1000 (the end not covered by the active electrode layer 132) may extend outside the chamber sidewall 112 to contact one or more conductive connections of the reader device 190. The conductive dowel 1000 may be substantially shaped as a cylinder with rounded edges.

[0153] The conductive dowels 1000 can be insert molded or glued to the chamber sidewall 112 .

[0154] In some embodiments, the active electrode layer 132 is a precious metal layer. For example, the active electrode layer 132 can be a platinum layer, a gold layer, or a combination or composite thereof. The platinum or gold layer can be deposited or otherwise applied to the conductive dowel 1000 via sputter deposition (e.g., physical vapor deposition (PVD) sputter deposition), evaporative deposition, or electrodeposition.

[0155] The active electrode layer 132 can have an active electrode layer thickness of at least 50 nm. In certain embodiments, the active electrode layer 132 can have an active electrode layer thickness of at least 400 nm. When the active electrode layer 132 is formed from platinum or gold, the active sensor 106 can be used to measure or monitor the ORP of a sample.

[0156] In an alternative embodiment, the platinum layer deposited on the conductive dowel 1000 can be modified using a surface modification technique to make the platinum layer pH-sensitive. For example, an oxygen plasma treatment can be used to oxidize the platinum layer to form a platinum oxide (PtO) layer. The platinum oxide layer thus formed responds to hydrogen ions and can be used as a pH-sensitive layer. In this embodiment, the active sensor 106 can be used to measure or monitor the pH of a sample.

[0157] FIG. 11 is a graph showing the time course of the oxidation-reduction potential (ORP) of three samples containing E. coli ATCC 25922 measured using three different sensors. As shown in FIG. 11, one sensor is a conventional ORP probe commonly used in diagnostic settings (e.g., a commercially available ORP probe sold by Mettler-Toledo, LLC). The other two sensors are embodiments of the sensor device 100 disclosed herein, one with an active sensor 106 comprised of a platinum active electrode layer deposited by evaporative deposition, and the other with an active sensor 106 comprised of an electroplated platinum active electrode layer. The active sensor 106 in each embodiment is coupled to at least a portion of the chamber sidewall 112 at a window opening 114 formed along the chamber sidewall 112. In these embodiments, the active sensor 106 is positioned so that no portion of the active sensor 106 extends into the chamber cavity 109 of the sample chamber 108. When each of the two sensor devices 100 was placed in the reader device 190, the change in ORP was measured by the reader device 190.

[0158] As shown by the three E. coli growth curves, the two sensor devices 100 performed similarly to the commercially available ORP probe, with acceptable variations in signal response.

[0159] FIG. 12 is a graph showing the change in pH over time measured for four samples containing different starting concentrations of E. coli ATCC 25922 using a sensor device 100 disclosed herein having an active sensor 106 comprising a tantalum oxide / tantalum pentoxide (TaO) active electrode layer.

[0160] The measured growth curve of E. coli shows a classic growth pattern in which the bacteria have an initial lag phase, followed by a logarithmic phase and finally a stationary phase, as shown in Figure 12. The pattern and shape of the curve are due to the cellular activity of the active E. coli in the sample.

[0161] 13A is a perspective view illustrating a reader device 190 configured to measure solution properties of a sample in the sample chamber 108 of the sensor device 100. The reader device 190 is capable of measuring solution properties of the sample based on a potential difference measured between the active sensor 106 (more specifically, the active electrode layer 132) and the reference sensor 122 (more specifically, the reference electrode material 149) when the active sensor 106 and the reference sensor 122 are electrically coupled via a conductive connection or interface within the reader device 190. The reader device 190 can function as a voltmeter or another type of high-impedance amplifier or source meter to measure the relative change in equilibrium potential at the interface between the electrode layers in fluid contact with a sample containing electroactive redox species or charged ions.

[0162] The growth and metabolism (or lack thereof) of infectious pathogens within a sample can alter the amount of electroactive redox species and H+ ions, thereby changing the solution properties of the sample. For example, the amount of electroactive redox species in a sample can change as a result of cellular activity caused by the infectious pathogen. More specifically, the growth and metabolism (or lack thereof) of infectious pathogens within a sample can alter the amount of energy carriers, such as nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FADH2), and therefore the amount of oxygen and electron donors.

[0163] The reader device 190 can measure the oxidation-reduction potential (ORP) of a sample when the active electrode layer 132 of the sensor device 100 is made of a material that is sensitive to the oxidation-reduction potential (ORP), such as platinum (Pt) or gold (Au). Furthermore, the reader device 190 can measure the pH of a sample when the active electrode layer 132 of the sensor device 100 is made of a material that is sensitive to pH, such as a metal oxide layer.

[0164] 13A illustrates that the reader device 190 can comprise a reader housing 1300 configured to house certain functional elements of the reader device 190, including a main controller 1301 (see, e.g., FIG. 13C), a signal readout control section 1303 (see, e.g., FIGS. 14, 15A, and 15B), a thermal control module 1305 (see, e.g., FIGS. 13B, 13C, and 13D), and an aeration control module 1307 (see, e.g., FIGS. 13B and 13C). The reader housing 1300 can also expose a touchscreen display 1302 configured to display measurement results and allow a user to input commands into the reader device 190.

[0165] The lid 1304 or cover of the reader device 190 can be opened or lifted to reveal a container receiving space 1306 (see, e.g., FIG. 13B ) configured to house or receive the sensor device 100 for analysis or interrogation by the reader device 190.

[0166] 13B is a partial cutaway view showing the reader device 190 with the sensor device 100 loaded with a sample within the reader device 190. When the sensor device 100 is positioned within the container-receiving space 1306, the reference electrode contact 1308 of the reader device 190 can be positioned or moved to contact the reference electrode material 149 positioned on the container cap 116 of the sensor device 100 (see, e.g., FIG. 1D ). Additionally, when the sensor device 100 is positioned within the container-receiving space 1306, the active electrode contact 1310 of the reader device 190 can be positioned or moved to contact the conductive substrate layer or conductive contact of the active sensor 106 (e.g., either the conductive substrate 406 of FIGS. 4B-4D or the conductive contact layer 502 of FIG. 5A ).

[0167] In some embodiments, the reference electrode contact 1308 and the active electrode contact 1310 may be comprised of one or more conductive pogo or spring-loaded pins, conductive leaf contacts, or a combination thereof. More specifically, the conductive pogo pins or leaf contacts may be formed from copper, nickel, stainless steel, or alloys thereof.

[0168] The reference electrode contact 1308 and the active electrode contact 1310 may be electrically coupled to a signal readout control 1303. The signal readout control 1303 may be comprised of one or more processors, chip sets, or chip modules programmed to convert and read out signals obtained from the active sensor 106 and the reference sensor 122 of the sensor device 100.

[0169] FIG. 13B also illustrates that the reader device 190 can comprise a thermal control module 1305 and an aeration control module 1307. The thermal control module 1305 can be configured to incubate the sample-filled sensor device 100. The thermal control module 1305 can incubate the sensor device 100 by heating at least a portion of the sensor device 100 via a heating block 1318 (see, e.g., FIG. 13D). In some embodiments, the heating block 1318 can heat a lateral side of the sample chamber 108 facing the active sensor 106. In certain embodiments, the heating block 1318 can partially surround or cradle the sample chamber 108 to heat the sensor device 100.

[0170] In some embodiments, the heating block 1318 may be formed in part from aluminum, while in other embodiments, the heating block 1318 may be formed in part from another type of thermally conductive metallic material.

[0171] The sensor device 100 can be heated to an incubation temperature of about 30°C to 40°C (e.g., about 35°C plus or minus 2°C). The sensor device 100 can be incubated for an incubation period, which can range from 15 minutes to over 48 hours. The incubation period can be adjusted depending on the type of infectious agent suspected in the sample.

[0172] In some embodiments, the thermal control module 1305 may be controlled by the main controller 1301 (see, e.g., FIG. 13C) of the reader device 190. In other embodiments, the thermal control module 1305 may be controlled by another controller or module within the reader device 190 or by the signal readout controller 1303.

[0173] In some embodiments, a nutrient or stimulus solution can be introduced into the sample chamber 108 prior to incubation in the sensor device 100. For example, the nutrient solution can be a solution containing Bactotryptone, yeast extract, beef extract, cation-adjusted Mueller-Hinton broth (CAMHB), starch, acid hydrolysate of casein, calcium chloride, magnesium chloride, sodium chloride, blood or hemolyzed horse serum (LHB), a CAMHB-LHB mixture, glucose, or a combination thereof. When the sample comprises a bodily fluid, the nutrient solution can be used to counteract the buffering effects of ions or substances contained in the sample.

[0174] The aeration control module 1307 may be configured to aerate the sample in the sample chamber 108 by pumping a gas 162 (see, e.g., FIG. 1D ) into the chamber cavity 109 containing the sample. The gas 162 may be pumped into the sample chamber 108 through aeration ports 160 formed along the bottom of the sample chamber 108 (see, e.g., FIG. 1D ).

[0175] Aerating the sample increases the oxygen supply to infectious pathogens within the sample, ensuring a faster growth rate of the infectious pathogens. Additionally, aerating the sample can also detach infectious pathogens from the interior walls of the sample chamber 108, which can inhibit biofilm formation.

[0176] 13C is a perspective view showing a portion of the reader device 190 with the reader housing 1300 removed. As shown in FIG. 13C, the aeration control module 1307 is capable of delivering gas 162 via a gas delivery conduit 1312 that connects the aeration control module 1307 to the sensor device 100. In some embodiments, at least one section of the gas delivery conduit 1312 can be positioned around or wrapped around the base or bottom of the reader device 190.

[0177] 13D shows an enlarged view of an embodiment in which a gas nozzle 1314 is connected to the bottom of the sensor device 100 to aerate the sample in the sample chamber 108. The gas nozzle 1314 can be located at the end or distal end of the gas delivery conduit 1312.

[0178] 13D, the gas nozzle 1314 can connect to the aeration port 160 at the bottom of the sample chamber 108 via a nozzle interface 1316. In some embodiments, the nozzle interface 1316 can be an O-ring. In other embodiments, the nozzle interface 1316 can be another type of gasket or fluid-tight interface.

[0179] In some embodiments, the gas 162 can be ambient air (e.g., the air in a laboratory, clinical environment, or testing facility). In other embodiments, the gas 162 consists of a combination of pressurized oxygen, carbon dioxide, nitrogen, and argon. Aerating the sample can accelerate the growth of microbial populations within the sample by providing an oxygen-rich environment within the sample chamber 108.

[0180] The aeration control module 1307 is capable of pumping gas 162 into the sample chamber 108 at a constant flow rate of approximately 1.0 mL / min to 10.0 mL / min.

[0181] In some embodiments, the aeration control module 1307 may be controlled by the master controller 1301 (see, e.g., FIG. 13C). In other embodiments, the aeration control module 1307 may be controlled by another controller or module within the reader device 190, or by the signal readout control 1303. For example, the amount of gas 162 (e.g., ambient air) pumped or directed into the sample chamber 108 may be indicated by a change in the solution properties of the sample, or the lack of such a change, as sensed by the reader device 190.

[0182] 14 illustrates a method 1400 for manufacturing a sensor device 100 for measuring solution properties of a sample. The method 1400 can include, in step 1402, cleaning a conductive substrate 406 (e.g., a sheet of stainless steel such as 316SS) with an acid and base treatment.

[0183] The conductive substrate 406 can first be cleaned with a series of acid and base treatments to remove impurities or surface contaminants (e.g., free iron). Such treatments can be performed with nitric acid (10%) followed by ammonium hydroxide (175 mM), isopropyl alcohol (99%), or acetone. In other embodiments, the conductive substrate 406 can be cleaned and descaled using other acids, bases, alcohols, solvents, or other chemicals.

[0184] The method 1400 may further include, at step 1404, depositing an adhesion material on one side of the cleaned conductive substrate 406 until an adhesion layer 408 is formed on the cleaned conductive substrate 406. In some embodiments, the adhesion layer 408 may be deposited by a sputter deposition technique, such as physical vapor deposition (PVD). In some embodiments, the adhesion layer 408 may be a layer of chromium (Cr). Chromium may be selected because it bonds with the chromium contained in the stainless steel of the conductive substrate 406. In other embodiments, the adhesion layer 408 may be a layer of gold (Au) or nickel (Ni).

[0185] Step 1404 may also include depositing an adhesion material (eg, Cr, Au, or Ni) until the adhesion layer 408 is at least 20 nm thick.

[0186] The method 1400 may further include depositing an active electrode material on the adhesion layer 408 until the active electrode layer 132 is formed on the adhesion layer 408 in step 1406. The active electrode layer 132 may be a precious metal layer, such as a platinum layer or a gold layer, when the sensor device 100 is used as an ORP sensor. Depositing the active electrode layer 132 may include depositing the active electrode material (e.g., Pt) using sputter deposition (e.g., PVD), evaporative deposition, or electrodeposition. In certain processes, such as evaporative deposition, the conductive substrate 406 may be pre-cleaned with argon (Ar) plasma etching in a vacuum. In other embodiments, the active electrode material may be applied using ink screen printing.

[0187] Step 1406 may also include depositing the active electrode material until the active electrode layer 132 has a thickness of at least 50 nm. Step 1406 may further include depositing the active electrode material until the active electrode layer 132 has a thickness of at least 400 nm. Applicant has discovered that this minimum thickness is necessary to prevent the formation of nano-sized holes in the active electrode layer 132 that would allow a fluid (e.g., a fluid sample) to pass through the active electrode layer 132 and contact other layers of the active sensor 106 (thereby adversely affecting measurements). Furthermore, when an antimicrobial material (e.g., copper or nickel) is used as the conductive substrate 406, an active electrode layer 132 having a minimum thickness of 400 nm can function as a barrier to protect microorganisms in the fluid sample.

[0188] In certain embodiments, the adhesion layer 408 can be deposited in a vacuum chamber, and the active electrode layer 132 can be deposited subsequent to the adhesion layer 408 in the same vacuum chamber.

[0189] Alternatively, step 1406 can include depositing a metal layer and surface modifying the metal layer to form a metal oxide layer. For example, step 1406 can include depositing a platinum layer and oxidizing the platinum layer to form a platinum oxide (PtO) layer that functions as the active electrode layer 132. The active electrode layer 132 can be a metal oxide layer (e.g., platinum oxide or tantalum oxide) when the sensor device 100 is used as a pH sensor.

[0190] The method 1400 may further include, at step 1408, singulating the conductive substrate 406 covered by the adhesive layer 408 and the active electrode layer 132. The conductive substrate 406 covered by the adhesive layer 408 and the active electrode layer 132 may be singulated by laser cutting, metal shearing, hot wire cutting, die cutting, stamping, or sawing. The conductive substrate 406 covered by the adhesive layer 408 and the active electrode layer 132 may be singulated to obtain active sensors 106 sized to cover window openings 114 formed along the chamber sidewalls 112 of the sample chamber 108 (see, e.g., FIGS. 1B-1D and 2 ).

[0191] The method 1400 may further include, at step 1410, coupling the active sensor 106 to at least a portion of the chamber sidewall 112. The active sensor 106 may be coupled to at least a portion of the chamber sidewall 112 such that no portion of the active sensor 106 extends into the chamber cavity 109 within the sample chamber 108, and the active electrode layer 132 faces the chamber cavity 109 such that any sample within the chamber cavity 109 can be in fluid contact with the active electrode layer 132 through at least a portion of the chamber sidewall 112 surrounding the window opening 114. The active sensor 106 may be coupled to at least a portion of the chamber sidewall 112 such that the active sensor 106 (including the active electrode layer 132) is positioned radially outward from an inward-facing or cavity-facing side of the chamber sidewall 112, and such that a lateral side 136 of the active sensor 106 is not in fluid communication with the chamber cavity 109.

[0192] In some embodiments, bonding the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may further include applying a bead of adhesive 138 to a portion of the chamber sidewall 112 within a recess 134 formed along the chamber sidewall 112 surrounding the window opening 114, pressing or placing the active sensor 106 onto the bead of adhesive 138 within the recess 134, and curing the adhesive 138.

[0193] In an alternative embodiment, coupling the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may include insert molding the active sensor 106 into the chamber sidewall 112 while forming the sample chamber 108 by injection molding.

[0194] In a further alternative embodiment, bonding the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may include locally melting (e.g., by ultrasonic welding) a portion of the chamber sidewall 112 surrounding the window opening 114, pressing or placing the active sensor 106 onto the melted portion of the chamber sidewall 112, and allowing the melted portion of the chamber sidewall 112 to cool to affix the active sensor 106 to the chamber sidewall 112.

[0195] 15 illustrates yet another method 1500 of manufacturing a sensor device 100 for measuring solution properties of a sample. The method 1500 can include, in step 1502, providing a non-conductive printed circuit (PCB) substrate 500 (see, e.g., FIG. 5A).

[0196] The method 1500 may further include depositing an active electrode material on one side of the non-conductive PCB substrate 500 until an active electrode layer 132 is formed on the non-conductive PCB substrate 500 in step 1504. Step 1504 may further include depositing the active electrode material until the active electrode layer 132 has a thickness of at least 50 nm. Step 1504 may further include depositing the active electrode material until the active electrode layer 132 has a thickness of at least 400 nm. After the depositing step, the active electrode layer 132 may be electrically coupled to a conductive contact or conductive contact layer 502 of the non-conductive PCB substrate 500 by a conductive via 504 extending through the non-conductive PCB substrate 500.

[0197] The active electrode layer 132 can be a precious metal layer, such as a platinum layer or a gold layer, when the sensor device 100 is used as an ORP sensor. Depositing the active electrode layer 132 can include depositing the active electrode material (e.g., Pt) using sputter deposition (e.g., PVD), evaporative deposition, or electrodeposition.

[0198] The method 1500 may further include, at step 1506, singulating the non-conductive PCB substrate 500 covered by the active electrode layer 132 to obtain active sensors 106 sized to cover the window openings 114 formed along the chamber sidewalls 112 of the sample chamber 108. The non-conductive PCB substrate 500 covered by the active electrode layer 132 may be singulated by laser cutting, metal shearing, hot wire cutting, die cutting, stamping, or sawing. The active sensors 106 may consist of at least one conductive via 504 extending through the PCB substrate 500.

[0199] The method 1500 may further include, in step 1508, coupling the active sensor 106 to at least a portion of the chamber sidewall 112. The active sensor 106 may be coupled to at least a portion of the chamber sidewall 112 such that no portion of the active sensor 106 extends into the chamber cavity 109 within the sample chamber 108, and the active electrode layer 132 faces the chamber cavity 109 such that any sample within the chamber cavity 109 can be in fluid contact with the active electrode layer 132 through at least a portion of the chamber sidewall 112 surrounding the window opening 114. The active sensor 106 may be coupled to at least a portion of the chamber sidewall 112 such that the active sensor 106 (including the active electrode layer 132) is positioned radially outward from an inward-facing or cavity-facing side of the chamber sidewall 112, and such that a lateral side 136 of the active sensor 106 is not in fluid communication with the chamber cavity 109.

[0200] In some embodiments, bonding the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may further include applying a bead of adhesive 138 to a portion of the chamber sidewall 112 within a recess 134 formed along the chamber sidewall 112 surrounding the window opening 114, pressing or placing the active sensor 106 onto the bead of adhesive 138 within the recess 134, and curing the adhesive 138.

[0201] In an alternative embodiment, coupling the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may include insert molding the active sensor 106 into the chamber sidewall 112 while forming the sample chamber 108 by injection molding.

[0202] In a further alternative embodiment, bonding the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may include locally melting (e.g., by ultrasonic welding) a portion of the chamber sidewall 112 surrounding the window opening 114, pressing or placing the active sensor 106 onto the melted portion of the chamber sidewall 112, and allowing the melted portion of the chamber sidewall 112 to cool to affix the active sensor 106 to the chamber sidewall 112.

[0203] 16 illustrates yet another method 1600 of fabricating a sensor device 100 for measuring solution properties of a sample. The method 1600 may include, at step 1602, providing a non-conductive polymer substrate 700 including a plurality of through-holes 702. The method 1600 may further include, at step 1604, depositing a conductive contact layer 502 on one side of the polymer substrate 700. Depositing the conductive contact layer 502 may include depositing a conductive material (e.g., Au) on the polymer substrate 700 using sputter deposition (e.g., PVD), evaporative vapor deposition, or electrodeposition.

[0204] The method 1600 may further include, in step 1606, depositing an active electrode layer 132 on another side of the polymer substrate 700. Depositing the active electrode layer 132 may include depositing an active electrode material (e.g., Pt) on the polymer substrate 700 using sputter deposition (e.g., PVD), evaporative deposition, or electrodeposition.

[0205] One end of each of the through-holes 702 may be covered by the active electrode layer 132, and the other end of each of the through-holes 702 may be covered by the conductive contact layer 502. The active electrode layer 132 may be electrically coupled to the conductive contact layer 502 via a conductive coating covering the lateral sides of the through-holes 702 after the deposition step.

[0206] The method 1600 may further include, at step 1608, singulating the non-conductive polymer substrate 700 covered by the active electrode layer 132 and the conductive contact layer 502 to obtain active sensors 106 sized to cover the window openings 114 formed along the chamber sidewalls 112 of the sample chamber 108. The non-conductive polymer substrate 700 covered by the active electrode layer 132 and the conductive contact layer 502 may be singulated by laser cutting, metal shearing, hot wire cutting, die cutting, stamping, or saw cutting. The active sensors 106 may include at least one through-hole 702 extending through the non-conductive polymer substrate 700.

[0207] The method 1600 may further include, at step 1610, coupling the active sensor 106 to at least a portion of the chamber sidewall 112. The active sensor 106 may be coupled to at least a portion of the chamber sidewall 112 such that no portion of the active sensor 106 extends into the chamber cavity 109 within the sample chamber 108, and the active electrode layer 132 faces the chamber cavity 109 such that any sample within the chamber cavity 109 can be in fluid contact with the active electrode layer 132 through at least a portion of the chamber sidewall 112 surrounding the window opening 114. The active sensor 106 may be coupled to at least a portion of the chamber sidewall 112 such that the active sensor 106 (including the active electrode layer 132) is positioned radially outward from an inward-facing or cavity-facing side of the chamber sidewall 112, and such that a lateral side 136 of the active sensor 106 is not in fluid communication with the chamber cavity 109.

[0208] In some embodiments, bonding the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may further include applying a bead of adhesive 138 to a portion of the chamber sidewall 112 within a recess 134 formed along the chamber sidewall 112 surrounding the window opening 114, pressing or placing the active sensor 106 onto the bead of adhesive 138 within the recess 134, and curing the adhesive 138.

[0209] In an alternative embodiment, coupling the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may include insert molding the active sensor 106 into the chamber sidewall 112 while forming the sample chamber 108 by injection molding.

[0210] In a further alternative embodiment, bonding the active sensor to at least a portion of the chamber sidewall 112 of the sample chamber 108 may include locally melting (e.g., by ultrasonic welding) a portion of the chamber sidewall 112 surrounding the window opening 114, pressing or placing the active sensor 106 onto the melted portion of the chamber sidewall 112, and allowing the melted portion of the chamber sidewall 112 to cool to affix the active sensor 106 to the chamber sidewall 112.

[0211] Although multiple embodiments have been described, those skilled in the art will appreciate that various changes and modifications can be made to the present disclosure without departing from the spirit and scope of the embodiments. The elements of the systems, devices, apparatus, and methods shown in connection with any embodiment are exemplary for a particular embodiment and can be used in combination with other embodiments within the present disclosure, or in other manners. For example, the steps of any method depicted in the figures or described in this disclosure do not require the particular order or sequential order shown or described to achieve the desired results. In addition, other step operations may be provided, or steps or operations may be deleted or omitted from a described method or process, to achieve the desired results. Furthermore, any component or part of any apparatus or system described in or depicted in the figures in this disclosure may be removed, deleted, or omitted to achieve the desired results. Additionally, certain components or parts of systems, devices, or apparatuses shown or described herein have been omitted for the sake of brevity and clarity.

[0212] Accordingly, other embodiments are within the scope of the following claims, and the specification and / or drawings may be regarded in an illustrative rather than a restrictive sense.

[0213] Each of the individual variations or embodiments described and illustrated herein has individual components and elements that can be readily separated or combined with the elements of any other variation or embodiment. Modifications may be made to adapt a particular situation, material, composition of matter, process, process act(s), or process step(s) to the objective, spirit, or scope of the present invention.

[0214] Methods recited herein may be carried out in any order of the recited events which is logically possible, and in the order recited, and additional steps or operations may be provided or steps or operations may be omitted to achieve desired results.

[0215] Furthermore, when a range of values ​​is specified, all intervening values ​​between the upper and lower limits of that range, and any other stated or intervening values ​​within that stated range, are intended to be encompassed within the scope of the invention. Additionally, any element of an aspect of the invention can be defined and claimed independently or in combination with any one or more elements described herein. For example, a description of a range of 1 to 5 should be considered to disclose subranges of 1 to 3, 1 to 4, 2 to 4, 2 to 5, 3 to 5, etc., as well as individual numbers within that range, e.g., 1.5, 2.5, etc., and whole or partial increments therebetween.

[0216] All pre-existing subject matter (e.g., publications, patents, patent applications) referred to herein is incorporated herein in its entirety, except to the extent that such subject matter may conflict with the subject matter of the present invention, in which case the present disclosure shall take precedence. The referenced items are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such material by virtue of prior invention.

[0217] Reference to a singular item includes the possibility of a plural of the same item. More specifically, as used in this specification and the appended claims, the singular forms "a," "an," "said," and "the" include plural references unless the context clearly dictates otherwise. It should be further noted that the claims may be drafted to exclude any element. Accordingly, this statement is intended to serve as a prerequisite for using exclusive terms such as "solely" and "only" in connection with the recitation of claim elements, or for using "negative" limitations. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0218] In understanding the scope of the present disclosure, the term "comprise" and its derivatives, as used herein, are intended to be open-ended, specifying the presence of stated features, elements, components, groups, integers, and / or steps, but not excluding the presence of other, unstated features, elements, components, groups, integers, and / or steps. The same applies to phrases of similar meaning, such as "comprise," "have," and their derivatives. Furthermore, the terms "part," "section," "portion," "member," "element," and "component," when used in the singular, can refer to both one part or multiple parts. As used herein, the directional terms "front, rear, above, below, vertical, horizontal, downward, transverse, and longitudinal," and any other similar directional terms, refer to the location of a device or apparatus or the direction of a device or apparatus that is translated or moved. Finally, as used herein, terms indicating degrees such as "substantially," "about," "approximately," and the like, refer to reasonable deviations from the stated value such that the end result is not appreciably or substantially altered (e.g., deviations of up to plus or minus 0.1%, plus or minus 1%, plus or minus 5%, or plus or minus 10%, as such variations are reasonable).

[0219] The present disclosure is not intended to be limited in scope to the particular forms defined, but is intended to cover alternatives, modifications, and equivalents of the variations or embodiments described herein. Moreover, the scope of the present disclosure fully encompasses other variations or embodiments that may become obvious to those skilled in the art in light of the present disclosure.

Claims

1. a sample container including a sample chamber, the sample chamber consisting of a chamber sidewall surrounding a chamber cavity configured to receive a sample; a reference sensor including a reference electrode material and a wick, the wick being in fluid communication with the sample chamber such that at least a portion of the sample is drawn by the wick toward the reference electrode material; an active sensor formed from a conductive substrate partially covered by an active electrode layer, the active sensor being coupled to at least a portion of the chamber sidewall at a window opening formed along the chamber sidewall, wherein no portion of the active sensor extends into the chamber cavity; Equipped with the active electrode layer faces the chamber cavity such that the sample in the chamber cavity can be in fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening; A sensor device for measuring solution properties of a sample, wherein the solution properties of the sample are determined based on a potential difference measured between the active sensor and the reference sensor when the reference sensor and the active sensor are electrically coupled to a reader.

2. 10. The sensor device of claim 1, wherein the solution property measured is redox potential and the active electrode layer is a platinum layer.

3. 10. The sensor device of claim 1, wherein the solution property being measured is pH and the active electrode layer comprises a platinum oxide layer and a tantalum oxide layer.

4. The sensor device of claim 1 , wherein the conductive substrate is stainless steel.

5. The sensor device of claim 1 , wherein the active electrode layer has an active electrode layer thickness of about 50 nm to 500 nm.

6. The sensor device of claim 5 , wherein the active sensor further comprises an adhesive layer between the conductive substrate and the active electrode layer, the adhesive layer having an adhesive layer thickness of 5 nm to 50 nm.

7. 7. The sensor device of claim 6, wherein the ratio of the thickness of the adhesive layer to the thickness of the active electrode layer is about 1:10 to 1:

20.

8. The sensor device of claim 6 , wherein the adhesion layer is a chromium layer.

9. The sensor device of claim 1 , wherein the active electrode layer has an active electrode layer thickness of at least 50 nm.

10. 10. The sensor device of claim 1, wherein the active sensor is insert molded into the chamber sidewall while the sample container is formed by injection molding.

11. 10. The sensor device of claim 1, wherein the active sensor is press molded into the chamber sidewall after the sample container is formed by injection molding.

12. 2. The sensor device of claim 1, wherein the chamber sidewall comprises a recess surrounding the window opening and formed along an outer side surface of the chamber sidewall, and the active sensor is adhered to at least a portion of the chamber sidewall within the recess via an adhesive.

13. 2. The sensor device of claim 1, wherein the active sensor includes an active electrode side, a conductive substrate side opposite the active electrode side, and a plurality of side surfaces, the side surfaces being covered by at least one of the chamber sidewall and an adhesive to prevent the side surfaces from contacting the sample.

14. 10. The sensor apparatus of claim 1, wherein the sample chamber is formed in part from at least one of polyoxymethylene, polyamide, polyethylene, acrylonitrile-butadiene-styrene, polycarbonate, and polypropylene.

15. The sensor apparatus of claim 1 , wherein the reference electrode material is a cured or hardened silver-silver chloride ink deposited or otherwise applied to the wick proximal end of the wick.

16. a sample container including a sample chamber, the sample chamber consisting of a chamber sidewall surrounding a chamber cavity configured to receive a sample; a reference sensor including a reference electrode material and a wick, the wick being in fluid communication with the sample chamber such that at least a portion of the sample is drawn by the wick toward the reference electrode material; an active sensor formed of a non-conductive printed circuit (PCB) substrate partially covered with an active electrode layer, the active electrode layer being electrically coupled to a plurality of conductive contacts of the PCB substrate with conductive vias extending through the PCB substrate, the active sensor being coupled to at least a portion of the chamber sidewall with a window opening formed along the chamber sidewall, no portion of the active sensor extending into the chamber cavity, the active sensor facing the chamber cavity such that the sample within the chamber cavity can fluidly contact the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening; Equipped with A sensor device for measuring solution properties of a sample, wherein the solution properties of the sample are determined based on a potential difference measured between the active sensor and the reference sensor when the reference sensor and the active sensor are electrically coupled to a reader.

17. The sensor device of claim 16 , wherein the PCB substrate is a flexible PCB substrate.

18. The sensor apparatus of claim 16 , wherein a portion of the PCB substrate is formed from polyimide.

19. The sensor apparatus of claim 16, wherein a portion of the PCB substrate is formed from FR-4 composite material.

20. The sensor apparatus of claim 16 , wherein the conductive vias are formed from copper.

21. 17. The sensor device of claim 16, wherein the solution property measured is redox potential and the active electrode layer is a platinum layer or a gold layer.

22. 17. The sensor device of claim 16, wherein the active electrode layer has an active electrode layer thickness of at least 50 nm.

23. 17. The sensor apparatus of claim 16, wherein the active sensor is insert molded into the chamber sidewall while the sample container is formed by injection molding.

24. 17. The sensor apparatus of claim 16, wherein the solution property being measured is pH and the active electrode layer is a metal oxide layer.

25. 17. The sensor device of claim 16, wherein the chamber sidewall comprises a recess surrounding the window opening and formed along an outer side of the chamber sidewall, and the active sensor is adhered to at least a portion of the chamber sidewall within the recess via an adhesive.

26. 17. The sensor device of claim 16, wherein the active sensor comprises an active electrode side covered by the active electrode layer, a conductive contact side opposite the active electrode side and made of the conductive contact, and a plurality of lateral sides, the lateral sides being covered by at least one of the chamber side wall and an adhesive to prevent the lateral sides from contacting the sample.

27. a sample container including a sample chamber, the sample chamber consisting of a chamber sidewall surrounding a chamber cavity configured to receive a sample; a reference sensor including a reference electrode material and a wick element, the wick element in fluid communication with the sample chamber such that at least a portion of the sample is drawn by the wick element toward the reference electrode material; an active sensor formed from a non-conductive polymer substrate including a through-hole, wherein one side of the polymer substrate and one end of the through-hole are covered with an active electrode layer, and the other side of the polymer substrate and the other end of the through-hole are covered with a conductive layer, the active electrode layer being electrically coupled to the conductive layer via a conductive coating covering multiple sides of the through-hole; Equipped with the active sensor is coupled to at least a portion of the chamber sidewall at a window opening formed along the chamber sidewall, no portion of the active sensor extends into the chamber cavity, and the active electrode layer faces the chamber cavity such that a sample in the chamber cavity can be in fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening; A sensor device for measuring solution properties of a sample, wherein the solution properties of the sample are determined based on a potential difference measured between the active sensor and the reference sensor when the reference sensor and the active sensor are electrically coupled to a reader.

28. 28. The sensor device of claim 27, wherein the active electrode layer is a platinum layer.

29. 28. The sensor device of claim 27, wherein the conductive layer is a gold layer.

30. 28. The sensor apparatus of claim 27, wherein the active electrode layer has an active electrode layer thickness of at least 50 nm.

31. 28. The sensor device of claim 27, wherein the active sensor is a rectangular component having a width dimension of about 100 μm to 6.0 mm and a length dimension of about 100 μm to 6.0 mm.

32. 28. The sensor apparatus of claim 27, wherein the through-holes have a diameter of about 10 μm to 100 μm.

33. 28. The sensor apparatus of claim 27, wherein the conductive coating covering the lateral sides of the through-hole is a platinum coating.

34. 28. The sensor apparatus of claim 27, wherein the conductive coating covering the lateral sides of the through-hole is a gold coating.

35. 28. The sensor apparatus of claim 27, wherein the active sensor is insert molded into the chamber sidewall while the sample container is formed by injection molding.

36. 28. The sensor apparatus of claim 27, wherein the active sensor is press molded into the chamber sidewall after the sample reservoir is formed by injection molding.

37. 28. The sensor device of claim 27, wherein the chamber sidewall comprises a recess surrounding the window opening and formed along an outer side of the chamber sidewall, and the active sensor is adhered to at least a portion of the chamber sidewall within the recess via an adhesive.

38. a sample container including a sample chamber, the sample chamber consisting of a chamber sidewall surrounding a chamber cavity configured to receive a sample; a reference sensor including a reference electrode material and a wick element, the wick element in fluid communication with the sample chamber such that at least a portion of the sample is drawn by the wick element toward the reference electrode material; an active sensor formed from a conductive dowel partially covered by an active electrode layer, the active sensor being coupled to at least a portion of the chamber sidewall at a window opening formed along the chamber sidewall, the portion of the conductive dowel covered by the active electrode layer extending into the chamber cavity so that the sample in the chamber cavity can be in fluid contact with the active electrode layer, and the end of the conductive dowel not extending into the chamber cavity extending outward from the chamber sidewall; Equipped with A sensor device for measuring solution properties of a sample, wherein the solution properties of the sample are determined based on a potential difference measured between the active sensor and the reference sensor when the reference sensor and the active sensor are electrically coupled to a reader.

39. 39. The sensor apparatus of claim 38, wherein the solution property measured is redox potential and the active electrode layer is a platinum layer.

40. 39. The sensor apparatus of claim 38, wherein the conductive dowel is formed in part from stainless steel.

41. 39. The sensor apparatus of claim 38, wherein the active sensor is insert molded into the chamber sidewall while the sample container is formed by injection molding.

42. 39. The sensor apparatus of claim 38, wherein the active sensor is adhered to a portion of the chamber sidewall surrounding the window opening.

43. 39. The sensor apparatus of claim 38, wherein the conductive dowel is substantially cylindrical in shape with rounded edges.

44. 39. The sensor apparatus of claim 38, wherein the active electrode layer has an active electrode layer thickness of at least 50 nm.

45. cleaning the conductive substrate with an acid and base treatment; depositing an adhesive layer on one side of the conductive substrate; depositing an active electrode layer on the adhesion layer; singulating the conductive substrate covered with the adhesive layer and the active electrode layer to obtain active sensors sized to cover window openings formed along chamber sidewalls of a sample chamber; coupling the active sensor to at least a portion of the chamber sidewall; whereby no portion of the active sensor extends into a chamber cavity within the sample chamber and the active electrode layer faces the chamber cavity such that any sample within the chamber cavity can make fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening.

46. 46. ​​The method of claim 45, wherein cleaning the conductive substrate further comprises treating the conductive substrate with ammonium hydroxide, isopropyl alcohol, or acetone after treating the conductive substrate with nitric acid.

47. 46. ​​The method of claim 45, wherein singulating the conductive substrate further comprises laser cutting, metal shearing, hot wire cutting, die cutting, stamping, or sawing the conductive substrate.

48. Coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber includes: applying a bead of adhesive to a portion of the chamber sidewall within a recess formed along the chamber sidewall surrounding the window opening; pressing the active sensor against the bead of adhesive in the recess; allowing the adhesive to cure; 46. ​​The method of claim 45, further comprising:

49. 46. ​​The method of claim 45, wherein coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber further comprises insert molding the active sensor into the chamber sidewall while the sample chamber is formed by injection molding.

50. Coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber includes: locally melting a portion of the chamber sidewall surrounding the window opening; pressing the active sensor against the melted portion of the chamber sidewall; cooling the melted portion of the chamber sidewall and affixing the active sensor to the chamber sidewall; 46. ​​The method of claim 45, further comprising:

51. 46. ​​The method of claim 45, wherein depositing the active electrode layer comprises depositing the active electrode material that makes up the active electrode layer until the active electrode layer has a thickness of at least 50 nm.

52. 52. The method of claim 51, wherein the active electrode material is platinum when the solution property being measured is the oxidation-reduction potential (ORP) of the sample.

53. 46. ​​The method of claim 45, wherein depositing the active electrode layer comprises depositing the active electrode material comprising the active electrode layer using sputter deposition, evaporative deposition, electrodeposition, or ink screen printing.

54. 54. The method of claim 53, wherein depositing the adhesion layer comprises depositing the adhesion material that makes up the adhesion layer using sputter deposition.

55. 55. The method of claim 54, wherein the adhesion layer is first deposited in a vacuum chamber and the active electrode layer is subsequently deposited in the vacuum chamber.

56. 52. The method of claim 51, wherein the active electrode material is a metal oxide when the solution property being measured is the pH of the sample.

57. 57. The method of claim 56, wherein the metal oxide is platinum oxide, and the platinum oxide covers a platinum layer deposited on the adhesion layer.

58. providing a non-conductive printed circuit (PCB) substrate; depositing an active electrode layer on one side of the PCB substrate, the active electrode layer being electrically coupled to a plurality of conductive contacts of the PCB substrate after the depositing step by conductive vias extending through the PCB substrate; singulating the PCB substrate covered by the active electrode layer to obtain active sensors sized to cover window openings formed along a chamber sidewall of a sample chamber, the active sensors including at least one conductive via extending through the PCB substrate; coupling the active sensor to at least a portion of the chamber sidewall; whereby no portion of the active sensor extends into a chamber cavity within the sample chamber and the active electrode layer faces the chamber cavity such that any sample within the chamber cavity can make fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening.

59. 60. The method of claim 58, wherein depositing the active electrode layer comprises depositing the active electrode material comprising the active electrode layer using sputter deposition, evaporative deposition, and electrodeposition.

60. 60. The method of claim 58, wherein depositing the active electrode layer comprises depositing the active electrode material that makes up the active electrode layer until the active electrode layer has a thickness of at least 50 nm.

61. 60. The method of claim 59, wherein the active electrode material is platinum or gold when the solution property being measured is the oxidation-reduction potential (ORP) of the sample.

62. 62. The method of claim 61, wherein the conductive contacts are formed in part from gold.

63. Coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber includes: applying a bead of adhesive to a portion of the chamber sidewall within a recess formed along the chamber sidewall surrounding the window opening; pressing the active sensor against the bead of adhesive in the recess; allowing the adhesive to cure; 59. The method of claim 58, further comprising:

64. 59. The method of claim 58, wherein coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber further comprises insert molding the active sensor into the chamber sidewall while the sample chamber is formed by injection molding.

65. Coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber includes: locally melting a portion of the chamber sidewall surrounding the window opening; pressing the active sensor against the melted portion of the chamber sidewall; cooling the melted portion of the chamber sidewall and affixing the active sensor to the chamber sidewall; 59. The method of claim 58, further comprising:

66. providing a non-conductive polymeric substrate having a plurality of through holes; depositing a conductive layer on one side of the polymer substrate; depositing an active electrode layer on another surface of the polymer substrate, wherein one end of the through-hole is covered by the active electrode layer and the other end of the through-hole is covered by the conductive layer, and the active electrode layer is electrically coupled to the conductive layer after the depositing step via a conductive coating covering multiple lateral sides of the through-hole; singulating the polymer substrate covered with the active electrode layer and the conductive layer to obtain active sensors sized to cover a window opening formed along a chamber sidewall of a sample chamber, the active sensors including at least one through-hole covered with the active electrode layer and the conductive layer; and bonding the active sensor to at least a portion of the chamber sidewall, whereby no portion of the active sensor extends into a chamber cavity within the sample chamber and the active electrode layer faces the chamber cavity such that any sample within the chamber cavity can make fluid contact with the active electrode layer through at least a portion of the chamber sidewall surrounding the window opening.

67. 67. The method of claim 66, wherein depositing the active electrode layer comprises depositing the active electrode material comprising the active electrode layer using sputter deposition, evaporative deposition, and electrodeposition.

68. 67. The method of claim 66, wherein depositing the active electrode layer comprises depositing the active electrode material that makes up the active electrode layer until the active electrode layer has a thickness of at least 50 nm.

69. 68. The method of claim 67, wherein the active electrode material is platinum when the solution property being measured is the oxidation-reduction potential (ORP) of the sample.

70. 67. The method of claim 66, wherein depositing the conductive layer comprises depositing a conductive material on the other side of the polymeric substrate.

71. 71. The method of claim 70, wherein the conductive material is gold.

72. Coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber includes: applying a bead of adhesive to a portion of the chamber sidewall within a recess formed along the chamber sidewall surrounding the window opening; pressing the active sensor against the bead of adhesive in the recess; allowing the adhesive to cure; 67. The method of claim 66, further comprising:

73. 67. The method of claim 66, wherein coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber further comprises insert molding the active sensor into the chamber sidewall while the sample chamber is formed by injection molding.

74. Coupling the active sensor to at least a portion of the chamber sidewall of the sample chamber includes: locally melting a portion of the chamber sidewall surrounding the window opening; pressing the active sensor against the melted portion of the chamber sidewall; cooling the melted portion of the chamber sidewall and affixing the active sensor to the chamber sidewall; 67. The method of claim 66, further comprising:

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