Systems and boxes for determining biological conditions

Abstract

The subject matter of this disclosure relates to systems, kits, and methods for determining biological conditions, and more specifically, to flow cytometry systems and methods for determining biological conditions.

Description

Technical Field

[0001] The subject matter of this disclosure relates to systems and methods for determining biological conditions, and more specifically, to flow cytometry systems and methods for determining biological conditions.

[0002] background

[0003] Many medical conditions can be diagnosed most reliably through laboratory testing of bodily samples. These tests are typically performed in laboratory facilities located away from where the patient receives treatment.

[0004] It is known that the turnaround time for diagnostic tests can be as long as 30-120 minutes. Often, the time wasted waiting for lab results can lead to further deterioration of a patient's condition, sometimes even death. In some cases, doctors are forced to act without lab results. This can result in inappropriate treatment of the patient. Therefore, reducing the time required to detect biological conditions in samples taken from patients, and providing means to perform appropriate tests outside of laboratory facilities, is associated with more appropriate treatment by doctors and, in some cases, lower mortality rates.

[0005] One drawback of current testing methods is the need for sample pretreatment before introducing the sample into the analytical equipment for measurement. This pretreatment requires reliable and accurate training for laboratory personnel. Furthermore, such pretreatment is typically time-consuming, requires additional laboratory instruments and tools, poses significant biohazard risks, and introduces multiple sources of error into the test results.

[0006] Overview

[0007] In one embodiment, the present invention provides a method for analyzing biological samples, the method comprising:

[0008] Introduce biological samples into the cartridge;

[0009] Insert the box into the analytical instrument;

[0010] Begin measurement;

[0011] Obtain the results.

[0012] According to this aspect and in one embodiment, the method of the present invention eliminates the need for manual preprocessing. According to this aspect and in one embodiment, preprocessing is performed automatically within the cassette.

[0013] The method of the present invention provides a rapid, simple, safe, accurate and reliable diagnostic means that can be performed at any location and can be performed immediately after drawing, for example, a blood sample from a patient.

[0014] According to this aspect and in one embodiment, after a blood sample is drawn from the patient, the sample or a portion thereof is injected into a cartridge, which is conveniently inserted into the instrument. Measurements are quickly initiated using a touch display, and diagnostic results are obtained automatically.

[0015] According to this aspect and in one embodiment, no manual or semi-manual pretreatment of the sample is required before it is introduced into the analytical device for measurement. Therefore, no high level of training is required for laboratory personnel, enabling on-site operation by emergency medical assistants, medical technicians, etc. The method of the present invention eliminates the need for cumbersome pretreatment, thereby saving valuable time. No additional pretreatment laboratory instruments and tools are required, eliminating the biohazard risks from benchtop pretreatment. The method of the present invention also eliminates sources of error caused by manual pretreatment.

[0016] In one embodiment, the optical-mechanical analysis instrument and the inserted cartridge are used for analysis according to the method of the invention (see...). Figure 1 In one embodiment, the optical-mechanical instrument includes the following:

[0017] a. A mechanical device for operation on a cassette to induce pretreatment and processing inside the cassette;

[0018] b. An optical device for detecting optical signals from the processed sample in the box;

[0019] c. A processing device for processing optical signals obtained from a sample;

[0020] d. Interface / display / input-output connection / remote communication features used for operation and / or for result display / result transmission.

[0021] In one embodiment, the processing of optical data obtained from the sample includes computer processing. In one embodiment, the computer processing includes using an algorithm. In one embodiment, the computer processing / algorithm of the present invention is novel and is designed to process optical results to generate various qualitative and / or quantitative information related to biological samples. The information provided by the measurements of the present invention can be tailored to various user needs. The computer processing in the embodiments of the present invention is designed according to user needs to generate information such as the identity of certain blood cells in a population, the percentage of certain blood cells in the total cell count, the ratio of two or more different cell types, cell concentration, etc.

[0022] In one embodiment, the novel cartridge of the present invention is designed such that pretreatment and processing are performed automatically within the cartridge using external mechanical devices provided by the system. According to this aspect and in one embodiment, pretreatment refers to the mixing and binding of cells with antibodies, and processing refers to the dilution / lysis of the pretreated sample. Once the biological sample is introduced into the cartridge, no additional liquid is introduced into the cartridge for processing. In embodiments of the invention, the ability to perform pretreatment within the cartridge is characterized by the presence of pre-packaged dry reagents in channels / chambers within the cartridge. Once a liquid sample is introduced into the cartridge and comes into contact with the dry reagents, the dry reagents are dissolved or dispersed into the liquid sample. For example, if the dry reagents include antibodies and the sample is a blood sample, the antibodies dissolve into the blood sample and bind to the corresponding cells in the sample. Further processing within the cartridge may include sample / cell dilution and / or lysis. In some embodiments, dilution / lysis involves liquid pre-placed in a chamber within the cartridge or pre-attached to a blister pack of the cartridge prior to sample introduction.

[0023] In addition to antibodies, the dry reagents pre-prepared in the kit may also include fluorescent beads. The fluorescent beads are used for control and calibration. For example, in one embodiment, the optical fluorescence obtained from the beads indicates the volume of the sample transmitted through the optical path. This can then be used to assess the cell concentration in the same volume of sample. The amount of reference beads in the dry reagent can be controlled. Furthermore, in one embodiment, the fluorescent reference beads can be used to test and calibrate the optical signals obtained by the optical detectors of the instrument's optical system.

[0024] In one embodiment, the box includes the following features:

[0025] Sample manipulation elements, used for sample introduction and pretreatment;

[0026] At least one bellows is used to induce fluid movement in a channel / chamber, fluid movement into a channel / chamber, or fluid movement from a channel / chamber.

[0027] At least one reactant chamber comprising a liquid solution for processing;

[0028] At least one mixing chamber for promoting mixing;

[0029] At least one fluid path for transporting the processed sample to the optical analysis area.

[0030] In one embodiment, the sample manipulation element includes:

[0031] Sample loading port; and

[0032] Mixed channels.

[0033] In one embodiment, the sample loading port and / or mixing channel, or a portion thereof, comprises a dry reagent. In one embodiment, the sample loading port is connected to the mixing channel. According to one aspect of the subject matter of this disclosure, and in one embodiment, a cartridge is provided for preparing biological samples for testing and for delivering the samples to an interrogation zone for testing, the cartridge being formed with a single cavity comprising:

[0034] Two blowers, each blower including a flexible dome;

[0035] A sample manipulation element configured to receive a biological sample therein;

[0036] One or more mixing chambers;

[0037] One or more reactant zones, each reactant zone comprising liquid reactants for selective delivery to one of the mixing chambers;

[0038] The query area includes the read area;

[0039] Multiple fluid paths, each providing fluid communication between at least two other elements of the cavity; and

[0040] Ventilation openings, including openings leading to the environment;

[0041] When the sample manipulation element is in the sealed position, the cavity is only in fluid communication with the environment through the vent.

[0042] In one embodiment, the sample manipulation element includes a seal configured to bring the sample manipulation element from an open position to a sealed position.

[0043] The sample manipulation unit may include one or more dry reagents. Each dry reagent may include one or more reagents selected from the group consisting of antibodies and fluorescent beads.

[0044] In one embodiment, two blowers are each located at one end of the cavity.

[0045] In one embodiment, the antibody is a target antibody, i.e., an antibody that specifically binds to a target antigen on a cell. In one embodiment, the antibody is linked to a colored moiety, a fluorescent moiety, or a combination thereof. According to this aspect and in one embodiment, the antibody is a cell staining agent. In some embodiments, the colored and / or fluorescent moiety on the antibody serves as a cell “staining agent” because its color / absorption / fluorescence signal can be optically monitored. In one embodiment, the dry reagent includes a positive control that identifies the antibody or a negative control that identifies the detection moiety. In one embodiment, the dry reagent includes a chemical indicator and / or a biological indicator.

[0046] The sample manipulation unit may include a mixing channel with a serpentine pathway.

[0047] The box may include two mixing chambers, with each end of the mixing channel connected to one of the mixing chambers via a fluid path spanning between the mixing chamber and the mixing channel.

[0048] In one embodiment, each end of the mixing channel is directly connected to one of the mixing chambers, without any additional fluid path.

[0049] At least one blower can be connected to one of the mixing chambers via a fluid path spanning between the blower and the mixing chamber.

[0050] At least one blower can be connected to the query area via a fluid path spanning between the blower and the query area.

[0051] Each reactant zone may include blister packs containing liquid reactants.

[0052] Each blister pack may include a flexible dome and a backing opposite the flexible dome, wherein each reactant area includes a socket, and a corresponding blister pack is hermetically received in the socket, the socket including a piercing element supported on the backing.

[0053] The box can be configured such that pressing down the dome increases the internal pressure of the blister pack, causing the backing to rupture.

[0054] The liquid reactants may include one or more items selected from the group consisting of diluents, pyrolytic agents, and reference substances. At least one of the fluid paths may include a serpentine channel. Brief description of the attached diagram

[0056] To better understand the subject matter disclosed herein and to illustrate how the subject matter can be implemented in practice, embodiments will now be described by way of non-limiting example only with reference to the accompanying drawings, in which:

[0057] Figure 1 A system based on the subject matter of this disclosure is illustrated schematically;

[0058] Figure 2: Figure 2A and Figure 2B They are Figure 1 Front and rear perspective views of an example of the box in the system shown;

[0059] Figure 3 yes Figure 1 A front perspective view of another example of the box of the system shown;

[0060] Figure 4 shows an example of an insert (plug) for a sample manipulation element; Figure 4A This is the front view; Figure 4B It is a rear view; Figure 4C It is an isometric view; Figure 4D This is a view assembled with the main frame;

[0061] Figure 5 shows an example of an insert (plug) for a sample manipulation element; Figure 5A It is a pre-expanded isometric view; Figure 5B Decompose the isometric view later.

[0062] Figure 6 is an example of the general part of the main skeleton; Figure 6A It is a general-purpose main framework; Figure 6B It is a main skeleton with two insertion areas, each of which can accommodate different components or no components.

[0063] Figure 7 shows an example of a main skeleton with two insertion areas: insertion area 1 (mixing channel) and insertion area 2 (loading port or sample manipulation element); Figure 7A and Figure 7B These are the front view and the rear view, respectively.

[0064] Figure 8 shows an example of a main skeleton with two insertion areas: insertion area 1 (loading port or sample manipulation element) and insertion area 2 (blank); Figure 8A This is the front view. Figure 8B This is the rear view.

[0065] Figure 9: Figure 9A This is a simplified three-dimensional internal front view of a system for detecting biological conditions according to an embodiment of the present invention;

[0066] Figure 9B This is a simplified three-dimensional internal rear view of a system for detecting biological conditions according to an embodiment of the present invention.

[0067] Detailed description

[0068] The subject matter of this disclosure can be more readily understood by referring to the following detailed description, which forms part of this disclosure. It should be understood that the subject matter of this disclosure is not limited to the specific products, methods, conditions, or parameters described and / or shown herein, and the terminology used herein is used only by way of example to describe particular embodiments and is not intended to limit the subject matter of this disclosure.

[0069] The subject matter of this disclosure provides a system, such as a flow cytometry system, and a method for biological assays. This system provides rapid sample processing and accurate results, enabling accurate and rapid diagnosis of biological conditions. The system is compact and easy to operate on-site as needed. The system includes a cartridge for initial sample manipulation and a cartridge for further sample processing. In some embodiments, the system further includes a spectromechanical device. This spectromechanical device is configured to receive the cartridge and perform mechanical manipulation of the cartridge's elements to facilitate sample processing within the cartridge. The spectromechanical device is also configured to perform optical measurements on the processed sample. The spectromechanical device processes the data obtained through optical measurements and provides results indicative of biological conditions. The results may be qualitative, quantitative, or both. The composition of a sample assayed according to the subject matter of this disclosure can reflect the biological condition of the subject.

[0070] Unless otherwise specified herein, scientific and technical terms used in connection with this application shall have the meanings commonly understood by one of ordinary skill in the art. Furthermore, unless the context requires otherwise, singular terms shall include plural forms, and plural terms shall include singular forms. As used above and throughout this disclosure, unless otherwise stated, the following terms and abbreviations shall be understood to have the following meanings.

[0071] In this disclosure, the singular forms “a,” “an,” and “the” include plural references, and references to a particular numerical value include at least that particular value unless the context clearly indicates otherwise. Thus, for example, a reference to “compound” refers to one or more such compounds and their equivalents known to those skilled in the art, etc. The term “a plurality” as used herein means more than one. When indicating a range of values, another embodiment includes from one particular value and / or to another particular value.

[0072] Similarly, when a value is expressed as an approximation using the antecedent “about,” it should be understood that the specific value forms another embodiment. All ranges are inclusive and composable. In the context of this disclosure, using “about” for a quantity means that the quantity is within ±20% of the quantity, or preferably within ±10% of the quantity, or more preferably within ±5% of the quantity.

[0073] In some embodiments, the terms “subject” and “patient” are used interchangeably herein and refer to an animal, such as a human, to which treatment according to the pharmaceutical composition of the subject matter of this disclosure is administered. In some embodiments, the terms “subject” and “patient” are used interchangeably herein and refer to an animal, such as a human, to which diagnostic tests are performed and / or from which samples are collected for diagnostic tests according to the subject matter of this disclosure. The term “subject” as used herein refers to both human and non-human animals. The terms “non-human animal” and “non-human mammal” are used interchangeably herein and include all vertebrates, such as mammals, such as non-human primates (especially higher primates), sheep, dogs, rodents (e.g., mice or rats), guinea pigs, goats, pigs, cats, rabbits, cattle, horses, and non-mammals, such as reptiles, amphibians, chickens, and turkeys. The formulations described herein can be used to diagnose / treat any suitable mammal, including primates (such as monkeys and humans), horses, cattle, cats, dogs, rabbits, and rodents (such as rats and mice). In one embodiment, the mammal to be diagnosed / treated is a human. A human can be anyone of any age. In one embodiment, a human is an adult. In another embodiment, the person is a child. According to any method of the subject of this disclosure, and in one embodiment, the subject is a human. In another embodiment, the subject is a non-human primate. In another embodiment, the subject is a rodent, specifically a mouse in one embodiment and a rat in another. In another embodiment, the subject is a dog, cat, cow, horse, rabbit, or pig. In yet another embodiment, the subject is a mammal.

[0074] In one embodiment, the analytical instrument of the present invention is a spectral-mechanical device (SMD) or includes a spectral-mechanical device. In some embodiments, the spectral-mechanical device is referred to as a "system". In other embodiments, the SMD is referred to as an optomechanical system or device, or as an instrument or analytical instrument, or as an optomechanical analytical instrument.

[0075] In some embodiments, pretreatment refers to mixing the sample with dry reagents. In one embodiment, pretreatment refers to mixing the sample with cells or other types of antibodies / labels for binding to the sample. Treatment in embodiments of the invention refers to any further processing of the sample after a pretreatment step. For example, in embodiments of the invention, dilution / lysis and mixing involving dilution / lysis are considered treatments or further treatments. Such treatments occur after a pretreatment step. In one embodiment, pretreatment and treatment together are referred to as a treatment.

[0076] like Figure 1As shown, a system (generally indicated as 10) is provided for analyzing liquid biological samples from a subject, for example using flow cytometry and / or by any other suitable method for performing one or more assays. System 10 includes a cartridge 12 for containing the biological sample and a spectromechanical device (SMD) 14 for facilitating manipulation of the biological sample within the cartridge and performing analysis on the cartridge.

[0077] Biological samples may include bodily fluid samples, such as blood, serum, plasma, urine, saliva, cerebrospinal fluid, serous fluid, peritoneal fluid, and / or synovial fluid. According to some embodiments, biological samples may include solids, such as a hair, a tooth, a piece of cartilage, a piece of skin, a piece of bone, and / or a piece of soft tissue.

[0078] Box 12 is configured to receive a biological sample therein and to facilitate optional pretreatment steps, processing steps (e.g., processing of a pretreated biological sample), and to position the processed biological sample at a query point where analysis can be performed on the processed biological sample at SMD 14. In the specification and appended claims, the term "biological sample" will be used to refer to a biological sample at any stage of processing.

[0079] Therefore, as Figure 2A and Figure 2B As shown, the box 12 includes a support panel 16 defining opposing front sides 16a and rear sides 16b, and carrying a sample manipulation element 18, a first blower 20a and a second blower 20b, a first mixing chamber 22a and a second mixing chamber 22b, a first reactant zone 24a, a second reactant zone 24b and a third reactant zone 24c, a vent 25, and a query zone 26. (In this specification, basic reference numerals without suffixes may be used to uniformly denote all elements indicated thereby; thus, for example, the term "blower 20" may be used to uniformly denote the first blower 20a and the second blower 20b, etc.). Furthermore, various fluid paths are formed in the support panel 16, fluidly connecting the elements carried by the support panel, as described below.

[0080] As described above, at least some of the elements carried by the support panel 16, such as the blower 20, mixing chamber 22, query area 26 and fluid path, can be formed as recesses in the rear side 16b, which protrude into the material of the support panel 16 toward the front side 16a of the support panel 16.

[0081] The housing 12 also includes a back plate (not shown) disposed on the rear side 16b of the support panel 16. The back plate is positioned at least aligned with the elements carried by the support panel 16 as described above, particularly with those elements protruding into the support panel material. The back plate is sealed to the rear side 16b of the support panel 16, thereby isolating the elements from external fluids of the housing 12 in the rearward direction. The back plate can be of any suitable design, including but not limited to rigid materials and adhesive films, with necessary modifications made without departing from the scope of the subject matter disclosed herein.

[0082] It should be understood that although the support panel 16 is described above as “carrying” various elements of the box 12, this is done for ease of description only, and the terminology used in this specification and the appended claims should be interpreted in its broadest sense, that is, including elements connected thereto, formed therein, etc., unless the context clearly indicates otherwise. For example, the blower 20 may include elements connected to the support panel 16, and the mixing chamber 22 may be defined by a structure formed in the material of the support panel.

[0083] The sample manipulation element 18 is configured to receive a biological sample therein and optionally to facilitate sample pretreatment therein. Therefore, the sample manipulation element 18 includes a mixing channel 28 formed therein, which spans between a first orifice 30a and a second orifice 30b (the positions of the first orifice 30a and the second orifice 30b are...). Figure 2B The instructions indicate that, and Figure 2B (As shown in the figure). The mixing channel 28 may include a serpentine passage 32 in which a receiving socket 34 is formed, for example for receiving a biological sample. The receiving socket 34 may be part of the serpentine passage 32, but has a wider width, and the receiving socket 34 may indicate where the biological sample should be placed, for example to ensure the proper functioning of the system 10, and / or to facilitate the placement of the biological sample therein.

[0084] At least a portion of the upper surface 36 of the sample manipulation element 18 may be flat, for example to facilitate the placement of the seal ( Figure 2A and Figure 2B A thin element (not shown) with an adhesive surface (such as a sticker) is sealed and attached to the upper surface, thereby bringing the sample manipulating element 18 into a sealed position. According to some examples, a continuous portion completely surrounding the upper surface 36 of the serpentine passage 32 may define a single plane, thereby facilitating contact between a planar seal and the surface.

[0085] like Figure 3As shown, according to some examples, the sample manipulation element 18 includes a sample port 34a for receiving a biological sample, and the cartridge 12 includes a corresponding plug 34b constituting a seal, for example, the plug 34b being formed together with the support panel 16 and configured to align with the sample port. The plug 34b is configured to seal against the sample port 34a, thereby moving the sample manipulation element 18 into a sealed position. According to these embodiments, the mixing channel 28 is isolated from the environment except through the sample port 34a and the vent 25.

[0086] like Figure 2B As best shown, the first orifice 30a is in fluid communication with a first sample mixing fluid path 38a spanning between the sample manipulation element 18 and the first mixing chamber 22a, and the second orifice 30b is in fluid communication with a second sample mixing fluid path 38b spanning between the sample manipulation element 18 and the second mixing chamber 22b. Since the sample manipulation element 18 is disposed between the two mixing chambers 22, the serpentine pathway 32 of the sample manipulation element 18 can be used to mix biological samples with reactants within the serpentine pathway of the sample manipulation element.

[0087] The sample manipulation element 18 may be provided with one or more reagents (e.g., dry reagents) arranged within the mixing channel 28 (e.g., within the serpentine pathway 32). These reagents may be configured for pretreatment of biological samples. Therefore, the reagents may include antibodies, target antibodies, positive control identification antibodies, negative control identification detection portions, target signal reference compositions, reference labeling compositions, cell staining agents, chemical indicators, biological indicators, and / or any other suitable reagents. The reagents may be provided on the surface of the reagent block, bound to a solid carrier, bound to / deposited on the walls / surfaces of the channel, or provided by any other suitable means.

[0088] It should be understood that although the terms "reagent" and "reactant" are used interchangeably, in this specification and appended claims, the term "reagent" is used to refer to a substance disposed within the sample manipulation element 18, and the term "reactant" is used to refer to a substance disposed within the reactant region 22. This distinction is made solely for clarity of this disclosure and should not be considered restrictive. In particular, substances that a person skilled in the art would classify as reagents may be included in the term "reactant," and conversely, substances that a person skilled in the art would classify as reactants may be included in the term "reagent." Furthermore, for the purposes of this disclosure, a single substance may be considered both "reactant" and "reagent," depending on its location.

[0089] Each blower 20 includes a flexible dome 40 (e.g., in the form of a hemispherical or spherical cap) projecting from the front side 16a of a support panel 16 and a back plate portion forming a planar base of the blower 20. The dome is formed such that it remains within its elastic limit by full depression, i.e., the dome can be fully depressed and returns to its original shape after the depressurizing force is removed. The dome 40 may be formed as part of the support panel 16 or may include a separate element connected to the support panel 16. The first blower 20a is in fluid communication with a blower-mixing fluid path 42a spanning between the first blower and the first mixing chamber 22a, and the second blower 20b is in fluid communication with a blower-query fluid path 42b spanning between the second blower and the query zone 26.

[0090] Each reactant zone 24 includes blister packs 44 (one of which is in...) Figure 2A (Seen in a separated configuration), and each reactant zone 24 is configured to selectively deliver its contents to one of the mixing chambers 22. Each blister pack 44 includes a flexible dome 46 (e.g., a flexible dome in the form of a hemispherical or spherical cap) and a backing material (not shown) opposite the flexible dome 46 and made of a thin layer of material (e.g., metal foil). Each reactant zone 24 also includes a socket 48 in the front side 16a of the support panel 16, for example, the socket 48 being a recess formed in the front side 16a of the support panel 16, which defines a base surface 50 and sealably receives a corresponding one of the blister packs 44 therein, such that the backing material is spaced apart from the base surface 50. Each base surface 50 includes a piercing element 52 projecting toward the front side 16a of the support panel 16. An outlet 54 including a through-hole is formed in each socket 48, for example adjacent to the bottom end of the socket 48. Figure 2B As can be seen most clearly, each outlet 54 is in fluid communication with a reactant-mixing fluid path 56, which spans between one of the corresponding reactant zone 24 and mixing chamber 22.

[0091] Each blister pack 44 may include one or more reactants, for example in liquid form, for delivery of biological samples into one of the mixing chambers 22. The reactant may be a diluent, lysis agent, reference material, or any other suitable reactant, and / or a combination thereof.

[0092] Each puncturing element 52 can be configured to be supported on the backing of the respective blister pack 44 when the blister pack 44 is received within the socket 48, such that the puncturing element 52 will not puncture the backing of the blister pack 44 until the internal pressure of the blister pack 44 increases, for example, by pressing down the dome 46, causing the backing to rupture. The support of the puncturing element 52 on the backing of the blister pack 44 can create a rupture zone on the blister pack, which is associated with the increased likelihood of rupture when the dome 46 is pressed down. Therefore, the puncturing element 52 can be positioned near its respective outlet 54 so that reactants released from the blister pack 44 upon rupture can immediately flow into the corresponding reactant-mixture path 56.

[0093] Vent 25 is in fluid communication with the environment and facilitates the movement of biological samples within the chamber when blower 20 is depressed and / or released. Therefore, vent 25 may be positioned above the remaining elements of the chamber (i.e., in the orientation of cartridge 12 when cartridge 12 is inserted into SMD 14) and in fluid communication with the remaining elements of the chamber, for example, connected to one of the mixing chambers 22 via a ventilation fluid path 25a.

[0094] like Figure 2B Most clearly visible is the query area 26, which is configured to facilitate the analysis of biological samples via the SMD 14. Therefore, the query area includes a read fluid path 58, which includes a narrow read region 60. The read region 60 can be designed to limit the number of particles passing through it simultaneously, for example, wide enough to allow only a single particle to pass through at a time for SMD 14 analysis. Furthermore, the material of the query area 26, particularly the material in the region of the read region 60, can be made of a material that is transparent to the wavelength of light emitted by the SMD 14 during analysis.

[0095] As described above, one end of the fluid reading path 58 is in fluid communication with the second blower 20b via the blower-query fluid path 42b. The other end of the fluid reading path 58 is in fluid communication with the second mixing chamber 22b via the mixing-query fluid path 62. At least a portion of the mixing-query fluid path 62 may form a serpentine channel 64. It should be noted that the geometry of the query fluid path can be varied and can be adjusted to any desired geometry suitable for a particular cartridge design. For example, in one embodiment, the fluid path 62 does not include a serpentine channel.

[0096] It should be understood that when the sample manipulation element 18 is in its sealed position, for example as described above, the elements of the aforementioned support panel 16 constitute a single cavity, which forms a closed system (i.e., isolated from the environment) such that nothing can be further introduced into the cavity except for the vent 25 necessary to allow compression of the blower 20, including but not limited to air or other fluids. Therefore, movement of the biological sample within the cavity can be achieved entirely through mechanical interaction with the elements of the support panel 16, including but not limited to action on the blower 20, as described below. (The blister pack 44 can be considered a sub-chamber within the cavity because the contents of the blister pack 44 are isolated from the environment and are introduced into the rest of the cavity during operation.) Therefore, no external pump or other means of introducing air into the cavity is required to move the biological sample within the cavity. In one embodiment, no external air pump is used to directly push air into the box.

[0097] SMD 14 can be any suitable device for interfacing with Box 12, such as those specified in US 2013 / 0102087, US2014 / 0170678, US 2014 / 0170680, US 2014 / 0287435, US 2015 / 0132776, US 2015 / 0293095, US 2015 / 0309049, US 2015 / 0330971, US 2016 / 0146793, US 2017 / 0350888, US 2017 / 0370914, US 2018 / 0231532, US 2018 / 0299443, US 2018 / 0306698 and / or US The devices described in one or more of 2020 / 0080928, the entire contents of which are incorporated herein by reference. Specifically, the SMD 14 includes elements configured to selectively interact with the blowers 20 and reactant zones 24, for example, selectively depressing and / or releasing each blower 20 and reactant zone 24 when the cartridge 12 is inserted into the SMD. The SMD 14 may also include optical units (e.g., including a light source, photon counter, and / or integrator) configured to facilitate analysis of a portion of a biological sample within the readout area 60 of the query area 26.

[0098] The SMD 14 may also include a processor to guide its operation. The processor may also perform one or more steps of analysis, such as being configured to receive data associated with multispectral emission signals detected by the optical unit, process that data, and output results, for example, related to a medical condition. Each multispectral emission signal may be associated with a biomarker.

[0099] In use, the biological sample is introduced into the receiving socket 34 formed within the sample manipulation element 18, and then the receiving socket 34 is sealed, for example, by providing a seal on the receiving socket, as described above. The cartridge 12 is inserted into the SMD 14, for example, into a slot designed for this purpose (see [link to SMD]). Figure 9A (Box 910). SMD 14 selectively interacts with the elements of the box to facilitate optional pretreatment, processing, and analysis of biological samples, as described below, for example.

[0100] In an optional pretreatment step, the first blower 20a and / or the second blower 20b are selectively depressed and released by the SMD 14. When the biological sample is within the closed system including the blowers 20, the interaction with the blowers causes the biological sample to move within the serpentine pathway 32 of the sample manipulation element 18, thereby contacting and dissolving the dry reagent within the mixing channel 28; the only opening in this closed system is the vent 25, which is necessary to allow the blowers to be depressed and released. The first blower 20a and the second blower 20b can be depressed alternately to move the biological sample back and forth within the serpentine pathway 32, thereby promoting the mixing of the biological sample with the dry reagent.

[0101] It should be understood that the description of blower 20 being “pressed down” or “released” in this document is not limited to being fully pressed down or released; each blower may be partially pressed down or released in order to move a predetermined amount of biological sample within the chamber.

[0102] The interaction between SMD 14 and blower 20 can be performed to control the mixing sequence of the biological sample with more than one reagent. For example, the first blower 20a can be depressed to move a predetermined amount of the biological sample within the mixing channel 28, allowing the biological sample to reach and dissolve the first dry reagent. The first blower 20a and the second blower 20b can then be depressed alternately to move the biological sample and the first reagent back and forth within a portion of the serpentine pathway 32, thereby achieving sufficient mixing and time for the reagent to act on the biological sample. Subsequently, the first blower 20a can be depressed to further move the biological sample along the mixing channel 28, allowing the biological sample to reach and dissolve the second dry reagent. It is understood that the sequence of interactions between SMD 14 and blower 20 can be performed to achieve any suitable sequence for mixing the biological sample with the dry reagent within the sample manipulation element 18, as necessary.

[0103] In one or more processing steps, the biological sample is selectively moved into one or both of the mixing chambers 22. For example, to move the biological sample from the sample manipulation element 18 to the first mixing chamber 22a, the second blower 20b is depressurized, thereby creating positive pressure behind it. The SMD 14 can selectively facilitate the delivery of the contents of one or more blister packs 44 into one of the mixing chambers 22 by depressing one or more blister packs 44, thereby achieving rupture in the backing of the blister pack, as described above, releasing the liquid reactants therein. The reactants released from the blister pack 44 upon rupture flow into the corresponding reactant-mixing fluid path and through that path to the corresponding mixing chamber 22.

[0104] During the query step, the processed biological sample is moved into the query area 26 through the interaction between the SMD 14 and the blower 20. The SMD 14 can be adjusted in terms of the degree of indentation on the blower 20 to adjust the position of the biological sample within the reading area 60 of the query area 26 as needed.

[0105] In some embodiments, any one of two or more blowers can induce movement of the liquid sample between components of the cartridge. Each blower can be pressed and depressed in a progressive manner to induce liquid to move back and forth through channels, to / from the chamber, or toward the optical query area. According to this aspect and in one embodiment, the cartridge includes only one blower. In one embodiment, two blowers work together to drive the liquid back and forth within the cartridge, as described herein. In one embodiment, one blower pushes the liquid in one direction while the other blower pushes the liquid in the opposite direction. In one embodiment, one or two blowers drive the liquid to induce mixing, and only one of the two blowers drives the liquid toward the optical query area. In one embodiment, pressing and depressing one blower is sufficient to move the liquid back and forth within the components of the cartridge. In one embodiment, alternating pressing and / or depressing of two blowers is used to move the liquid back and forth within the components of the cartridge. In one embodiment, progressively pressing / depressing one or more blowers determines how far the liquid will travel in a particular channel / component of the cartridge.

[0106] Figure 4 illustrates an embodiment of the cartridge, in which a reagent sample plug (RSP) is attached to the cartridge after manufacturing. This RSP is as follows: Figure 4A (Front view) and Figure 4B (Rear view shown). The sample loading chamber is as follows. Figure 4AAs shown (element 1). In one embodiment, the reagent is dried within the sample loading chamber (1). In one embodiment, the reagent is dried within a raised area (shown on the left, as part of a serpentine channel). In one embodiment, the dried reagent may be dried in both areas or within the serpentine channel. Any combination of positions of the dried reagent within the sample manipulation element is included in embodiments of the invention. Figure 4B The diagram shows the upper interface (element 2) with the box and the lower interface (element 3) with the main frame of the box. Figure 4D A cassette with a sample insert connected to the cassette is shown. According to this aspect and in one embodiment, the RSP is not part of the cassette mold. Instead, the RSP is added as an additional element and connected to the mold. Once the RSP and mold are connected, the cassette is ready for sample insertion. In this configuration, in some embodiments, an external pump is present that draws the sample into the cassette and influences mixing in the serpentine channels. In other embodiments, mixing or sample delivery through the channels of the sample plug is achieved by at least one of the blowers described herein. In embodiments, the term "backbone" is used for cassette use. It should be noted that the sample manipulation element can be located anywhere on the cassette. The sample manipulation element can be manufactured during the mold stage along with other parts of the cassette, or it can be added and fixed to the cassette afterwards. The sample manipulation element can be connected between two mixing chambers, or it can be connected to only one mixing chamber in a one- or two-mixing-chamber configuration. Figure 5A and Figure 5B These are pre-expanded isometric views and post-expanded isometric views, showing the area where the RSP connects to the box and the associated mating parts. Figure 6 illustrates an embodiment of the invention where the box mold design has two zones. Each zone can accommodate different features or no features. Any mold production can include one or more features as needed. Figure 6A A general main frame that can support additional features in different zones is shown. For example, Figure 6B Zones 1 and 2 are shown. In zone 1, a sample manipulation element can be placed (when producing the mold), or a mixing channel without a sample insertion port can be formed. In zone 2, a sample manipulation element can be formed (when producing the mold), or this zone can be left blank. It should be noted that additional zones can be formed as part of the mold. Features such as sample manipulation elements can be manufactured with the mold and can be added in subsequent mold production. Figure 7A and Figure 7B The back and front of the box are shown, with section 1 including a mixing channel and section 2 including a sample manipulation element. Figure 8 shows an example of a main frame with two insertion sections; insertion section 1 (loading port or sample manipulation element) and insertion section 2 (blank); Figure 8A This is the front view. Figure 8B This is the rear view.

[0107] Figure 9 shows an example of the system (analytical instrument) of the present invention. Figure 9A In the middle, left-side view 920 shows an ITX computer 922, a Galil motor controller 924, an electronic power supply 926, a box 910 inserted into a cartridge handling unit (CHU) 928, and a forward scattering detector 930. Figure 9B The right-side view 940 shows the reader optics 942, the data acquisition board 944, and the general-purpose electronic printed circuit board 946.

[0108] In one embodiment, the present invention provides a system and method for biological assays, the system comprising a cartridge as described herein for performing assays therein, the cartridge being adapted to contain at least one reagent adapted to react with a sample. In one embodiment, the system of the present invention includes a mechanical controller comprising urging means adapted to apply force from the outside to components of the cartridge, such as blister packs and blowers. In one embodiment, the system includes an optical reader adapted to detect a sample and a processor adapted to receive data from the optical reader and process the data to provide assay results.

[0109] In one embodiment, the present invention provides a system for testing biological samples, the system comprising:

[0110] As described herein, a kit for performing tests therein includes at least one reagent suitable for reacting with a sample;

[0111] Mechanical controller, comprising:

[0112] At least one first force-applying device is adapted to apply force to a blower to induce fluid movement along a first direction and / or along a second direction;

[0113] At least one second force-applying device adapted to apply force to one or more blister packs to release the diluted / pyrolyzed solution;

[0114] An optical reader adapted to detect the sample; and

[0115] A processor adapted to receive data from an optical reader and process the data to provide test results.

[0116] In addition, according to an embodiment of the present invention, the box also includes an alignment device adapted to align the reading channel on the box with the optical components of the system for optical detection of the sample in the box by the system's optical reader.

[0117] In one embodiment, the method of the present invention includes the following steps:

[0118] Load the sample into the sample manipulation element;

[0119] Assemble the sample components into the box;

[0120] Insert the box into the system;

[0121] Mix the contents of the sample manipulation element (mix the sample with the dry reagent);

[0122] Extract the sample from the sample element into the mixing chamber;

[0123] Processing and testing begin.

[0124] The order of steps can be changed or modified as described in the embodiments of the invention. For example, in an embodiment where the sample element is part of a box mold, the step of fixing the element to the box is not required. In one embodiment, sample mixing and extraction are performed at least partially in parallel.

[0125] For example, other features of the system are described in WO 2014 / 097287, the entire contents of which are incorporated herein by reference.

[0126] In one embodiment, the channel or fluid path in the cartridge has a cross-sectional dimension ranging from 200 μm to 1 mm. In another embodiment, the channel in the cartridge has a rectangular cross-section with a width ranging from 200 μm to 1 mm and a depth ranging from 200 μm to 1 mm. In yet another embodiment, the channel has a cross-section with a width of 800 μm and a depth of 600 μm.

[0127] In one embodiment, the reagent is dried within the channel before the insert is connected to the cartridge. After drying, the insert is connected to the cartridge. All further operations (sample introduction, sample flow through the channel with the dried reagent, and processing) are performed by a blower. No external pump is required.

[0128] In one embodiment, the query region is the area including a readout channel through which fluorescently labeled cells pass (with or without additional fluorescent beads). In one embodiment, the readout channel is transparent to the excitation wavelength and also transparent within the wavelength range of fluorescence emitted from the cells. In one embodiment, transparency at a specific wavelength is complete transparency. In one embodiment, partial transparency of the channel at a specific wavelength is sufficient to emit / detect signals passing through the channel.

[0129] The processing in embodiments of the present invention may involve chemical / biological processing in some embodiments, and computer processing of signals in other embodiments. In one embodiment, sample processing refers to both chemical and optical processing. The meaning of the term "processing" is clear from the context of the embodiments described herein. In one embodiment, the reading area of ​​the cartridge is an optical reading area. According to this aspect and in one embodiment, the sample in the reading area is illuminated by light from a light source, and in response, the sample emits light toward at least one detector. The light (optical) signal obtained by the detector is processed to produce a result indicating the composition of the sample. This result may be qualitative, quantitative, semi-quantitative, or any combination thereof.

[0130] In one embodiment, staining refers to the mixing and / or binding of an antibody (or other conjugate described herein) with cells or particles / species / fragments present in the sample. In one embodiment, staining refers to a pretreatment step or part thereof. In one embodiment, staining refers to the association of a fluorescent label with cells or other species in the sample. In one embodiment, staining or pretreatment refers to the process of attaching or binding an antibody (or similar compound) already linked to a fluorophore to cells (or other particles / species in the sample). According to this aspect and in one embodiment, the fluorescent antibody bound to the cells enables optical detection of the cells.

[0131] In some embodiments, the sample manipulation element is referred to as a sample processing unit. In some embodiments of the invention, the terms "zone," "area," "region," and "channel" are interchangeable and can refer to the same feature.

[0132] In one embodiment, only one such fluid path is needed, rather than multiple fluid paths, in addition to the channel for the sample manipulation element, each fluid path providing fluid communication between at least two other elements of the cavity. According to this aspect and in one embodiment, the fluid path connects the mixing chamber to the readout channel.

[0133] It should be noted that the orientation of the components in the box shown in the figure is illustrative. Various components of the box can be positioned in many different orientations within the box. Because the various components of the box are connected via fluid paths, the components can be assembled in many different configurations, and the fluid paths (channels) can be designed to connect the components as needed. All such orientations are included in the embodiments of the invention.

[0134] In one embodiment, the system adjusts the time and / or speed of pressing / depressing the blower 20 to adjust the time / speed of liquid flow within the elements of the cartridge.

[0135] In one embodiment, the cartridge does not include sample manipulation elements. According to this aspect and in one embodiment, pretreatment is performed outside the cartridge, and the pretreated sample is injected directly into the mixing chamber or a fluid channel connected to the mixing chamber for further processing. According to this aspect and in one embodiment, pretreatment includes adding fluorescent beads to the sample.

[0136] It should be noted that the connection between the reactant zone and the mixing chamber shown in the figure is illustrative. In embodiments, each reactant zone may be connected to each mixing chamber as needed for a specific application. In one embodiment, the cartridge comprises 1 to 3 blister packs. In one embodiment, the blister pack filler is used only for dilution / lysis. In one embodiment, the blister pack filler is not used for antibody / reference beads. In one embodiment, when more than one blister pack is present, different blister packs / more than one blister pack can be used to achieve different dilution ratios as needed. In one embodiment, two or more blister packs comprise the same solution, and in one embodiment, two or more blister packs comprise different solutions. In one embodiment, two or more blister packs comprise solutions containing the same components but at different concentrations.

[0137] In one embodiment, inserts for sample loading and serpentine dry reagent channels (sample manipulation elements) are added to the cartridge after it is molded.

[0138] In embodiments of the present invention, the sample manipulation element insert is considered to be part of the box.

[0139] In one embodiment, the sample loading area / port and the serpentine dry reagent channel (sample manipulator element) are manufactured as part of the cassette mold. The sample manipulator element is part of the cassette mold according to this embodiment.

[0140] In embodiments of the present invention, the sample manipulation element insert is considered to be part of the box.

[0141] In one embodiment, after the sample is introduced into the sample manipulation element, a sticker seal is used to seal the opening of the sample loading port. Other sealing options are provided in embodiments of the invention, such as... Figure 3 The plug 34b is such as a plastic cap, screw cap, adhesive, rubber seal, etc., known in the art. In one embodiment, a "co-molded plug" (e.g.) Figure 3 Element 34b) is used to seal the opening after the sample is introduced / loaded through the opening (e.g., element 34a).

[0142] In one embodiment, a sample loading port is present in the dry reagent insert (see, for example, the raised area in Figure 4a, element 1). In one embodiment, the cartridge is referred to as the main skeleton. In one embodiment, the sample manipulation element is referred to as the reagent sample plug (RSP).

[0143] In one embodiment, an external pump draws the sample through a channel in the sample manipulation element for further processing. In other embodiments, no external pump is required, and the sample is drawn into a cartridge by a blower through a channel in the sample manipulation element for further processing.

[0144] In one embodiment, the component that moves the liquid through the cartridge is not operated by an external pump. In one embodiment, the cartridge does not include a valve. In one embodiment, the cartridge is valveless.

[0145] In one embodiment, a blower is used to move the liquid through the components of the box, reducing system complexity and allowing for easy handling because no external pump is connected to the channels in the box.

[0146] In one embodiment, the reagent is dried within the channel of the sample handling element before the sample processing insert is connected to the cartridge. According to this embodiment, after drying, the insert is connected to the cartridge. In one embodiment, all further operations (sample extraction, flow through the channel containing the dried reagent, and processing) are performed by a blower. No external pump is required. In one embodiment, staining is induced by a blower, and no external pump is required.

[0147] In one embodiment, the present invention provides a testing method, comprising:

[0148] Provide the box as described above;

[0149] Introduce biological samples into the box;

[0150] Insert the box into the optical-mechanical system as described herein;

[0151] Use at least one force-applying device in the system to operate at least one blower in the box to induce sample flow through the dry reagent channel of the sample manipulation element;

[0152] At least one force-applying device in the system is used to operate at least one blower in the box to induce the stained (pretreated) sample to flow from the sample manipulation element to at least one mixing chamber;

[0153] Use at least one force-applying device in the system to burst at least one bubble in the box to induce the dilution / lysis solution to flow from the reactant zone to at least one mixing chamber;

[0154] Use at least one force-applying device in the system to operate at least one blower in the box to induce the dilution / lysis solution to mix with the pretreated sample in at least one mixing chamber;

[0155] At least one force-applying device in the system is used to operate at least one blower in the box to induce the treated solution to flow from at least one mixing chamber to a query area including a reading channel;

[0156] The system's light source illuminates the readout channel, and the light signal obtained from the readout channel is detected by at least one detector of the system.

[0157] The system's processor processes the detected light signals to obtain analysis results.

[0158] The embodiments of the boxes and systems described herein are applicable to embodiments of the methods of the present invention described herein.

[0159] In one embodiment, the system of the present invention does not include a heater. In one embodiment, the system of the present invention does not include a temperature control device. In one embodiment, the system of the present invention does not include a thermometer.

[0160] In addition to antibodies (and optional reference beads), one or more dry reagents of the present invention may also include chemicals such as salts, stabilizers, pH-adjusting materials, ionic strength-adjusting materials, and other compounds / materials compatible with the sample and / or conducive to maintaining the sample under conditions suitable for handling and measurement. According to this aspect and in one embodiment, the dry reagent comprises a mixture.

[0161] In one embodiment, the sample comprises cells. In one embodiment, the cells are blood cells. In one embodiment, the cells comprise red blood cells, white blood cells, or a combination thereof. In one embodiment, the cells comprise stem cells. In one embodiment, the sample comprises cell debris. In one embodiment, the sample comprises particles. In one embodiment, the sample comprises an antigen. In one embodiment, the sample comprises cells, and the cells comprise cell surface markers. In one embodiment, the surface marker is an antigen or comprises an antigen. In one embodiment, in the kit assay of the present invention, an antigen in the sample (e.g., an antigen on the cell surface) binds to an antibody in a dry reagent. In one embodiment, a fluorescent tag on the antibody is used to identify the cell. Different cell surface markers may bind to different antibodies. In one embodiment, different antibodies comprise different fluorescent tags. According to this aspect and in one embodiment, a particular cell will bind to a different antibody with a different fluorescent tag. In some embodiments, the fluorescence wavelength and fluorescence intensity from a particular cell indicate the cell type.

[0162] In one embodiment, the assay of the present invention is a flow cytometry assay. In one embodiment, the determination / detection of a biological condition is referred to as sample analysis or sample assay, or analysis or assay or determination of a medical condition. In one embodiment, for simplicity, the reactant region is referred to as a blister. In one embodiment, the antibody is a fluorescent antibody. In one embodiment where the term "cell" is used, the cell is a biological cell. Embodiments involving cells may also involve other particles, cell debris, proteins, antigens, biomolecules, cellular components, viruses, bacteria, microorganisms, or components thereof. All such embodiments are included in the present invention. In one embodiment, the particle is a cell, a bead, or any of the kinds described above.

[0163] In one embodiment, the reference bead is a fluorescent bead, which may also be referred to as a microsphere, nanosphere, fluorescent sphere, or a combination of these terms. In one embodiment, fluorophore and fluorescent dye are interchangeable terms. In one embodiment, the fluorescence obtained from the cells / particles of the present invention is direct, indirect, or any combination thereof. Embodiments of the invention described for therapeutic purposes may also relate to diagnostics. In one embodiment, the terms "tag," "probe," and "marker" are interchangeable. The mixing chamber of the present invention can take many shapes, sizes, and geometries, all of which are included in the embodiments of the invention. In one embodiment, the present invention provides cell staining in an automated cassette. In one embodiment, not all reactant zones include blister packs. In one embodiment, at least some reactant zones do not include blister packs. In one embodiment, at least some reactant zones include blister packs. In one embodiment, the positions of the zones / chambers / elements / components on the cassette can be varied, as long as the required connections between any two or more zones / chambers / elements / components are maintained.

[0164] In one embodiment, the raised area in the sample manipulation element is used to place dry reagents, introduce samples, or both. In one embodiment, the assay of the present invention is a white blood cell assay. In one embodiment, the sample before processing is a whole blood sample.

[0165] In one embodiment, the assay of the present invention is completed within 30 minutes or 1 hour from the time the sample is introduced into the sample manipulation element. In another embodiment, the assay of the present invention is completed within 30 minutes or 1 hour from the time the cartridge is inserted into the system.

[0166] Those skilled in the art will readily understand that many changes, variations, and modifications can be made without departing from the scope of the currently disclosed subject matter.

Claims

1. A cartridge for preparing a biological sample for an assay and for carrying the sample to a query area for the assay, characterized by, The cartridge is formed with a single cavity, the cavity comprising: a sample manipulation element configured to receive the biological sample therein via a sample port, the sample manipulation element comprising one or more reagents; a seal configured to seal the sample port; two air blowers configured to induce controlled movement of liquid within the cartridge, each of the air blowers comprising a flexible dome; one or more mixing chambers; one or more reagent zones, each reagent zone comprising a liquid reagent for selective delivery to one of the mixing chambers, wherein the one or more mixing chambers are in fluid communication with the one or more reagent zones; a query zone comprising a reading area configured to receive the biological sample; a plurality of fluidic pathways, each fluidic pathway providing fluid communication between at least two elements of the cavity; and a vent connected to one of the mixing chambers via a vent fluidic pathway, the vent comprising an opening to the environment; wherein, when the sample manipulation element is in its sealed position, the cavity is in fluid communication with the environment only via the vent; an insertion zone configured to receive the sample manipulation element.

2. The cartridge of claim 1, wherein, At least one of the one or more reagents comprises an antibody.

3. The cartridge of claim 2, wherein, The antibody is conjugated to a fluorescent label.

4. The cartridge of claim 3, wherein, The one or more reagents comprise fluorescent beads.

5. The cartridge of claim 1, wherein, The sample manipulation element comprises a mixing channel having a serpentine pathway.

6. The cartridge of claim 5, wherein, Each end of the mixing channel is connected to one of the mixing chambers by a fluidic pathway spanning between the mixing channel and the mixing chamber.

7. The cartridge of claim 1, wherein, At least one of the air blowers is connected to one of the mixing chambers by a fluidic pathway spanning between the air blower and the mixing chamber.

8. The cartridge of claim 1, wherein, At least one of the air blowers is connected to the query zone by a fluidic pathway spanning between the air blower and the query zone.

9. The cartridge of claim 1, wherein, Each of the reagent zones comprises a blister pack containing the liquid reagent.

10. The cartridge of claim 9, wherein, Each of the blister packs comprises a flexible dome and a backer opposite the flexible dome, and each of the reagent zones comprises a receptacle in which a respective one of the blister packs is sealingly received, the receptacle comprising a piercing element supported on the backer.

11. The cartridge of claim 10, wherein, The cartridge is configured such that depressing the dome increases the internal pressure of the blister pack, such that rupture of the backer occurs.

12. The cartridge of claim 1, wherein, The liquid reagent comprises one or more selected from the group comprising a diluent, a lysis agent, and a reference substance.

13. The cartridge of claim 1, wherein, At least one of the plurality of fluidic pathways comprises a serpentine channel.

14. The cartridge of claim 1, wherein, The one or more reagents are dry reagents.

15. The cartridge of claim 1, wherein, The seal is a co-molded plug.

16. The cartridge of claim 1, wherein, The seal is a sticker seal.

17. The cartridge of claim 1, wherein, The seal is a plastic lid or rubber seal.

18. The cartridge of claim 1, wherein, The seal is a screw cap.

19. The cartridge of claim 1, wherein, The seal is a plug.

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