CARTRIDGE FOR LAB-ON-CHIP APPLICATIONS

Description

The present invention relates to a cartridge for sample preparation and molecular analysis. Specifically, the present invention relates to the field of so-called Lab-On-Chip (LOC) devices, in which a single cartridge comprises structures designed to perform at least some sample processing steps for the purpose of extracting and analyzing molecules. State of the art Lab-On-Chip (LOC) systems are well-known, particularly LOC systems that rely on cartridges (e.g., disposable cartridges) inserted into a machine that analyzes the substances contained within them, typically after pretreatment. These systems are of great importance to health, a value that is increasing over time as the number of analyses can be easily performed by a patient independently or with the assistance of non-experts. Specifically, the above systems enable the analysis of biological molecules, such as nucleic acids, proteins, lipids, polysaccharides, etc. These analyses involve a variety of operations starting from a raw material, such as a blood or saliva sample, or a sample collected via a nasal swab (nasal sample). These operations may include varying degrees of sample pretreatment, lysis, purification, amplification, and analysis of the resulting product. The analysis methods vary depending on the target biological molecule to be analyzed or detected. Furthermore, LOC systems can also be used for the purification of non-biological samples, such as water samples, and for the analysis of non-biological molecules. Sample processing and analysis require specialized reagents. For example, nucleic acid analyses, such as those based on Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), Polymerase Chain Reaction (PCR), Ligase Chain Reaction (LCR), Strand-Displacement Amplification (SDA), Transcription-Mediated Amplification (TMA), Rolling-Circle Amplification (RCA), Loop-Mediated Isothermal Amplification (LAMP), and the like, require specialized enzymes to perform amplification and / or target recognition steps.As another example, in proteomic analyses such as Enzyme-Linked Immunosorbent Assay (ELISA), antibodies and enzymes suitable for selectively binding a target analyte of interest are required for detection. Reagents can be stored in liquid or solid form (e.g., freeze-dried, dehydrated, or desiccated) depending on their nature. Solid reagents (hereinafter referred to as dry reagents), such as freeze-dried enzymes in the case of a CRISPR-based assay, require dry storage to prevent unwanted hydration. Indeed, hydration of dry reagents must occur during analysis or sample pretreatment at specific, predetermined steps depending on their function. Premature and unwanted hydration would accelerate the degradation of dry reagents, thus reducing their shelf life. A well-known solution to prevent premature hydration of dried reagents is to store them in specialized chambers created within the LOC cartridge. The LOC cartridge is then packaged within a sealed outer container or pouch that houses a desiccant (e.g., a sachet containing silica beads or silica gel) along with the cartridge. The container can also be filled with dry inert gases, such as nitrogen (N2) or argon (Ar). The cartridge is then stored inside the sealed package with the desiccant for its entire shelf life, until it is used. However, moisture can (accidentally) enter the package and penetrate the cartridge, or it can be present inside the cartridge, for example, from before the packaging stage, hydrating and deteriorating the dried reagents. Another possible cause of hydration, and therefore deterioration, of dried reagents can arise during the early stages of the analysis. For example, during the CRISPR-based analysis mentioned above, a lysis step is required to extract nucleic acids from the cells present in a biological sample, which is typically diluted in an aqueous buffer. The lysis step can be performed by heating the biological sample in the presence of specific reagents, thereby causing part of the aqueous buffer to evaporate. The resulting vapor can then reach the dried reagents, hydrating them and possibly degrading them before their intended use. Therefore, there is a need to provide a cartridge that allows for more effective and safe storage of dry reagents until their use. Summary According to the present invention, a cartridge for Lab-On-Chip applications, as defined in claim 1, is provided. Brief description of the drawings For a better understanding of the present invention, some embodiments thereof are now described, purely by way of non-limiting example, with reference to the attached drawings, in which: - Figure 1 schematically illustrates an embodiment of a Lab-On-Chip (LOC) system 1 according to an embodiment of the present invention; - Figure 2A schematically illustrates a fluidic module A in a top plan view according to an embodiment of the present invention; - Figure 2B illustrates the fluidic module A in an exploded view; - Figures 3A-3B schematically illustrate the paths followed by air and liquids inside the fluidic module A of Figure 2A, during an exemplary analysis; - Figure 4A schematically illustrates a portion of a reservoir module B of cartridge 2, including a buffer container; - Figure 4B illustrates the swab container of Figure 4A housing a swab; - Figure 5A illustrates a different fluidic module A and reservoir module B according to an embodiment of the present invention; - Figure 5B illustrates a detail of a mechanical junction between the fluidic module A and the reservoir module B of Figure 5A; - Figure 6 illustrates another different fluidic module A and reservoir module B according to another embodiment of the present invention; - Figure 7 illustrates the fluidic module A in an exploded view according to another embodiment of the present invention. Description of preferred embodiments of the invention Figure 1 schematically illustrates a non-limiting embodiment of a Lab-On-Chip (LOC) system 1. The LOC system 1 of the illustrated embodiment is configured to perform biological sample preparation steps and to perform, for example, CRISPR analysis of a biological sample. However, the present invention is not limited to this application and can be used for various analyses such as Polymerase Chain Reaction (PCR), Loop-mediated Isothermal Amplification (LAMP), proteomic analysis, as well as analysis of non-biological samples or even other applications. During the analysis, the biological or non-biological samples are eluted into a liquid or fluid medium, whether water-based or oil-based, or any other liquid solution. The LOC system 1 comprises a cartridge 2 connectable to a control machine 3 (the control machine 3 not being part of the present invention) via a first fluid connection 4 and, optionally, a second fluid connection 4'. For example, the control machine 3 is described in US2019 / 0201897. In one embodiment of the present invention, the cartridge 2 is a disposable cartridge. The control machine 3 is designed to control the movement of fluids within the cartridge 2 and to perform sample processing and analysis steps. For example, in one embodiment of the present invention, the control machine 3 performs thermal heating steps necessary for CRISPR analysis to be performed on the biological samples within the cartridge. According to the present invention, the cartridge 2 comprises a fluidic module A and a reservoir module B, fluidically connected to each other through a third fluidic connection 6 and mechanically coupled to each other. The fluidic module A can be connected to the control machine 3 through the first fluidic connection 4. Reservoir module B can be connected to control machine 3 via the second fluid connection 4'. Alternatively, the second fluid connection 4' is omitted and reservoir module B is connected to the external environment via a fluid connection (not shown) sealed by a wettability-proof membrane configured to allow air to flow through it. In one embodiment, the first fluidic connection 4 has a first portion 4a belonging to the fluidic module A and a second portion 4b belonging to the control machine 3. The first portion 4a and the second portion 4b are designed to mate and seal together. For example, the first portion 4a and the second portion 4b are male and female pneumatic connections, respectively (or vice versa). In an alternative embodiment, the connection type of the first fluidic connection 4 includes a needle configured to fit into a rubber seat (e.g., 3D printed). Specifically, the needle is part of the control machine 3 and the rubber seat is part of the fluidic module A, arranged within the cavity 40 of Figure 2A. The second fluid connection 4' is formed similarly to the first fluid connection 4 and comprises corresponding features (in particular a first portion 4a', corresponding to the portion 4a, and a second portion 4b', corresponding to the portion 4b). The third fluidic connection 6 has a first portion 6a belonging to the fluidic module A and a second portion 6b belonging to the reservoir module B. The first portion 6a and the second portion 6b are designed to mate and seal together. For example, the first portion 6a and the second portion 6b are male and female Luer Slip connections, respectively (or vice versa). In one embodiment, the fluidic module A includes a solid body 5, e.g., generally parallelepiped or substantially quadrangular in shape, which includes the first portion 4a of the first fluidic connection 4, the first portion 6a of the third fluidic connection 6, a fluidic circuit 8, a pneumatic circuit 10, and a storage chamber 12, configured to store (and in one embodiment, actually store) desiccants, e.g., in the form of a sachet containing silica beads or silica gel. The fluid circuit 8 includes a fluid channel 14a, a fluid channel 14b, a first valve 15, a lysis chamber 16 and one or more reaction chambers 18(1)-18(n). The fluid channel 14a connects the first portion 6a of the third fluid connection 6 to the lysis chamber 16. The fluid channel 14b connects the lysis chamber 16 to the reaction chambers 18(1)-18(n). The valve 15 is located in the fluid channel 14b. The opening and closing of the first valve 15 can be controlled by the control machine 3 in a manner known per se. The valve 15 may be one of: a magnetic valve, a mechanical one-way valve, an elastomeric duckbill valve, a pneumatic valve, etc. The pneumatic circuit 10 comprises air paths 11a, 11b, 11c(1)-11c(n) and a second valve 17. The air path 11a connects the first portion 4a of the first fluid connection 4 to the storage chamber 12, the air path 11b connects the storage chamber 12 to the lysis chamber 16 and the air paths 11c(1) 11c(n) connects the storage chamber 12 to the reaction chambers 18(1)-18(n) respectively. The second valve 17 is located in the air path 11b. The opening and closing of the second valve 17 can be controlled by the control machine 3 in a manner known per se. The valve 17 can be one of: a magnetic valve, a mechanical one-way valve, an elastomeric duckbill valve, a pneumatic valve, etc. The reservoir module B includes: a solid body 7 of generally parallelepiped or substantially quadrangular shape, housing the second portion 6b of the third fluidic connection 6; a buffer container or buffer storage tube 22 configured to contain or store (and in one embodiment, actually store) buffers and liquid reagents and adapted to house a buffer 24 having a head or tip suitable for collecting biological samples; a fluidic channel 26 configured to fluidically connect the buffer container 22 to the second fluidic connection 4' or to the external environment, according to the respective embodiments mentioned above; a fluidic channel 28; and, when present, the first portion 4a' of the second fluidic connection 4'. The buffer container 22 is connected to the second portion 6b of the third fluid connection 6 via the fluid channel 28. Figure 2A illustrates the fluidic module A in a triaxial system with orthogonal x, y, and z axes, in a top plan view on the xz plane. Figure 2B schematically illustrates the fluidic module A in an exploded view, in the triaxial system with orthogonal x, y, and z axes. The solid body 5 of the fluidic module A is formed from three portions (illustrated in the exploded view in Figure 2B), including a first portion 50, a second portion 51, and a third portion 52. The first portion 50 has a front face 50a and a back face 50b opposite each other along the y-axis, a top side 50c and a bottom side 50d opposite each other along the z-axis, a first lateral side 50e, and a second lateral side 50f opposite each other along the x-axis. The front face 50a is connected to the back face 50b by the top side 50c, the bottom side 50d, the first lateral side 50e, and the second lateral side 50f. The second portion 51 has a front face 51a and a rear face 51b opposite each other along the y-axis, a top side 51c and a bottom side 51d opposite each other along the z-axis, a first lateral side 51e and a second lateral side 51f opposite each other along the x-axis. The front face 51a is connected to the rear face 51b by the top side 51c, the bottom side 51d, the first lateral side 51e, and the second lateral side 51f. The third portion 52 has a front face 52a and a rear face 52b opposite each other along the y-axis, a top side 52c and a bottom side 52d opposite each other along the z-axis, a first lateral side 52e and a second lateral side 52f opposite each other along the x-axis. The front face 52a is connected to the rear face 52b by the top side 52c, the bottom side 52d, the first lateral side 52e, and the second lateral side 52f. The second portion 51 is joined to the first portion 50 at the front face 50a of the first portion 50; and the third portion 52 is joined to the first portion 50 at the rear face 50b of the first portion 50. When portions 50-52 are joined together, the rear face 51b of the second portion 51 faces directly onto the front face 50a of the first portion. 50, the front face 52a of the third portion 52 directly faces the rear face 50b of the first portion 50. The front face 51a of the second portion 51 forms a front face 5a of the solid body 5; the rear face 52b of the third portion 52 forms a rear face 5b of the solid body 5; the upper sides 51c, 50c and 52c form, as a whole, an upper side 5c of the solid body 5; the lower sides 51d, 50d and 52d form, as a whole, a lower side 5d of the solid body 5; the first lateral sides 51e, 50e and 52e and the second lateral sides 51f, 50f and 52f form, as a whole, a first lateral side 5e and, respectively, a second lateral side 5f of the solid body 5. Portions 50-52 are joined together, for example, by glue or thermal welding, and may have gaskets and sealing means (not shown) to prevent the leakage of liquids to the outside and to ensure the separation of the various channels from each other and isolation from the external environment. In one embodiment, the second portion 51 is a transparent tape or film. The first portion 50 and the third portion 52 are, for example, made of a plastic or polymeric material or a biocompatible material suitable for carrying out the aforementioned biological analyses. During use, when the cartridge 2 is connected to the drive machine 3, the lower face 5d of the solid body 5 faces the drive machine 3 and the upper face 5c faces away from the drive machine 3. The third portion 52 includes a cavity 40 at the lower side 52d and on the front face 52a, having a main extension along the second z-axis, towards the upper side 52c. The first portion 50 includes a first through hole 110a extending through the first portion 50 from the rear face 50b to the front face 50a, parallel to the first y-direction. The first portion 50 further includes a second through hole 113a extending through the first portion 50 from the rear face 50b to the front face 50a, parallel to the first y-direction. The first and second through holes 110a, 113a are spaced apart from each other, particularly along the z-axis. When the third portion 52 is joined to the first portion 50, the cavity 40 forms, at one end, an opening 40a in the lower side 5c of the solid body 5 and, at an opposite end, the cavity 40 faces the through hole 110a. The opening 40a and the cavity 40 form, at least in part, the first portion 4a of the first fluid connection 4 of Figure 1. The first portion 50 further includes a first air recess 112a at the front face 50a, located proximal to the lateral side 5f and having a main extension along the z-axis. The first through hole 110a connects the cavity 40 to the first air recess 112a. When the second portion 51 is joined to the first portion 50, the first air recess 112a forms part of the first air path 11a. The first air recess 112a extends from the through hole 110a, parallel to the z-axis, toward the upper side 50c, to the second through hole 113a. The opening 40a, the cavity 40, the first through hole 110a, the first air recess 112a, and the second through hole 113a are located in a portion of the solid body 5 proximal to the lateral side 5f. The third portion 52 includes a second air recess 114a at the front face 52a. When portions 50 and 52 are joined together, the second through-hole 113a connects the first air recess 112a to the second air recess 114a. When the third portion 52 is joined to the first portion 50, the second air recess 114a forms a further portion of the first air path 11a. The second air recess 114a extends parallel to the x-axis towards the lateral side 50e, reaching a third through hole 115a that extends through the first portion 50 into the interior of the storage chamber 12. Thus, the interior of the storage chamber 12 is in air connection with the opening 40a, particularly when the second portion 51 is coupled to the first portion 50 and protects the interior of the storage chamber 12 from the external environment. A portion of the second air path 11b is formed by a third air recess 110b extending at the front face 50a parallel to the x-axis. The third air recess 110b extends away from the storage chamber 12 on an opposite side of the storage chamber 12 from the second air recess 114a. The third portion 52 includes a fourth air recess 112b at the front face 52a, extending primarily along the x-axis toward the lateral side 52e. The third air recess 110b is connected to the fourth air recess 112b by a through hole 111b formed through the first portion 50. The first portion 50 includes a valve hole 17a, which is a through hole connecting the rear face 50b to the front face 50a. The fourth air recess 112b ends at the valve hole 17a of the first portion 50. The valve hole 17a houses the valve 17. The first portion 50 also includes a lysis chamber 16, where a lysis step is performed during use, to extract nucleic acids from the cells present in the biological sample. The first portion 50 also includes a recess 114b at the front face 50a, forming a further portion of the second air path 11b extending primarily along the z-axis, toward the lower side 50d, and connecting the valve port 17a to the lysis chamber 16. A fluidic inlet port 14a' connects an upper portion of the lysis chamber 16 to the fluidic channel 14a. The fluidic channel 14a extends at the front face 52a of the third portion 52, parallel to the x-axis, from the fluidic inlet port 14a' toward the first lateral face 5e of the solid body 5, terminating at the first portion 6a of the fluidic connection 6. The first portion 6a includes a through hole 60a that is in fluidic continuity with the fluidic channel 14a. In particular, in the embodiment illustrated in Figure 2A, the first portion 6a is a male Luer Slip connector. The first portion 50 further includes the fluid recesses 140a and 140c, a valve hole 15a, a through hole 141 and a plurality of (here, three) reaction chambers 18(1), 18(2), 18(3). The first portion 50 includes a fluid recess 140b. Fluid recess 140a extends at the front face 50a, starting from a lower portion (opposite the upper portion along the z-axis) of the lysis chamber 16 and extending primarily parallel to the z-axis toward the upper side 50c of the first portion 50, terminating at the valve hole 15a. The valve hole 15a is a through-hole connecting the front face 50a and the rear face 50b and housing the valve 15. Fluid recess 140b extends at the front face 52a of the third portion 52, with a main extension that is substantially parallel to the x-axis. When the portions 50 and 52 are joined together, fluid recess 140b faces at one end toward the valve hole 15a and at the opposite end toward the through-hole 141.The fluidic recess 140c extends at the front face 50a of the first portion 50, from the through hole 141 towards the lower side 50d of the first portion 50, where it forms three branches 140c', 140c'' and 140c''', each of which reaches a respective reaction chamber 18(1), 18(2) and 18(3). The fluid recess 140a, the valve hole 15a, the fluid recess 140b, the through hole 141 and the fluid recess 140c together form the fluid channel 14b of Figure 1. When portions 50-52 are joined together, the lysis chamber 16 and the reaction chambers 18(1), 18(2) and 18(3) are sealed on one side by the second portion 51 and on the other, opposite, side by the third portion 52. In one embodiment, the third portion 52 houses two heaters 9, 13 (of a type known per se). When the first portion 50 and the third portion 52 are joined together, one heater 9 is arranged at the lysis chamber 16 and the other heater 13 is arranged at the reaction chambers 18(1), 18(2) and 18(3). The heater 9 is controllable by the control machine 3 to perform heating steps of the lysis chamber, for example at a temperature of 95 °C for a time of 5 minutes in order to extract nucleic acids from the cells present in the biological sample during the exemplary CRISPR analysis.The heater 13 is controllable from the control machine 3 to maintain the reaction chambers 18(1)-18(3) at a fixed temperature, for example 37 °C. The temperatures reached by heaters 9 and 13 may be different from the examples above, depending on the specific analysis being performed. The first portion 50 also includes a plurality of (here, three) fluid recesses 110c(1), 110c(2), 110c(3), a plurality of (here, six) through holes 111c(1)-111c(6) and a plurality of (here, three) air recesses 112c(1), 112c(2) and 112c(3). The third portion 52 also includes a recess 80, for example of rectangular shape, housing a membrane 81 permeable to gases and vapours and impermeable to liquids. The membrane 81 is for example a layer of material impermeable to liquids (wet-proof fabric) but permeable to air and vapour (breathable fabric), for example a PTFE membrane. As an alternative to a membrane 81, an equivalent permeable film (e.g. a perforated film or a permeable PDMS layer) may be used. The fluid recesses 110c(1), 110c(2), 110c(3) extend in correspondence with the front face 50a of the first portion 50 with a main extension substantially parallel to the z axis. The fluid recesses 110c(1), 110c(2) and 110c(3) start from the respective upper portions of the reaction chambers 18(1), 18(2) and 18(3) and reach, respectively, the through holes 111c(1), 111c(2) and 111c(3). Through holes 111c(1), 111c(2) and 111c(3) connect the front face 50a with the rear face 50b of the first portion 50. Through holes 111c(4), 111c(5) and 111c(6) are located at a distance along the z-axis from through holes 111c(1), 111c(2) and 111c(3), such that through holes 111c(4), 111c(5) and 111c(6) are closer to the upper side 50c of the first portion 50 than through holes 111c(1), 111c(2) and 111c(3).Furthermore, through holes 111c(4), 111c(5) and 111c(6) are substantially aligned along the z-axis with through holes 111c(1), 111c(2) and 111c(3) respectively. The air cavities 112c(1), 112c(2) and 112c(3) extend in correspondence with the front face 50a of the first portion 50, with a main extension substantially parallel to the z-axis, respectively connecting the through hole 111c(4) with the storage chamber 12, the through hole 111c(5) with the storage chamber 12 and the through hole 111c(6) with the storage chamber 12. When the first portion 50 and the third portion 52 are joined together, the recess 80 faces the through holes 111c(1)-111c(6), so that the membrane 81 fluidly connects the through holes 111c(1)-111c(3) with the through holes 111c(4)-111c(5). The fluid recesses 110c(1)-110c(3), the through holes 111c(1)-111c(3), the membrane 81 in the cavity 80, the through holes 111c(4)-111c(6) and the air recesses 112c(1)112c(3), together, form the air paths 11c(1)11c(n) of Figure 1, connecting the reaction chambers 18(1)-18(n) to the storage chamber 12. Reaction chambers 18(1)-18(n) are configured to store (and, in one embodiment, store) dehydrated or lyophilized reagents prior to their use for sample analysis. In the non-limiting example described herein, reaction chambers 18(1)-18(n) are configured to store (and, in one embodiment, store) lyophilized enzymes required to perform the steps of the CRISPR assay. In another non-limiting example (not described), reaction chambers 18(1)-18(n) are configured to store (and, in one embodiment, store) lyophilized antibodies and enzymes required to perform the steps of an ELISA assay in a manner known per se.In another non-limiting example (not described), reaction chambers 18(1)-18(n) are configured to store (and, in one embodiment, do store) freeze-dried antibodies immobilized on magnetic beads necessary to perform protein biosensing steps in a manner known per se. In another non-limiting example (not described), reaction chambers 18(1)-18(n) are configured to store (and, in one embodiment, do store) freeze-dried reagents necessary to perform PCR assay steps in a manner known per se. It is apparent that other applications are possible, wherein reaction chambers 18(1)-18(n) may be configured to store (and, in one embodiment, do store) specific freeze-dried, or dried, or desiccated, or solid reagents to be utilized in steps of an assay performed by system 1 during use. During the storage period of the fluidic module A of the cartridge 2, when the storage chamber 12 stores the desiccant and the reaction chambers 18(1)-18(n) store the dry, or desiccated, or freeze-dried, or solid reagents (referred to as dry reagents hereinafter), the desiccant prevents the hydration of said reagents. Preventing hydration improves the shelf life of the dried reagents and therefore of the fluidic module A in general, compared to a case where the desiccant is placed in an external package, which in turn contains the fluidic module A. Indeed, the presence of the desiccant inside the storage chamber 12 allows capturing water vapor that may (even by chance) reach the fluidic module A from an external environment before the packaging of the fluidic module A or during the storage period of the fluidic module A. When using cartridge 2, the desiccant in storage chamber 12 further protects the dried reagents from hydration during bioanalysis steps when liquids flow through fluidic module A, but these reagents must still be kept dry. Figure 3A schematically illustrates the paths followed by air and liquids within the fluidic module A shown in Figures 1 and 2A, for example, during the aforementioned CRISPR analysis performed on the biological sample. In the illustrated phase of the analysis, fluidic module A is connected to reservoir module B (not shown) via fluidic connection 6, and fluidic module A is connected to control machine 3 (not shown) via fluidic connection 4. The drive machine 3, in the step of Figure 3A, opens valve 17 and closes valve 15. Additionally, the drive machine 3 draws air from the fluidic module A through the fluidic connection 4, generating an air flow 30 along the air path 11a. The air flow 30 draws air from the storage chamber 12, generating a negative pressure within the storage chamber 12. The negative pressure in the storage chamber 12 draws air from the lysis chamber 16 along the air path 11b and opens valve 17, resulting in an air flow 32. The air flow 32 generates a negative pressure in the lysis chamber 16. The negative pressure in the lysis chamber 16 draws liquids (in the illustrated example the biological sample to be analyzed plus buffers and liquid reagents) from the reservoir module B through the fluidic connection 6 and the fluidic channel 14a, generating a liquid flow 34.The liquid flow 34 at least partially fills the lysis chamber 16. In the described embodiment, the control machine 3 then controls the heater 9 to perform a heating step. During the heating step, the liquid in the lysis chamber 16 is heated to a high temperature, e.g., in the range 90-99°C, e.g., °C, and held at said temperature for e.g., 3-10 minutes, e.g., 5 minutes, to extract nucleic acids from cells in biological samples. During the heating step, some of the liquid evaporates. The evaporated liquids, in the form of vapor, exit the lysis chamber 16 via air path 11b. The vapor is, at least in part, captured by the desiccant in the storage chamber 12 and optionally, at least in part, extracted by the control machine 3 via air path 11a, thereby preventing premature hydration of the dried reagents stored in the reaction chambers 18(1)18(n). Figure 3B illustrates a next step in the analysis. In the step of Figure 3B, the control machine 3 closes valve 17 and opens valve 15. Additionally, the control machine 3 draws air from the fluidic module A through the fluidic connection 4, generating air flow 30 through the air path 11a. The air flow 30 draws air from the storage chamber 12, generating negative pressure within the storage chamber 12. The negative pressure in the storage chamber 12 draws air from the reaction chambers 18(1)-18(3) along the air paths 11c(1)-11c(3), generating air flows 36(1)-36(3), respectively, in the air paths 11c(1)-11c(3). Air flows 36(1)-36(3) generate negative pressures in reaction chambers 18(1)-18(3).Negative pressures in reaction chambers 18(1)-18(3) draw liquids from the lysis chamber 16 through fluid path 14b and through the open valve 15, generating a liquid flow 38. The liquid flow 38 separates into three streams 38', 38'', and 38''', reaching and at least partially filling the reaction chambers 18(1)-18(3), respectively, and hydrating the dry reagents stored in said reaction chambers. In the described embodiment, the control machine 3 then controls the heater 13 to maintain a fixed temperature, e.g., 37 °C, in the reaction chambers 18(1)-18(3) to perform a step of the CRISPR assay in a manner known per se. During this phase, liquids can completely fill the reaction chambers 18(1)-18(3) and partially fill the air paths 11c(1)-11c(3) up to the membrane 81.The membrane 81 (being, as mentioned, wettability-proof) prevents liquids from reaching the storage chamber 12, emptying the reaction chambers 18(1)-18(3). In this way, the membrane 81 also prevents contact between the liquids and the desiccant stored inside the storage chamber 12, thus avoiding unwanted absorption and any possible contamination (or degradation) of the sample to be analyzed that may result from contact of said sample with the desiccant. Note that during this phase, the valve 17 remains closed, preventing liquids from flowing into the storage chamber 12 along the air path 11b. Figure 4A illustrates a portion of one embodiment of the reservoir module B of the cartridge 2, including the buffer container 22. The swab container 22 is, in one embodiment, made of a biocompatible plastic material. In a non-limiting example, the swab container 22 is made of polypropylene and is manufactured using technological processes known in the art for the manufacturing of medical packaging. The buffer container 22 is substantially in the form of a hollow cylinder, terminating in a generally round bottom wall 22a at one end, a generally round opening 22b at an opposite end, and a solid side wall 22c connecting the bottom wall 22a to the opening 22b. The side wall 22c includes an internal surface 220 facing the inside of the hollow cylinder and an external surface 222, opposite the internal surface 220, facing the outside of the cylinder. In the illustrated embodiment, sidewall 22c includes a portion 224 proximal to opening 22b, portion 224 including a male thread 223 on outer surface 222. In another embodiment (not shown) a female thread is present on the internal surface 220 in portion 224 instead of the male thread 223 on the external surface 222. A portion 226 of the buffer container 22, proximal to the bottom wall 22a, houses, on the inner surface 220, a plurality of projections or fins 225(1)-225(5). The fins 225(1)-225(5) project inwardly from the inner surface 220 radially toward the main axis of the cylinder. In one embodiment, the fins 225(1)-225(5) extend longitudinally along the cylinder. In one embodiment, the fins 225(1)-225(5) reach the bottom wall 22a; in another embodiment, the fins 225(1)-225(2) are remote from the bottom wall 22a. The buffer container 22 further includes a projection 227 extending outward from a subportion of the portion 226 at the outer surface 222, specifically near the bottom wall 22a. The projection 227 terminates in a surface 227a, such as a flat surface. A through-hole 228 extends from the surface 227a into the interior of the cylinder, connecting the surface 227a to the inner surface 220. In one embodiment, the protrusion 227 and the through hole 228 together form the second portion 6b of the fluid connection 6. Specifically, the protrusion 227 and the through hole 228 form a female Luer Slip connector. Prior to use, the through hole 228 may be sealed by a frangible membrane 229. The frangible membrane 229 is configured to rupture when the reservoir module B is coupled to the fluidic module A, fluidically connecting the interior of the buffer container 22 with the fluidic channel 14a. A portion 221 of the buffer container 22 between portion 226 and portion 224 is more flexible and elastic, e.g., thinner, than portions 224 and 226, so that only portion 221 bends or deforms inward under the negative pressure generated by drive machine 3 to draw liquids from module B (generating the previously mentioned liquid flow 34). By modulating the pressure generated by drive machine 3, it is possible to adjust the degree of deformation of portion 221 and even reverse the direction of flow 34 (as well as flows 38), if required. For example, the thicker portions 224 and 226 have a thickness in the range of 1 to 1.5 mm, and the thinner portion 221 has a thickness in the range of 0.4 to 1 mm, specifically 0.5 to 0.6 mm. Figure 4B illustrates swab container 22 housing swab 24. In the illustrated embodiment, swab 24 is coupled, or integral, with a female threaded cap 240. Female threaded cap 240 is matable to thread 223. When swab 24 is inserted into swab container 22 and female threaded cap 240 is threaded onto male thread 223, the interior of swab container 22 is effectively isolated from the outside environment and the swab tip is squeezed by fins 225(1)-225(5), releasing previously collected biological material from the tip. Biological material released by the squeezing action elutes into a liquid buffer or liquid medium contained within swab container 22. The buffer or liquid medium may be manually inserted into the swab container or may be pre-stored in the swab container during manufacture of the swab. In another embodiment (not shown) when there is a female thread on the internal surface 220 instead of the male thread 223 on the external surface 222 of the swab container, the swab 24 is coupled to, or integral with, a male threaded cap. Figures 5A-5B illustrate a different embodiment of the present invention. In Figures 5A-5B, elements of the fluidic module A and / or the reservoir module B that are in common with the fluidic module A and / or the reservoir module B of Figures 1-4 are indicated with the same reference numerals and are not further described. In the embodiment illustrated in Figure 5A5B, the fluidic module A further includes an integrated buffer container 22'; the fluidic module A and the buffer container 22' are monolithic or a single piece. Referring to Figure 2A, and as shown in Figure 5A, the buffer container 22' extends as a continuation of the lateral side 5e. To accommodate the buffer container 22' at the lateral side 5e, ensuring a proper fluid connection with the fluid circuit 8, the first portion 6a of the fluid connection 6 is, in the embodiment of Figure 5A, formed laterally of the buffer container 22', fluidly coupled to the buffer container as further described below. That is, the buffer container 22' extends between the first portion 6a of the fluid connection 6 and the through hole 14a'. The lateral side 5e of the solid body 5 is, in this embodiment, a lateral side of the buffer container 22'. The buffer container 22' includes a recess 200, an opening 206 at one end of the recess 200 for inserting the buffer, and a through hole 202 forming an entry point for a buffer solution, during use. The recess 200 extends at the front face 50a of the first portion 50, with a major dimension substantially parallel to the z-axis between the upper side 50c (where the opening 206 is present) and the lower side 50d of the first portion 50 (without reaching the lower side 50d). When the first portion 50 and the second portion 51 are joined together, the recess 200 forms the swab container 22'. The through hole 202 connects the front face 50a to the rear face 50b of the first portion 50. The first portion 6a of the fluid connection 6 is formed on a physical support protruding from the lateral side 5e of the buffer container 22'. The through hole 202 is fluidically connected to the first portion 6a. The buffer container 22' is fluidically connected to the lysis chamber 16 via the fluidic channel 14a. In one embodiment, a plastic cap 208 is attached to the solid body 5 via a flexible mechanical joint 207, the cap 208 being adapted to seal the opening 206 when the swab is housed in the swab container 22'. In another embodiment (not shown), a cap adapted to seal the opening 206 is integral with a swab at one end of a rod, or shaft, of said swab. In yet another embodiment, a cap adapted to seal the opening 206 is detachable from the solid body 5 and the shaft of the swab. In one embodiment, the fluidic module A further includes one or more protruding mechanical connectors (hereinafter also referred to as pins) 210 and one or more protruding guides 212, for example in the form of planes inclined with respect to the surface of the front face 5a of the solid body 5. A pin 210 is shown in Figures 5A and 5B. Pin 210 includes a body 210a and a head 210b and is, for example, mushroom-shaped or T-shaped. Body 210a of pin 210 extends at the first lateral side 5e of the swab container 22', away from the first lateral side 5e with a major dimension parallel to the x-axis. Head 210b extends at one end of body 210a opposite another end of body 210a that is physically coupled to lateral side 5e. In one example, the major dimension of head 210b is transverse to the major dimension of body 210a, such that head 210b protrudes laterally, or protrudes laterally, from body 210a. The lateral prominences of the head 210b form, as further explained below, an interlocking mechanism that is used to maintain the reservoir module B in a fixed position, when the reservoir module B is coupled to the fluidic module A in the embodiment of Figure 5A. The protruding guide 212 similarly extends at the first lateral side 5e of the body 5, away from the pin 210. The protruding guide 212 serves to assist in the alignment of the reservoir module B, when the reservoir module B is to be coupled to the fluidic module A in the embodiment of Figure 5A. The protruding guide 212 also functions as a locking means to prevent the detachment of the reservoir module B from the fluidic module A (and vice versa). The solid body 7 of the tank module B has a front face 7a and a rear face 7b opposite each other along the y-axis, a top side 7c and a bottom side 7d opposite each other along the z-axis, a first lateral side 7e and a second lateral side 7f opposite each other along the x-axis. The front face 7a is physically connected to the rear face 7b by the top side 7c, the bottom side 7d, the first lateral side 7e and the second lateral side 7f. The solid body 7 includes one or more openings or slots or housings 710 adapted to physically mate with the one or more pins 210 of the fluidic module A. For example, the opening 710 is formed as a trench at the front face 7a and extends from the first lateral side 7e to the second lateral side 7f. The opening 710 has dimensions that coincide with the body 210a, so that the body 210a can be inserted inside the opening 710 and the prominences of the head 210b come into contact with the first lateral side 7e. The body 210a may serve as a pivot point to facilitate alignment and mating of the reservoir module B with the fluidic module A. The solid body 7 also includes one or more protruding guides 712 (e.g. one or more inclined planes) on the second lateral side 7f, configured to mate with the corresponding side of the fluidic module A, to mate with the protruding guides 212. As a consequence of the above-described embodiment, when fluidic module A and reservoir module B are coupled together, pin 210 engages in the housing formed by opening 710, creating a mechanical joint between fluidic module A and reservoir module B (see Figure 5B). Further, when fluidic module A and reservoir module B are coupled together, protruding guide 712 is pressed onto protruding guide 212 until protruding guide 712, bending protruding guide 212, snaps underneath protruding guide 212. In other words, guides 212 and 712 function as a snap-action mechanism. The mechanical connection provided by the pin 210 when engaged in the opening 710 and by the protruding guides 212, 712, allows for precise alignment and avoids unwanted mutual movements between the two modules A and B. In another embodiment (not shown) the one or more pins 210 are on the reservoir module B and the one or more openings 710 are on the fluidic module A. In the embodiment of Figure 5A, the solid body 7 of the reservoir module B includes: an air inlet 714 (forming at least in part the first portion 4a' of the second connection 4', previously described), a reservoir chamber 716, a through hole 718, a through hole 720, an air channel 722, a through hole 724, a through hole 726, and a fluid channel 728. The air inlet 714 is formed by a cavity extending at the bottom side 7d of the solid body. The through hole 718 connects the air inlet 714 to the air channel 722. The air channel 722 extends at the front face 7a of the solid body 7, connecting the through hole 718 to the through hole 720. The through hole 720 connects the air channel 722 to the reservoir chamber 716. The through hole 720 is located in a region of the reservoir chamber 716 that cannot be reached, during use, by the liquid solution / buffer contained within the reservoir. The reservoir chamber 716 extends at the rear face 7b of the solid body 7. The reservoir chamber 716 is sealed, for example, by a plastic film or wall bonded or glued to the rear surface 7b of the solid body 7. Through hole 724 connects reservoir chamber 716 to fluid channel 728. Fluid channel 728 extends at the front face 7a of solid body 7, connecting through hole 724 to through hole 726. Second through hole 726 connects fluid channel 728 to second portion 6b of fluid connection 6. The through hole 718, the air channel 722, the through hole 720, the through hole 724, the fluid channel 728 and the through hole 726 are sealed for example by a transparent tape or film or by a plastic wall joined or glued at the front face 7a of the solid body 7, protecting them from the external environment and preventing liquid leaks. When fluidic module A and reservoir module B are mated together, the alignment provided by pin 210 with opening 710 and protruding guides 212, 712 further ensures proper mating between first portion 6a and second portion 6b of fluidic connection 6. A frangible membrane or removable cap may be provided to seal second portion 6b to prevent fluid leakage prior to use. The frangible membrane may be configured to rupture when second portion 6b is mated to first portion 6a. Figure 6 shows a further embodiment of the present invention. In Figure 6, the reservoir module B is the same as already described and shown for Figure 5A. However, unlike Figure 5A, in Figure 6 the lysis chamber 16 extends to the upper side 5c of the solid body 5 and is configured (shaped) to accommodate the buffer 24. In other words, the buffer container 22' and the lysis chamber 16 are formed by a common recess fluidically connected to the reaction chambers 18(1)-18(N) via the fluidic channel 14b. According to a further embodiment of the present invention, shown in Figure 7, the solid body 5 of the fluidic module A is formed by three portions (illustrated in the exploded view in Figure 7), including the first portion 50, the second portion 51 and the third portion 52. Different from the previously described embodiments, in the embodiment of Figure 7 the second portion 51 and the third portion 52 include respective tapes or films. In particular, the third portion 52 may be formed by a plurality of parts, in particular including a portion 52' of a film or tape and one or more portions 52' ' of semiconductor material (e.g., silicon), glued and / or mechanically connected to the first portion 50. In this embodiment, the elements previously described as part of the third portion 52 are instead formed on the rear face 50b of the first portion 50. The front face 50a of the first portion 50 is as previously described. The elements formed at the rear face 50b of the first portion 50 include, in particular, the second air recess 114a, the fourth air recess 112b, the fluid channel 14a, the fluid recess 140b, the recess 80 (housing the membrane 81). Furthermore, the cavity 40 extends at the lower face 50d of the first portion 50. The one or more portions 52'' of semiconductor material, forming part of the third portion 52, form the heaters 9, 13. Note that fluidic module A and reservoir module B of cartridge 2, in all disclosed embodiments, are adapted to store, prior to use, dry reagents (in module A) and liquid buffers / reagents (in module B). More specifically, fluidic module A is configured to store dry reagents in a humidity-controlled environment, as provided by the presence of the desiccant in the storage chamber 12; reservoir module B is configured to store liquids in the reservoir chamber 716 or in the buffer container 22 (depending on the respective embodiments). Fluidic module A is coupleable to reservoir module B. Since fluidic module A is a separate physical entity from reservoir module B, the present invention allows the two modules A and B to be manufactured according to respective separate processes, even in different environments or manufacturing facilities.For example, the fabrication and packaging of fluidic module A can be performed in a controlled environment with low relative humidity, reducing potential sources of contamination or unwanted hydration of the dried reagents. The fabrication and packaging of reservoir module B, on the other hand, can be performed in an environment with more relaxed constraints, thus limiting production costs. Furthermore, different technologies can be used for the fabrication of fluidic module A compared to the fabrication of reservoir module B.

Claims

CLAIMS 1. Cartridge (2) for Lab-On-Chip applications, comprising: - a first module (A) including: at least one desiccant storage chamber (12), a dry reagent storage chamber (18(1)-18(N)), a fluid circuit (8), fluidically connected to the dry reagent storage chamber (18(1)18(N)), a first air path (11c(1)-11c(N)) connecting the dry reagent storage chamber (18(1)-18(N)) to the desiccant storage chamber (12) and comprising a membrane (81) which is impermeable to liquids and permeable to gases and vapours, the membrane (81) being disposed in the first air path (11c(1)-11c(N)) in such a way as to prevent the flow of liquids from the dry reagent storage chamber (18(1)-18(N)) to the desiccant storage chamber (12); and - a second module (B) including a container (22; 22';716) having an internal cavity for containing a liquid solution, wherein: the first module (A) and the second module (B) have a respective first and second fluidic connector (6a, 6b) adapted to be fluidically coupled to each other, the first connector (6a) of the first module (A) is fluidically coupled to the fluidic circuit (8) and the second connector (6b) of the second module (B) is fluidically coupled to the container (22; 22'; 716).; 2. A cartridge according to claim 1, further comprising one or more desiccants in the desiccant storage chamber (12) and one or more dry reagents in the dry reagent storage chamber (18(1)-18(N)).

3. Cartridge according to any of the preceding claims, wherein the first module (A) comprises a first layer (50) and a second layer (52) coupled to each other, the first air path (11c(1)-11c(N)) comprising: a first portion (110c(1)-110c(N)) in the first layer (50), fluidly connected to the dry reagent storage chamber (18(1)-18(N)); a second portion (80) in the second layer (52); a third portion (112c(1)-112c(N)) in the first layer (50), fluidly connected to the desiccant storage chamber (12); a first through hole (111c(1)-111c(N)) extending through the first layer (50), fluidly connected to the first portion (110c(1)-110c(N)) of the first air path and air-coupled to the second portion (80) of the first air path via the membrane (81);and a second through hole (111c(N+1)-111c(2N)) extending through the first layer (50), fluidly connected to the third portion (112c(1)-112c(N)) of the first air path and air-coupled to the second portion (80) of the first air path via the membrane (81).; 4. Cartridge according to claim 1 or claim 2, wherein the first module (A) comprises a first layer (50) having a first face and a second face opposite to each other, the first air path (11c(1)-11c(N)) comprising: a first portion (110c(1)-110c(N)) at the first face (50a) of the first layer (50), fluidly connected to the dry reagent storage chamber (18(1)-18(N)); a second portion (80) at the second face (50b) of the first layer (50); a third portion (112c(1)-112c(N)) at the first face (50a) of the first layer (50), fluidly connected to the desiccant storage chamber (12);a first through hole (lllc(l)-lllc(N)) extending through the first layer (50) from the first to the second face (50a, 50b), fluidly connected to the first portion (ll0c(l)-ll0c(N)) of the first air path and air-coupled to the second portion (80) of the first air path via the membrane (8l); and a second through hole (lllc(N+l)-lllc(2N)) extending through the first layer (50) from the first to the second face (50a, 50b), fluidly connected to the third portion (ll2c(l)-ll2c(N)) of the first air path and air-coupled to the second portion (80) of the first air path via the membrane (8l).; 5. Cartridge according to claim 4, wherein the first module (A) comprises a second layer (51) and a third layer (52), wherein the second layer (51) includes a tape or film coupled to the first face (50a) of the first layer (50), configured to seal the fluid circuit (8), the desiccant storage chamber (12) and the dry reagent storage chamber (18(1)18(N)), and wherein the third layer (52) includes at least one tape or film coupled to the second face (50b) of the first layer (50) and at least one heating element (9; 13) of semiconductor material coupled to the second face (50b) of the first layer (50).

6. Cartridge according to any of the preceding claims, wherein the first module (A) and the second module (B) have respective mechanical coupling means (210, 710) for mechanically coupling the first module (A) to the second module (B).

7. Cartridge according to any of the preceding claims, wherein the first and second fluidic connectors (6a, 6b) are Luer connectors.

8. A cartridge according to any preceding claim, wherein: the container (22; 22') is a sample collection container comprising a swab (24) having a tip, a shaft coupled to the tip, and a cap (240) coupled to the shaft, the container has a threaded opening (22b) for inserting the swab (24) into the cavity, the cap (240) of the swab (24) is a threaded cap (240) adapted to seal the cavity when the swab (24) is inserted into the container, and the threaded cap is screwed onto the threaded opening, the container includes a first portion (221) of deformable material configured to undergo elastic deformation when internal pressure is exerted.

9. A cartridge according to claim 8, wherein the container (22; 22') includes a second portion (226) provided with protrusions within the cavity such that, when the swab is inserted into the container, the tip contacts the protrusions, the second portion (226) being configured to sustain said internal pressure without deformation.

10. A cartridge according to claim 9, wherein the second fluidic connector (6b) includes a through hole (228) through the container (22; 22') at said second portion (226), said elastic deformation being caused, during use, by pumping the liquid solution out of the container.

11. A cartridge according to any preceding claim, further comprising a second air path (11a) connecting by air the desiccant storage chamber to an outlet opening (40a) of the cartridge, said outlet opening (40a) being configured to be connected to external control means for air intake.

12. Cartridge according to any of claims 1-4 and claims 6-11, further comprising a sealing layer (51) coupled to the first module (A) for sealing the fluid circuit (8), the desiccant storage chamber (12) and the dry reagent storage chamber (18(1)-18(N)).

13. Cartridge according to any preceding claim, wherein the fluidic circuit (8) includes: a reaction chamber (16); a first fluidic channel (14a) fluidically connecting the first fluidic connector (6a) to the reaction chamber (16); and a second fluidic channel (14b) fluidically connecting the reaction chamber (16) to the dry reagent storage chamber (18(1)-18(N)).