Pyrotechnic cell disruption apparatus

Abstract

To provide a technique capable of improving disruption of cells contained in a fluid sample as compared with the traditional art.SOLUTION: This pyrotechnic cell disruption apparatus is provided with: a pyrotechnic charge that is configured to be ignited, and to be burnt upon ignition; and a pressure chamber that is configured to house a cell-containing fluid sample therein and to be pressurized when the pyrotechnic charge is ignited and burnt.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a pyrotechnic cell disruption device and a pyrotechnic cell disruption method. [Background technology]

[0002] Cell disruption, also known as cell lysis, is used to disrupt the outer boundary or cell membrane of a cell to release intracellular material, such as DNA, RNA, proteins, or organelles, from the cell. This release of intracellular material is important for various types of molecular diagnostics. To name a few, such diagnostics may include pathogen detection platforms, immunoassays for point-of-care diagnostics, protein purification for studying protein function and structure, cancer diagnostics, drug screening, mRNA transcriptome determination, and analysis of the individual or complex composition of specific proteins, lipids, and nucleic acids.

[0003] Patent Documents 1, 2, etc. disclose point-of-care devices for detecting nucleic acids. The point-of-care devices disclosed in Patent Documents 1, 2, etc. are adapted to receive a sample and include an extraction chamber containing a lysis solution for extracting and lysing the sample and a heater, and dissolve the sample in the extraction chamber to release nucleic acids. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2016 / 004539 [Patent Document 2] International Publication No. 2017 / 117666 Summary of the Invention [Problem to be solved by the invention]

[0005] Small sample sizes, especially when collected from humans or animals, have many advantages. They not only allow for less invasive sampling methods but also reduce the risk of contaminating the environment. Small sample sizes also allow for greater automation. Furthermore, (smaller) samples are easier to transport within the device, with less risk of sample contamination before testing, less risk of environmental contamination, and safe disposal after testing is complete. Furthermore, small sample sizes are more suitable for complying with various regulatory standards for clinical testing. Because of the lower risk associated with small sample sizes, handling these samples may require somewhat fewer trained technicians. Small sample sizes also work well for disposable point-of-care devices (POCs), where size and unit cost are important.

[0006] Other types of samples can be plants and fungi. For example, farmers may want to know which pathogens or bioengineered agents are affecting their crops. Food safety and testing are other areas where small sample sizes are appropriate.

[0007] Microfluidic technology involves the handling and manipulation of very small volumes of fluid, such as microliters, and offers various advantages, such as small reagent volumes, high specific surface area (surface-to-volume ratio), low cost, and easy handling of small volumes of fluid suitable for cell analysis. Microfluidic devices have also been proposed for cell disruption. However, cell disruption with very small volumes of fluid presents various challenges, one of which is how to increase pressure. Some cell disruption methods may require increasing and releasing pressure for a short period of time to optimize efficiency. However, cell disruption using a hydraulic press has not been widely reported. If done, it can take time to build up pressure and require large equipment.

[0008] Another factor is heat: while excessive heat over a long period of time may be undesirable as it may damage proteins that are desired to be extracted from inside the cells, short-term exposure to heat in combination with pressure may have an additional positive effect on cell disruption.

[0009] Additionally, cell disruption by thermal lysis requires significant power, and chemical lysis uses strongly alkaline materials such as KOH. Chemical lysing reagents may require an additional buffering / neutralization step before the lysed sample is suitable for downstream analysis. [Means for solving the problem]

[0010] According to one aspect of the present invention, a pyrotechnic cytolysis device includes a pyrotechnic charge configured to be ignited and to combust upon ignition, and a pressure chamber including an interior space configured to contain a fluid sample containing cells and to be pressurized upon ignition and combustion of the pyrotechnic charge. [Effects of the Invention]

[0011] The technology disclosed herein can provide a technology that can improve the disruption of cells contained in a fluid sample compared to conventional techniques. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 shows a schematic cross-sectional view of a first embodiment of a first pyrotechnic cytolysis device before pressurization. [Figure 2] FIG. 2 shows a schematic cross-sectional view of the second embodiment of the first pyrotechnic cytolysis device after pressurization. [Figure 3] FIG. 3 shows a schematic cross-sectional view of the first embodiment of the second pyrotechnic cytolysis device before pressurization. [Figure 4] FIG. 4 shows a schematic cross-sectional view of the second embodiment of the second pyrotechnic cytolysis device after pressurization. [Figure 5] FIG. 5 shows a schematic cross-sectional view of a first embodiment of the third pyrotechnic cytolysis device before pressurization. [Figure 6]FIG. 6 shows a schematic cross-sectional view of the second embodiment of the third pyrotechnic cytolysis device after pressurization. [Figure 7] FIG. 7 shows a schematic cross-sectional view of a first embodiment of the fourth pyrotechnic cytolysis device before pressurization. [Figure 8] FIG. 8 shows a schematic cross-sectional view of the second embodiment of the fourth pyrotechnic cytolysis device after pressurization. [Figure 9] FIG. 9 shows an exploded view of a pyrotechnic cell disruption device according to the fifth embodiment. [Figure 10] FIG. 10 shows a schematic cross-sectional view of a pyrotechnic cell disruption device according to the fifth embodiment. [Figure 11] FIG. 11 shows a perspective view of a pyrotechnic cell disruption device according to the sixth embodiment. [Figure 12] FIG. 12 shows a schematic cross-sectional view of a pyrotechnic cell disruption device according to the sixth embodiment. [Figure 13] FIG. 13 shows a schematic cross-sectional view of a pyrotechnic cell disruption device including a sample tip according to the seventh embodiment. [Figure 14] FIG. 14 shows a cross-sectional view and a plan view similar to FIG. 13, but without the clamp in the first embodiment before pressure is applied. [Figure 15] FIG. 15 shows a cross-sectional view and a plan view as in FIG. 13, but without the clamps, in the second embodiment after pressure application. [Figure 16] FIG. 16 shows a schematic cross-sectional view of a pyrotechnic cytolysis device including a sample tip with downstream processing according to an eighth embodiment. [Figure 17] FIG. 17 shows a cross-sectional view and a plan view similar to FIG. 16, but without the clamp in the first embodiment before pressure is applied. [Figure 18] FIG. 18 shows a cross-sectional view and a plan view as in FIG. 16, but without the clamp in the second embodiment after pressure application. [Figure 19] FIG. 19 shows an exploded view of the pyrotechnic cell disruption device according to the ninth embodiment. [Figure 20] FIG. 20 shows a schematic cross-sectional view of a pyrotechnic cell disruption device according to the ninth embodiment. [Figure 21]FIG. 21 shows a cross-sectional view of only the tip shown in FIGS. 19 and 20 to illustrate the first step of the cell disruption method. [Figure 22] FIG. 22 shows a cross-sectional view of only the tip shown in FIGS. 19 and 20 to illustrate the second step of the cell disruption method. [Figure 23] FIG. 23 shows a cross-sectional view of a tenth embodiment similar to FIGS. 19-22, but in which the chip is an integral part of the lower housing. [Figure 24] FIG. 24 shows a plan view of the lower housing part forming the chip in the tenth embodiment shown in FIG. [Figure 25] FIG. 25 shows a cross-sectional view of the tip only to illustrate the first step of the cell disruption method. [Figure 26] FIG. 26 shows a cross-sectional view of the tip only to illustrate the second step of the cell disruption method. [Figure 27] FIG. 27 shows a schematic cross-sectional view of a pyrotechnic cell disruption device including a sample tip according to the eleventh embodiment. [Figure 28] FIG. 28 shows a cross-sectional view and a plan view similar to FIG. 27, but without the clamp in the first embodiment before pressure is applied. [Figure 29] FIG. 29 shows a cross-sectional view and a plan view as in FIG. 27, but without the clamp in the second embodiment after pressure application. DETAILED DESCRIPTION OF THE INVENTION

[0013] The pyrotechnic cell lysis device disclosed in each embodiment described below includes a pyrotechnic charge (explosive) configured to be ignited and combust upon ignition, and a pressure chamber configured to contain a fluid sample containing cells and to be pressurized upon ignition and combustion of the pyrotechnic charge. The pyrotechnic cell lysis method disclosed in each embodiment includes containing a fluid sample containing cells in a pressure chamber configured to be pressurized by the pyrotechnic charge, and pressurizing the fluid sample contained in the pressure chamber by igniting and combusting the pyrotechnic charge.

[0014] In this way, by using the fuel gas of the pyrotechnic charge as the main pressure source for pressurizing the fluid sample, cell disruption can be achieved in an extremely short time. This allows cell disruption to be achieved in an extremely short time compared to pressurizing the fluid sample using a hydraulic press. Furthermore, unlike the thermal lysis method, cells are not exposed to excessive heat for a long period of time, which can prevent cell damage. Furthermore, unlike the chemical lysis method, which uses chemical substances, cell disruption can be achieved without using chemical substances that are difficult to handle.

[0015] The pyrotechnic cell disruption device according to the present disclosure may also include an initiator (igniter) having a housing for accommodating a pyrotechnic charge. The initiator may be, for example, an initiator for activating a vehicle airbag, and controls the ignition of the pyrotechnic charge by receiving a supply of operating power from an external power source. The pyrotechnic charge may be, but is not limited to, a ZPP (zirconium potassium perchlorate), ZWPP (zirconium tungsten potassium perchlorate), THPP (titanium hydride potassium perchlorate), lead tricinate, etc.

[0016] The fluid sample accommodated in the pressure chamber according to the present disclosure is not particularly limited as long as it is a fluid sample containing cells, for example, a cell suspension in which cells are dispersed in a liquid. The cells contained in the fluid sample are not particularly limited, and may be cells collected from a human or animal, plant cells, fungi, or other cells.

[0017] In the present disclosure, the volume (scale, size) of the fluid sample accommodated in the pressure chamber is not particularly limited, but for example, an extremely small volume on the order of microliters (μL) can be accommodated. For example, the volume of the fluid sample may be between 10 μL and 500 μL. Alternatively, for example, 20 μL, 50 μL, 100 μL, 150 μL, 200 μL, or 300 μL may be used as the upper or lower limit of the volume of the fluid sample. Of course, the volume of the fluid sample can be on the order of milliliters (mL) or larger.

[0018] The number of cells contained in the fluid sample is also not particularly limited. For example, the number of cells contained in the fluid sample is 1×10 2 cells / cm 3 More than 1×10 9 cells / cm 3 It may be the following: For example, 1×10 3 cells / cm 3 , 1×10 4 cells / cm 3 , 1×10 5 cells / cm 3 , 1×10 6 cells / cm 3 , 1×10 7 cells / cm 3 , 1×10 8 cells / cm 3 The upper limit of the number of cells contained in the fluid sample. Alternatively, it may be used as a lower limit.

[0019] The pyrotechnic cell lysis device and pyrotechnic cell lysis method according to the present disclosure ignite and burn a pyrotechnic charge to pressurize a fluid sample containing cells contained in a pressure chamber and lyse the cells in a very short time, and also significantly shorten the time that the cells contained in the fluid sample are exposed to heat during pressurization. In the pyrotechnic cell lysis device and pyrotechnic cell lysis method according to the present disclosure, the time that the cells are exposed to heat during pressurization of the fluid sample is not particularly limited, but may be, for example, 0.1 ms (milliseconds) or more and 500 ms or less. Furthermore, the time that the cells are exposed to heat during pressurization of the fluid sample may be, for example, 1 ms, 10 ms, 20 ms, 30 ms, 40 ms, 50 ms, 60 ms, 70 ms, 80 ms, 90 ms, 100 ms, 200 ms, 300 ms, or 400 ms. may be used as the upper or lower limit of the time for which the cells are exposed to heat when pressurized. Furthermore, the time from the start of operation of the pyrotechnic cell disruption device according to the present disclosure to the completion of the disruption process of the cells contained in the fluid sample (disruption process duration) is not particularly limited, but may be, for example, 0.1 ms or more to 1 s ( For example, any of 1 ms, 10 ms, 100 ms, and 500 ms may be used for the crushing process. It may be used as an upper or lower limit for the duration of the process.

[0020] The pressure chamber of the pyrotechnic cytolysis device according to the present disclosure can contain, in addition to the fluid sample, any material other than the fluid sample. The material other than the fluid sample contained in the pressure chamber can be water, any other liquid, or other material such as a water-absorbing polymer, which can be filled into the pressure chamber. In this way, when the pyrotechnic charge is ignited and burned to pressurize the pressure chamber, the pressure shock wave generated can be smoothed.

[0021] The pyrotechnic cytolysis device according to the present disclosure may include an orifice through which the fluid sample contained in the pressure chamber flows after the fluid sample is pressurized by combustion gas from a pyrotechnic charge. The orifice is a minute passage through which the pressurized fluid sample flows in the pressure chamber. The orifice may be, for example, a precision orifice formed as a channel having an orifice diameter and a flow path length several times longer than the orifice diameter. The diameter of the orifice can be set to a size that allows the flow of the fluid sample and can apply sufficient shear stress to the fluid sample during flow. The diameter of the orifice can be set to different sizes depending on the size, number, type, etc. of cells contained in the fluid sample, but is, for example, 1 μm or more and 500 μm or more. Furthermore, for example, any of 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 60 μm, 70 μm, 80 μm, 90 μm, 100 μm, 200 μm, 300 μm, and 400 μm may be adopted as the upper or lower limit of the diameter of the orifice.

[0022] The pyrotechnic cell disruption device according to the present disclosure may include a pressure release unit that releases pressure from the pressure chamber after pressurizing the fluid sample contained in the pressure chamber with combustion gas from the pyrotechnic charge. The pressure release unit may include a pressure release valve, rupture disc, or the like that opens the pressure chamber to the outside space. The pressure value at which the pressure release valve opens or the rupture disc ruptures is not particularly limited, but may be set to a pressure value of 20,000 psi or more and 50,000 psi or less, for example. Also, for example, 25,000 psi, 30,000 psi, 35,000 psi, 40,000 psi, or 45,000 psi. may be used as the upper or lower limit of the pressure value at which the pressure release valve opens or the pressure value at which the rupture disc bursts.

[0023] Hereinafter, embodiments of the present disclosure will be described with reference to the drawings. Note that the configurations and combinations thereof in each embodiment are merely examples, and additions, omissions, substitutions, and other modifications of the configurations are possible as appropriate within the scope of the present invention. The present disclosure is not limited by the embodiments, but is limited only by the claims.

[0024] First Embodiment 1 shows a schematic cross-sectional view of a first embodiment of a first pyrotechnic cytolysis device before pressurization. This first pyrotechnic cytolysis device includes a cylinder body 1 having a piston 2 that forces a fluid sample 3 through a precision orifice 4 and releases (discharges) the sample to atmospheric pressure. The precision orifice 4 is formed within a separate part 10 that is bonded or pressed into a pressure chamber outlet such as a release (discharge) channel 9. The pressure source is not hydraulic, but rather comes from igniting a pyrotechnic charge 5. The pyrotechnic cytolysis device further includes a cap 6 that allows the device to be opened or closed, for example, to fill the device with a fluid sample 3 or to include the pyrotechnic charge 5.

[0025] In the first embodiment, which shows the device before pressurization, a low-pressure space 7 of the pressure chamber 15 may be provided between the piston 2 and the fluid sample 3. After ignition of the pyrotechnic charge 5, the device assumes the second embodiment shown in FIG. 2. Here, the low-pressure space 7 shown in FIG. 1 transforms into a high-pressure space 8 with a smaller volume and higher pressure. Alternatively, no space may be provided between the piston and the sample. The low-pressure space 7 may also be filled with a variety of different materials, such as liquids, as well as other materials, such as water-absorbing polymers. The latter may smooth the pressure shock wave to some extent, helping to achieve the desired cell disruption results. The sample can be loaded after removing the cap and piston, but it is also possible to load the sample through the release channel 9, especially after removing the insert containing the precision orifice 4.

[0026] The precision orifice 4 generates shear stress on the cells as they pass through it. The cells are then rapidly depressurized after passing through the precision orifice 4. The sample size can be as small as, for example, 150 μL, or as small as, for example, 10 μL. Larger sample sizes, such as 1 milliliter, are also possible. Given that the sample size is typically very small, the release time through the precision orifice 4 is very short. The gas, including the gas in the low-pressure space 7 that is rapidly compressed after ignition of the pyrotechnic charge 5, is heated by the compression very quickly and is exposed to that heat for only a very short time prior to its release through the precision orifice 4. This time is short enough to prevent damage to the cellular material, yet long enough to aid in the cell disruption process.

[0027] The pyrotechnic charge is contained within a separate pyrotechnic charge chamber housing 16, which also contains a first piston 12. Such pyrotechnic charge chamber housing 16 may include a first cylinder chamber 13 containing the pyrotechnic charge 5 and the first piston 12. The first piston 12 is movable relative to the first cylinder chamber 13 under pressure generated by igniting and burning the pyrotechnic charge 5. The first piston 12 is connected to a second piston, here the aforementioned piston 2, which is disposed within a pressure chamber 15. The piston 2 is disposed within the pressure chamber 15 so as to form one wall of the pressure chamber 15. The first cylinder chamber 13 is disposed within the pyrotechnic charge chamber housing 16, at least a portion of which is contained within the pressure chamber 15.

[0028] As described above, the first pyrotechnic cell lysis device D1 (see FIGS. 1 and 2) according to the first embodiment includes a pyrotechnic charge 5 (explosive) configured to be ignited and burn upon ignition, a pressure chamber 15 (low-pressure space 7 / high-pressure space 8) configured to accommodate a fluid sample 3 and to be pressurized upon ignition and combustion of the pyrotechnic charge 5, and an open channel 9 connecting the pressure chamber 15 (low-pressure space 7 / high-pressure space 8) to an external space. The open channel 9 has a precision orifice 4 configured as a passage for the fluid sample 3. The first pyrotechnic cell lysis device D1 includes a cylinder body 1 having the pressure chamber 15 therein. The cylinder body 1 has, for example, a cylindrical shape with a bottom and an open end at the top, as shown in FIGS. 1 and 2, and a cap 6 is detachably attached to the open end. The cylinder body 1 may also be a housing member having a cylindrical shape with a bottom. Furthermore, the pyrotechnic charge chamber housing 16 is, for example, integrally fixed to the inner surface of the cap 6. In the example shown in FIGS. 1 and 2, the fluid sample 3 is placed on the bottom of the cylinder body 1, and the fluid sample 3 is contained in the pressure chamber 15.

[0029] When the first pyrotechnic cell disruption device D1 is activated, ignition control of the pyrotechnic charge 5 is performed, for example, the pyrotechnic charge 5 is burned. Here, the first pyrotechnic cell disruption device D1 includes an initiator (igniter) for controlling ignition of the pyrotechnic charge 5, and the pyrotechnic charge 5 and a housing that holds it may form part of the initiator. The initiator may further include, for example, a wire or the like that is connected to an external power source, and can control ignition of the pyrotechnic charge 5 by receiving a supply of operating power from the external power source.

[0030] As described above, in the first pyrotechnic cell disruption device D1, the pyrotechnic charge 5 and the first piston 12 are accommodated in the first cylinder chamber 13 formed in the pyrotechnic charge chamber housing 16, at least a portion of which is accommodated in the cylinder body 1. Therefore, the pyrotechnic charge chamber housing 16 can also be referred to as the "first cylinder chamber housing" that forms the first cylinder chamber 13. As shown in FIG. 1, in the first mode (before pressurization) of the first pyrotechnic cell disruption device D1, the pyrotechnic charge 5 is accommodated in the upper region of the first cylinder chamber 13, and at least a portion of the first piston 12 is accommodated in the lower region thereof. As shown in FIG. 1, before operation of the first pyrotechnic cell disruption device D1, the pyrotechnic charge 5 is accommodated in the first cylinder chamber 13 defined by the pyrotechnic charge chamber housing 16, which is arranged separately from the cylinder body 1 of the pressure chamber 15. Here, the first cylinder chamber 13 can be identified as a "pyrotechnic charge chamber" that houses the pyrotechnic charge 5.

[0031] Here, the first piston 12 includes a head portion 121 and a rod portion 122 extending downward from the head portion 121 and integral with the head portion 121. A rod insertion hole 161 is formed through the bottom of the pyrotechnic charge chamber housing 16. With the rod portion 122 of the first piston 12 inserted into the rod insertion hole 161, the lower end of the rod portion 122 is connected to a piston 2 (second piston) housed in the pressure chamber 15 (outside the first cylinder chamber 13). With the head portion 121 housed in the pyrotechnic charge chamber housing 16, the first piston 12 is configured to be movable, for example, along the up-down direction of the first cylinder chamber 13. Furthermore, the diameter of the head portion 121 of the first piston 12 may be larger than the diameter of the rod insertion hole 161, so that the head portion 121 does not fall out of the first cylinder chamber 13.

[0032] Within the pressure chamber 15, the region sandwiched between the bottom of the cylinder body 1 and the piston 2 (second piston) is also referred to as the "second cylinder chamber." The second cylinder chamber in the pressure chamber 15 forms a low-pressure space 7 in the first mode before pressurization (before activation) of the first pyrotechnic cell disruption device D1 (see FIG. 1), and forms a high-pressure space 8 in the second mode after pressurization (after activation) (see FIG. 2).

[0033] During operation of the first pyrotechnic cell disruption device D1, for example, when the initiator controls the ignition of the pyrotechnic charge 5, the pyrotechnic charge 5 burns, generating combustion gas, which increases the pressure in the first cylinder chamber 13. As a result, the head portion 121 of the first piston 12 is pressed under the pressure generated by the combustion of the pyrotechnic charge 5, causing the first piston 12 to move downward relative to the first cylinder chamber 13. Accordingly, the piston 2 (second piston) connected to the rod portion 122 of the first piston 12 also moves downward (toward the bottom of the cylinder body 1) in conjunction with this. As a result, the volume of the second cylinder chamber in the pressure chamber 15 decreases, and the second cylinder chamber changes from a low-pressure space 7 to a high-pressure space 8. As a result, the fluid sample 3 contained in the second cylinder chamber (here, high-pressure space 8) of the pressure chamber 15 is pressurized, and cells (for example, cell outer shells (cell membranes, cell walls, etc.)) contained in the fluid sample 3 can be crushed. Note that in this specification, the terms "low-pressure space 7 / high-pressure space 8" refer to the same space within the pressure chamber 15. In other words, before the pyrotechnic cell crushing device D1 is activated (before pressurization), it is formed as a low-pressure space 7, and after the device is activated (after pressurization), the low-pressure space 7 changes to a high-pressure space 8, and this also applies to the following embodiments.

[0034] Furthermore, the fluid sample 3 contained in the pressure chamber 15 (high-pressure space 8) pressurized by the piston 2 (second piston) as described above is forced into the release (discharge) channel 9 communicating with the high-pressure space 8 and is discharged to the outside of the device through the release channel 9. The fluid sample 3 that passes through the release channel 9 is collected in a collection container (not shown), for example, under atmospheric pressure. Because the precision orifice 4 has a very small diameter, shear stress is generated when the fluid sample 3 passes through the precision orifice 4, and this shear stress acts on the fluid sample 3. In this embodiment, the shear stress generated when passing through the precision orifice 4 is applied to cells contained in the fluid sample 3, thereby effectively lysing the cells. Furthermore, the fluid sample 3 that has passed through the precision orifice 4 is suddenly depressurized by being exposed to atmospheric pressure. As a result, the fluid sample 3 suddenly expands, and the expansion pressure promotes lysing of the cells contained in the fluid sample 3. In this manner, the first pyrotechnic cell disruption device D1 can disrupt cells contained in the fluid sample 3 and separate intracellular substances such as DNA, RNA, proteins, or organelles from the cells, thereby enabling various cell analyses, cell diagnoses, etc.

[0035] Furthermore, with the first pyrotechnic cell lysis device D1, the pressure source for pressurizing the fluid sample 3 can be obtained by igniting and burning the pyrotechnic charge 5 rather than by hydraulic means. In this way, the combustion gas generated by ignition of the pyrotechnic charge 5 (gunpowder) operates the pistons (first piston 12 and second piston), instantaneously forming a high-pressure space 8 within the pressure chamber 15, rapidly pressurizing the fluid sample 3 and releasing the pressurized fluid sample 3 to the outside through the precision orifice 4. This enables efficient cell lysis in a short time. Furthermore, with the first pyrotechnic cell lysis device D1, the fluid sample 3 can be pressurized in a short time as described above, thereby preventing the cells contained in the fluid sample 3 from being exposed to high temperatures for a long period of time. This effectively prevents damage to the cells contained in the fluid sample 3.

[0036] Furthermore, according to the first pyrotechnic cell lysis device D1, a pyrotechnic charge 5 (explosive) is used as the pressure source for pressurizing the fluid sample 3, so that efficient cell lysis can be achieved with a small pressure source. As a result, the pyrotechnic cell lysis device can be made compact. Furthermore, the pyrotechnic charge 5 (explosive) used as the pressure source for pressurizing the fluid sample 3 can be said to be a material suitable for precise control of the generated pressure (output). Therefore, even if the size of the fluid sample 3 contained in the pressure chamber 15 is on the order of microliters, pressure control can be performed with high precision when pressurizing the cells contained in the fluid sample 3. This makes it possible to achieve a high degree of accuracy in pressure control when pressurizing the cells contained in the fluid sample 3, compared to conventional cell lysis devices (for example, French press ( This allows for a reduction in sample size, which was previously not possible with the conventional pyrotechnic cytolysis device (registered trademark). Thus, according to this embodiment, the fluid sample 3 can be made extremely small, enabling minimally invasive cell collection, particularly when collecting cells from humans or animals. Furthermore, the small sample size reduces the risk of environmental contamination and enables more advanced automation. Furthermore, the small sample size makes it easier for non-experts to handle. Furthermore, a small sample size works well for disposable point-of-care (POC) devices, where size and cost are important, and therefore a pyrotechnic cytolysis device suitable for point-of-care applications can be provided.

[0037] Second Embodiment FIG. 3 shows a schematic cross-sectional view of the first embodiment of the second pyrotechnic cytolysis device before pressurization, and FIG. 4 shows a schematic cross-sectional view of the second embodiment of the second pyrotechnic cytolysis device after pressurization. The embodiment shown in FIGS. 3 and 4 is essentially a variant of the embodiment shown in FIGS. 1 and 2. Similar elements are designated by the same reference numerals. The embodiment according to FIGS. 3 and 4 does not include a piston, but instead includes a deformable diaphragm 14 that divides the low-pressure space 7 and the corresponding high-pressure space 8 into two parts. As a result, the pressure chamber 15 is at least partially defined by the deformable diaphragm 14, which is configured to deform under the pressure generated by the pyrotechnic charge 5 upon ignition and combustion of the pyrotechnic charge 5, as shown in FIG. 4. The diaphragm 14 reduces the volume of the pressure chamber upon deformation, thereby pressurizing the pressure chamber upon deformation.

[0038] The pyrotechnic charge chamber housing 16 includes a predetermined break (break point) 17 at the lower end of the pyrotechnic charge pressure relief channel 18. A fluid sample 3 can be inserted into the device shown in Figures 3 and 4 by splitting the housing into two parts at the diaphragm, removing the diaphragm, and assembling the housing holding the diaphragm in place.

[0039] An advantage of the embodiment shown in Figures 3 and 4 is that the diaphragm 14 hermetically seals the fluid sample 3 from the pyrotechnic charge 5, thereby preventing by-products of the pyrotechnic charge 5 from mixing with the fluid sample 3 after ignition. The diaphragm is preferably made of metal, but other materials, such as PE, PP, or other plastically deformable polymers, may also be used. Another option is a composite structure such as coated fabric or polymer-coated glass, where the polymer coating holds the glass together even if the glass breaks. A bellows-like structure may also be useful as a diaphragm. A further option is to use a mesh screen that helps trap particles and protect the sample from debris but does not provide a pressure barrier, which is essentially a combination of the embodiments shown in Figures 3 and 5.

[0040] As described above, in the second pyrotechnic cell disruption device D2 according to the second embodiment (see FIGS. 3 and 4), the pyrotechnic charge 5 is accommodated in a separate pyrotechnic charge chamber 11 separated from the pressure chamber 15 formed inside the cylinder body 1. The pyrotechnic charge chamber 11 is formed inside the pyrotechnic charge chamber housing 16, and as shown in FIG. 3, a pressure release channel 18 is provided at the bottom of the pyrotechnic charge chamber housing 16. The pressure release channel 18 is a channel 18 for releasing the pressure generated inside the pyrotechnic charge chamber housing 16 when the pyrotechnic charge 5 is ignited and burned to the pressure chamber 15 in which the diaphragm 14 is disposed. In the second pyrotechnic cell disruption device D2, the pressure chamber 15 is divided into two spaces by the diaphragm 14. Of the pressure chamber 15 partitioned by the diaphragm 14, the space on the pyrotechnic charge chamber housing 16 side is referred to as the "first internal space S1," and the space on the bottom side of the cylinder body 1 on which the fluid sample 3 is placed is referred to as the "second internal space S2." Note that the first internal space S1 of the pressure chamber 15 does not include the internal space of the pyrotechnic charge chamber housing 16. As shown in Figures 3 and 4, the fluid sample 3 is The pressure chamber 15 is accommodated in a second internal space S2. The release channel 9 having the precision orifice 4 is connected to the second internal space S2 of the pressure chamber 15, and the second internal space S2 can be communicated with the external space via the release channel 9 having the precision orifice 4.

[0041] In the second pyrotechnic cell disruption device D2, in a pre-activation (pre-pressurization) state (first embodiment shown in FIG. 3 ), the pressure release channel 18 of the pyrotechnic charge chamber housing 16 does not communicate with the first internal space S1 of the pressure chamber 15, and is blocked by a break 17 formed in the bottom of the pyrotechnic charge chamber housing 16 facing the first internal space S1 (pressure chamber 15). Note that the pressure release channel 18 may be formed, for example, as a recess formed in the bottom of the pyrotechnic charge chamber housing 16, or the break 17 may be formed by a portion that is thinned by forming the pressure release channel 18. In this case, the break 17 in the pyrotechnic charge chamber housing 16 is formed as a weakened portion that is weaker than other portions. The break 17 in the pyrotechnic charge chamber housing 16 is arranged facing the first internal space S1.

[0042] When the second pyrotechnic cell disruption device D2 is activated, the pyrotechnic charge 5 is ignited and burned. The combustion gas generated by the pyrotechnic charge 5 increases the pressure in the pyrotechnic charge chamber 11, causing the rupture portion 17 to burst. As a result, the pyrotechnic charge chamber 11 and the first internal space S1 of the pressure chamber 15 communicate with each other via the pressure release channel 18, and the combustion gas from the pyrotechnic charge 5 is introduced into the first internal space S1. As a result, as shown in FIG. 4, the diaphragm 14 deforms from the first internal space S1 toward the second internal space S2. Compared to the state shown in FIG. 3, the volume of the first internal space S1 increases and the volume of the second internal space S2 decreases. As a result, the pressure in the second internal space S2 containing the fluid sample 3 increases, changing it from a low-pressure space 7 to a high-pressure space 8. 3, the combustion gas of the pyrotechnic charge 5 flows in through the pressure release channel 18, and therefore, the first internal space S1 also changes from a low-pressure space 7 to a high-pressure space 8 as the second pyrotechnic cell disruption device D2 is activated. In this embodiment, as in the first embodiment, the cap 6 is detachably provided on the upper end opening of the cylinder body 1, and a pyrotechnic charge chamber housing 16 that accommodates the pyrotechnic charge 5 can be fixed inside the cap 6.

[0043] As described above, the second pyrotechnic cell lysis device D2 is activated, pressurizing the fluid sample 3 contained in the second internal space S2 of the pressure chamber 15. As a result, cells contained in the fluid sample 3 can be lysed. Furthermore, as described above, the release (discharge) channel 9 including the precision orifice 4 is connected to the second internal space S2 of the pressure chamber 15, so the pressurized fluid sample 3 is forced into the release (discharge) channel 9. The shear stress generated when the fluid sample 3 passes through the precision orifice 4 is applied to the fluid sample 3, thereby promoting lysis of the cells contained in the fluid sample 3. After passing through the precision orifice 4, the fluid sample 3 is exposed to atmospheric pressure when released outside the device, causing rapid decompression and expansion, further promoting cell lysis. The fluid sample 3 subjected to cell lysis in this manner can be collected in a collection container, as in the first embodiment. The second pyrotechnic cell lysis device D2 of this embodiment provides the same effects as the first pyrotechnic cell lysis device D1. When the diaphragm 14 is made of a metal material, the material may be, for example, aluminum, cast iron, or stainless steel.

[0044] Third Embodiment Figure 5 shows a schematic cross-sectional view of the first embodiment of the third pyrotechnic cytolysis device before pressurization, and Figure 6 shows a schematic cross-sectional view of the second embodiment of the third pyrotechnic cytolysis device after pressurization. This third embodiment is very similar to the second embodiment, except that the diaphragm 14 is omitted entirely. Thus, a separate pyrotechnic charge chamber housing 16 opens directly into the pressure chamber upon ignition and combustion of the pyrotechnic charge. This embodiment allows for the use of a separate pyrotechnic charge chamber housing 16 directly into the pressure chamber during ignition and combustion of the pyrotechnic charge. This simplifies sample loading and minimizes sealing and moving parts, but exposes the sample to pyrotechnic charge by-products after ignition. However, depending on the specific sample and the desired intracellular molecules of interest, this may be acceptable if the intracellular molecules do not chemically interact with the by-products or are contaminated to such an extent that proper detection / diagnosis is compromised. Chemical interaction with the pyrotechnic charge by-products can also be used advantageously if the by-products enable / enhance cell lysis. Certain chemicals and surfactants can be used alone to lyse cells. Furthermore, heat generated by the pyrotechnic charge can also be advantageous in enabling / enhancing cell lysis. Mechanical disruption can also be utilized in this device by adding hard beads or other projectiles / agitators to the reaction chamber. As with the embodiments of Figures 1-4, similar reference numbers are used for similar elements in this third embodiment compared to the first and second embodiments.

[0045] In the third pyrotechnic cell rupture device D3 (see FIGS. 5 and 6) configured as described above, the pressure chamber 15 is not divided by a diaphragm or the like. The pressure chamber 15 is formed as a low-pressure space 7 before the third pyrotechnic cell rupture device D3 is activated (before pressurization). After the third pyrotechnic cell rupture device D3 is activated (before pressurization), the internal pressure of the pressure chamber 15 increases, changing it from the low-pressure space 7 to a high-pressure space 8. As is clear from FIG. 5, the structure of the pyrotechnic charge chamber housing 16 that houses the pyrotechnic charge 5 before the third pyrotechnic cell rupture device D3 is the same as that of the second pyrotechnic cell rupture device D2, and the pyrotechnic charge chamber housing 16 is provided with a pressure release channel 18 and a rupture portion 17. The pressure release channel 18 does not communicate with the internal space (low-pressure space 7) of the pressure chamber 15 before the third pyrotechnic cell rupture device D3 is activated (before pressurization). Then, as the device is activated, the pyrotechnic charge 5 is ignited, and the fragile rupture portion 17 ruptures under the pressure of the combustion gases generated by the combustion of the pyrotechnic charge 5. This causes the pyrotechnic charge chamber housing 16 (pyrotechnic charge chamber 11) to open directly to the pressure chamber 15 (low-pressure space 7), and the pressure within the pyrotechnic charge chamber housing 16 is released (discharged). This changes the pressure chamber 15 containing the fluid sample 3 from the low-pressure space 7 to the high-pressure space 8, pressurizing the fluid sample 3 and rupturing the cells contained in the fluid sample 3. Furthermore, the fluid sample 3 is forced into the release (discharge) channel 9, where it is subjected to shear stress as it passes through the precision orifice 4, thereby accelerating cell rupture. The fluid sample 3 that has passed through the release (discharge) channel 9 expands rapidly when released under atmospheric pressure, further accelerating cell rupture, and is then collected, for example, in a collection container. The third pyrotechnic cell disruption device D3 in this embodiment can also provide the same effects as the first and second pyrotechnic cell disruption devices D1 and D2.

[0046] <Fourth embodiment> FIG. 7 shows a schematic cross-sectional view of a first embodiment of the fourth pyrotechnic cytolysis device before pressurization, and FIG. 8 shows a schematic cross-sectional view of a second embodiment of the fourth pyrotechnic cytolysis device after pressurization. In contrast to the embodiment shown in FIGS. 1-6, the embodiment shown in FIGS. 7-8 does not have a precision orifice 4 but includes a hermetically sealed pressure chamber 15 that forms a low-pressure space 7 before ignition of the pyrotechnic charge, while the exact same space forms a high-pressure space 8 after ignition of the pyrotechnic charge, the latter embodiment shown in FIG. 8. The sample size can be similar to that shown in FIGS. 1-6, e.g., 1 mL, 150 μL, or 10 μL. In this embodiment, the fluid sample 3 is provided in a separate sample container 19. Furthermore, in this embodiment, the container includes a pressure release portion 21 equipped with a pressure release valve 20. The container 19 is sufficiently flexible so that it can collapse under pressure and pressurize the fluid sample 3. For example, the container may be formed as a sample pouch (see, for example, Figure 9). Alternatively, if the container 19 is gas-free and is filled entirely with the fluid sample 3, the support of the fluid sample 3 may prevent the container from collapsing, but the container should still be flexible enough to pressurize the sample. Apart from containing the sample, the container 19 may also contain a different material, for example a water-absorbing polymer, which may increase the integrity of the container when exposed to the pressure shock wave resulting from the combustion of the pyrotechnic charge 5. Conversely, if desired, the container can be configured to burst under pressure from a pyrotechnic charge. Also, one or more samples (pouches) can be processed simultaneously in the same chamber.

[0047] After rapid pressurization upon ignition and combustion of the pyrotechnic charge 5, the fluid sample 3 is exposed to a sudden pressure increase due to the gas pressure generated by the pyrotechnic charge 5, and the pressure release valve 20 is then opened at a time effective for cell disruption. The opening of the pressure release valve 20 also typically occurs quickly, causing a rapid pressure drop over time, facilitating cell membrane rupture due to cell expansion. The heat generated by the pyrotechnic charge 5 can help make the cell membrane vulnerable to rupture, thus aiding cell disruption. Various other physical and chemical conditions resulting from the combustion of the pyrotechnic charge 5 can contribute to cell disruption. The composition of the charge (explosive) itself can be applied to adjust the rate of gas and heat generation, depending in some cases on the initiator and gas generator combination. Waste heat can be used for other purposes within the point-of-care (POC) detection system (amplification and detection). Waste gas pressure can be used to move the sample from one location (stage) to another or stored for use in the POC device.

[0048] The container 19 concept helps comply with various regulatory standards for clinical trials because the operator is protected from exposure to the sample. Depending on the material from which the container 19 is made and the amount of sample that fills the container, the container may either burst under pressure or remain intact.

[0049] The fluid sample 3 can be removed from the device by opening the cap 6 or, if the container 19 is configured to rupture, by releasing it via the pressure release valve 20. Such release of the disrupted cell sample can be achieved by turning (rotating) the device and using some residual pressure to release the sample.

[0050] The advantage of this hydrostatic shock is that it minimizes seals and moving parts, and the housing can be spring loaded or elastically deformed to create a resonant pressure wave at a specific frequency and amplitude, increasing the extent of cell lysis before the pressure is released.

[0051] The use of pyrotechnic cell disruption generates sufficient shear stress and provides rapid enough decompression that the sample does not need to be forced through a precision orifice 4 in this embodiment. This offers significant advantages over simply replacing bulky equipment such as a prior art French press with a pyrotechnic cell disruption. Specifically, the entire sample preparation and assay can be performed in a single chamber of the device, i.e., a hydrostatic process. This device has a simpler design and fewer parts. This not only reduces development and manufacturing costs, but also reduces the risk of mechanical failure and contamination. This may also reduce sample waste, thereby reducing the required sample volume and / or improving the assay signal.

[0052] The fourth pyrotechnic cytolysis device D4 (see Figures 7 and 8) configured as described above comprises a pyrotechnic charge 5 configured to be ignited and burn upon ignition, a pressure chamber 15 (low pressure space 7 / high pressure space 8) that contains a fluid sample 3 and is configured to be pressurized upon ignition and combustion of the pyrotechnic charge 5, and a pressure release section 21 (pressure release valve 20) that releases pressure from the pressure chamber 15 after the pressure chamber 15 is pressurized by ignition and combustion of the pyrotechnic charge 5. As is clear from Figure 7, the structure of the pyrotechnic charge chamber housing 16 that contains the pyrotechnic charge 5 before operation (before pressurization) of the fourth pyrotechnic cytolysis device D4 is the same as that of the second and third pyrotechnic cytolysis devices D2 and D3, and the pyrotechnic charge chamber housing 16 is provided with a pressure release channel 18 and a rupture section 17, and the pyrotechnic charge 5 is ignited and burned. The pyrotechnic charge 5 is formed in a pyrotechnic charge chamber housing 16 configured to explode upon combustion and is contained in a separate pyrotechnic charge chamber 11 separated from the pressure chamber 15. In the fourth pyrotechnic cell rupture device D4, similar to the third pyrotechnic cell rupture device D3, the pyrotechnic charge 5 is ignited during operation, and the rupture portion 17 is ruptured by the pressure of the combustion gas generated, causing the pyrotechnic charge chamber housing 16 (pyrotechnic charge chamber 11) to open directly to the pressure chamber 15 (low-pressure space 7). As a result, the combustion gas of the pyrotechnic charge 5 flows from the pyrotechnic charge chamber housing 16 into the pressure chamber 15 in which a container 19 holding (accommodating) the fluid sample 3 is disposed, changing the interior of the pressure chamber 15 from the low-pressure space 7 to the high-pressure space 8. This pressurizes the fluid sample 3 contained in the container 19, allowing the cells contained in the fluid sample 3 to be ruptured.

[0053] In the fourth pyrotechnic cell lysis device D4, the container 19 is formed, for example, as a flexible pouch that can be filled (contained) with the fluid sample 3, and the pouch is configured not to burst when exposed to the pressure generated when the pyrotechnic charge is ignited and burned. This allows the fluid sample 3 filled therein to be suitably pressurized and cells to be lysed when the fourth pyrotechnic cell lysis device D4 is activated without destroying the container 19.

[0054] Furthermore, in the fourth pyrotechnic cell lysis device D4, the pressure release unit 21 includes a pressure release valve 20. The pressure release valve 20 may be, for example, a valve element that automatically opens when a predetermined time has elapsed since the ignition of the pyrotechnic charge 5 during operation of the fourth pyrotechnic cell lysis device D4. Alternatively, the pressure release valve 20 of the pressure release unit 21 may be a valve element that automatically opens when the pressure in the pressure chamber 15 (high-pressure space 8) containing the container 19 rises to a predetermined pressure during operation of the fourth pyrotechnic cell lysis device D4. The pressure setting value at which the pressure release valve 20 automatically opens is not particularly limited, but may be set to open when exposed to a pressure of at least 20,000 psi. The pressure setting value at which the pressure release valve 20 automatically opens can be appropriately set depending on, for example, the amount of fluid sample 3 contained in the container 19, the type of cells contained in the fluid sample 3, etc. Alternatively, the pressure release valve 20 may be a valve element that can be manually opened. The pressure relief portion 21 may also include a rupture disc that ruptures when exposed to a predetermined pressure, for example, the rupture disc may be configured to rupture when exposed to a pressure of at least 20,000 psi.

[0055] As described above, when the pressure release valve 20 is opened while the fluid sample 3 contained in the pressure chamber 15 (high-pressure space 8) is pressurized under high pressure, a sudden drop in pressure occurs in the high-pressure space 8. As a result, cells contained in the fluid sample 3 suddenly expand, and large shear stress acts on the cells, which can promote cell disruption. The material of the container 19 that fills (contains) the fluid sample 3 is not particularly limited, and may be a flexible pouch. Alternatively, the container 19 may be formed of a hard material such as resin or glass. In this case, when the combustion gas of the pyrotechnic charge 5 changes the pressure chamber 15 from the low-pressure space 7 to the high-pressure space 8, the container 19 may be crushed or ruptured, thereby pressurizing the fluid sample 3 inside.

[0056] Fifth Embodiment FIG. 9 shows an exploded view of the fifth pyrotechnic cell disruption device, and FIG. 10 shows a schematic cross-sectional view of the fifth pyrotechnic cell disruption device. The pressure release section 21 is formed as a rupture disk assembly 22 including a rupture disk 23 as shown in FIG. 10. This rupture disk quickly releases pressure in a controlled manner and within the time frame required to rupture the rupture disk 23. This rupture disk is designed to allow for such a time frame required to achieve cell disruption. Also, the sample pouch 24 is provided within a sample cage 25. This cage 25 is not necessary, but it helps to handle the sample pouch 24 carefully and makes it easier to insert the sample pouch into the pressure chamber 15. The cage also prevents the sample pouch from accidentally blocking the pressure relief vent after the pyrotechnic charge is detonated. If this were to occur, the sample could potentially be aerosolized and blown into the surroundings. This is further mitigated by the embodiment shown in Figures 11 and 12, which positions the sample outside the direct path between the gases generated by the pyrotechnic charge and the vent. Assembly of the device is accomplished by threading the pressure chamber 15, containing the sample pouch 24 inserted within the cage 25, together with the gasket 26 and cap 6, with high-strength bolts 27 or other clamping methods. The pyrotechnic charge is contained within an initiator 28 inserted coaxially within the pressure chamber 15. Ignition of the pyrotechnic charge can be achieved by connecting the initiator 28 via its wires 29 to a power source that provides sufficient voltage to ignite the pyrotechnic charge within the initiator 28.

[0057] The fifth pyrotechnic cell rupture device D5 in the fifth embodiment is a more specific realization of the concept of the fourth pyrotechnic cell rupture device D4 described in Figures 7 and 8. In the fifth embodiment, like elements to those in the fourth embodiment are designated by like reference numerals. As shown in Figure 10, the pressure chamber 15 has a pressure vessel 150, and a hollow portion is provided extending through the pressure vessel 150 in the vertical direction of the pressure vessel 150. An initiator 28 is attached to the portion where the hollow portion opens to the bottom of the pressure vessel 150 so as to airtightly close the opening. As shown in Figure 10, the pyrotechnic charge chamber housing 16 of the initiator 28 is disposed facing the interior of the pressure vessel 150.

[0058] Reference numeral 153 in FIG. 9 denotes a cap fastening portion of the pressure vessel 150. In the example shown in FIG. 9, the cap fastening portion 153 is located on the upper side of the pressure vessel 150, and allows the cap 6 to be detachably attached. Threaded holes for inserting high-strength bolts 27 are formed in the cap fastening portion 152, the gasket 26, and the cap 6. By sandwiching the gasket 26 between the pressure vessel 150 and the cap 6 and screwing the high-strength bolts 27 inserted into each threaded hole, the cap 6 can be fastened integrally to the cap fastening portion 153 of the pressure vessel 150. Conversely, the cap 6 can be detached from the pressure vessel 150 by removing the high-strength bolts 27. Attaching the cap 6 to the pressure vessel 150 forms an airtight pressure chamber 15 inside the pressure vessel 150.

[0059] The pressure chamber 15 in this embodiment extends in the vertical direction (axial direction) of the pressure vessel 150 and includes a sample container accommodating section 151 and a pressure channel 30 that are coaxially connected. As shown in FIGS. 9 and 10 , the sample container accommodating section 151 opens to the top of the pressure vessel 150, and the lower end of the sample container accommodating section 151 is connected to the upper end of the pressure channel 30. The pressure channel 30 and the sample container accommodating section 151 are, for example, cylindrical hollow sections. The diameter (cross-sectional area) of the sample container accommodating section 151 is slightly larger than the diameter (cross-sectional area) of the pressure channel 30. A sample container mounting section 152 is formed by a radially extending step provided between the connection (boundary) of the sample container accommodating section 151 and the pressure channel 30. A single sample pouch 24 or a sample pouch 24 with a cage 25 attached can be mounted on the sample container mounting section 152. This allows the sample pouch 24 to be stably accommodated within the pressure chamber 15. The cage 25 is, for example, a basket-like member capable of holding a flexible sample pouch 24. Even when the cage 25 is attached to the sample pouch 24, a portion of the sample pouch 24 remains exposed. Although this embodiment describes an example in which a fluid sample is accommodated in a flexible sample pouch 24, another container to be filled with a fluid sample may be placed on the sample container placement portion 152 and accommodated in the sample container accommodation portion 151.

[0060] A pyrotechnic charge chamber 11 for accommodating a pyrotechnic charge 5 is formed inside the pyrotechnic charge chamber housing 16 of the initiator 28. The pyrotechnic charge chamber housing 16 is disposed in the pressure channel 30. The pyrotechnic charge chamber housing 16 is, for example, The fifth pyrotechnic cell disruption device D5 may be formed of a cup member that can be ruptured by the energy generated when the pyrotechnic charge 5 is ignited and burned. The cup member may be formed of a thin-walled metal member such as aluminum. This allows the pyrotechnic charge chamber 11 to open directly to the pressure channel 30 in the pressure chamber 15 by the energy generated when the pyrotechnic charge 5 is ignited and burned, and allows the combustion gas of the pyrotechnic charge 5 to be introduced into the pressure channel 30. The pressure channel 30 functions as an air passage for supplying the combustion gas of the pyrotechnic charge 5 to the sample container holder 151 when the initiator 28 is activated. Note that before activation of the fifth pyrotechnic cell disruption device D5, the pressure chamber 15 (pressure channel 30 and sample container holder 151) is formed as a low-pressure space 7.

[0061] As shown in Figures 9 and 10, the sample container receptacle 151 is open on the top surface of the pressure vessel 150, and is configured so that the sample container receptacle 151 is covered by the cap 6 when the cap 6 is attached to the pressure vessel 150. Also, as shown in Figures 9 and 10, the cap 6 is provided with a pressure release section 21. The pressure release section 21 includes a rupture disk assembly 22 and a pressure release vent passage 61. The pressure release vent passage 61 is a vent passage formed to penetrate the cap 6 in the axial direction. The rupture disk assembly 22 includes a pressure release vent passage 22A connected to the pressure release vent passage 61 and a rupture disk 23 arranged to block (block) the pressure release vent passage 22A midway. The pressure release vent passage 22A extends vertically through the rupture disk assembly 22. When the rupture disk 23 ruptures after the pressure chamber 15 is pressurized by the ignition and combustion of the pyrotechnic charge 5, the pressure release vents 61, 22A connect the sample container housing 151 to the external space, releasing the pressure in the pressure chamber 15 to the outside. The rupture disk 23 is configured to rupture when the primary pressure, i.e., the pressure in the pressure chamber 15, rises to a predetermined pressure. The pressure at which the rupture disk 23 ruptures can be appropriately set depending on, for example, the amount of fluid sample 3 filled in the sample pouch 24 and the type of cells contained in the fluid sample 3. As shown in FIG. 10 , when the cap 6 is attached to the pressure vessel 150, the pressure channel 30, the sample container housing 151, and the pressure release vents 61, 22A are coaxially arranged. More specifically, when the cap 6 is attached to the pressure vessel 150 , the pressure channel 30 , the sample vessel receptacle 151 , and the pressure release vent passages 61 and 22 A are coaxially arranged so as to be aligned in a straight line passing through the central axis of the pressure vessel 150 .

[0062] In the fifth pyrotechnic cell disruption device D5 configured as described above, when the initiator 28 is activated to ignite the pyrotechnic charge 5, combustion gas generated by the combustion of the pyrotechnic charge 5 increases the internal pressure of the pyrotechnic charge chamber 11, causing the pyrotechnic charge chamber housing 16 (e.g., a cup member) to rupture. As a result, the combustion gas of the pyrotechnic charge 5 is introduced from the pyrotechnic charge chamber 11 into the pressure channel 30 of the pressure chamber 15. The combustion gas of the pyrotechnic charge 5 is then introduced into the sample container receptacle 151 of the pressure chamber 15, which is coaxially connected to the pressure channel 30, causing a sudden increase in pressure in the sample container receptacle 151, in which the sample pouch 24 is accommodated. This changes the sample container receptacle 151 from the low-pressure space 7 to the high-pressure space 8. A portion of the sample pouch 24 is exposed even when the cage 25 is attached. This allows the sample pouch 24 to be exposed to the high pressure inside the sample container storage section 151, which has been transformed into a high-pressure space 8. As a result, the fluid sample 3 filled in the sample pouch 24 is suddenly pressurized, and cells contained in the fluid sample 3 can be ruptured. Furthermore, when the pressure in the sample container storage section 151 rises to a predetermined pressure, the rupture disk 23 ruptures, and the sample container storage section 151 is quickly depressurized. As a result, the cells contained in the fluid sample 3 filled in the sample pouch 24 suddenly expand, and for example, a large shear stress acts on the cells, further promoting cell rupture. In the above example, the pressure release vent path 22A of the pressure release section 21 is blocked by the rupture disk 23 (rupture disk), but this may be replaced with the pressure release valve 20 described in the fourth embodiment. That is, pyrotechnics The pressure release valve 20 may be automatically opened when a predetermined time has elapsed since the ignition of the charge 5, or when the pressure in the pressure chamber 15 has risen to a predetermined pressure.

[0063] In the fifth pyrotechnic cytolysis device D5, the pressure channel 30 and the sample container housing 151 are coaxially arranged. Therefore, when the initiator 28 is activated to combust the pyrotechnic charge 5, combustion gas is smoothly introduced into the sample container housing 151 via the pressure channel 30, enabling rapid lysis of cells contained in the fluid sample 3 filled in the sample pouch 24. Furthermore, in the fifth pyrotechnic cytolysis device D5, the pressure channel 30, the sample container housing 151, and the pressure release vents 61 and 22A are coaxially arranged when the cap 6 is attached to the pressure vessel 150. Therefore, when the rupture disk 23 ruptures after the initiator 28 is activated, combustion gas can be smoothly released from the pressure chamber 15 (the sample container housing 151 and the pressure channel 30) to the outside through the pressure release vents 61 and 22A. This allows the pressure chamber 15 to be depressurized in a shorter time. As a result, the cells contained in the fluid sample 3 in the sample pouch 24 expand more rapidly, further promoting cell disruption.

[0064] Sixth Embodiment FIG. 11 shows a perspective view of a sixth embodiment of a pyrotechnic cell disruption device, and FIG. 12 shows a schematic cross-sectional view of the sixth embodiment. This sixth embodiment is very similar to the fifth embodiment. However, the initiator 28 is arranged laterally and is connected to the low-pressure / high-pressure spaces 7 and 8 in the radial direction of the pressure chamber 15 via a pressure channel 30. Compared with the coaxial approach shown in FIGS. 9 and 10, the advantage of the lateral approach shown in FIGS. 11 and 12 is that, after ignition of the pyrotechnic charge, the sample pouch 24 in the sample cage 25 is not directly exposed to the gas flow from the initiator 28. Furthermore, the pressure channel 30 can cross the spaced-apart low-pressure / high-pressure spaces 7 and 8 beyond one axial end of the sample pouch 24 / sample cage 25 combination. Meanwhile, the other axial end of the sample pouch 24 / sample cage 25 combination can rest on the bottom of the pressure chamber 15. Apart from avoiding direct exposure of the sample pouch 24 and sample cage 25 combination to the gas flow, the pressure created by the pyrotechnic charge also forces the sample pouch 24 and sample cage 25 combination down towards the bottom of the pressure chamber 15. This holds the sample pouch 24 and sample cage 25 combination securely in place and avoids undesired movement of the sample pouch 24 and sample cage 25 combination.

[0065] The sixth pyrotechnic cell lysis device D6 according to the sixth embodiment is a modification of the fifth pyrotechnic cell lysis device D5. In the sixth embodiment, like elements to those in the fifth embodiment are designated by like reference numerals. The sixth pyrotechnic cell lysis device D6 includes a pressure vessel 150 having a cylindrical shape with a bottom, and a pressure chamber 15 formed therein. The pressure chamber 15 extends in a first direction and includes a pressurized space 155 and a pressure channel 30 that branches off from the pressurized space 155 in a second direction different from the first direction and connects to the pressurized space 155. In the example shown in FIGS. 11 and 12, the pressurized space 155 extends vertically (axially, first direction) of the pressure vessel 150, and the pressure channel 30 extends horizontally (radially) of the pressure vessel 150, branching off perpendicularly from the pressurized space 155. The pressure chamber 15 is formed as a low-pressure space 7 before the sixth pyrotechnic cell disruption device D6 is activated, and is formed as a high-pressure space 8 after the sixth pyrotechnic cell disruption device D6 is activated.

[0066] The sixth pyrotechnic cell rupture device D6 also includes a pressure release section 21 in the cap 6, similar to the fifth pyrotechnic cell rupture device D5. The pressure release section 21 includes a rupture disk assembly 22 and a pressure release vent passage 61. The pressurized space section 155 is located proximal to the pressure release section 21. 150. The pressure vessel 150 has a proximal end and a distal end, with the distal end 155B being positioned at the bottom 157 of the pressure vessel 150. Meanwhile, the proximal end of the pressurized space 155 opens to the top surface of the pressure vessel 150, and is configured so that the pressurized space 155 is covered by the cap 6 when the cap 6 is attached to the pressure vessel 150. Furthermore, when the cap 6 is attached to the pressure vessel 150, the proximal end of the pressurized space 155 is configured to be connected to the pressure release vent path 61. For example, the pressurized space 155 and the pressure release vent path 61 are arranged coaxially through the central axis of the pressure vessel 150.

[0067] The pressurized space 155 includes a sample container accommodating section 156 for accommodating the sample pouch 24 between the connecting section 155C with the pressure channel 30 and the distal end. In other words, the pouch holding section 156 refers to the area of ​​the pressurized space 155 below the connecting section P1. In this embodiment, the sample pouch 24 with the cage 25 attached can be placed on the bottom 157 of the pressure vessel 150. Of course, a single sample pouch 24 may also be placed on the bottom 157 of the pressure vessel 150. The height of the sample container accommodating section 156 is greater than or equal to the height of the sample pouch 24, so that when the sample pouch 24 is accommodated in the sample container accommodating section 156, the sample pouch 24 does not protrude toward the connecting section P1. In other words, the sample pouch 24 can be accommodated in the sample container accommodating section 156 so that the upper end of the sample pouch 24 is located below the connecting section P1 with the pressure channel 30 in the pressurized space 155. 11 and 12, the initiator 28 is attached to the side of the pressure vessel 150, and is disposed laterally so that the pyrotechnic charge chamber housing 16 of the initiator 28 faces the inside of the pressure channel 30. In other words, the pyrotechnic charge 5 of the initiator 28 is disposed in the pressure channel 30.

[0068] The sixth pyrotechnic cell lysis device D6 configured as described above has the following additional effect compared to the effects described for the fifth pyrotechnic cell lysis device D5. That is, according to the sixth pyrotechnic cell lysis device D6 of this embodiment, the combustion gas generated by the ignition and combustion of the pyrotechnic charge 5 when the initiator 28 is activated is supplied to the pressurized space 155 via the pressure channel 30, causing the pressurized space 155 to change from the low-pressure space 7 to the high-pressure space 8. At this time, the pressure of the combustion gas flowing downward from the connection part P1 into the sample container housing 156 presses the sample pouch 24 with the sample cage 25 attached toward the bottom of the pressure vessel 150, allowing the pressurized sample pouch 24 to be stably held in the sample container housing 156. Furthermore, since the upper end of the sample pouch 24 is located below the connection part P1 in the pressurized space part 155, the sample pouch 24 is prevented from being directly exposed to the combustion gas flow from the pyrotechnic charge 5.

[0069] When the rupture disk 23 in the rupture disk assembly 22 bursts, the pressure chamber 15 communicates with the external space, and the pressure in the pressure chamber 15 is released (vented) to the outside through the pressure release vent channels 61 and 22A. Since most of the gas released from the pressure chamber 15 to the outside through the pressure release vent channels 61 and 22A does not pass through the sample container receptacle 156, the sample pouch 24 can be stably placed on the bottom of the pressure vessel 150 when the pressure in the pressure chamber 15 is released. This prevents, for example, the sample pouch 24 from moving upward from the sample container receptacle 156 or the pressure release vent channel 61 from being blocked by the sample pouch 24, thereby preventing the pressure chamber 15 from being quickly and smoothly depressurized. While the present embodiment describes an example in which a fluid sample is accommodated in the flexible sample pouch 24, other containers filled with a fluid sample may also be accommodated in the sample container receptacle 156.

[0070] Seventh Embodiment 13 to 15 show a pyrotechnic cell disruption device including a sample tip according to the seventh embodiment. As shown in FIG. 13, a clamp 31 holds a fluid sample 3 in a first recess 33 formed in a substrate 34. The chip 32 holds the fluid sample 3 and has a second recess 35 formed in the substrate 34, which defines an expansion chamber 36. The pyrotechnic charge chamber housing 16 can be formed integrally with one of the clamps 31 or can be provided separately. The pressure release channel 18 can be centrally located above the fluid sample 3 when the chip is in the clamped position between the clamps 31. The pressure chamber 15 is sealed from the environment by the clamps and / or the pyrotechnic charge chamber housing 16, or a combination thereof. A precision orifice 4 connects the pressure chamber 15 and the expansion chamber 36 to each other. Such a precision orifice 4 can be created, for example, by a groove in the substrate 34 that is closed by a clamp to form a closed channel between the pressure chamber 15 and the expansion chamber 36. A thin film can be adhered to the chip 34 to seal the precision orifice 4, the expansion chamber 36, and the pressure chamber 15. The film may be provided with small holes or weakened portions for pressurization by a pyrotechnic charge. Other types of orifices may also be employed, such as orifices extending completely through the substrate 34.

[0071] Figure 14 shows the state before ignition, while Figure 15 shows the state after ignition and movement of fluid sample 3 from pressure chamber 15 into expansion chamber 36. This shows that fluid sample 3, shown in black in Figure 14, is dispersed over expansion chamber 36, which collects material from within the disrupted cells, in Figure 15.

[0072] As described above, the seventh pyrotechnic cell disruption device D7 according to the seventh embodiment (see FIGS. 13 to 15 ) includes a substrate 34, a pressure chamber 15 formed by a first recess 33 formed in the substrate 34, and a tip 32 (pyrotechnic cell disruption tip) including an expansion chamber 36 formed by a second recess 35 formed in the substrate 34. In this embodiment, similar elements to those in the previous embodiments are designated by similar reference numerals. In the example shown in FIG. 13 , the first recess 33 (pressure chamber 15) and the second recess 35 (expansion chamber 36) are formed so as to open on the upper surface of the substrate 34, and the first recess 33 (pressure chamber 15) and the second recess 35 (expansion chamber 36) are connected by a precision orifice 4 (first channel). The precision orifice 4 may be formed as an open groove that opens on the upper surface of the substrate 34. In the example shown in FIG. 13 , the upper surface of the substrate 34 is covered with a thin top layer film 46. For example, the top layer film 46 may be adhered to the upper surface of the substrate 34, and the top layer film 46 may seal the first recess 33, the second recess 35, and the opening grooves for forming the precision orifice 4, thereby sealing the pressure chamber 15, the expansion chamber 36, and the precision orifice 4 from the outside. Here, the top layer film 46 and the bottom layer film 47 described below may be made of various polymer films, and may be formed, for example, by compounding or laminating polypropylene (PP), polyethylene (PE), or other thermoplastic resins, and films that have been given heat shrinkability, hydrophilicity, or hydrophobicity as necessary can be used.

[0073] The seventh pyrotechnic cell disruption device D7 is further clamped by a pair of clamps 31 respectively disposed on the upper (top) and lower (bottom) portions of the tip 32 (pyrotechnic cell disruption tip). The pair of clamps 31 are, for example, high-strength clamps having rigidity, and can be detachably attached to the tip 32. The seventh pyrotechnic cell disruption device D7 further includes a pyrotechnic charge chamber housing 16 forming a pyrotechnic charge chamber 11, a pyrotechnic charge 5 housed in the pyrotechnic charge chamber 11, etc., and as shown in FIG. 13, the pyrotechnic charge chamber housing 16 is disposed above the first recess 33 (pressure chamber 15) in the tip 32 (substrate 34). The pyrotechnic charge chamber housing 16 may be formed integrally with the clamp 31 that clamps the upper side of the tip 32, or may be provided separately.

[0074] As in the second to fourth embodiments, a pressure release chamber is provided at the bottom of the pyrotechnic charge chamber housing 16. The pyrotechnic charge chamber housing 16 has a pressure release channel 18 and a break 17 formed therein. The pressure release channel 18 is formed as a recess that opens to the outside of the pyrotechnic charge chamber housing 16, and the pressure release channel 18 is positioned at the center of the first recess 33 (pressure chamber 15) in the chip 32 (substrate 34). The pyrotechnic charge chamber housing 16 is also positioned so that the pressure release channel 18 is in close contact with the top film 46.

[0075] FIG. 14 shows a cross-sectional view and a plan view of the seventh pyrotechnic cytolysis device D7 in a first mode before pressurization (before ignition of the pyrotechnic charge 5) with the clamp removed. FIG. 15 shows a cross-sectional view and a plan view of the seventh pyrotechnic cytolysis device D7 in a second mode after pressurization (after ignition of the pyrotechnic charge 5) with the clamp removed. Each of FIGS. 14 and 15 shows a cross-sectional view at the top and a plan view at the bottom. The bottom plan views of FIGS. 14 and 15 show the top surface of the substrate 34 through the top film 46. Before activation of the seventh pyrotechnic cytolysis device D7, a fluid sample 3 is contained in the pressure chamber 15 of the chip 32. In FIG. 14, the fluid sample 3 contained in the pressure chamber 15 is filled in black. When the seventh pyrotechnic cytolysis device D7 is activated, the pyrotechnic charge 5 is ignited, and combustion gas is generated as the pyrotechnic charge 5 burns. This increases the pressure in the pyrotechnic charge chamber 11, causing the rupture portion 17 of the pyrotechnic charge chamber housing 16 to burst (crack), thereby connecting the pressure release channel 18 of the pyrotechnic charge chamber housing 16 to the pyrotechnic charge chamber 11. As a result, the pressure in the pyrotechnic charge chamber 11 ruptures the portion of the top layer film 46 facing the pressure release channel 18, allowing combustion gas to flow into the pressure chamber 15 in the tip 32. This rapidly pressurizes the pressure chamber 15 in the tip 32, rupturing the cells contained in the fluid sample 3 stored in the pressure chamber 15. Note that a small hole or a weakened portion may be previously provided in the top layer film 46 at the portion facing the pressure release channel 18. This makes it easier to introduce the combustion gas of the pyrotechnic charge 5 into the pressure chamber 15 when the seventh pyrotechnic cell rupture device D7 is activated.

[0076] Furthermore, the pressure chamber 15 in the chip 32 is pressurized by the combustion gas of the pyrotechnic charge 5, forcing the fluid sample 3 into the precision orifice 4. The fluid sample 3 then moves through the precision orifice 4 to the expansion chamber 36, where it is retained (collected). As the fluid sample 3 passes through the precision orifice 4, a large shear stress acts on the cells contained in the fluid sample 3, promoting cell disruption. The expansion chamber 36 has a larger volume than the pressure chamber 15, and the fluid sample 3 is decompressed when it flows from the pressure chamber 15 into the expansion chamber 36 through the precision orifice 4. This causes the fluid sample 3 to rapidly expand when it flows into the expansion chamber 36 through the precision orifice 4, further promoting cell disruption. The fluid sample 3 that has undergone cell disruption in this manner is collected in the expansion chamber 36. The chip 32 in this embodiment may also be provided with a vent 48 that connects the expansion chamber 36 to the outside. The vent 48 can be formed, for example, by a groove opening on the upper surface of the substrate 34 and an opening in the top layer film 46 formed at a position overlapping the groove. The vent 48 allows ventilation to the outside through the opening in the top layer film 46 and the gap between the top layer film 46 and the clamp 31, and atmospheric pressure can be introduced into the expansion chamber 36. By placing the expansion chamber 36 under atmospheric pressure in this way, the fluid sample 3 transferred from the pressure chamber 15 to the expansion chamber 36 can be depressurized and expanded more rapidly, thereby more efficiently lysing cells contained in the fluid sample 3.

[0077] In this embodiment, a reagent may be added to the fluid sample 3 collected in the expansion chamber 36, and a chemical reaction may be carried out in the expansion chamber 36. The reagent added to the fluid sample 3 is a reagent for causing a chemical reaction in the cells after disruption contained in the fluid sample 3, and may be, for example, a reagent for achieving a reaction related to polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), or any other isothermal amplification. 2 may further include a detection chamber formed in the form of a third recess in the substrate 34, and the expansion chamber 36 and the detection chamber may be connected by a channel. After a reagent is added to the fluid sample 3 in the expansion chamber 36 to carry out a chemical reaction, the fluid sample 3 after the reaction process may be released from the expansion chamber 36 to the detection chamber through the channel. Also, multiple types of reagents may be added to the fluid sample 3 in the expansion chamber 36.

[0078] Eighth Embodiment Figures 16-18 show an eighth embodiment, very similar to Figures 13-15, that allows downstream processing of disrupted cells on a chip. Similar elements are designated by the same reference numerals used in Figures 13-15. This eighth embodiment can be applied, for example, as a point-of-care (POC) device. The first recess 33 in the substrate has a more elongated shape in this embodiment, as shown in Figure 17, while the expansion chamber 36 has an elliptical shape. The expansion chamber is followed by a downstream reaction chamber 37 containing reagents 38, for example, for performing a polymerase chain reaction (PCR). An optional vent 39 may also be provided, which can be configured to only partially release pressure so that residual pressure is maintained. This vent 39 facilitates filling of the chamber 37. It may be a hydrophobic vent, so that once the chamber is filled, the vent no longer acts as a vent in a hydrostatic lock (hydrolock) state, maintaining residual pressure behind the liquid. Finally, there may be a valve 40 that is opened after the PCR reaction is completed, allowing the processed sample to move into a detection chamber 41 .

[0079] As described above, the eighth pyrotechnic cell lysis device D8 according to the eighth embodiment (see FIGS. 16 to 18) is a modified version of the seventh pyrotechnic cell lysis device D7 described in FIGS. 13 to 15. FIG. 16 shows a schematic cross-sectional view of the eighth pyrotechnic cell lysis device D8, FIG. 17 shows a cross-sectional view and a plan view of the device in a first state before pressurization (before ignition of the pyrotechnic charge 5) with the clamp removed, and FIG. 18 shows a cross-sectional view and a plan view of the device in a second state after pressurization (after ignition of the pyrotechnic charge 5) with the clamp removed. Each of FIGS. 17 and 18 shows a cross-sectional view in the upper part and a plan view in the lower part. The plan views in the lower parts of FIGS. 17 and 18 show the top surface of the substrate 34 through the top layer film 46. In this embodiment, like elements are designated by like reference numerals as compared to the previous embodiments.

[0080] The chip 32 of the eighth pyrotechnic cell disruption device D8 includes, in addition to the first recess 33 (pressure chamber 15), the second recess 35 (expansion chamber 36), and the precision orifice 4, a reaction chamber 37 formed as a third recess on the upper surface of the substrate 34, and a detection chamber 41 formed as a fourth recess on the upper surface of the substrate 34. The reaction chamber 37 is connected to the expansion chamber 36 through a second channel 61. The detection chamber 41 is connected to the reaction chamber 37 through a third channel 62. As shown in FIG. 17 , the downstream reaction chamber 37 is located downstream of the expansion chamber 36, and the detection chamber 41 is located further downstream of the reaction chamber 37. The second channel 61 and the third channel 62 may be formed, for example, by grooves opening into the upper surface of the substrate 34. A top layer film 46 is adhered to the top surface of the substrate 34 of the chip 32, sealing the pressure chamber 15, expansion chamber 36, precision orifice 4, reaction chamber 37, detection chamber 41, second channel 61, and third channel 62. In the example shown in FIG. 17, the top surface of the chip 32 is provided with a vent 39 communicating with the third channel 62 and a valve 40 located downstream of the vent 39 in the third channel 62. The vent 39 can be formed, for example, by a recess opening into the top surface of the substrate 34 and an opening in the top layer film 46 formed at a position overlapping the recess. The vent 39 allows ventilation between the expansion chamber 36 and the outside through the opening in the top layer film 46 and the gap between the top layer film 46 and the clamp 31, and the expansion chamber 36, reaction chamber 37, detection chamber 41, second channel 61, and third channel 62 are vented. 7, etc., can be exposed to the atmosphere (outside). When valve 40 is open, atmospheric pressure is also introduced into detection chamber 41 through vent hole 39.

[0081] In the eighth pyrotechnic cell lysis device D8 configured as described above, the fluid sample 3, in which cells have been lysed by ignition and combustion of the pyrotechnic charge 5, is collected in the expansion chamber 36 and then sequentially transferred to the subsequent reaction chamber 37 and detection chamber 41. In this embodiment, the chip 32 is provided with a vent 39, which facilitates transfer of the fluid sample 3 from the pressure chamber 15 to the expansion chamber 36 and reaction chamber 37. Furthermore, the reaction chamber 37 in this embodiment contains a reagent 38 for causing a chemical reaction with the lysed cells contained in the fluid sample 3. For example, the reagent 38 is a reagent for performing a polymerase chain reaction (PCR). After the PCR reaction is completed in the reaction chamber 37, the valve 40 is opened, allowing the fluid sample 3 containing the cells reacted with the reagent 38 to be transferred to the detection chamber 41 via the third channel 62. In this embodiment, the reagent 38 contained in the downstream reaction chamber 37 is not limited to a reagent for achieving a polymerase chain reaction (PCR), but may be, for example, a reagent for performing a reaction related to loop-mediated isothermal amplification (LAMP) or any other isothermal amplification. Furthermore, the reaction chamber 37 may contain multiple types of reagents.

[0082] Ninth Embodiment FIG. 19 shows an exploded view of a ninth embodiment of a pyrotechnic cytolysis device based on the concept shown in FIGS. 13-15. FIG. 19 shows more structural details. The clamp 31 is formed as an upper housing 42, which houses the initiator 28 at one end and has a pressure release portion 21 at the other end, which includes a threaded barb connector 43 and, for example, a 3 / 16-inch diameter ID tube 44. The tip assembly 45 includes the tip 32, a top film 46, and a bottom film 47. The top film 46 and the bottom film 47 sandwich and seal the tip 32 containing the sample disposed in the first recess 33. Additionally, the tip assembly may be provided with a vent 48.

[0083] When the chip assembly 45 is installed, it is sandwiched between the silicone gasket 49, the upper housing 42, the lower housing 50, and the gasket 49. The chip assembly 45 is held together by positioning pins in holes in diagonally opposite corners of the upper and lower housings 42 and 50.

[0084] FIG. 20 shows a schematic cross-sectional view of the ninth embodiment, showing the pressure relief channel 18, the vent channel 51 for relieving pressure after combustion of the pyrotechnic charge.

[0085] As shown in FIGS. 21 and 22, the sample is forced down through downward channel 52, travels along precision orifice 4 formed in the manner of a grooved channel in chip 32 sealed by gasket 49, and then travels up channel 54 into expansion chamber 36. Alternatively, precision orifices not formed in chip channels can be used. Such precision orifices can be formed from other suitable materials, such as sapphire, ruby, glass, or polymers, and can be glued or pressed into a recess in the chip. FIG. 22 shows a second embodiment after combustion of the pyrotechnic charge, with fluid sample 3 located at the bottom of expansion chamber 36. Here, the sample currently being processed contains disrupted cells with their contents released for further processing with reagents and eventual detection.

[0086] As described above, the ninth pyrotechnic cell disruption device D9 (see FIGS. 19 to 22) according to the ninth embodiment is a more specific structure of the seventh pyrotechnic cell disruption device D7. In the present embodiment, like elements as compared with the above-described embodiments are designated by like reference numerals. As shown in Figures 19 and 20, the ninth pyrotechnic cytolysis device D9 includes a tip assembly 45 including a tip 32 comprising a substrate, a top film 46 covering the upper surface of the tip 32, and a bottom film 47 covering the lower surface of the tip 32. Figures 21 and 22 are schematic cross-sectional views of the tip assembly 45 in the ninth pyrotechnic cytolysis device D9, with Figure 21 showing a first state before the pyrotechnic charge is burned and Figure 22 showing a second state after the pyrotechnic charge is burned.

[0087] As in the seventh embodiment, the chip 32 according to the ninth embodiment has a pressure chamber 15 formed by a first recess 33 on the upper surface of the substrate, and an expansion chamber 36 formed by a second recess 35. Also, as shown in Fig. 19, the chip 32 is provided with an air vent 48 communicating with the expansion chamber 36.

[0088] 21 and 22, the pressure chamber 15 and the expansion chamber 36 are connected via a first channel 63. The first channel 63 includes a precision orifice 4, a downward channel 52, a lateral channel 53, an upward channel 54, and the like. In the example shown in FIGS. 21 and 22, the upper end of the precision orifice 4 is connected to the bottom of the first recess 33 so that the pressure chamber 15 and the precision orifice 4 are in communication with each other, and the downward channel 52 connected to the lower end of the precision orifice 4 extends to the lower surface of the chip 32. Meanwhile, the upward channel 54 of the first channel 63 has its upper end connected to the bottom of the second recess 35 that forms the expansion chamber 36, and extends downward from the bottom of the second recess 35 so that its lower end reaches the lower surface of the chip 32. The downward channel 52 and the upward channel 54 of the first channel 63 may be formed, for example, by holes extending from the lower surface of the substrate 34 toward the upper surface. One end of the lateral channel 53 is connected to the lower end of the downward channel 52, and the other end is connected to the lower end of the upward channel 54. The lateral channel 53 may be formed, for example, by a groove channel that opens to the lower surface of the chip 32.

[0089] In this embodiment, the first recess 33 (pressure chamber 15) and the second recess 35 (expansion chamber 36) that open to the upper surface of the chip 32 can be sealed from the outside by covering the upper surface of the chip 32 with the top layer film 46. In addition, the first channel 63 can be sealed from the outside by covering the lower surface of the chip 32 with the bottom layer film 47.

[0090] As shown in FIGS. 19 and 20, the ninth pyrotechnic cell disruption device D9 has a pair of rigid clamps 31 formed as a housing. That is, the pair of clamps 31 includes an upper housing 42 as an upper clamp and a lower housing 50 as a lower clamp. In the example shown in FIG. 19, the upper housing 42 (upper clamp) and the lower housing 50 (lower clamp) are formed as housings having a substantially rectangular parallelepiped shape. However, the shapes of the upper housing 42 and the lower housing 50 are not particularly limited. Here, the upper surface side of the lower housing 50 (lower clamp) has a tip recess 50A capable of accommodating a tip assembly 45 including a tip 32. In addition, the lower surface of the upper housing 42 (upper clamp) forms a substantially flat clamp surface 42A facing the loading area of ​​the tip 32. In addition, holes for fitting connecting pins 64 are provided at diagonally opposite corners of the upper housing 42 and the lower housing 50, respectively.

[0091] As shown in Figures 19 and 20, the initiator 28 is attached to one side of the upper housing 42 (upper clamp), and the threaded barb connector 43 of the pressure release part 21 is attached to the opposite side. The initiator 28 is connected to the pyrotechnic charge chamber housing 16 that forms the pyrotechnic charge chamber 11, the pyrotechnic charge 5 housed in the pyrotechnic charge chamber 11, and the pyrotechnic charge 5 housed in the pyrotechnic charge chamber 11. , wire 29, etc. The initiator 28 is fixed to the upper housing 42 so that the pyrotechnic charge chamber housing 16 is accommodated inside the upper housing 42 and the wire 29 is exposed to the outside. One end of a pressure release channel 18 is connected to the pyrotechnic charge chamber housing 16 of the initiator 28. The pressure release channel 18 is formed, for example, by a metal conduit, and the other end is joined to the clamping surface 42A of the upper housing 42 from the inside. The opening on the other end side of the pressure release channel 18 communicates with the outside of the upper housing 42 through a gas outlet 42B, which is an opening formed in the clamping surface 42A. The pressure release channel 18 is, for example, previously connected to the pyrotechnic charge chamber 11 of the initiator 28, or is cleaved by the combustion energy of the pyrotechnic charge 5, so that combustion gas of the pyrotechnic charge 5 is released from the gas outlet 42B of the clamping surface 42A of the upper housing 42 when the initiator 28 is activated. The open end of the pressure release channel 18 and the gas outlet 42B of the clamping surface 42A may be positioned at the center of the first recess 33 (pressure chamber 15) in the chip 32.

[0092] 20 , an ID tube 44 and a ventilation channel 51 are connected to the threaded barb connector 43 of the pressure release section 21. The ID tube 44 is a hollow tube and is disposed outside the upper housing 42. The ventilation channel 51 is formed, for example, by a metal conduit. One end of the ventilation channel 51 is connected to the threaded barb connector 43, and the other end is joined to the clamp surface 42A of the upper housing 42 from the inside. The opening on the other end of the ventilation channel 51 communicates with the outside of the upper housing 42 through a vent port 42C, which is an opening formed in the clamp surface 42A. The interior of the threaded barb connector 43 is hollow, and ventilation paths are formed inside the ventilation channel 51, the threaded barb connector 43, and the ID tube 44.

[0093] When the ninth pyrotechnic cell lysis device D9 configured as described above is assembled, the tip assembly 45 is housed in the tip recess 50A of the lower housing 50. At this time, as shown in Fig. 19, the tip assembly 45 is housed in the tip recess 50A while being sandwiched between a pair of gaskets 49. After the tip assembly 45 sandwiched between the pair of gaskets 49 is housed in the tip recess 50A in this manner, the upper housing 42 and the lower housing 50 are fixed together using the connecting pin 64, thereby assembling the ninth pyrotechnic cell lysis device D9. However, the connecting structure of the upper housing 42 and the lower housing 50 is not particularly limited.

[0094] When the initiator 28 of the ninth pyrotechnic cell lysis device D9 is activated, the pyrotechnic charge 5 is ignited and burned, and the resulting combustion gas is released from a gas outlet 42B formed in a clamping surface 42A of the upper housing 42 through the pyrotechnic charge chamber 11 and the pressure release channel 18. Here, the top film 46 of the chip assembly 45 and a gasket 49 arranged on the upper surface side of the chip assembly 45 are formed with openings for venting the combustion gas released from the gas outlet 42B to the pressure chamber 15 of the chip 32. As a result, the combustion gas from the pressure release channel 18 flows into the pressure chamber 15, causing the pressure chamber 15 to be suddenly pressurized, and cells contained in the fluid sample 3 held in the pressure chamber 15 to be lysed.

[0095] The fluid sample 3 pressurized in the pressure chamber 15 is transferred to the expansion chamber 36 through the precision orifice 4 of the first channel 63, the downward channel 52, the lateral channel 53, and the upward channel 54 in that order. When the fluid sample 3 flows through the precision orifice 4, the cells contained in the fluid sample 3 are subjected to a large shear stress, which promotes cell rupture. An opening is formed in the top film 46 of the chip assembly 45 and the gasket 49 arranged on the upper surface side of the chip assembly 45 at a position overlapping the vent 48, and a vent 42C formed in the clamping surface 42A of the upper housing 42 also overlaps the vent 48. The expansion chamber 36 of the chip assembly 45 is positioned so that the expansion chamber 36 overlaps the pressure chamber 15. As a result, the expansion chamber 36 of the chip assembly 45 is vented through the vent hole 48 and the pressure release section 21 (vent channel 51, threaded barb connector 43, and ID tube 44), and atmospheric pressure is introduced. Therefore, the fluid sample 3 transferred from the pressure chamber 15 to the expansion chamber 36 through the first channel 63 is depressurized in the expansion chamber 36. As a result, the cells contained in the fluid sample 3 rapidly expand, further accelerating cell lysis. In this manner, the fluid sample 3 after cell lysis is collected in the expansion chamber 36 of the chip assembly 45 (chip 32).

[0096] Of course, the upper housing 42 and the lower housing 50 in this embodiment are detachable. After operation (use) of the ninth pyrotechnic cell lysis device D9, the upper housing 42 and the lower housing 50 are separated by, for example, removing the connecting pin 64, thereby exposing the tip assembly 45 (tip 32) housed in the tip recess 50A of the lower housing 50. The fluid sample 3 after cell lysis processing can then be recovered from the expansion chamber 36 by, for example, peeling off the top film 46 covering the upper surface of the tip 32. Also in this embodiment, as described in the seventh embodiment, reagents may be added to the fluid sample 3 collected in the expansion chamber 36, and various reaction processes may be performed.

[0097] Tenth Embodiment Figures 23-26 show an embodiment very similar to Figures 19-22. In this embodiment, the tip is replaced by a laser-cut plastic or glass tip with a precision orifice 4 machined into the tip shape. The tip is therefore essentially an integral part of the lower housing 50. This embodiment does not include designated venting, but venting is achieved through the gap between the upper housing 42 and the lower housing 50.

[0098] FIG. 23 shows a cross-sectional view of a tenth pyrotechnic cell lysis device D10 according to a tenth embodiment. The tenth pyrotechnic cell lysis device D10 is a modified version of the ninth pyrotechnic cell lysis device D9. In this embodiment, like elements are designated by like reference numerals as compared to the previous embodiments. In the tenth pyrotechnic cell lysis device D10, the tip 32 is formed into a tip shape by processing the upper surface of the lower housing 50 using laser cutting technology, and is realized in a manner in which the tip 32 is integrated into the lower housing 50. FIG. 24 is a plan view of the tip formation region on the upper surface of the lower housing 50 where the tip 32 is formed. The tip formation region of the lower housing 50 can be formed, for example, from a polymeric material (such as plastic) or glass. 25 and 26 are schematic cross-sectional views of the tip 32 (the tip forming area in the lower housing 50) in the tenth pyrotechnic cell disruption device D10, with FIG. 25 showing a first state before the pyrotechnic attachment is burned, and FIG. 26 showing a second state after the pyrotechnic attachment is burned.

[0099] The chip 32, which is formed integrally with the chip formation region of the lower housing 50, is provided with a pressure chamber 15 formed by a first recess 33 and capable of accommodating a fluid sample 3 before cell disruption processing, and an expansion chamber 36 formed by a second recess 35, and the pressure chamber 15 and the expansion chamber 36 are connected via a first channel 63 including a precision orifice 4. As in the seventh embodiment, the expansion chamber 36 in the chip 32 has a volume sufficiently larger than that of the pressure chamber 15, and when the fluid sample 3 enters the expansion chamber 36 from the pressure chamber 15, the fluid sample 3 is subjected to reduced pressure, causing the cells contained in the fluid sample 3 to rapidly expand.

[0100] The upper housing 42 of the tenth pyrotechnic cell disruption device D10 is provided with the initiator 28 and the pressure release channel 18 as in the ninth embodiment, but is not provided with the pressure release section 21. Also, as in the ninth embodiment, the upper housing 42 and the lower housing The housings 50 each have holes for receiving connecting pins, which can be used to connect the upper and lower housings 42 and 50 together and can also be separated from each other after the device is operated. The upper surface of the chip 32 integrated into the lower housing 50 may be covered with a top film 46. In this case, the top film 46 may have small holes or weakened portions formed therein to allow combustion gas released from the gas outlet 42B of the clamping surface 42A of the upper housing 42 to vent to the pressure chamber 15. When assembling the upper housing 42 to the lower housing 50, a gasket 49 may be interposed between the upper surface of the chip 32 integrated into the lower housing 50 and the clamping surface 42A of the upper housing 42. In this case, an opening may be formed in the gasket 49 to supply combustion gas released from the gas outlet 42B of the clamping surface 42A to the pressure chamber 15.

[0101] The operation of the tenth pyrotechnic cell lysis device D10 configured as described above is basically the same as that of the ninth pyrotechnic cell lysis device D9 according to the ninth embodiment. That is, when the initiator 28 is activated, the pyrotechnic charge 5 is ignited and burned, and the resulting combustion gas is supplied to the pressure chamber 15 of the chip 32 through the pressure release channel 18. The fluid sample 3 contained in the pressure chamber 15 is suddenly pressurized, thereby lysing the cells contained in the fluid sample 3. The pressurized fluid sample 3 in the pressure chamber 15 is then transferred to the expansion chamber 36 through the first channel 63, which includes the precision orifice 4. As the fluid sample 3 flows through the precision orifice 4, the cells contained in the fluid sample 3 are subjected to a large shear stress, thereby facilitating cell lysis. When the fluid sample 3 flows from the first channel 63 into the expansion chamber 36, which has a large volume, the cells contained in the fluid sample 3 are suddenly expanded by the reduced pressure, further facilitating cell lysis. In this way, the fluid sample 3 containing the cells after the lysing process is collected in the expansion chamber 36. As in the ninth embodiment, the tip 32 may be provided with a vent hole 48 communicating with the expansion chamber 36, and atmospheric pressure may be introduced into the expansion chamber 36 through the vent hole 48.

[0102] Eleventh Embodiment Figures 27-29 show schematic cross-sectional views of a pyrotechnic cytolysis device including a sample tip according to an eleventh embodiment, with Figure 28 showing the tip in a first state before pressurization and Figure 29 showing the tip in a second state after pressurization. This embodiment is similar to the embodiment shown in Figures 13-15, but does not include a precision orifice 4. Thus, this embodiment is based on the same hydrostatic impact pressure wave concept as the embodiment described in Figures 5 and 6. Elements similar to those described in Figures 13-15 are designated by the same reference numerals.

[0103] This eleventh embodiment according to Figures 27-29 essentially replicates the concept of the third embodiment according to Figures 5 and 6 in chip form. Although only one recess 33 for holding a fluid sample 3 is shown, chips with multiple recesses 33 for multiple different samples can be mounted on a single chip. Multiple pyrotechnic charges 5 can be applied simultaneously on multiple sample chips, or a single pyrotechnic charge housing can be moved between multiple samples and reloaded while moving between samples.

[0104] The eleventh pyrotechnic cell disruption device D11 shown in Figures 27 to 29 comprises a substrate 34, a tip 32 (pyrotechnic cell disruption tip) having a pressure chamber 15 formed by a first recess 33 formed in the substrate 34. The eleventh pyrotechnic cell disruption device D11 further comprises a pair of clamps 31 arranged on the upper (top) and lower (bottom) sides of the tip 32 (pyrotechnic cell disruption tip) respectively to clamp the tip 32, a pyrotechnic charge chamber housing 16 forming a pyrotechnic charge chamber 11, and a pyrotechnic charge 5 housed in the pyrotechnic charge chamber 11. The tip 32 also has a thin top layer film 46 covering the upper surface of the substrate 34, and the top layer film 46 protects the first recess 33 (pressure chamber 15) of the substrate 34 from the outside. It can be sealed. Note that Figure 28 is a schematic cross-sectional view and a plan view of the eleventh pyrotechnic cell lysis device D11 before activation (before pressurization). In Figure 28, the clamp 31 is omitted, and the cross-sectional view is shown in the upper part and the plan view is shown in the lower part. Also, Figure 29 is a schematic cross-sectional view and a plan view of the eleventh pyrotechnic cell lysis device D11 after activation (after pressurization). In Figure 29, the clamp 31 is omitted, and the cross-sectional view is shown in the upper part and the plan view is shown in the lower part. Note that the plan views of Figures 28 and 29 show the top surface of the substrate 34 through the top layer film 46.

[0105] When the eleventh pyrotechnic cell lysis device D11 configured as described above is activated, the pyrotechnic charge 5 is ignited, and the pyrotechnic charge 5 burns, generating combustion gas. This increases the pressure in the pyrotechnic charge chamber 11, causing the rupture portion 17 of the pyrotechnic charge chamber housing 16 to rupture, and the pressure release channel 18 to communicate with the pyrotechnic charge chamber 11. As a result, the pressure in the pyrotechnic charge chamber 11 is released, causing the portion of the top layer film 46 facing the pressure release channel 18 to rupture, allowing combustion gas to flow into the pressure chamber 15 in the tip 32. Alternatively, a small hole may be formed in advance in the portion of the top layer film 46 facing the pressure release channel 18, and combustion gas may be supplied to the pressure chamber 15 through the small hole. The pressure chamber 15, to which the combustion gas from the pyrotechnic charge 5 is supplied, is rapidly pressurized, resulting in rupture of the cells contained in the fluid sample 3 contained in the pressure chamber 15. The eleventh pyrotechnic cell disruption device D11 may also include the precision orifice 4 and pressure release section 21 described in the above-described embodiment.

[0106] The following additional notes are provided regarding the above embodiment. (Appendix 1) a pyrotechnic charge configured to be ignited and to burn upon ignition; a pressure chamber configured to contain a fluid sample containing cells and to be pressurized upon ignition and combustion of the pyrotechnic charge; A pyrotechnic cell disruption device comprising: (Appendix 2) a pressure chamber outlet connecting the pressure chamber to an external space; the pressure chamber outlet has an orifice that applies shear stress to the fluid sample when the fluid sample flows through the orifice; 10. A pyrotechnic cell disruption device as described in Appendix 1. (Appendix 3) 3. The pyrotechnic cell disruption device of claim 2, wherein the orifice is formed in a separate part that is bonded or press-fit into the pressure chamber outlet. (Appendix 4) 4. A pyrotechnic cell disruption device as described in any one of appendixes 1 to 3, wherein the pyrotechnic charge is contained in a separate pyrotechnic charge chamber separated from the pressure chamber. (Appendix 5) 5. The pyrotechnic cell disruption device of claim 4, wherein the pyrotechnic charge chamber is formed in a pyrotechnic charge chamber housing having a rupture portion that ruptures upon ignition and combustion of the pyrotechnic charge. (Appendix 6) A pyrotechnic cell disruption device as described in Appendix 5, wherein the rupture portion is formed by a weak portion in which the portion of the pyrotechnic charge chamber housing facing the pressure chamber is weaker than other portions. (Appendix 7) A pyrotechnic cell disruption device as described in Appendix 6, wherein the fragile portion is formed by thinning the thickness of the pyrotechnic charge chamber housing compared to other portions. (Appendix 8) The pyrotechnic charge chamber opens directly to the pressure chamber upon rupture of the rupture portion. A pyrotechnic cell disruption device according to any one of appendices 5 to 7. (Appendix 9) a first cylinder chamber containing the pyrotechnic charge; a first piston at least partially contained within the first cylinder chamber and movable relative to the first cylinder chamber under pressure generated by ignition and combustion of the pyrotechnic charge; a second piston disposed within the pressure chamber and connected to the first piston; Further provided with Upon ignition and combustion of the pyrotechnic charge, the second piston moves in conjunction with the first piston to pressurize the fluid sample contained in the pressure chamber. 4. The pyrotechnic cell disruption device of any one of appendices 1 to 3. (Appendix 10) 10. The pyrotechnic cell disruption device of claim 9, wherein the first cylinder chamber is formed within a first cylinder chamber housing that is at least partially contained within the pressure chamber. (Appendix 11) a diaphragm dividing the pressure chamber into a first interior space into which combustion gases of the pyrotechnic charge are introduced upon ignition and combustion of the pyrotechnic charge, and a second interior space into which the fluid sample is accommodated; When the pyrotechnic charge is ignited and burned, the combustion gas is introduced into the first internal space, causing the diaphragm to deform, and the volume of the second internal space to decrease, thereby pressurizing the fluid sample contained in the second internal space. 9. The pyrotechnic cell disruption device of any one of appendices 1 to 8. (Appendix 12) a pressure chamber outlet connecting the second interior space to an exterior space; the pressure chamber outlet has an orifice that applies shear stress to the fluid sample when the fluid sample flows through the orifice; 12. The pyrotechnic cell disruption device of claim 11. (Appendix 13) a pressure release section that releases pressure from the pressure chamber after the pressure chamber is pressurized by ignition and combustion of the pyrotechnic charge; 13. The pyrotechnic cell disruption device of any one of appendices 1 to 12. (Appendix 14) 14. The pyrotechnic cell disruption device of claim 13, wherein the pressure release section has a valve body. (Appendix 15) 15. The pyrotechnic cell disruption device of claim 14, wherein the valve body is a pressure relief valve that opens under a predetermined pressure. (Appendix 16) 14. The pyrotechnic cell disruption device of claim 13, wherein the pressure release section has a rupture disc that ruptures under a predetermined pressure. (Appendix 17) 17. A pyrotechnic cell lysis device according to any one of claims 13 to 16, wherein the pressure chamber accommodates a sample container into which the fluid sample is filled. (Appendix 18) 18. The pyrotechnic cell disruption device of claim 17, wherein the sample container is a flexible pouch. (Appendix 19) 19. The pyrotechnic cell disruption device of claim 17 or 18, wherein the pressure chamber includes a pressure channel in which the pyrotechnic charge is disposed, and a sample container receiving portion coaxially connected to the pressure channel and for receiving the sample container. (Appendix 20) A pyrotechnic cell disruption device as described in Appendix 19, wherein a sample container mounting section for mounting the sample container is formed at the connection section of the sample container storage section with the pressure channel. (Appendix 21) 21. A pyrotechnic cell lysis device as described in Appendix 20, wherein the cross-sectional area of ​​the sample container accommodating portion is larger than the cross-sectional area of ​​the pressure channel, and the sample container mounting portion is formed by a step formed between the sample container accommodating portion and the pressure channel. (Appendix 22) a pressure vessel having the pressure chamber formed therein and the sample vessel accommodating portion opening on the top surface; a cap in which the pressure release portion is installed and which can be attached to the pressure vessel so as to cover the upper surface of the pressure vessel; Equipped with the pressure release section has a pressure release vent passage that communicates the sample container receptacle with an external space after the pressure chamber is pressurized by ignition and combustion of the pyrotechnic charge; 22. The pyrotechnic cell disruption device of any one of appendices 19 to 21. (Appendix 23) 23. The pyrotechnic cell lysis device of claim 22, wherein the pressure channel, the sample vessel receptacle, and the pressure release vent are coaxially arranged when the cap is attached to the pressure vessel. (Appendix 24) the pressure release vent path is blocked by a rupture disc or a valve body, and when the rupture disc ruptures or the valve body is opened, the sample container accommodation portion communicates with an external space. 24. The pyrotechnic cell disruption device of claim 22 or 23. (Appendix 25) the pressure chamber includes a pressurized space portion extending in a first direction and having a proximal end and a distal end relative to the pressure release portion, and a pressure channel branching from the pressurized space portion in a second direction different from the first direction and connected to the pressurized space portion, the pyrotechnic charge is disposed in the pressure channel; the pressurized space portion has a sample container accommodating portion for accommodating the sample container between the connection portion with the pressure channel and the distal end; 19. The pyrotechnic cell disruption device of claim 17 or 18. (Appendix 26) 27. The pyrotechnic cell disruption device of claim 25, wherein the first direction and the second direction are perpendicular to each other. a pressure vessel having a bottom, the pressure chamber formed therein, and a proximal end of the pressurized space portion opening on an upper surface; a cap in which the pressure release portion is installed and which can be attached to the pressure vessel so as to cover the upper surface of the pressure vessel; Equipped with the pressurized space portion extends along the vertical direction of the pressure vessel, and the distal end is positioned at the bottom of the pressure vessel, so that the sample vessel can be placed on the bottom. 27. The pyrotechnic cell disruption device of claim 25 or 26. (Appendix 28) the pressure release section has a pressure release vent passage that communicates the pressurized space section with an external space after the pressure chamber is pressurized by ignition and combustion of the pyrotechnic charge; the proximal end of the pressurized space is connected to the pressure release vent channel; 28. The pyrotechnic cell disruption device of any of appendices 25 to 27. (Appendix 29) 29. The pyrotechnic cell disruption device of claim 28, wherein the pressurized space and the pressure release vent are coaxially arranged when the cap is attached to the pressure vessel. (Appendix 30) the pressure release vent path is blocked by a rupture disc or a valve body, and when the rupture disc ruptures or the valve body is opened, the sample container accommodation portion communicates with an external space. 30. The pyrotechnic cell disruption device of claim 28 or 29. (Appendix 31) a chip having a substrate; the pressure chamber is formed by a first recess provided on the surface of the substrate; 10. A pyrotechnic cell disruption device as described in Appendix 1. (Appendix 32) a film covering the surface of the substrate; 32. The pyrotechnic cell disruption device of claim 31, wherein the first recess is covered by the film to seal the pressure chamber. (Appendix 33) a pyrotechnic charge chamber housing for containing the pyrotechnic charge; the pyrotechnic charge chamber housing is disposed above the pressure chamber; 32. The pyrotechnic cell disruption device of claim 31. (Appendix 34) the pyrotechnic charge chamber housing has a rupture portion that ruptures upon ignition and combustion of the pyrotechnic charge; The breaking portion is disposed so as to face the pressure chamber. 34. The pyrotechnic cell disruption device of claim 33. (Appendix 35) an expansion chamber formed by a second recess in the surface of the substrate; a first channel provided in the substrate connecting the pressure chamber and the expansion chamber; Further provided with the first channel has an orifice that applies shear stress to the fluid sample when the fluid sample flows through the first channel; 35. The pyrotechnic cell disruption device of any one of appendices 31 to 34. (Appendix 36) the expansion chamber has a larger volume than the pressure chamber, and the fluid sample is subjected to a reduced pressure as it flows from the pressure chamber through the orifice into the expansion chamber; 36. The pyrotechnic cell disruption device of claim 35. (Appendix 37) 37. The pyrotechnic cell disruption device of claim 35 or 36, wherein the expansion chamber is open to the outside. (Appendix 38) a reaction chamber formed by a third recess provided on the surface of the substrate, the reaction chamber containing a reagent for reacting cells contained in the fluid sample; a second channel provided in the substrate connecting the expansion chamber and the reaction chamber; 38. The pyrotechnic cell disruption device of any of claims 35 to 37, further comprising: (Appendix 39) 39. The pyrotechnic cell disruption device of claim 38, wherein the reaction chamber is open to the outside. (Appendix 40) a detection chamber formed by a fourth recess provided in the surface of the substrate; a third channel provided in the substrate and connecting the reaction chamber and the detection chamber; 40. The pyrotechnic cell disruption device of claim 38 or 39, further comprising: (Appendix 41) 41. The pyrotechnic cell disruption device of any one of claims 31 to 40, further comprising a pair of clamps for clamping the tip. (Appendix 42) 42. The pyrotechnic cell disruption device of claim 41, wherein the pair of clamps includes an upper clamp having a substantially flat clamping surface facing the tip loading area and a lower clamp having a tip recess configured to accommodate the tip. (Appendix 43) 43. The pyrotechnic cytolysis device of claim 42, wherein the upper clamp and the lower clamp each have a housing configuration. (Appendix 44) containing a fluid sample containing cells in a pressure chamber configured to be pressurized by a pyrotechnic charge; pressurizing the fluid sample contained in the pressure chamber by igniting and burning the pyrotechnic charge; 10. A pyrotechnic cell disruption method comprising: (Appendix 45) 45. The pyrotechnic cell disruption method of claim 44, further comprising maintaining pressure in the pressure chamber resulting from combustion of the pyrotechnic charge for a period of time. (Appendix 46) 46. ​​The pyrotechnic cell disruption method of claim 44 or 45, further comprising releasing the fluid sample pressurized in the pressure chamber through an orifice into an external space, and applying shear force to the fluid sample as it passes through the orifice. (Appendix 47) 47. The pyrotechnic cell disruption method of claim 46, wherein the external space is an expansion chamber that receives the fluid sample that has passed through the orifice, and expands the fluid sample as it is received in the expansion chamber. (Appendix 48) 48. The pyrotechnic cell disruption method of claim 47, further comprising adding a reagent to the fluid sample in the expansion chamber to react the fluid sample with the reagent. (Appendix 49) 48. The pyrotechnic cell disruption method of claim 47, further comprising moving the fluid sample from the expansion chamber into a reaction chamber containing a reagent, wherein the fluid sample reacts with the reagent in the reaction chamber. (Appendix 50) 50. The pyrotechnic cell disruption method of claim 49, further comprising reacting the fluid sample with the reagent in the reaction chamber and then releasing the fluid sample from the reaction chamber into a detection chamber. (Appendix 51) 51. A pyrotechnic cell disruption method according to any one of claims 48 to 50, wherein the reaction using the reagent is a polymerase chain reaction (PCR) or a loop-mediated isothermal amplification (LAMP). (Appendix 52) 52. A pyrotechnic cell lysis method according to any one of claims 44 to 51, wherein the pyrotechnic cell lysis method does not include lysing cells contained in the fluid sample using a chemical.

[0107] Each feature disclosed herein may be combined with any other feature disclosed herein. [Explanation of symbols]

[0108] D1~D11: Pyrotechnic cell disruption equipment 1. Cylinder body 2. Piston 3. Sample 4. Precision Orifice 5. Pyrotechnic charges 6···Cap 7. Low-pressure space 8. High-pressure space 11. Pyrotechnic charge chamber 14. Diaphragm 15. Pressure chamber 16. Pyrotechnic charge chamber housing 20 Pressure relief valve 28 Initiator 31. Clamp 32 chips 33 First recess 34... Circuit board 35...Second recess 36 Expansion chamber 37. Reaction chamber 38···Reagents 41 Detection chamber

Claims

1. a pyrotechnic charge configured to be ignited and to burn upon ignition; a pressure chamber configured to contain a fluid sample containing cells and to be pressurized upon ignition and combustion of the pyrotechnic charge; a sample container disposed in the pressure chamber and filled with the fluid sample; a pressure relief valve for releasing pressure from the pressure chamber after the pressure chamber has been pressurized by ignition and combustion of the pyrotechnic charge, thereby expanding cells contained in the fluid sample; Equipped with Pyrotechnic cell disruption device.

2. 10. The pyrotechnic cytolysis device of claim 1, wherein the sample container is flexible.

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