Inductively coupled plasma based atomic analysis system and method

Abstract

The present invention relates to systems and methods for inductively coupled plasma (ICP) analysis, including mass cytometry. Inductively coupled plasma (ICP) analyzers use an ICP torch to generate a plasma that atomizes and ionizes a sample. Analysis of the atomic ions can be performed by atomic analysis, such as mass spectrometry (MS) or atomic emission spectrometry (AES). Particle-based ICP analysis includes analysis of particles, such as cells, beads, or laser ablation plumes, by atomizing and ionizing the particles in an ICP torch prior to atomic analysis. In mass cytometry, the mass tags of the particles are analyzed by mass spectrometry, such as ICP-MS. The systems and methods of the present application include one or more of an ICP load coil including annular fins, particle suspension sample introduction fluidics, and an ICP analyzer.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application is a continuation of U.S. Provisional Patent Application No. 63 / 080,672, filed September 18, 2020. U.S. Provisional Patent Application No. 63 / 081,172, filed September 21, 2020, filed May 5, 2021 This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 184,521 filed on Dec. 1, 2002, and is hereby incorporated by reference for all purposes. The entire disclosure of which is incorporated herein by reference.

[0002] The technical field of this application is the field of inductively coupled plasma (ICP) analysis, including mass cytometry. The present invention relates to a system and method for [Background technology]

[0003] Inductively coupled plasma (ICP) analyzers use an ICP torch to atomize and inject the sample. The atomic ions are analyzed by mass spectrometry (MS) or atomic emission spectrometry (AES). This can be done by atomic analysis such as atomic emission spectroscopy (AES). Particle-based ICP analysis includes: Cell analysis by atomizing and ionizing particles in an ICP torch prior to atomic analysis. This includes the analysis of particles such as particles, beads, or laser ablation plumes. In mass cytometry, particle mass tags are analyzed by mass cytometry such as ICP-MS. will be done. Summary of the Invention

[0004] The systems and methods of the present application include one or more removable ICP torch holder accessories. The ICP load coil includes an annular fin, a particle suspension sample introduction fan, and an external ignition device. Luidix and their ICP analyzers.

[0005] This specification refers to the following accompanying drawings, in which like reference numbers in different drawings refer to the same or similar figures: or a similar component. [Brief explanation of the drawings]

[0006] [Figure 1] FIG. 1 is a schematic diagram of a standard suspension mass cytometry workflow. [Figure 2] FIG. 1 is a diagram of a standard suspension mass cytometry system. [Figure 3] FIG. 1 is a diagram of a standard ICP torch. [Figure 4] 1 is a diagram of an exemplary ICP torch box in the present application. [Figure 5] FIG. 5 is an exploded view of the ICP torch box and upstream atomization chamber of FIG. [Figure 6] FIG. 5 is a cross-sectional view of the ICP torch box of FIG. 4. [Figure 7] FIG. 1 illustrates an exemplary torch holder assembly of the present application. [Figure 8] FIG. 8 is an exploded view of the torch holder assembly of FIG. [Figure 9] FIG. 1 is a circuit diagram of an exemplary external ignition device of the present application. [Figure 10] 1 is a diagram of an exemplary sample loop of the sample introduction device of the present application. [Figure 11] FIG. 11 is a diagram of the exemplary sample loop shown in FIG. 10. [Figure 12] FIG. 10 is a diagram of the sample introduction fluidics during sample loading. [Figure 13] FIG. 1 is a diagram of the sample introduction fluidics during sample injection. [Figure 14] 8 is an exemplary cross-sectional view of the torch holder assembly of FIG. 7. DETAILED DESCRIPTION OF THE INVENTION

[0007] Certain aspects and configurations of the present disclosure may be used in mass cytometry as further described herein. A system and method for inductively coupled plasma (ICP) analysis, including: Inductively Coupled Plasma (ICP) Systems, Samples, and Methods

[0008] Inductively coupled plasma (ICP) is a plasma created by electromagnetic induction (i.e., by a time-varying magnetic field). ICP is a type of plasma source that is energized by an electric current generated by the plasma. Industrial-scale applications of are in micromachining (e.g., etching or cleaning) ) or waste treatment. Such applications involve the generation of plasma within the ICP torch. It cannot be used with ICP load coils and cannot operate under atmospheric conditions. and / or not of a scale suitable for atomic analysis of the sample (e.g., the plasma generated is (The size can be at least larger than the plasma of the ICP analyzer.) The physical characteristics of industrial ICP are different from those of ICP analysis using an ICP torch, and are different from those of the present disclosure. In this specification, the ICP analyzer is used with an ICP torch. Systems and methods are discussed.

[0009] For an overview of ICP mass spectrometry (ICP-MS), see Montaser and Akbar, Inductively Coupled Plasma Mass Spectrometers. The Quantitative Analysis System, John Wiley & Sons, 1998, includes a discussion of vortex flow and ignition. The considerations for the material introduction and ICP torch are similar to those for atomic emission spectroscopy (AES). Also known as optical emission spectroscopy, which is also within the scope of this application Atomic spectroscopy as used herein is the same as atomic analysis (ICP-M Suitable samples may include atomic mass spectrometry (e.g., S) or ICP-AES. In certain embodiments, biological samples include biological material and / or contaminants. Fluids containing substances (e.g., metal toxins) or cells (e.g., in suspension or in tissue sections) or particles such as beads (used to assay biological materials). do. Mass cytometry system and method

[0010] An aspect of this application is the detection of mass tags within cells or beads by mass cytometry. This includes an ICP torch system and method for mass cytometry. are described in U.S. Patent Application Publication No. 2005 / 0218319, U.S. Patent Application Publication No. 201 No. 6 / 0195466 and U.S. Patent Application Publication No. 2019 / 0317082 and the like, which are incorporated by reference in their entireties. The laser absorbance generated from suspended particles (e.g., cells or beads) or tissue sections The particles may be generated from a solid sample, such as a suspension plume. In cytometry, a suspension of cells or beads containing mass tags is analyzed by atomic mass spectrometry. The analysis was performed using laser ablation (LA) ICP-MS. Lee discloses U.S. Patent Application Publication No. 20160056031 and U.S. Patent Application Publication No. 20160056032. No. 0140287953, which is incorporated herein by reference. Imaging mass cytometry by LA-ICP-MS was described by Giesen, Charlotte et al. “Highly multiplexed imaging of tumor tissues with subcellular resolution by mas s cytometry.” (Nature methods 11.4 (2014): 417-422).

[0011] Mass tags can be used to bind affinity reagents (e.g., antibodies, oligonucleotides, avidin, or target For example, the tag may be a metal tag attached to a target biomolecule (e.g., another bioparticle that specifically binds to the target biomolecule). For example, metal nanoparticles or metal chelating polymers can be attached (e.g., covalently attached) to affinity reagents. Suitable mass tags may be prepared by combining them together and then applying them to the sample. No. 0072250 and U.S. Patent Application No. 20080003616. , which are incorporated by reference in their entireties. In certain embodiments, some mass tags are , are not bound to affinity reagents such as metallodrugs or histochemical stains.

[0012] Figure 1 shows a schematic of a typical mass cytometry workflow, and Figure 2 shows the FIG. 1 is a diagram of a standard mass cytometry system used in such a workflow. The suspension mass cytometry workflow involves labeling cells with mass tags. (See the cells labeled with the labeled antibody in the upper left of Figure 1.) Particles such as beads are introduced into the sample held in the sample holder by the sample introduction fluidics. The material is transferred from a container (e.g., a tube) through a nebulizer and into a nebulization chamber. The atomization chamber is a chamber that surrounds the droplets released from the nebulizer with a carrier gas (i.e. Unlike many other nebulization chambers, the suspension The nebulization chamber of the mass cytometry system directs particles into the ICP instead of the waste outlet. A typical atomization chamber suitable for mass cytometry is the US No. 2013 / 0181126, the entire contents of which are incorporated by reference. The ICP torch box, as further described herein, incorporates an ICP torch. The particles pass through the ICP plasma of the ICP torch, where they are atomized and ionized. The vacuum differential forces the ionized atoms from the plasma into the sample cone (and one or more The ion optics of the mass analyzer are, for example, For example, the mass tag is configured to filter ions at If you have heavy metals (such as transition elements or lanthanides) or their isotopes (deflation a mass filter (such as a quadrupole or high frequency quadrupole) acts as a high pass mass filter, It can be configured to remove ions below a certain mass. Argon dimer is 80 amu, so a high-pass mass filter must be at least 80 amu. Such high-pass filters can remove ions with a cutoff of 100 u. The mass analyzer is then used as a time-of-flight detector (i.e., TOF-M). Suitable mass analyzers include TOF-MS or magnetic sector MS. In certain embodiments, the mass analyzer includes a quadrupole MS (QMS) The ions of the mass tags detected by the mass analyzer can be other analyzers such as The mass tag indicates the presence of the target of the affinity reagent to which it specifically binds. Compared to the detection tag of fluorescent tags (showing overlap of Because there is relatively little crosstalk between channels, many mass tags can be distinguished by individual particles. In certain embodiments, a plurality of distinguishable mass tags (e.g., at least 20, At least 30, or at least 40 mass tags) are detected in a single cell event.

[0013] In certain embodiments, the system includes a filter disposed at the inlet to the nebulizer. The filter can pass single cells, but can also pass e.g., To reduce clogging in the nebulizer (such as at the tip of the nebulizer), The filter is made of nylon mesh and prevents particles from passing through the nebulizer. The filter may comprise a mesh, such as a mesh of a polymeric material, or any material suitable for straining cells. Smaller than the inner diameter of the nebulizer channel, e.g., smaller than 80% of the inner diameter of the channel For example, a nebulizer can pass 20 or less particles. 0 microns or less, 150 microns or less, or 100 microns or less, for example, 50 microns or less The filter can have an inner diameter of 100 to 150 microns. To allow particles to pass through the nebulizer, the particles must be larger than 25 microns, 30 microns, or Cutoff greater than Kron, greater than 80 microns, or greater than 100 microns As shown in FIG. 2, the filter can be used in conjunction with a nebulizer and a sample introduction sieve. The filter may be located at the interface between the sample introduction fluidic device and the sample introduction fluidic device. The sample loop may be in close proximity to the sealed coupling of the sample loop and the nebulizer. The inner diameter of the loop may be, for example, at least twice as large, or smaller than, the inner diameter of the nebulizer channel. For example, the inner diameter of the sample loop should be at least 0.2 mm, at least 0.5 mm, at least 1 mm, or at least 2 mm. The sealed coupling and / or filter can be easily removed, e.g. to replace the filter. Additionally, the device may be detachable from the nebulizer.

[0014] The workflow and system for mass cytometry have been described above, but laser ablation To transport the plume particles to the mass analyzer, a laser ablation system is used. It is understood that such sampling systems can be an alternative to sample introduction fluidics. ICP Torch Box

[0015] Generally, an ICP torch box consists of the ICP torch body, the outer tube of the torch body, and Optionally, the ICP torch box further includes an ICP load coil disposed adjacent the ICP torch box. The ICP torch may include an igniter and / or a gas supply manifold. The body includes at least an inner tube and an outer tube, the inner tube being the innermost The inner tube and the intermediate tube may be disposed around the inner tube. If the outer tube is separable, they may be separated into a separate inner tube body and an outer tube. The embodiments of the present invention may be described as belonging to any of the tube bodies described herein. Any embodiment of the ICP torch assembly (e.g., a removable ICP torch holder assembly) may be used. special features such as an external ignition device, an ICP load coil, and / or a gas supply manifold. The present invention includes an ICP torch box, including a specific apparatus and method for using the apparatus.

[0016] As shown in FIG. 3, an exemplary conventional torch uses a single external gas (i.e., plasma gas ) inlet, or possibly two inlets, can provide the vortex. The gas inlet may extend from the torch tube and may be of the same material.

[0017] An exemplary torch box of the present application is shown in FIG. 4, which shows the outer torch tube. The device has two high-voltage transformers that output AC output voltages to two electrodes located outside the device. The external ignition device includes a circuit board that supports the torch box. , as shown in Figure 5. Gas supply manifold that holds the removable torch holder assembly Similar to the Nifold, the ICP load coil with annular fins is shown on the left side of Figure 5. The atomization chamber is shown upstream of the torch box and is the inner chamber of the torch assembly. An injector extends into the tube.

[0018] Figure 6 is a cross-sectional view of the torch assembly. Most of the components are injectors and The annular fins of the ICP load coil extend radially around the axis of the torch. The torch holder is shown thinner than the inner and outer torch bodies. and is shown disposed within the gas supply manifold. As discussed further in, gas flows from the manifold through the torch holder to the inner trough. It flows into the torch or outer torch.

[0019] FIG. 7 shows an enlarged view of the torch holder assembly of FIG. 5. FIG. 8 shows the inner tube base. an inner torch body having an inner tube connected to the outer tube base; 1 is an exploded view of an exemplary torch holder assembly including an outer torch body having a tube. The inner base and the outer base are received by the torch holder, and the torch holder and and can be detached from their respective bases.

[0020] As described in one or more embodiments herein, an ICP torch box The torch holder assembly may provide vortex flow and / or be removable. As described herein, for example, an outer torch body and optionally The inner torch body is removable from the torch holder. Alternatively, or in addition, Additionally, the outer tube may be detachable from the outer tube base and / or may be The tube may be detachable from the inner tube base. Additionally, the torch holder assembly can be removed (e.g., by twisting) from the torch box. and / or by pulling from behind) and the outer It can be removed without having to remove the ignition electrode from the tube.

[0021] As described in one or more embodiments herein, an ICP torch box The external ignition device is provided on the outside of the outer tube and upstream of the ICP coil. The external ignition device may be located on the opposite side of the outer torch tube. As further described herein, an external ignition device may be used. The mechanism can be by electrical discharge, such as a dielectric barrier discharge. External ignition devices are especially useful for ICP-MS analysis where the ignition electrode of a torch is not suitable. Allows for detachable torch design and operation. Any of the aspects discussed herein ( The external ignition device (or embodiments) may be used within an ICP torch box to incorporate the features discussed herein. Can be combined with any embodiment(s) of the torch holder assembly .

[0022] In certain embodiments, an ICP torch box comprises an ICP load coil of the present application. The ICP load coil of any embodiment(s) discussed herein may be The torch holder assembly of any embodiment(s) discussed, and / or any The embodiment may be combined with an external ignition electrode of the embodiment (or embodiments).

[0023] Optionally, any of the ICP torch boxes described above may further comprise any of the ICP torch boxes described herein. Optionally, a gas supply manifold (or manifolds) may further be included.

[0024] In certain embodiments, the atomizing chamber and / or injector (from the atomizing chamber) The tube extending from the torch box may be described as part of the atomizing chamber. The bar may be described as part of the sample introduction system to the torch box. The mist chamber injector is located in the inner tube (i.e., the innermost tube) of the ICP torch. The sample (e.g., aerosolized particles) is delivered concentrically into the inner tube (the It may be inserted into the tube.

[0025] In certain embodiments, the ICP torch box is part of an ICP analyzer. For example, The sample cone of the mass spectrometer is generated from the sample by the ICP plasma of the ICP torch. In such a case, the ionized atoms may be arranged to receive the ionized atoms. The disclosed external ignition device is a torch holder assembly as described herein. This allows for a removable design of the battery.

[0026] This embodiment uses a removable ICP torch holder assembly, a dielectric barrier discharge an external ignition device for igniting the plasma, and / or an ICP load coil with annular fins. The ICP load coil is used for 3D plasma generation. May be linted.

[0027] In a specific embodiment, an inductively coupled plasma (ICP) torch box comprises: can.

[0028] (1) an inner torch body having an inner tube connected to an inner tube base; an outer torch body having an outer tube connected to a torch base, and a removable inner A torch holder for holding a torch base and a removable external torch base. Detachable Inductively Coupled Plasma (ICP) Torch Holder Assembly

[0029] (2) An ICP load coil arranged around an outer tube, the cylindrical coil has an annular fin, the annular fin being located at least once around the cylindrical coil. The ICP load coil is primarily made of copper, which has a lower oxidation rate than copper. ICP load coil with metal or alloy

[0030] (3) An ignition device for igniting a plasma in an inductively coupled plasma (ICP) torch, comprising: The ignition device includes an oscillator, a first high voltage transformer coupled to the first electrode, and a second high voltage transformer coupled to the second electrode. The ignition device includes a circuit having a second high voltage transformer, and the ignition device generates plasma by dielectric barrier discharge. The first electrode and the second electrode are configured to ignite the ignition source. , located within 5 mm of the outer tube, an ignition device Torch Holder Assembly

[0031] Various embodiments of the present application, as exemplified in the manner discussed below, Such aspects relate to any operable combination of They can be combined in a single step.

[0032] The torch assembly embodiment is similar to an ICP torch box having a torch assembly. The present invention relates to a removable ICP torch holder assembly and a method of use. The torch can be removed by moving the torch holder backward (e.g., to remove the torch during operation). the direction of gas flow through the torch box (e.g., In certain embodiments, the gas supply manifold may be detachable from the work box. The removable torch is secured by a gas source manifold inside the ICP torch box. may be twisted before removing the torch assembly (e.g., remove the pin on the removable torch). (the valve may be twisted from its locked position on the gas flow manifold). In certain embodiments, the ICP torch box may include an ICP torch acetate as described in The electrode is not inserted into the outer tube of the assembly, which prevents the torch assembly from being inserted into the torch box. External points to make the assembly removal step easier (removal in a single step) This removal allows for easy maintenance of the ICP torch (e.g., This can facilitate cleaning or replacement of one or more torch tubes.

[0033] Alternatively, or in addition, the torch holder of the removable ICP torch assembly At least the outer tube base is connected to the outer tube (and Similarly, the torch holder may be configured to be The torch base may be configured to receive an internal torch base coupled to the inner tube. Alternatively, the torch holder may be attached to the inner tube and / or an inner tube coupled to the inner tube. May be permanently attached to the tube base (e.g. machined as the same part) The torch assembly consists of an inner tube and The inner tube base and / or the outer tube may be configured to be coaxially aligned. Or an O-ring to provide an airtight seal between the base of the outer tube and the torch holder In certain embodiments, the inner tube base and the outer tube base may be Threads on one of the inner and outer torch bases that screw into the other torch base. They may directly reference each other so that they are irremovably connected (e.g., they are fabricated as the same part). or glued together), or joined directly to each other by fasteners. The inner and outer bases, which directly reference each other, may be configured as a single unit. to remove it from the torch holder.

[0034] The inner tube and / or the outer tube described in the embodiments herein may be made of glass, stone, or the like. The inner tube described in the embodiments herein may be made of a material such as stainless steel or ceramic. The tube base and / or outer tube base may be made of a different material such as metal (e.g., aluminum alloy). The material may be:

[0035] In certain embodiments, the outer torch tube is The outer torch tube can be removed from the outer torch base for replacement or replacement. The outer torch tube can be damaged by chipping, heating, or precipitation during operation. Less expensive than an external torch base, this removable mechanism reduces maintenance costs Similarly, in certain embodiments, the outer torch tube , can be removed from the outer torch base for maintenance (e.g., cleaning or replacement) The outer torch tube can be damaged by chipping, heating, or settling during operation. The outer torch tube is less expensive than the outer torch base, and this installation The removable mechanism reduces maintenance costs.

[0036] The torch assembly is connected to the torch accelerant where the external gas (also known as plasma gas) It can be configured to be able to enter the outer tube of the yellowtail.

[0037] Previously, one or sometimes two holes were used to introduce the vortex. If so, the bore must be made of the same material as the outer torch tube (e.g., glass, quartz, or ceramic). The torch tube may be defined by a tube of the same design and disposed to the side of the torch tube. Removal of the touch assembly (e.g., from the gas supply manifold) by / or complicating removal of the outer torch tube from the torch holder of the torch assembly Furthermore, such a design may be unable to accommodate several such holes. I can't.

[0038] In certain embodiments, the outer torch body of the torch assembly comprises an inner tube and an outer tube. The outer gas is arranged and oriented to direct the outer gas into the annular region between the inner and outer tubes, and the inner gas is swirled. The holes may have a radius of 0.5 mm or more, such as 0.5 mm or more, and may have a radius of 0.5 mm or more, such as 0.5 mm or more. It may be symmetrical in the direction of the arrow or may be oriented obliquely (i.e., by the outer tube). (The bore may not extend radially outward from the axis of the barrel defined by the outer torch body.) The outer torch body and the inner torch body may be attached to the torch holder. Thus, when held (e.g., by their respective bases), the holes in the outer torch bases The gas inlet (e.g., port) on the torch holder is connected to the inner and outer tubes. The torch holder may allow fluid communication to the annular region between the torch holder and the gas supply. The gas supply manifold is in turn held by the external gas inlet (e.g. It provides fluid communication between the gas inlet (e.g., a gas port) and the external gas inlet of the torch holder. In this embodiment, the external torch base, torch holder, and gas supply manifold are all It is mainly made of metal.

[0039] Alternatively, or in addition, the torch holder may contain an internal gas (known as an auxiliary gas). providing fluid communication for internal gases to pass from the gas supply manifold to the internal torch body; Alternatively, or in addition, the gas supply manifold may A carrier gas (also known as a gas or aerosol gas) is supplied to the gas supply manifold. The carrier gas flows from the carrier gas inlet (e.g., port) of the atomizer and flows through the atomization chamber. Thus, the gas supply manifold may provide fluid communication with the carrier gas, the internal gas, and the may have separate ports for one or more of the internal and external gases Each port may be coupled to a gas source, such as a gas source containing argon and / or helium. In certain embodiments, the external gas may comprise argon. Alternatively, the external gas may comprise nitrogen. The gas may include gases other than argon or helium, such as:

[0040] The fluid communication described above allows the tube to be moved away from the torch holder assembly (e.g. For example, it may be positioned (without being directly connected to the torch holder or external torch body) and the torch The holder can be more easily removed from the torch box (e.g., its gas supply manifold). can be removed.

[0041] In certain embodiments, the removable ICP torch holder assembly includes an inner tube. an inner torch body having an inner tube connected to a torch base; and an outer tube connected to a torch base. an outer torch body having an outer tube and a base for receiving the inner and outer torch bases; a torch holder configured to receive the inner tube and the outer tube; The torch holder defines an annular region when holding the inner torch base and the outer torch base. do.

[0042] In certain embodiments, the outer tube is separate from the outer tube base and / or the inner The tube separates from the inner tube base.

[0043] The torch holder may have an external gas inlet, the external gas inlet being in fluid communication with the annular region. The outer tube base is adapted to provide fluid communication between the external gas inlet and the annular region. The pores may define three or more holes (e.g., six holes) arranged in such a manner that three or more holes The upper holes are oriented to create a vortex flow. The vortex flow does not dissipate and only a small gas flow is required. If desired, the hole may be located close to the plasma. For example, the hole may be located near the inner torch. The outer tube may be tapered within 2.5 cm of the tube exit. Instead, the outer tube defines a taper that directs the outer gas toward the plasma. Accelerate.

[0044] The inner tube base and outer tube base can be removed from the torch holder. or may be referenced to each other (e.g., inner tube base and outer tube The bases are permanently joined together and the inner and outer torch bases are threaded. (These components are configured to be connected by fasteners such as

[0045] The torch holder is designed to reduce precipitation and / or clogging in the injector. A heat conducting element is provided that is positioned to heat the injector.

[0046] In certain embodiments, a metal or alloy, such as an aluminum alloy, is used as the main component of the torch holder. The outer tube must have holes to allow the electrodes to extend into the outer tube. stomach.

[0047] The torch box is a gas supply manifold configured to accept a torch holder. For example, the torch holder may be mounted within a gas supply manifold. The torch holder may include a pin for aligning its position in the This allows the torch holder to be removed from the gas supply manifold. The holder can define an external gas inlet, the external gas inlet providing a gas supply to the annular region and the gas It is in fluid communication with the outer gas flow region of the supply manifold.

[0048] In certain embodiments, the inductively coupled plasma (ICP) torch assembly comprises an inner tube an outer tube; and an outer tube base connected to the outer tube. The outer tube and the outer tube define an annular region, and the outer tube base allows external gas to pass through the annular region. and defining three or more holes arranged to allow the flow of the fluid through the holes, the three or more holes being is directed to generate

[0049] 9 is a cross-sectional view of an exemplary torch holder assembly. As shown, The torch holder holds the torch base, which is a single piece (e.g., a plastic a continuous piece of plastic, or an outer base and an inner base permanently attached to each other) Alternatively, the base may comprise separate inner and outer bases that can be separated from one another. By having an inner base that is separable from the outer base, cleaning or For replacement, the inner torch body (inner base and inner tube) can be separated from the outer torch body. For example, the outer torch can be used to heat or deposit material from the plasma. Therefore, more regular maintenance is required.

[0050] As shown in FIG. 9, the torch base may include one or more external gas inlets (e.g., 6, 8). At least two, at least three, at least four, at least five, such as 10, 20, 30, 40, 50, 60, 70, 80, 90, 10 ... Two external gas inlets may be provided, each connected to a gas supply manifold (not shown). ) into the annular space between the inner and outer tubes of the torch-hold assembly. The external gas is directed through the space between the inner and outer tubes. The holes may be located directly adjacent to the outer tube so that injection may be performed directly between the holes. are angled to provide a vortex flow.

[0051] In certain embodiments, the outer tube and inner tube are permanently attached to the torch base. (e.g. outer tube to outer torch base, inner tube to inner torch base) For example, the outer tube may be secured by adhesive (e.g., a heat-resistant adhesive). The outer wall of the outer tube and the torch base (e.g., the outer The gap between the torch base and the adhesive allows the two to bond together during heating (e.g. (e.g., while the torch is running) to allow the outer tube and / or adhesive to expand. In certain embodiments, the gap is greater than 0.2 mm, e.g., 2 mm. , greater than 0.5 mm, or greater than 1 mm. Due to irremovable joints and / or gaps, the outer tube is attached to the torch base without an interference fit. The length of overlap between the outer tube and the torch base is The vortex of the external gas injected into the outer torch by the hole is then generated by the torch. It may be short, so that there is a minimum distance before the Zuma that it dissipates or decreases. For example, it may be shorter than 10 mm or shorter than 5 mm. external ignition

[0052] The ICP load coil is capable of sustaining the plasma within the ICP torch and is typically used for testing. An ignition electrode, such as a Lacoil electrode, creates a process described as an arc or spark. The spark from the ignition electrode is used to ignite the plasma. by a conductive element such as an ICP coil (e.g., a grounded conductive element) that is in contact with the Yes. ICP torches operate at atmospheric pressure (e.g., between 0.9 atm and 1.1 atm). In ICP-AES, the plasma is directly analyzed by spectroscopy. Therefore, the electrode must be located downstream of the outer torch tube (e.g., downstream in the gas flow through the torch). However, in ICP-MS, the ignition electrode is located at the electrode where the plasma is maintained. To avoid sparking with the sample cone and any conductive parts in close proximity to the sample, , disposed adjacent to the outlet of the outer tube to receive the ionized atoms into the mass analyzer. The sample cone placed on the torch may obstruct the ignition electrode of the vortex. This problem can be solved externally (for example, as explained in U.S. Pat. No. 4,766,287). This has been previously solved by inserting the ignition electrode through the torch tube. However, the torch assembly can be maintained by inserting the electrode through the torch body. for service (e.g., for cleaning or replacing one or more torch tubes) Thus, in certain embodiments, the ease of removal may be hindered. The external ignition device and method of the present application may include a removable torch as further described herein. can be combined with

[0053] External ignition allows for the attachment of a metal assembly for the insert / holder and gas supply. The inventors have discovered that the proximity of metal parts within the torch assembly allows for a removable torch. Therefore, the ignition method for the mass cytometer using an internal spark electrode is not available, and the spark The sample will fly towards the rear of the base of the torch, not towards the sample collection interface. The external electrodes were arranged so that they would not get in the way and were close to the plasma space. The first successful experiments used a single electrode, and the discharge was generated by the torch tube. The AC pulse was generated using a Tesla coil on the outside of the torch. The inventors also found that the use of symmetrical outputs can also be useful in forming plasmas. In this study, we identified a higher energy voltage AC generator, which is easily manufactured. This provided repeatable and reliable ignition for the

[0054] External ignition devices are used to generate electrical currents such as electrostatic barrier discharges in inductively coupled plasma (ICP) torches. The plasma is ignited by an electric discharge. The external ignition device applies an AC current to at least one electrode. The power supply may be configured to provide:

[0055] The exemplary external breakdown circuit shown in FIG. 9 uses a low frequency (100 Hz) modulator to Alternating current with a frequency of approximately 30 kHz, varying within a range of + / - 2 kHz around the center value The modulated signal is then fed to two transformers T1 and T2. The transformers T1 and T2 are amplified by a power bridge circuit that drives the respective Opposite voltages in the circuit so that the output voltages are out of phase (180 degrees out of phase with each other) Transformers T1 and T2 are connected in polarity to operate close to their natural frequencies and maximize efficiency. The large voltage is amplified to 30 kV and then connected to the ignition electrode attached to the torch assembly. It is noteworthy that the voltage is 30 kV (e.g., anywhere in the range of 1 kV to 100 kV). A transformer using a different frequency than the one used in the previous example is combined with an oscillator operating at a similar frequency. The intermediate frequency of the oscillator is approximately the resonant frequency of the amplifier transformers T1 and T2. The frequency was chosen to be 30 kHz (in principle, they are identical to each other, but the manufacturing (May be affected by tolerances).

[0056] Due to manufacturing tolerances, the actual resonant frequency of the transformer cannot be precisely specified at 30 kHz. However, the actual resonant frequency of the transformer must be adjusted to achieve the maximum output voltage for ignition. It is important to drive the transformer with a sufficient AC current.

[0057] To ensure that the resonant frequency of the transformer is matched, a triangular ramp signal is applied to the control input of the 30kHz oscillator. A 100Hz modulator is used to apply a signal, which keeps the modulator frequency at approximately 28k. The frequency is constantly swept from 100Hz to 32kHz. At each match, the transformer output voltage applied to the ignition electrode increases momentarily to 30 kV. A discharge is initiated in the argon gas column inside the torch. Once initiated, the frequency drops to the resonant peak. The voltage required to maintain the discharge is 100% for argon gas. This is significantly lower than the voltage required to initiate a discharge within the

[0058] The ignition circuit is supplied with a constant voltage of 24Vdc from the host device. ON / OFF is triggered by a signal from the Zuma management system.

[0059] In this particular embodiment shown in FIG. 9 and described above, all internals of the ignition module The operation is fixed and no external software control is required other than on-off operation. However, in other applications, the computer can be used to control the output voltage, modulation frequency, and range. It is desirable to have physical computer interfaces that provide such control. Interfaces can be easily added.

[0060] Exemplary alternatives to FIG. 9 include one or more of the following:

[0061] Using different amplifier transformers, the intermediate oscillators can be set to different frequencies.

[0062] (For example, by providing a trimming potentiometer in the circuit board, No low frequency modulation (if the frequency can be exactly matched to the transformer resonance)

[0063] Using only one transformer, the output is single-ended with the second electrode grounded. For example, the second electrode may not be present at all, and the grounded end of the discharge may be connected to the other grounded ends of the torch. The metal part may be provided with a protective layer.

[0064] The on / off control line can be omitted and the on / off operation can be controlled by 24V if desired. It could be controlled by switching the DC

[0065] It is not intended to be limiting to the above alternatives, but rather to include a number of possible variations. It is intended to illustrate a few aspects.

[0066] In certain embodiments, the external ignition device comprises at least one electrode. The fire device consists of a single externally coupled insulated element (e.g., torch tube) in the path of the discharge. The ignition mechanism can be described as a dielectric barrier discharge (DBD). As described herein, the torch tube is positioned opposite the The two electrodes will provide ignition by dielectric barrier discharge (DBD). A discharge (DBD) is an electrical discharge between two electrodes separated by an insulating dielectric. In the above-described embodiment of the two electrodes arranged on both sides of the torch tube, The tube is an insulating dielectric and the discharge crosses the flow of gas through the torch. The mechanism can be described as an electrolytic breakdown discharge (or electrical discharge) and is to metal parts such as the base of the torch tube (e.g., the grounded part of an ICP torch) Examples of discharges: In some cases, the mechanism of ignition can be explained as an electrostatic discharge. Cut.

[0067] In some embodiments, the mechanism of ignition by the external ignition electrode is as a glow discharge (e.g., However, as used herein, As such, such glow discharges exist at or near atmospheric pressure, and the plasma is at a constant pressure. An external ignition device and operation are required that are different from those used when ignited by an external ignition electrode. The plasma density generated by the arc discharge is much lower than that of the glow discharge. It may be approximated as Zuma density.

[0068] External ignition as described herein includes arc discharge, high voltage breakdown, DC current, Tesla current, and / or direct contact of the ignition electrode with the gas to be ignited in the plasma; This is different from multiple firing schemes.

[0069] In certain embodiments, the external ignition device that ignites the plasma within the ICP torch comprises an oscillator and a first high voltage transformer circuit coupled to the first electrode, and optionally coupled to the second electrode; and a second high voltage transformer connected to the ignition device, the ignition device igniting the plasma by dielectric barrier discharge. The plasma is configured to ignite under atmospheric conditions (e.g., within 10% of 1 atm) or The external ignition device can be ignited at a temperature higher than at least 100 Torr. configured to provide an AC output voltage to at least one electrode, such as the second electrode, e.g. For example, a first high voltage transformer and a second high voltage transformer are connected in a circuit with opposite polarities. The device applies a voltage between the first and second electrodes ranging from 2 kV to 100 kV, and from 5 kV to 50 kV. Provide a voltage difference of at least 1 kV, such as from 20 kV to 40 kV. The maximum output voltage of the high voltage transformer in the circuit can be from 10 kV to 40 kV. In certain embodiments, the maximum voltage of the high voltage transformer of the circuit is The voltage is less than 1 kV, such as less than 200 kV. The frequency of the current voltage is from 5kHz to 100kHz or from 20kHz to 40kHz. The AC output power and / or its frequency is lower than atmospheric pressure (e.g. This is sufficient to ignite a plasma at temperatures within 10% of 1 atm.

[0070] The external ignition device generates a plasma, for example, via an electrical breakdown discharge caused by the breakdown of a dielectric barrier. The plasma may be ignited adjacent to the first electrode. the capacitance between the portion of the outer torch body adjacent to the second electrode and the portion of the outer torch body adjacent to the second electrode. It can be ignited by the amount.

[0071] In certain embodiments, the external ignition device is activated by sparking or arcing (e.g., Not configured or operable to ignite a plasma by, for example, arc discharge The external ignition device does not have to include a Tesla coil.

[0072] The device may comprise two electrodes, e.g., an outer electrode of an ICP torch. The outer toe is positioned within 5 mm or 3 mm of the wall. The two high voltage transformers may be placed outside the switch. The output voltages of the two high voltage transformers are in opposite phase. Two high voltage transformers are connected to the circuit with opposite polarity so that the voltage The circuit may have a maximum power output of up to 100 kHz. Further comprising a voltage modulator configured to match the natural resonant frequency of each of the voltage transformers. The circuit has a constant AC output voltage and frequency. At least one of the numbers is controllable by a computer external to the external ignition device.

[0073] The device can be part of an ICP torch, and the electrode can be an outer torch body of the ICP torch. The ICP torch may have an internal torch body and an external At least a portion of the torch body is coaxial with the inner torch body and includes a first electrode and a second electrode. The poles are arranged so that an axis passing through the first electrode and the second electrode intersects a portion of the outer torch body. It has been done. ICP Load Coil

[0074] Aspects of the present application may be used to sustain a plasma (e.g., when operated with an AC current at RF frequencies). The ICP load coil includes an ICP load coil (ICP coil) for defining a cylinder. It can be cylindrical, meaning spiral. In most cases, the coil has three turns. The ICP load coil may be located within a torch box that surrounds the outer torch tube.

[0075] Standard tubular ICP load coils are made from copper and can be solid wire or hollow (tube) In contrast, the ICP load coil embodiment of the present application is made of a material other than copper. It may include other materials and shapes other than a solid wire or hollow tube. The ICP load coil materials and configurations discussed in the publication can reduce coil aging. For example, to reduce heating, oxidation, and / or deformation during operation to sustain a plasma. It can be reduced.

[0076] In certain embodiments, the ICP coil may be 3D printed. The wheels may be machined or cast.

[0077] In certain embodiments, the ICP coil can include annular fins. is understood to be a fin that extends and diverges from a central axis. When bent to form a coil, the wire deforms to expand (stretch) in this direction. It is difficult to machine metal because it may be required to , the ICP coil can have 3D printed annular fins. This allows for increased cooling of the ICP coil during operation (which also reduces oxidation). The annular fin should extend along at least one-eighth of a turn of the ICP coil. The inventors have found that continuous fins can extend along two or more turns. It was found that the efficiency of the coil is not reduced by the operating frequency, inductance, and plasma Metrics such as retention remain unchanged compared to designs with gaps in the annular fin. The absence (or infrequent gaps) in the Further structural stability (such as withstanding deformation during operation as described above) can be provided. Those skilled in the art will appreciate that current flows partially along the outside of a coil having continuous annular fins. This may reduce the efficiency of the ICP and interfere with the operation of the coil. Maybe.

[0078] The material and / or shape of the ICP coil may improve the structural stability of the coil and make it stronger. This may allow for a longer manufacturing process and / or longer lifespan. The P coil can reduce ICP efficiency by more than 10% without interruption (e.g. without reducing the signal detected by the mass analyzer by more than 10%, or at least 5,000 hours, at least 1 0,000 hours, at least 20,000 hours, or at least 50,000 hours of operation can withstand.

[0079] In certain embodiments, the ICP coil can be an alloy, such as an aluminum alloy. The alloy may be suitable for 3D printing. Alternatively, or in addition, the alloy may be , can have many additional properties compared to copper, which are described herein.

[0080] In certain embodiments, the ICP load coil with annular fins is a cylindrical coil. The fins may be continuous over at least one-eighth of a turn of the cylindrical coil, e.g. For example, the ICP load coil may be continuous along at least two turns of the coil. It may primarily comprise metals or alloys that oxidize at a lower rate than copper during or over the life of the equipment. The annular fin may be substantially planar. The inner portion of the ICP load coil may be The inner portion defines a circular cross section and the outer portion defines an annular flange. The inner part of the ICP load coil is at most larger than the outer part of the ICP load coil. The ICP load coil may have a small diameter. The ICP load coil may be solid. The ICP load coil may have a plurality of The annular fin may not be provided. One or more annular fins may be provided between the three turns of the ICP load coil. The annular fin may be located along the innermost portion of the annular fin to the outermost portion of the annular fin. The distance to the side portion may be at least twice the thickness of at least a portion of the annular fin. stomach.

[0081] In certain embodiments, the ICP coil is, for example, 5 mm thick (along the axis of the ICP coil barrel). It can be shorter than 10cm, such as cm long.

[0082] The metal or alloy may be an aluminum alloy. The metal or alloy may have a lower conductivity than copper. , a lower melting point than copper, or a lower ductility than copper. The coils are made by 3D printing, casting, or machining metal or alloys. The metal or alloy may be formed by forming an annular flange, as compared to copper. The alloy is one that reduces deformation compared to copper without the fin. and reducing deformation, for example, along the axis of the tube defined by the coil. The deformation is perpendicular to the axis of the tube defined by the coil. It is a shape.

[0083] The ICP load coil includes a first portion closest to the longitudinal axis of the cylindrical coil, The charge coil has a second portion furthest from the longitudinal axis of the tubular coil, and the first portion is The first thickness is characterized by a first axis that is parallel to the longitudinal axis and perpendicular to the first axis. The second portion is characterized by a second thickness, the second thickness being measured along the longitudinal axis The first thickness is greater than the second thickness, as measured perpendicular to a second axis parallel to the first thickness. Sample introduction

[0084] Aspects of the invention include particle (e.g., cell or bead) delivery systems or laser ablation systems. Sample introduction systems such as particle introduction systems for mass cytometry. The system includes a sample holder, a probe for accessing the sample in the sample holder, and / or Introduce the sample (e.g., a suspension of cells) into the spray chamber upstream of the ICP torch box. In certain embodiments, the method includes a number of components, such as sample introduction fluidics for , the sample holder can accommodate at least 4, at least 8, or at least 12 samples, The sample holder can be configured to hold multiple samples. The sample container may be movable (e.g., a carousel) for presentation. Thus, an embodiment of the present application is a method for automatically obtaining the cell suspension. Such systems include a sample introduction system for maintaining the Passive micromixers with interlocking semi-circular and omega-shaped micromixers by Deen et al. modules: Experiments and simulations.” (Micromachines 6.7 (2015): 953-968) The system shown includes components used in mixed applications in flow cytometry. In certain embodiments, the radius of the loop and the flow rate of the sample through the loop may The flow rate is low (i.e., turbulent).

[0085] In certain embodiments, the sample introduction fluidics are directly coupled to the probe and the nebulization chamber. The sample loop may include a sample loop in direct or indirect fluid communication with the sample loop. For example, the helix may define a cylinder. The axis of the cylinder defined by the helix may be aligned with the cell ( horizontal (e.g., axial) direction to reduce the portion of the loop where particles (e.g., granules or other particles) can settle. For example, within 15 degrees of perpendicular to the axis of gravity. The loop may be disposed around a cylindrical agitator that contacts the loop, the agitator being connected to a spray channel. The particles are kept suspended within the loop during transport to the chamber.

[0086] The sample introduction fluidics can be coupled to one or more reservoirs. The reservoir may include a waste reservoir, a cell capture solution reservoir, a cleaning solution reservoir, and / or a water reservoir. The sample introduction fluidics may include a reservoir and a sample tube (sample lumen). The device may include a valve system, such as a rotary valve system, for directing fluid flow from the pump. As explained in the specification, the sample is transferred to the nebulizer, the atomization chamber, and the injector. The particles are then introduced into the ICP plasma through the nozzle, and at this stage the particles in the solution (e.g., cells and and / or beads) are atomized and separated prior to analysis by an analyzer (e.g., a simultaneous mass analyzer). In certain embodiments, the injector is a heated injector. A description of the heated injector and cell capture solution is provided in U.S. Patent Application Publication No. 201903170 82, which is incorporated herein by reference. Therefore, the cell capture buffer may contain salts in solution, and the salts may be carbon-free and metal-free. (For example, there are no elements with an atomic weight greater than 0).

[0087] The sample introduction system comprises at least two syringes that alternate between cleaning and sample introduction functions. For example, the first pump may pump the sample, beads, washing solution, or cell capture solution. The second pump may pump the syringe into a waste bottle, It is a multi-port valve that connects to a cell capture solution reservoir and a switching valve. The function is to nebulize the sample, beads, solution, and / or cell capture solution from the sample loop. The cell capture solution is pushed directly from the reservoir bottle into the nebulizer. do.

[0088] The sample introduction system includes at least two rotary valves, such as a selection valve and a switching valve. The selector valve may be used to withdraw the probe, the reservoir, and / or the air bubble. A port may be provided to connect to the empty port. The position of the selector valve determines where the pump 1 is drawn from. The switching valve may determine whether to release the sample loop between a load position and an injection position. When the switching valve is in the load position, the sample loop is pumped through the fill pump. In the injection position, the sample loop is aligned with the nebulizer and its contents The material is pushed into the nebulizer via a second pump. The second pump may be a push pump.

[0089] FIG. 10 is an exemplary sample loop of the sample introduction device of the present application.

[0090] Figure 11 is a diagram of the agitator for the sample loop of Figure 10. The spiral of the sample loop of Figure 10 , which fits around the cylinder of the agitator of FIG.

[0091] The components of FIGS. 10 and 11 may be present in the sample introduction fluidics shown in FIG. .

[0092] The cell sample in the carousel may be aspirated via a first pump. For example, in FIG. As shown, the sample flows through the probe to the selection valve and the switching valve is in the load position. Once the sample is loaded into the sample loop, The switching valve may be switched to the inject position and the sample is injected into the first The sample capture solution may be pumped towards the nebulizer. The cell capture solution may be used as a carrier to push the cells towards the cell capture device and into the system. To prevent mixing at the interface between the sample and the liquid, an air bubble may be placed between them.

[0093] In certain embodiments, the sample loop is a microprocessor loop as described in the 2015 Micromachines article. To achieve a low Dean number, a tightly wound sample loop was used based on a narrow radius of curvature. The sample loop is aligned with the horizontal axis of rotation so that the flow of cells (or particles) The force may be positioned (at the bottom of the loop) to minimize settling time. A stirrer (vibrating rod) was used as an additional stimulus to prevent the settling of the sample in the sample loop. However, in contrast to its use in other applications, vibration is not intended to settle the material during transport. Simply add agitation to the flow to maintain cell suspension within the carrier fluid without allowing it to settle or clump. This allows summer students to periodically tap the sample loop. It works just like uploading a file, but without the cost.

[0094] The sample introduction device may be used in conjunction with an ICP analyzer, or alternatively, a flow cytometer. It may be used or combined with any particle-based analytical system such as a stomach.

[0095] The sample introduction device of the present application may include a sample loop, the sample loop comprising at least The sample loop is wrapped around the sample tube at least three times. The wrapping may be around a cylindrical stirrer. The spiral may be at most five turns long. The spiral may be at most 15 degrees from a direction perpendicular to the axis of gravity. The cylinder has an axis of

[0096] Sampling system and method of use (e.g., implemented by a computer storage medium) has built-in jam detection, automatic de-clogging (e.g., jam clearing) (unclogging) and / or shutdown can improve productivity.

[0097] In certain embodiments, the sample system comprises, as part of a pellet resuspension workflow: Also, run it before loading the sample. While mixing, check the probe line and fill pump. The pumps, working together, may function like a pipette, dispensing a fixed volume (e.g. For example, between 25 μL and 250 μL. Select a value (e.g., between 1 and 10) to determine how quickly this amount will appear in the sample tube. It is possible to specify how much stirring force is applied (e.g., 1 represents the least stirring force, 1 (0 represents the strongest stirring power). This parameter is set to the default value (e.g., 5).

[0098] During sample acquisition, the probe line may agitate the sample to prevent it from settling. The user can set how fast the probe line rotates (e.g. 0 means no mixing). Specify the agitation speed value (e.g., 0 to 9) to specify the agitation speed (where 9 is the fastest speed and 0 is the slowest). You may choose a time between 0 and 1000. This parameter should be set to the default value (e.g., 5). During acquisition, the inside and / or outside of the probe line can be adjusted by the user. When the pre-wash cycle is selected or executed by the computer readable medium, It may be washed.

[0099] The default sample loop vibration setting reduces the amount of cells settling in the sample loop. Helps

[0100] The device may further comprise a nebulizer in fluid communication with the sample loop. The atomizing chamber may be in fluid communication with the atomizing chamber, which passes the particles through the injector. The device may be configured to hold one or more samples. The sample holder may be configured to hold a plurality of tubes. For example, the sample holder is configured to cool multiple tubes. The nebulizer is inserted into the probe via the sample loop from the tube provided by the sample holder. a plurality of pumps, valves, reservoirs, and The device may include a probe and / or a tube. a pressure sensor that detects a drop in pressure in the air; and optionally, a control circuit that, when executed, controls the processor. and a computer readable medium storing a plurality of instructions for operating the sample guidance system. For example, several instructions can be given to automatically flush the probe when a blockage is detected in the probe. Reversing the pressure (or adjusting the pressure, e.g., increasing the pressure) and transferring the cells to the processor, thereby removing any clogging of the probe. The sample guidance system can be operated to resuspend the cells in the tube before delivery. can.

[0101] The computer readable medium of the present application may include an autosampler module, Operate the system described in the white paper to automate the clog detection and removal workflow. Aspects of the present application include one or more of the following automated method steps: (e.g., the sample described herein) introduction system or a mass cytometry system including such a sample introduction system The sample introduction system includes a computer readable medium (such as a computer readable medium) (as shown in FIGS. 12 and 13). There are at least two pressure sensors (such as a push pump and a fill pump) During sample injection, measurements may be taken at least once every 5-6 seconds. The pressure may be obtained from the push pump (once per minute). The average pressure of the measurements or a predetermined number of consecutive measurements may be obtained. Repeated measurements (e.g., five measurements) When the number of measurements (number of measurements or more) reaches a set threshold, a blockage may be detected (e.g., (This may be reported to the user). Thereafter, sample acquisition (including, for example, sample injection) It will be paused and an automatic jam clearing routine will be initiated.

[0102] The unclogging routine may involve switching from injection mode to load mode, which This allows the push pump to pump the nebulizer while keeping the sample in the sample loop undisturbed. The first attempt to clear the blockage should be performed after about 1-2 minutes. This may be done by pulsing the clog with heavy aspirate and flush for 2 minutes. If the clog is not removed, A discrimination workflow may then be run (where, for example, two pressure sensors indicate that a blockage is present). in the sample loop or downstream of the sample loop, e.g., in the nebulizer line or (This is used to determine whether the jam is in the During the unclogging procedure, the flow rate level and and / or pulse rate (e.g., sample introduction, such as in the probes described herein). In certain embodiments, the amount of fluidic energy (e.g., input by a user) may be increased. Any of the above-described methods for increasing the pressure, flow rate, or pulse rate of the clog removal pressure (applied) For example, the unclogging pressure may be applied in steps from 5 psi to 40 psi. Greater than 5 psi, such as between 10 psi and 20 psi, or between 10 psi and 20 psi In certain embodiments (e.g., by a user) A predetermined maximum number of clogging events (entered by good.

[0103] An automated workflow allows acquisition to resume once the blockage has been successfully cleared. If it cannot be removed using an automated workflow, acquisition can be stopped. The sample is then returned to the sample tube (if the acquisition template specifies a return sample). The jam may then be manually cleared (e.g., a user manually clears the jam). (The software may be instructed to perform ICP analyzer

[0104] The ICP analyzer of the present application is described herein and is compatible with atomic analyzers such as MS or AES. The analyzer may include an optional ICP torch box coupled to the analyzer. The analyzer may be a TOF-MS or a magnetic The ICP analyzer may be a simultaneous mass analyzer, such as a sector MS. The ICP analyzer may be an imaging mass spectrometer. The ICP analyzer may be an external gas flow of less than 20 L / min. ICP analyzers may be operated to provide a dielectric barrier discharge under atmospheric conditions. The ICP analyzer can then measure the mass of the tagged cells or tissue. In certain embodiments, ICP analysis may be used to analyze biological samples such as sections. The analyzer may be used to analyze non-biological samples. System monitoring equipment

[0105] In certain embodiments, the mass cytometry system or sample introduction system described herein The system may be configured to perform one or more of the settings and / or real-time measurements described herein. System monitoring equipment that is configured to put system performance data into a database, such as The device may further comprise a computer used to operate the system. may operate independently or may communicate with such a computer. or, in addition, a computer may be configured to perform the functions described herein for the device. The device may be a wireless device. The database may be stored in a separate geographically remote location. The device may include system performance data from the system. May communicate with the base.

[0106] In certain aspects, the settings are determined from any parameters or values ​​entered by the user. In certain embodiments, the real-time measurement may be a system internal measurement, or measurements made by environmental sensors (e.g., humidity, pressure, and / or Real-time internal measurements may include one or more blockage events. (e.g., direct data from a pressure sensor or frequency of clogging), (e.g., sample fluid, or nebulizer gas, carrier gas, internal gas, and / or external gas, etc. the flow rate of one or more gases described in the document, the flow rate of one or more components (e.g., torch) the temperature (in the assembly or load coil), the voltage of the mass detector in the system, the current in the cone (e.g., sampling cone and / or skimmer cone of an ICP torch) The frequency or power at which the ICP load coil operates may include one or more of the following: Mass signals from standard beads, mass detector signals (e.g., detector damage or wear, cell events) detector signal above a predetermined threshold that may indicate a defect), reservoir (e.g., gas tank) the amount of gas pressure in the tank (or dewar that supplies gas to the system), the operating temperature of the system, It may operate during working hours, etc.

[0107] A laser ablation system (e.g., for histology) The plume (e.g., instead of suspended cells or suspended beads) may be used in the mass cytometer of the present application. One associated with the laser ablation system if configured to transfer Alternatively, multiple settings and / or real-time measurements may be provided. For example, laser abrasion configuration (e.g., output power, operating voltage, operating frequency, operating mode, and / or power dissipation) , ablation chamber fluid parameters (e.g., the amount of fluid that can be delivered to the ablation plume) a carrier and / or sheath gas flow rate to achieve the desired concentration), a cytometer as described herein; Humidity inside the laser ablation fluidics, measured by a mass detector The ablation plume width, etc.

[0108] In certain embodiments, system execution from one or more systems running on the same system may be performed. The raw data can be used to identify preventative maintenance needed on the system. In that regard, the user may be notified of the need for such maintenance (e.g., (You may be prompted to schedule such maintenance.) Maintenance is , instrument adjustment, instrument cleaning (e.g., sample introduction fluid, torch body or its tube, or cleaning of the sample cone downstream of the ICP torch), or specific parts (e.g. Laser, mass detector, gas reservoir, torch body or its tube, nebulizer or The replacement may be of another element (such as another element of the sample introduction fluidics described herein). usefulness

[0109] Aspects of the present system and method extend the life of ICP components and facilitate user operation. This can make the operation easier and / or reduce maintenance or running costs. The embodiments of the ICP load coil described herein have improved life spans compared to conventional coils. The removable torches described herein allow for easy maintenance and low-cost replacement (e.g., Easy access to the torch tube, which can be separated from the larger assembly to be replaced The vortex flow in the torch assembly allows for the external heat required to sustain the plasma. The gas flow may be reduced. The external ignition electrode allows for a removable torch. and / or may have a longer life than an electrode inserted into the torch body. Generally, a longer lifespan means that the part will last less time before it breaks or is severely damaged. It may be at least twice the lifespan, or at least five times the lifespan (for example, They operate at less than 90% or less than 80% efficiency, and efficiency is the energy required to operate the device. (The energy, or the intensity of the signal detected by an analyzer downstream of the ICP). The sample introduction system described in increases user exit time and automates sample introduction for multiple samples. Enables implementation.

Claims

1. a removable ICP torch holder assembly; an ICP load coil having an annular fin; an external ignition device that ignites plasma by dielectric barrier discharge; An inductively coupled plasma (ICP) torch box comprising:

2. (a) a removable inductively coupled plasma (ICP) torch holder assembly; 、 an inner torch body including an inner tube coupled to an inner tube base; an outer torch body including an outer tube coupled to an outer tube base; A torch that holds a removable inner torch base and a removable outer torch base. with the course holder an inductively coupled plasma (ICP) torch holder assembly comprising: (b) an ICP load coil disposed about the outer tube; a cylindrical coil; the cylindrical coil has an annular fin; The annular fin is continuous over at least one-eighth of one turn of the cylindrical coil, and The ICP load coil is primarily comprised of a metal or alloy that has a lower oxidation rate during operation than copper. A CP load coil; (c) an ignition device for igniting a plasma in an inductively coupled plasma (ICP) torch; So, an oscillator; a first high voltage transformer coupled to the first electrode; a second high voltage transformer coupled to the second electrode; The ignition device is configured to ignite the plasma by a dielectric barrier discharge. 、 The first electrode and the second electrode are disposed on opposite sides of the outer tube. an ignition device including a circuit located within 5 millimeters of the bulb; An inductively coupled plasma (ICP) torch box comprising:

3. 1. A removable inductively coupled plasma (ICP) torch holder assembly comprising: an inner torch body including an inner tube coupled to an inner tube base; an outer torch body including an outer tube coupled to an outer tube base; a torch holder configured to receive the inner torch base and the outer torch base; and When the torch holder holds the inner torch base and the outer torch base, The inner tube and the outer tube define an annular region. ) Torch holder assembly.

4. The tow of claim 3 , wherein the outer tube is detached from the outer tube base. Chi holder assembly.

5. 5. The method of claim 3, wherein the inner tube is detachable from the inner tube base. Torch holder assembly.

6. The torch holder defines an external gas inlet, the external gas inlet communicating with the annular region.

6. The torch holder assembly of claim 3, wherein the torch holder assembly is in communication with the other torch holder.

7. The outer tube base provides fluid communication between the external gas inlet and the annular region. and defining three or more holes arranged to generate a vortex flow. The torch holder assembly of claim 3 , wherein the torch holder assembly is oriented.

8. 8. The torch holder assembly of claim 7, wherein the outer tube base defines six holes. Nburi.

9. 9. The method of claim 7 or 8, wherein the hole is within 2.5 cm of the outlet of the inner torch tube.

2. The torch holder assembly according to claim 1 .

10. 10. The tow of claim 3, wherein the outer tube is not tapered. Chi holder assembly.

11. The inner tube base and the outer tube base are respectively separated from the torch holder.

11. The torch holder according to any one of claims 3 to 10, which can be individually removed. assembly.

12. The inner tube base and the outer tube base are permanently joined to one another.

12. The torch holder assembly of claim 3, wherein

13. The inner tube base and the outer tube base are configured to be connected by a fastener.

13. The torch holder assembly according to claim 3, wherein the torch holder assembly is configured as follows:

14. The torch holder includes a heat conducting element positioned to heat the injector.

14. A torch holder assembly according to claim 3, comprising:

15. 15. The method according to claim 3, wherein metal is the main material of the torch holder. Torch holder assembly.

16. 16. The method according to claim 3, wherein the aluminum alloy is the main material of the torch holder. The torch holder assembly according to any one of claims 1 to 4.

17. The outer tube does not have holes for extending electrodes into the outer tube.

17. The torch holder assembly of any one of claims 3 to 16.

18. a gas supply manifold configured to receive the torch holder; 18. A torch holder assembly according to any one of claims 3 to 17, comprising:

19. The torch holder has a mounting hole for aligning its position within the gas supply manifold. a pin that releases the torch from the gas supply manifold when the torch holder is twisted.

20. The torch holder assembly of claim 18, wherein the torch holder is removable.

20. The torch holder defines an external gas inlet, the external gas inlet extending through the annular region; and in fluid communication with an external gas flow region of the gas supply manifold.

20. The torch holder assembly of claim 19.

21. An inner tube; An outer tube; an outer tube base connected to the outer tube, the inner tube and the outer tube define an annular region; The outer tube base is positioned to allow external gas to enter the annular region. the three or more holes being oriented to generate vortex flow. An inductively coupled plasma (ICP) torch assembly.

22. 22. The torch holder assembly of claim 21, wherein the outer tube does not have a taper. Li.

23. An external ignition device for igniting a plasma in an inductively coupled plasma (ICP) torch. The ignition device is an oscillator; a first high voltage transformer coupled to the first electrode; a second high voltage transformer coupled to the second electrode; The ignition device is configured to ignite the plasma by a dielectric barrier discharge. External ignition device.

24. 24. The external ignition device of claim 23, wherein the plasma is under atmospheric conditions.

25. The external ignition device is configured to provide an AC output voltage to the first electrode and the second electrode.

25. The external ignition device according to claim 23 or 24, configured as follows:

26. The first high voltage transformer and the second high voltage transformer are connected to the circuit with opposite polarities.

26. The external ignition device of claim 25.

27. The apparatus provides a voltage difference of at least 1 kV between the first electrode and the second electrode.

27. An external ignition device according to claim 25 or 26, configured to provide

28. 28. The external ignition device of claim 27, wherein the voltage difference is from 2 kV to 100 kV.

29. 29. The external ignition device of claim 28, wherein the voltage difference is from 5 kV to 50 kV.

30. The maximum output voltage of the first high voltage transformer and the maximum output voltage of the second high voltage transformer are 30. The method of claim 23, wherein the voltage is from 2 kV to 100 kV, respectively. External ignition device.

31. 31. A method according to any one of claims 25 to 30, wherein the frequency of the AC voltage is less than 1 MHz. The external ignition device described.

32. 32. The external ignition device of claim 31, wherein the frequency is from 5 kHz to 100 kHz. Place.

33. 33. The external ignition device of claim 32, wherein the frequency is from 20 kHz to 40 kHz. Place.

34. The AC output voltage and the frequency of the AC output are sufficient to ignite a plasma at atmospheric pressure.

34. An external ignition device according to any one of claims 25 to 33.

35. the voltage difference and the frequency are sufficient to ignite a plasma within the ICP torch; 35. An external ignition device according to any one of claims 25 to 34.

36. The external ignition device is configured to ignite a plasma by an electrolytic breakdown discharge.

55. An external ignition device according to any one of claims 23 to 54.

37. The external ignition device is configured to ignite a plasma by sparking.

37. An external ignition device according to any one of claims 23 to 36.

38. The external ignition device is not configured to ignite a plasma by an arc discharge.

38. An external ignition device according to any one of claims 23 to 37.

39. 39. Any one of claims 23 to 38, wherein the external ignition device ignites a plasma at atmospheric pressure. Item 1. An external ignition device according to item 1.

40. 24. The electrode of claim 23, wherein the electrode is positioned within 5 mm of an outer torch wall of the ICP torch.

40. An external ignition device according to any one of claims 1 to 39.

41. 41. The method of claim 23, wherein the external ignition device does not include a Tesla coil. The external ignition device described.

42. The two high voltage transformers are of opposite polarity so that their output voltages are of opposite phase.

42. An external ignition device according to any one of claims 23 to 41, electrically connected to the circuit.

43. The maximum voltage output of the two high voltage transformers is from 5 kV to 100 kV.

43. An external ignition device according to any one of claims 23 to 42.

44. The circuit is configured such that the oscillator is connected to the first high voltage transformer and the second high voltage transformer, respectively. a voltage modulator configured to periodically match the natural resonant frequency of each of the sensors; 44. An external ignition device according to any one of claims 23 to 43.

45. 45. Any one of claims 23 to 44, wherein the circuit is a fixed AC output voltage and frequency. Item 1. An external ignition device according to item 1.

46. At least one of the output voltage and the frequency of the output voltage is controlled externally to the external ignition device.

46. ​​The method of claim 23, wherein the method is controllable by a computer. External ignition device.

47. The ICP torch further includes an ICP torch, and the electrode is disposed outside an outer torch body of the ICP torch.

47. An external ignition device according to any one of claims 23 to 46, arranged

48. The plasma flows between the portion of the outer torch body adjacent the first electrode and the portion of the second electrode.

5. Ignition by capacitance between the pole and the portion of the outer torch body adjacent the pole.

8. The external ignition device according to claim 7.

49. The ICP torch includes an inner torch body, and at least a portion of the outer torch body is The inner torch body is coaxial with the first electrode and the second electrode. and an axis passing through the second electrode is disposed so as to intersect with the portion of the outer torch body.

49. The external ignition device of claim 47 or 48,

50. An external ignition device that ignites a plasma in an inductively coupled plasma (ICP) torch by means of an electrical discharge. an external ignition device configured to supply AC current to at least one electrode; Place.

51. A cylindrical coil is provided. the cylindrical coil includes an annular fin; The annular fin is continuous over at least one-eighth of one turn of the cylindrical coil. The ICP load coil is mainly made of a metal or alloy that has a lower oxidation rate than copper during operation. Including, an ICP load coil.

52. 52. The ICP load coil of claim 51, wherein the annular fin is substantially planar.

53. 53. The ICP negative electrode of claim 51 or 52, wherein the metal or alloy is an aluminum alloy. Load coil.

54. 54. Any one of claims 51 to 53, wherein the metal or alloy has a lower electrical conductivity than copper. Item 2. The ICP load coil according to item 1.

55. 55. Any one of claims 51 to 54, wherein the metal or alloy has a lower melting point than copper. The ICP load coil described in

56. 56. Any one of claims 51 to 55, wherein the metal or alloy has a lower ductility than copper. The ICP load coil described in

57. The ICP load coil is formed by 3D printing the metal or alloy.

57. An ICP load coil as described in any one of claims 51 to 56.

58. an inner portion of the ICP load coil is thicker than an outer portion of the ICP load coil; Item 58. An ICP load coil according to any one of items 51 to 57.

59. 6. The method of claim 5, wherein the inner portion defines a circular cross section and the outer portion is the annular fin.

8. An ICP load coil according to claim 8.

60. The ICP load coil includes a first portion closest to a longitudinal axis of the cylindrical coil, the CP load coil includes a second portion furthest from the longitudinal axis of the cylindrical coil; The first portion is characterized by a first thickness, the first thickness being a thickness parallel to the longitudinal axis. The second portion is characterized by a second thickness, measured perpendicular to the first axis. the second thickness is measured perpendicular to a second axis parallel to the longitudinal axis, and the first thickness is 60. The ICP negative electrode of any one of claims 51 to 59, wherein the thickness is greater than the second thickness. Load coil.

61. The inner portion of the ICP load coil has a maximum diameter greater than the outer portion of the ICP load coil.

61. An ICP load coil as described in any one of claims 51 to 60 having a small diameter.

62. The metal or alloy may be thicker than if the ICP load coil did not include the annular fin.

62. Any one of claims 51 to 61, wherein the alloy is an alloy that reduces deformation during operation compared to copper. Item 2. The ICP load coil according to item 1.

63. 62. The metal or alloy is an alloy that reduces deformation compared to copper. An ICP load coil according to any one of claims 1 to 4.

64. 64. Any of claims 51 to 63, wherein the shape of the ICP load coil reduces deformation during operation. An ICP load coil according to any one of claims 1 to 4.

65. the deformation is perpendicular to the axis of the tube defined by the coil; 65. The ICP load coil according to claim 64.

66. 65. The method of claim 64, wherein the deformation is along the axis of the tube defined by the coil. Or an ICP load coil as described in 65.

67. 67. The ICP load coil of any one of claims 51 to 66, wherein the ICP load coil is solid state. CP load coil.

68. One or more annular fins are present along the majority of the three turns of the ICP load coil.

54. An ICP load coil as described in any one of claims 51 to 53.

69. The annular fin is continuous over at least two turns of the ICP load coil.

69. An ICP load coil as described in any one of claims 51 to 68.

70. 70. Any of claims 51 to 69, wherein the ICP load coil does not include a plurality of fins.

3. An ICP load coil according to claim 1.

71. an inner portion of the ICP load coil is thicker than an outer portion of the ICP load coil; Item 71. An ICP load coil according to any one of items 51 to 70.

72. The volume of the plasma sustained by the ICP load coil is less than 10 square centimeters.

72. An ICP load coil according to any one of claims 51 to 71, wherein:

73. 73. Any one of claims 51 to 72, wherein the length of the ICP load coil is less than 5 cm. Item 2. The ICP load coil according to item 1.

74. The distance from the innermost portion of the annular fin to the outermost portion of the annular fin is:

74. The method of claim 51, wherein the thickness is at least twice the thickness of at least a portion of the annular fin. An ICP load coil according to any one of claims 1 to 4.

75. 3D printed ICP load coil with annular fins.

76. A method of making an ICP load coil, comprising 3D printing the ICP load coil. A method comprising:

77. 77. The method of claim 76, wherein the ICP load coil comprises an annular fin.

78. a sample loop; a cylindrical agitator; the sample loop is wound around the cylindrical stirrer at least three times; 。

79. 79. The method of claim 78, wherein the sample loop forms a helix comprising at least five turns. Sample introduction device.

80. 8. The method of claim 7, wherein the spiral defines a cylinder having an axis within 15 degrees of normal to the axis of gravity.

10. The sample introduction device according to 9.

81. a nebulizer in fluid communication with the sample loop, the nebulizer having a nebulization chamber; the atomizing chamber is in fluid communication with the bar, the atomizing chamber being configured to deliver particles to the injector.

81. The sample introduction device of any one of claims 78 to 80,

82. Claim 10 further comprising a sample holder configured to hold a plurality of tubes.

82. A sample introduction device according to any one of claims 78 to 81.

83. 9. The sample holder according to claim 8, wherein the sample holder is configured to cool the plurality of tubes.

3. The sample introduction device according to claim 2.

84. The probe is then guided through the sample loop to the tube provided by the sample holder. a plurality of ports configured to transport a suspension of cells to and out of the nebulizer; 84. Any one of claims 78 to 83, further comprising a pump, a valve, a reservoir, and a tube. The sample introduction device according to claim 1.

85. configured to detect a drop in pressure when the probe becomes clogged with one or more particles.

85. The sample introduction device of claim 84, further comprising a pressure sensor configured therein.

86. storing a plurality of instructions that, when executed, control a processor to operate the sample introduction system; 86. The sample introduction device of claim 85, further comprising a computer readable medium storing thereon.

87. The instructions automatically reverse flow when a blockage is detected in the probe.

88. The method of claim 86 or 87, further comprising: dislodging the probe by The sample introduction device.

88. The processor may also be configured to resuspend the cells in the tube before transferring the cells.

87. The sample introduction device of claim 86, which operates a sample introduction system.

89. A removable ICP torch assembly according to any one of claims 3 to 22; 50. An external ignition device according to any one of claims 23 to 49; 80. An ICP load coil according to any one of claims 51 to 79; An inductively coupled plasma (ICP) torch box comprising at least one of:

90. A removable ICP torch assembly according to any one of claims 3 to 22; 50. An external ignition device according to any one of claims 23 to 49; 80. An ICP load coil according to any one of claims 51 to 79; An inductively coupled plasma (ICP) torch box comprising at least two of:

91. A removable ICP torch assembly according to any one of claims 3 to 22; 50. An external ignition device according to any one of claims 23 to 49; and an ICP load coil according to any one of claims 51 to 79. Plasma (ICP) torch box.

92. A removable ICP torch assembly according to any one of claims 3 to 22. Inductively coupled plasma (ICP) torch box.

93. a gas supply manifold that receives the removable ICP torch; 93. The inductively coupled plasma (ICP) torch box of claim 92.

94. 50. An inductively coupled plasma comprising an external ignition device according to any one of claims 23 to 49. (ICP) Torch Box.

95. 80. An inductively coupled printer comprising an ICP load coil according to any one of claims 51 to 79. Zuma (ICP) Torch Box.

96. The ICP torch box has an annular flow separated by a complete rotation. and a gas flow mechanism adapted to provide a gas flow across said annular fin so as to pass between portions of the fin.

96. The ICP torch box of claim 95, wherein the ICP torch box is functionalized.

97. 97. The method of claim 96, wherein the gas flow is exhausted from the ICP torch assembly during operation. ICP torch box included.

98. A removable ICP torch assembly according to any one of claims 3 to 22; 50. An external ignition device according to any one of claims 23 to 49; An inductively coupled plasma (ICP) torch box comprising:

99. The first electrode and the second electrode of the external ignition device are connected to the removable ICP 99. The IC of claim 98, wherein the IC is disposed on either side of the outer tube of the torch assembly. P torch box.

100. The ICP torch assembly according to any one of the preceding claims; configured to detect ionized atoms produced by the ICP torch. and an inductively coupled plasma (ICP) analyzer.

101. 89. A sample introduction system according to any one of claims 78 to 88.

100. An ICP analyzer according to claim 100.

102. a nebulization chamber disposed between the sample introduction system and the ICP torch assembly; 102. The ICP analyzer of claim 101,

103. 103. The IC of any one of claims 100 to 102, wherein the analyzer is a mass analyzer. P analyzer.

104. 104. The ICP analyzer of claim 103, wherein the analyzer is a time-of-flight mass spectrometer.

105. The analyzer includes a high-pass filter with a cutoff of at least 80 amu.

105. The ICP analyzer of claim 104.

106. 104. The ICP analyzer of claim 103, wherein the analyzer is a quadrupole mass spectrometer.

107. 104. An ICP analyzer as described in claim 103, wherein the analyzer is a magnetic sector analyzer.

108. 108. Any of claims 103 to 107, wherein the ICP analyzer comprises a particle introduction system.

10. The ICP analyzer of claim 1.

109. 109. The ICP analyzer of claim 108, wherein the ICP analyzer is a mass cytometer. 。

110. 101. The ICP analyzer of claim 100, wherein the ICP analyzer is an imaging mass spectrometer.

111. 110. The ICP analyzer is a laser ablation ICP mass spectrometer.

1. An ICP analyzer according to claim 1.

112. 112. The ICP analyzer of claim 111, wherein the ICP analyzer is an imaging mass cytometer. Analyser.

113. 103. The method according to any one of claims 100 to 102, wherein the analyzer is an optical emission spectrometer. CP analyzer.

114. Induction analysis of a sample using an ICP analyzer according to any one of the above ICP system claims. Methods for inductively coupled plasma (ICP) analysis.

115. 115. The method of claim 114, further comprising providing an external gas flow of less than 20 liters per minute. How to do it.

116. 116. The method of claim 114 or 115, comprising igniting the plasma by glow discharge. method.

117. The plasma is generated by applying an AC output voltage to two external electrodes of the ICP torch. The method of claim 116, wherein the ignition is performed by

118. 118. The method of claim 117, wherein the AC output voltage is from 2 kV to 100 kV.

119. 118. A method according to claim 118, wherein the frequency of the AC output voltage is from 2 kHz to 100 kHz. The method described.

120. 120. A method according to any one of claims 116 to 119, wherein the plasma is ignited under atmospheric conditions. The method described.

121. 121. The method of any one of claims 114 to 120, wherein the sample is a suspension of cells. Law.

122. 122. The method of claim 121, wherein the cells are labeled with a metal-tagged antibody.

123. 121. The method of any one of claims 114 to 120, wherein the sample is a tissue section.

124. 124. The method of claim 123, wherein the tissue section is labeled with a metal-tagged antibody.

125. 121. The method of any one of claims 114 to 120, wherein the sample is not a biological sample. 。

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