Inductively coupled plasma based atomic analysis system and method

Abstract

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 the 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 claims the benefit of priority to 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, and U.S. Provisional Patent Application No. 63 / 184,521, filed May 5, 2021, the entire disclosures of which are incorporated herein by reference for all purposes.

[0002] The technical field of this application relates to systems and methods for inductively coupled plasma (ICP) analysis, including mass cytometry. [Background technology]

[0003] 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 the 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 cytometry, such as ICP-MS. Summary of the Invention

[0004] The systems and methods of the present application include one or more removable ICP torch holder assemblies, an external ignition device, an ICP load coil with annular fins, particle suspension sample introduction fluidics, and an ICP analyzer thereof.

[0005] This specification refers to the following accompanying drawings, in which like reference numbers in different drawings indicate like or similar elements. [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] Particular aspects and configurations of the present disclosure are systems and methods for inductively coupled plasma (ICP) analysis, including mass cytometry, as further described herein. Inductively Coupled Plasma (ICP) Systems, Samples, and Methods

[0008] Inductively coupled plasma (ICP) is a type of plasma source that is energized by an electric current generated by electromagnetic induction (i.e., by a time-varying magnetic field). Industrial-scale applications of ICP include micromachining (e.g., etching or cleaning) or waste treatment. Such applications cannot generate plasma within an ICP torch, cannot use an ICP load coil, cannot operate under atmospheric conditions, and / or are not at a scale suitable for atomic analysis of samples (e.g., the generated plasma can be at least larger than the plasma of an ICP analyzer). Thus, the physics of industrial ICP differ from ICP analysis using an ICP torch and may be outside the scope of embodiments of the present disclosure. Systems and methods using ICP torches, such as ICP analyzers, are discussed herein.

[0009] An overview of ICP mass spectrometry (ICP-MS) is provided in Inductively Coupled Plasma Mass Spectroscopy, edited by Montaser and Akbar, John Wiley & Sons, 1998, including a discussion of vortex flow and ignition. The discussion of sample introduction and the ICP torch is similar to atomic emission spectroscopy (AES), also known as optical emission spectroscopy, which is also within the scope of this application. As used herein, atomic spectroscopy is the same as atomic analysis and can include atomic mass spectrometry (such as ICP-MS) or ICP-AES. Suitable samples include biological samples, geological samples, and products. In certain embodiments, biological samples can be fluids containing biological material and / or contaminants (e.g., metal toxins), or particles such as cells (e.g., in suspension or in tissue sections) or beads (used to assay biological material). Mass cytometry system and method

[0010] Aspects of the present application include ICP torch systems and methods for mass cytometry, which is the detection of mass tags within cells or beads by mass cytometry. Mass cytometry is discussed in U.S. Patent Application Publication Nos. 2005 / 0218319, 2016 / 0195466, and 2019 / 0317082, the entire contents of which are incorporated by reference. Mass cytometry can be of suspended particles (e.g., cells or beads) or particles generated from solid samples, such as laser ablation plumes generated from tissue sections. In suspension mass cytometry, a suspension of cells or beads containing mass tags is analyzed by atomic mass spectrometry. Imaging mass cytometry by laser ablation (LA) ICP-MS is described in U.S. Patent Application Publication Nos. 20160056031 and 20140287953, the entire contents of which are incorporated by reference. Imaging mass cytometry using LA-ICP-MS is described by Giesen, Charlotte et al. in “Highly multiplexed imaging of tumor tissues with subcellular resolution by mass cytometry.” (Nature methods 11.4 (2014): 417-422).

[0011] The mass tag may be a metal tag attached to an affinity reagent (e.g., an antibody, oligonucleotide, avidin, or other bioparticle that specifically binds to a target biomolecule). For example, a metal nanoparticle or metal chelating polymer may be attached (e.g., covalently attached) to an affinity reagent and then applied to the sample. Suitable mass tags are described in U.S. Patent Application Nos. 20040072250 and 20080003616, which are incorporated by reference in their entireties. In certain embodiments, some mass tags are not attached to affinity reagents, such as metal-containing drugs or histochemical stains.

[0012] Figure 1 is a schematic diagram of a standard mass cytometry workflow, and Figure 2 is a diagram of a standard mass cytometry system used in such a workflow. The suspension mass cytometry workflow can include labeling cells with mass tags (see cells labeled with labeled antibodies in the upper left of Figure 1). Particles, such as cells and / or beads, are transported by sample introduction fluidics from a sample container (e.g., a tube) held in a sample holder, through a nebulizer, and into a spray chamber. The spray chamber passes a carrier gas (i.e., an aerosol spray) around droplets released from the nebulizer. Unlike many other spray chambers, the spray chamber of a suspension mass cytometry system passes particles to an ICP torch box rather than to a waste outlet. An exemplary spray chamber suitable for mass cytometry is discussed in U.S. Patent Application Publication No. 2013 / 0181126, which is incorporated by reference in its entirety. The ICP torch box, further described herein, includes an ICP torch. Particles pass through the ICP plasma of the ICP torch, where they are atomized and ionized. A vacuum differential directs ionized atoms from the plasma through a sample cone (and one or more additional cones) to a mass analyzer. The ion optics of the mass analyzer are, in some instances, configured to filter ions. For example, if the mass tag comprises a heavy metal (such as a transition element or lanthanide) or its isotope, a mass filter (such as a deflector or radio frequency quadrupole) can be configured to act as a high-pass mass filter and remove ions below a certain mass. Because argon dimers, typically found in ICP plasmas, are 80 amu, a high-pass mass filter can remove ions with a cutoff of at least 80 amu. Such a high-pass filter is particularly useful when the mass analyzer is a time-of-flight detector (i.e., TOF-MS). Suitable mass analyzers include simultaneous mass analyzers such as TOF-MS or magnetic sector MS. In certain embodiments, the mass analyzer can be another analyzer, such as a quadrupole MS (QMS).Ions of the mass tag detected by the mass analyzer indicate the presence of the target of the affinity reagent to which the mass tag specifically binds. Compared to detecting fluorescent tags (which exhibit spectral overlap), there is relatively little crosstalk between mass channels in mass cytometry, so many mass tags can be distinguished in individual particles. In certain embodiments, multiple distinguishable mass tags (e.g., at least 20, at least 30, or at least 40 mass tags) are detected in a single cellular event.

[0013] In certain embodiments, the system can include a filter disposed at the inlet to the nebulizer. The filter can allow single cells to pass but prevent clusters of cells from passing through the nebulizer, for example, to reduce clogging at the nebulizer (such as at the tip of the nebulizer). The filter can comprise a mesh, such as a nylon mesh, or any material suitable for straining cells. The filter can allow particles smaller than the inner diameter of the nebulizer channel, for example, smaller than 80% or smaller than 50% of the inner diameter of the channel, to pass through. For example, the nebulizer can have an inner diameter of 200 microns or less, 150 microns or less, or 100 microns or less, for example, from 50 microns to 150 microns. The filter can have a cutoff greater than 25 microns, greater than 30 microns, greater than 80 microns, or greater than 100 microns, so that particles smaller than the cutoff can pass through the nebulizer. As shown in FIG. 2, the filter can be at the interface between the nebulizer and the sample introduction fluidics. The filter may be proximate to the sealed coupling between the sample loop of the sample introduction fluidics and the nebulizer. The inner diameter of the sample loop can be, for example, at least two times or at least five times larger than the inner diameter of the nebulizer channel. For example, the inner diameter of the sample loop can 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 removable from the nebulizer, for example, to replace the filter.

[0014] Although a mass cytometry workflow and system is described above, it is understood that a sampling system, such as a laser ablation system, can replace the sample introduction fluidics to transport the laser ablation plume particles to the mass analyzer. ICP Torch Box

[0015] Generally, an ICP torch box includes an ICP torch body and an ICP load coil disposed around the outer tube of the torch body. Optionally, the ICP torch box can further include an igniter and / or a gas supply manifold. The ICP torch body includes at least an inner tube and an outer tube, although the inner tube can be an intermediate tube disposed around the innermost tube. If the inner tube and the outer tube are separable, they can be described as belonging to separate inner and outer tube bodies, respectively. Embodiments of the present invention include ICP torch boxes, including specific devices and methods using the devices, such as any aspect of the ICP torch assembly (e.g., a removable ICP torch holder assembly), ICP load coil, and / or gas supply manifold described herein.

[0016] An exemplary conventional torch can provide the vortex flow through a single outer gas (i.e., plasma gas) inlet, or possibly two inlets, as shown in Figure 3. The outer gas inlet can extend from the torch tube and can be of the same material.

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

[0018] Figure 6 is a cross-sectional view of the torch assembly. Most components follow a radial pattern around the axis of the injector and torch. The annular fins of the ICP load coil are shown thinner than their length. The torch holder is shown positioned around the inner and outer torch bodies and within a gas supply manifold. As discussed further herein, gas flows from the manifold through the torch holder and into the inner or outer torch.

[0019] Figure 7 shows an enlarged view of the torch holder assembly of Figure 5. Figure 8 shows an exploded view of an exemplary torch holder assembly including an inner torch body having an inner tube coupled to an inner tube base and an outer torch body having an outer tube coupled to an outer tube base. The inner and outer bases are received by the torch holder and are detachable from the torch holder and their respective bases.

[0020] As described in one or more embodiments herein, an ICP torch box can include a torch holder assembly that provides vortex flow and / or is removable. For example, the outer torch body and, optionally, the inner torch body, can be removable from the torch holder, as described herein. Alternatively, or in addition, the outer tube can be removable from the outer tube base, and / or the inner tube can be removable from the inner tube base. Alternatively, or in addition, the torch holder assembly can be easily removable from the torch box (e.g., by twisting and / or pulling back), allowing for removal without having to remove the ignition electrode from the outer tube.

[0021] As described in one or more embodiments herein, the ICP torch box includes an external ignition device. The external ignition device may be located outside the outer tube and upstream of the ICP coil. The external ignition device may include two electrodes located on opposite sides of the outer torch tube. As further described herein, the mechanism of the external ignition device can be an electrical discharge, such as a dielectric barrier discharge. In certain embodiments, the external ignition device enables the design and operation of a removable torch, particularly for ICP-MS analysis where a torch ignition electrode is not suitable. The external ignition device of any embodiment(s) discussed herein can be combined with the torch holder assembly of any embodiment(s) discussed herein within the ICP torch box.

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

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

[0024] In certain embodiments, the atomization chamber and / or injector (a tube extending from the atomization chamber) may be described as part of the torch box, while the atomization chamber may be described as part of a sample introduction system to the torch box. Generally, the atomization chamber injector provides sample (e.g., aerosolized particles) concentrically into the inner tube (i.e., the innermost tube) of the ICP torch and may be inserted into the inner tube during operation.

[0025] In certain embodiments, the ICP torch box is part of an ICP analyzer. For example, the sample cone of a mass analyzer may be positioned to receive ionized atoms generated from the sample by the ICP plasma of the ICP torch. In such cases, the external ignition devices described herein allow for a removable design of the torch holder assembly as described herein.

[0026] This embodiment includes an inductively coupled plasma (ICP) torch box with a removable ICP torch holder assembly, an external igniter that ignites the plasma by dielectric barrier discharge, and / or an ICP load coil with annular fins, which may be 3D printed.

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

[0028] (1) A removable inductively coupled plasma (ICP) torch holder assembly including an inner torch body having an inner tube coupled to an inner tube base, an outer torch body having an outer tube coupled to an outer tube base, and a torch holder that holds the removable inner torch base and the removable outer torch base.

[0029] (2) An ICP load coil disposed around an outer tube, the ICP load coil including a cylindrical coil having an annular fin, the annular fin being continuous for at least one-eighth of a turn of the cylindrical coil, the ICP load coil being comprised primarily of a metal or alloy having an oxidation rate lower than that of copper.

[0030] (3) An ignition device for igniting a plasma in an inductively coupled plasma (ICP) torch, the ignition device comprising a circuit having an oscillator, a first high-voltage transformer coupled to a first electrode, and a second high-voltage transformer coupled to a second electrode, the ignition device being configured to ignite the plasma by a dielectric barrier discharge, the first electrode and the second electrode being positioned outside an outer tube and within 5 millimeters of the outer tube. Torch Holder Assembly

[0031] Various embodiments of the present application relate to torch assemblies and their uses, as exemplified by the aspects discussed below. Such aspects may be combined in any operable combination.

[0032] Aspects of the present torch assembly include a removable ICP torch holder assembly, such as an ICP torch box having a torch assembly, and a method of use. The removable torch can be removed from the torch box (e.g., from the gas supply manifold of the torch box) by pulling the torch holder of the removable torch backward (e.g., opposite the direction of gas flow through the torch during operation). In certain aspects, the removable torch can be secured by a gas source manifold within the ICP torch box and can be twisted (e.g., twisting the pin of the removable torch from its locked position in the gas flow manifold) before removing the torch assembly. As described herein, in certain aspects, the ICP torch box includes an external ignition electrode such that no electrode is inserted into the outer tube of the ICP torch assembly, thereby facilitating removal of the torch assembly from the torch box (e.g., removal in a single step). This removal can facilitate maintenance of the ICP torch (e.g., cleaning or replacement of one or more torch tubes).

[0033] Alternatively, or in addition, the torch holder of a removable ICP torch assembly may be configured to receive (and thus be removable from) at least the outer tube base coupled to the outer tube. Similarly, the torch holder may be configured to receive the inner torch base coupled to the inner tube. Alternatively, the torch holder may be permanently attached (e.g., machined as the same part or glued together) to the inner tube and / or the inner tube base coupled to the inner tube. The torch assembly may be configured to coaxially align the inner and outer tubes. O-rings may be used to provide a gas-tight seal between the inner and / or outer tube bases and the torch holder. In certain embodiments, the inner and outer tube bases may permanently reference each other directly (e.g., machined as the same part or glued together), such as threads on one of the inner and outer torch bases screwing into the other torch base, or may be configured to join directly to each other with fasteners. The inner and outer bases, which reference each other directly, can be removed from the torch holder as a single unit.

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

[0035] In certain aspects, 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 precipitation during operation. The outer torch tube is less expensive than the outer torch base, and this removable feature can reduce maintenance costs. Similarly, in certain aspects, 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 precipitation during operation. The outer torch tube is less expensive than the outer torch base, and this removable feature can reduce maintenance costs.

[0036] The torch assembly can be configured to allow an external gas (also known as a plasma gas) to enter the outer tube of the torch assembly.

[0037] Previously, one hole, and sometimes two holes, were used to introduce the vortex flow. Often, the holes were defined by a tube of the same material as the outer torch tube (e.g., glass, quartz, or ceramic) and may be located on the side of the torch tube. Such designs can complicate removal of the touch assembly (e.g., from the gas supply manifold) and / or removal of the outer torch tube from the torch holder of the torch assembly. Furthermore, such designs cannot accommodate several such holes.

[0038] In certain embodiments, the outer torch body of the torch assembly may include a plurality of holes (e.g., three or more holes, such as six holes) positioned and oriented to direct external gas into the annular region between the inner and outer tubes, creating a vortex therein. The holes may be radially symmetric or obliquely oriented (i.e., not extending radially outward from the axis of the cylinder defined by the outer tube). The holes may be in the outer torch base of the outer torch body. When the outer torch body and the inner torch body are held by a torch holder (e.g., at their respective bases), the holes in the outer torch base can allow fluid communication from an external gas inlet (e.g., a port) in the torch holder to the annular region between the inner and outer tubes. The torch holder is in turn held by a gas supply manifold, providing fluid communication between the external gas inlet (e.g., a port) of the gas supply manifold and the external gas inlet of the torch holder. In certain aspects, the external torch base, the torch holder, and the gas supply manifold are all constructed primarily of metal.

[0039] Alternatively, or in addition, the torch holder may provide fluid communication for an internal gas (known as an auxiliary gas) to pass from a gas supply manifold to the internal torch body. Alternatively, or in addition, the gas supply manifold may provide fluid communication for a carrier gas (also known as an atomization chamber gas or an aerosol gas) to flow from a carrier gas inlet (e.g., port) in the gas supply manifold and into the atomization chamber. Thus, the gas supply manifold may have separate ports for one or more of the carrier gas, the internal gas, and the external gas. 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 include argon. Alternatively, the external gas may include a gas other than argon or helium, such as nitrogen.

[0040] The fluid communication described above allows the tube to be positioned away from the torch holder assembly (e.g., not directly coupled to the torch holder or external torch body), allowing the torch holder to be more easily removed from the torch box (e.g., its gas supply manifold).

[0041] In certain aspects, a removable ICP torch holder assembly includes an inner torch body having an inner tube coupled to an inner tube base, an outer torch body having an outer tube coupled to an outer tube base, and a torch holder configured to receive the inner torch base and the outer torch base, the inner tube and the outer tube defining an annular region when the torch holder holds the inner torch base and the outer torch base.

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

[0043] The torch holder may be an external gas inlet, the external gas inlet being in fluid communication with the annular region. The outer tube base may define three or more holes (e.g., six holes) arranged to provide fluid communication between the external gas inlet and the annular region, with the three or more holes oriented to create a vortex. The holes may be positioned near the plasma so that the vortex does not dissipate and only a small gas flow is required. For example, the holes may be within 2.5 cm of the outlet of the inner torch tube. The outer tube may not include a taper. Instead, the outer tube defines a taper, which accelerates the external gas toward the plasma.

[0044] The inner tube base and the outer tube base may each be removably separated from the torch holder or may be referenced to each other (e.g., the inner tube base and the outer tube base may be permanently joined to each other, and the inner torch base and the outer torch base may be configured to be connected by fasteners such as screws).

[0045] The torch holder includes a heat conducting element positioned to heat the injector to reduce precipitation and / or clogging within the injector.

[0046] In certain embodiments, a metal or alloy, such as an aluminum alloy, is the primary material of the torch holder. The outer tube does not have a hole for the electrode to extend into the outer tube.

[0047] The torch box may further include a gas supply manifold configured to receive the torch holder. For example, the torch holder can include a pin for aligning its position within the gas supply manifold, and the torch holder can be removed from the gas supply manifold by twisting the torch holder. The torch holder can define an external gas inlet, the external gas inlet in fluid communication with the annular region and the external gas flow region of the gas supply manifold.

[0048] In certain aspects, an inductively coupled plasma (ICP) torch assembly comprises an inner tube, an outer tube, and an outer tube base coupled to the outer tube, the inner tube and outer tube defining an annular region, the outer tube base defining three or more holes positioned to allow external gas to pass through the annular region, the three or more holes oriented to create a vortex flow.

[0049] FIG. 9 is a cross-sectional view of an exemplary torch holder assembly. As shown, the torch holder holds a torch base. The torch base can be a single piece (e.g., a continuous piece of plastic, or an outer base and an inner base permanently affixed together) or can include separate inner and outer bases that can be separated from one another. By having the inner base separable from the outer base, the inner torch body (inner base and inner tube) can be separated from the outer torch body for cleaning or replacement. For example, the outer torch requires more regular maintenance due to heating or material precipitation from plasma irradiation.

[0050] As shown in FIG. 9, the torch base can include one or more external gas inlets (e.g., at least two, at least three, at least four, at least five, such as six, eight, or ten external gas inlets), each communicating with a separate hole that directs external gas from a gas supply manifold (not shown) into the annular space between the inner and outer tubes of the torch-holding assembly. The holes can be positioned directly adjacent to the outer tube so that the external gas is injected directly into the space between the inner and outer tubes. The holes are angled to provide a vortex flow.

[0051] In certain embodiments, the outer tube and the inner tube may be permanently attached to the torch base (e.g., the outer tube to the outer torch base and the inner tube to the inner torch base). For example, the outer tube may be affixed to the outer torch base with an adhesive (e.g., a heat-resistant adhesive). A gap between the outer wall of the outer tube and the torch base (e.g., the outer torch base) allows the adhesive to bond the two and allows the outer tube and / or adhesive to expand during heating (e.g., while the torch is operating). In certain embodiments, the gap may have a width greater than 0.2 mm, greater than 0.5 mm, or greater than 1 mm, such as 2 mm. Such a permanent bond and / or gap allows the outer tube to be bonded to the torch base without an interference fit. The length of overlap between the outer tube and the torch base may be short so that the vortex of the external gas injected into the outer torch by the hole has a minimum distance to dissipate or reduce before the plasma sustained by the torch. This length may be, for example, less than 10 mm, or less than 5 mm. external ignition

[0052] An ICP load coil can sustain the plasma within the ICP torch, and an ignition electrode, such as a Tesla coil electrode, is typically used to ignite the plasma by a process described as arcing or sparking. The spark from the ignition electrode can be generated by a conductive element (e.g., a grounded conductive element) such as an ICP coil that is in close proximity to the plasma. ICP torches can operate at atmospheric pressure (e.g., between 0.9 and 1.1 atm). In ICP-AES, the plasma is directly analyzed by spectroscopy, so the electrode can be positioned downstream of the outer torch tube (e.g., downstream in the gas flow through the torch). However, in ICP-MS, the ignition electrode sparks using the sample cone rather than any conductive component in close proximity to where the plasma is sustained, and the sample cone, which is positioned in close proximity to the outlet of the outer tube, can obstruct the ignition electrode in the torch vortex. This problem has previously been solved by inserting an ignition electrode through the external torch tube (e.g., as described in U.S. Pat. No. 4,766,287). However, inserting the electrode through the torch body can prevent the torch assembly from being easily removed for maintenance (e.g., for cleaning or replacement of one or more torch tubes). Thus, in certain aspects, the external ignition devices and methods of the present application can be combined with the removable torches described further herein.

[0053] External ignition allows for a removable torch with a metal assembly for the insert / holder and gas supply. The inventors found that the proximity of metal parts within the torch assembly precluded the use of an internal spark electrode for ignition of the mass cytometer; the spark would fly toward the rear of the base of the torch rather than toward the sample collection interface. The external electrode placement positions the electrode out of the way and generates the discharge near the plasma volume. Initially successful experiments used a single electrode, with the discharge directed toward the metal base of the torch tube. AC pulses generated using a Tesla coil external to the torch can also be useful for plasma formation. The inventors also identified a higher-energy voltage AC generator with a symmetrical output. This provided reproducible and reliable ignition for easy manufacturing.

[0054] The external igniter ignites a plasma in the inductively coupled plasma (ICP) torch by an electrical discharge, such as an electrostatic barrier discharge. The external igniter may be configured to supply an AC current to at least one electrode.

[0055] The exemplary external breakdown circuit shown in FIG. 9 includes an oscillator that generates an AC signal having a frequency of approximately 30 kHz, modulated by a low-frequency (100 Hz) modulator within a range of ±2 kHz around the center point. The modulated signal is then amplified in a power bridge circuit that drives two transformers T1 and T2. Transformers T1 and T2 are connected with opposite polarity in the circuit so that their respective output voltages are antiphase (180 degrees out of phase with each other). Transformers T1 and T2 operate near their natural oscillations, effectively amplifying the maximum voltage to 30 kV, which is then applied to the ignition electrode attached to the torch assembly. Of note, transformers using frequencies other than 30 kV (e.g., anywhere from 1 kV to 100 kV) can be combined with oscillators operating at similar frequencies. The intermediate frequency of the oscillator was chosen to be 30 kHz, as this is approximately the resonant frequency of the amplifier transformers T1 and T2 (which in principle are identical to each other, but may be affected by manufacturing tolerances).

[0056] Due to manufacturing tolerances, the actual resonant frequency of the transformer cannot be specified exactly at 30 kHz, but it is important to drive the transformer with an AC current at its actual resonant frequency to achieve maximum output voltage for ignition.

[0057] To ensure that the transformer's resonant frequency is matched, a 100 Hz modulator is used that applies a triangular ramp signal to the control input of a 30 kHz oscillator, constantly sweeping the modulator's frequency from approximately 28 kHz to 32 kHz. Each time the transformer's natural frequency is matched momentarily, the transformer's output voltage applied to the ignition electrode momentarily increases to 30 kV, initiating a discharge in the argon gas column within the torch. Once initiated, the discharge continues even when the frequency passes the resonant peak. The voltage required to maintain the discharge is significantly lower than that required to initiate a discharge in argon gas.

[0058] The ignition circuit is supplied with a constant voltage of 24Vdc from the host device, and its operation is triggered to turn on / off by a signal from the plasma management system within the device.

[0059] In this particular embodiment shown in Figure 9 and described above, all internal operation of the ignition module was fixed, and no external software control was required other than on-off operation. However, in other applications, it may be desirable to have a computer control the output voltage, modulation frequency, range, etc. A physical computer interface providing such control could easily be 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] No low frequency modulation (e.g., if the oscillator frequency can be precisely matched to the transformer resonance by providing a trimming potentiometer on the circuit board)

[0063] Using only one transformer, the output is single-ended by grounding the second electrode. For example, the second electrode may be omitted entirely, and the grounded end of the discharge may be provided to another grounded metal part of the torch.

[0064] The on / off control lines could be omitted and the on / off operation could be controlled by switching 24Vdc as needed.

[0065] The above alternatives are not intended to be limiting, but instead to illustrate some aspects that may be varied.

[0066] In certain embodiments, the external ignition device includes at least one electrode. For example, the external ignition device can include a single external electrode paired with an insulating element (e.g., a torch tube) in the path of the discharge, and the mechanism of ignition can be described as a dielectric barrier discharge (DBD). Similarly, two electrodes positioned on opposite sides of the torch tube, as described herein, would provide ignition by a dielectric barrier discharge (DBD). A DBD is an electrical discharge between two electrodes separated by an insulating dielectric. In the above-described embodiment of two electrodes positioned on either side of the outer torch tube, the torch tube is the insulating dielectric, and the discharge crosses the flow of gas through the torch. More generally, the mechanism can be described as an electrolytic breakdown discharge (or electrical discharge), and examples of discharge to a metal part (e.g., a grounded part of an ICP torch) such as the ICP coil or the torch tube base are described. In some cases, the mechanism of ignition can be described as an electrostatic discharge.

[0067] In some embodiments, the mechanism of ignition by an external ignition electrode can be described as a glow discharge (e.g., as opposed to an arc discharge). However, as used herein, such a glow discharge exists at or near atmospheric pressure and requires different external ignition equipment and operation than if the plasma were ignited at constant pressure. The plasma density generated by an external ignition electrode can be the much lower of an arc discharge and can approximate the plasma density of a glow discharge.

[0068] External ignition as described herein is distinct from ignition schemes that use one or more of an arc discharge, a high voltage breakdown, a DC current, a Tesla coil, and / or direct contact between an ignition electrode and the gas to be ignited in the plasma.

[0069] In certain embodiments, an external ignition device for igniting a plasma in an ICP torch includes an oscillator, a first high-voltage transformer circuit coupled to a first electrode, and optionally, a second high-voltage transformer coupled to a second electrode, the ignition device configured to ignite the plasma by a dielectric barrier discharge. The plasma can be ignited at atmospheric conditions (e.g., within 10% of 1 atm) or at least above 100 Torr. The external ignition device is configured to provide an AC output voltage to at least one electrode, such as the first electrode and the second electrode, where, for example, the first high-voltage transformer and the second high-voltage transformer are connected in a circuit with opposite polarity. The device can provide a voltage difference of at least 1 kV between the first electrode and the second electrode, such as 2 kV to 100 kV, 5 kV to 50 kV, or 20 kV to 40 kV. The maximum output voltage of the high voltage transformer of the circuit is from 2 kV to 50 kV, such as from 10 kV to 40 kV. In certain embodiments, the maximum voltage of the high voltage transformer of the circuit is less than 1 MHz, such as less than 200 kV. In certain embodiments, the frequency of the AC voltage is less than 1 MHz, such as from 5 kHz to 100 kHz or from 20 kHz to 40 kHz. The AC output power and / or its frequency is sufficient to ignite a plasma at atmospheric pressure (e.g., within 10% of 1 atm).

[0070] The external ignition device may be configured or operative to ignite the plasma, for example, via an electrolytic breakdown discharge due to dielectric barrier breakdown. The plasma may be ignited by capacitance between a portion of the outer torch body adjacent the first electrode and a portion of the outer torch body adjacent the second electrode.

[0071] In certain aspects, the external igniter may not be configured or operative to ignite the plasma by sparking or by arcing (e.g., arc discharge). The external igniter may not include a Tesla coil.

[0072] The device may include two electrodes. The electrodes may be positioned outside the outer torch, for example, within 5 millimeters or 3 millimeters of the outer wall of the ICP torch. The two high-voltage transformers are connected to the circuit with opposite polarities so that their respective output voltages are in opposite phase. The two high-voltage transformers may have a maximum power output of 5 kV to 100 kV. The circuit further includes a voltage modulator configured to periodically tune the oscillator to the natural resonant frequency of each of the first and second high-voltage transformers. The circuit provides a constant AC output voltage and frequency. Alternatively, at least one of the output voltage and its frequency can be controlled by a computer external to the external ignition device.

[0073] The apparatus can be part of an ICP torch, and the electrode is disposed outside an outer torch body of the ICP torch. The ICP torch can include an inner torch body, at least a portion of the outer torch body being coaxial with the inner torch body, and the first and second electrodes are disposed such that an axis passing through the first and second electrodes intersects a portion of the outer torch body. ICP Load Coil

[0074] Aspects of the present application include an ICP load coil (ICP coil) for sustaining a plasma (e.g., when operated with an AC current at RF frequencies). The ICP load coil can be cylindrical, meaning it has a helical shape that defines a cylinder. Often, the coil has three turns. The ICP load coil can be located within a torch box that surrounds the outer torch tube.

[0075] Standard cylindrical ICP load coils are made from copper and are either solid wire or hollow (tube) in shape. In contrast, embodiments of ICP load coils in the present application can include materials other than copper and shapes other than solid wire or hollow tube. The ICP load coil materials and shapes discussed herein can reduce coil aging, for example, reducing heating, oxidation, and / or deformation during operation to sustain a plasma.

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

[0077] In certain embodiments, the ICP coil can include annular fins. An annular fin is understood to be a fin that diverges as it extends from the central axis. Such fins are difficult to form from machined metal because they may require deformation to diverge (stretch) in this direction when bent to form the coil. Thus, the ICP coil can have 3D-printed annular fins. Annular fins can increase cooling of the ICP coil during operation (and thereby reduce oxidation). An annular fin can extend along at least one-eighth of one turn of the ICP coil, and can extend along two or more turns. The inventors have found that continuous fins do not reduce the efficiency of the ICP coil. Metrics such as operating frequency, inductance, and plasma maintenance remain unchanged compared to designs with gaps in the annular fin. The absence (or infrequent gaps) of gaps in the annular fin can provide additional structural stability (e.g., to withstand deformation during operation as described herein). Those skilled in the art may be concerned that current will flow partially along the outside of a coil with continuous annular fins, thereby reducing the efficiency of the ICP and interfering with the operation of the coil.

[0078] The materials and / or geometry of the ICP coil can improve the structural stability of the coil, allowing for a more robust manufacturing process and / or longer lifespan. For example, the ICP coils of the present application can survive at least 5,000 hours, at least 10,000 hours, at least 20,000 hours, or at least 50,000 hours of operation (e.g., without interruption, without a greater than 10% decrease in ICP efficiency, without a greater than 10% decrease in signal detected by a mass analyzer, or without deviation from frequency or induction tolerances).

[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 can have many additional properties compared to copper, as described herein.

[0080] In certain embodiments, the ICP load coil with annular fins is a cylindrical coil. The annular fin may be continuous over at least one-eighth of a turn of the cylindrical coil, for example, along at least two turns of the coil. The ICP load coil may be primarily comprised of a metal or alloy that oxidizes at a lower rate than copper during operation or over the life of the device. The annular fin may be substantially planar. The inner portion of the ICP load coil is thicker than the outer portion of the ICP load coil. The inner portion defines a circular cross section, and the outer portion is an annular fin. The inner portion of the ICP load coil may have a minimum diameter that is larger than the outer portion of the ICP load coil. The ICP load coil may be solid. The ICP load coil may not include multiple annular fins. One or more annular fins may be present along most of the three turns of the ICP load coil. The distance from the innermost portion of the annular fin to the outermost portion of the annular fin may be at least twice the thickness of at least a portion of the annular fin.

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

[0082] The metal or alloy may be an aluminum alloy. The metal or alloy may have at least one of a lower electrical conductivity than copper, a lower melting point than copper, or lower ductility than copper. The ICP load coil may be formed by 3D printing, casting, or machining the metal or alloy. The metal or alloy is an alloy that reduces deformation compared to copper compared to when the ICP load coil does not include the annular fin. The metal or alloy is an alloy that reduces deformation compared to copper, for example, perpendicular to the axis of the cylinder defined by the coil and / or along the axis of the cylinder defined by the coil.

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

[0084] Embodiments of the present invention include sample introduction systems, such as particle (e.g., cell or bead) introduction systems or laser ablation systems. Particle introduction systems for mass cytometry include multiple components, such as a sample holder, a probe for accessing the sample in the sample holder, and / or sample introduction fluidics for introducing the sample (e.g., a suspension of cells) into a spray chamber upstream of an ICP torch box. In certain embodiments, the sample holder can be configured to hold multiple samples, such as at least 4, at least 8, or at least 12 samples. The sample holder can be movable (e.g., a carousel) to present samples to the probe. Automated acquisition of multiple samples can take several hours. Thus, embodiments of the present application include sample introduction systems for maintaining a suspension of cells. Such a system may include components used in flow cytometry mixing applications, such as the system described by Al-Halhouli, Ala'aldeen et al. in "Passive micromixers with interlocking semi-circle and omega-shaped modules: Experiments and simulations" (Micromachines 6.7 (2015): 953-968). In certain embodiments, the radius of the loop and the flow rate of the sample through the loop result in a low Dean number (i.e., turbulent flow).

[0085] In certain embodiments, the sample introduction fluidics may include a sample loop in direct or indirect fluid communication with the probe and the nebulization chamber. The sample loop may comprise multiple turns (e.g., a helix defining a cylinder). The axis of the cylinder defined by the helix may be oriented horizontally (e.g., within 15 degrees of perpendicular to the axis of gravity) to reduce the portion of the loop on which cells (or other particles) may settle. Optionally, the sample loop may further be disposed around a cylindrical agitator that contacts the loop, which maintains a suspension of particles within the loop during transport to the nebulization chamber.

[0086] The sample introduction fluidics can be coupled to one or more reservoirs. The reservoirs 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 valve system, such as a rotary valve system, that directs fluid flow from the reservoirs and the sample tubing (sample loop). As described herein, the sample is directed through a nebulizer, a spray chamber, and an injector to the ICP plasma, where stage particles (e.g., cells and / or beads) in solution are atomized and ionized prior to analysis by an analyzer (e.g., a simultaneous mass analyzer). In certain embodiments, the injector is a heated injector. A description of heated injectors and cell capture solutions is provided in U.S. Patent Application Publication No. 20190317082, incorporated herein by reference. In certain embodiments, the cell capture buffer may include a salt in solution, where the salt is carbon-free and metal-free (e.g., no elements with an atomic weight greater than 0).

[0087] The sample introduction system may include at least two syringes that alternate between wash and sample introduction functions. For example, a first pump may function to draw sample, beads, wash solution, or cell capture solution into the sample loop. A second pump is a multi-port valve that connects the syringes to a waste bottle, a cell capture solution reservoir, and a switching valve. Its primary function is to push sample, beads, solution, and / or cell capture solution from the sample loop to the nebulizer, and to push cell capture solution from the reservoir bottle directly to the nebulizer.

[0088] The sample introduction system may include at least two rotary valves, such as a selector valve and a switching valve. The selector valve may include a port connected to a probe, a reservoir, and / or a void port used to draw air bubbles. The position of the selector valve may determine where one pump draws from. The switching valve may switch the sample loop between a load position and an injection position. When the switching valve is in the load position, the sample loop may be filled via a fill pump. In the injection position, the sample loop is aligned with the nebulizer, and its contents are pushed into the nebulizer via a second pump. The first pump may be a fill 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 an agitator for the sample loop of Figure 10. The spiral of the sample loop of Figure 10 fits around the cylinder of the agitator of Figure 11.

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

[0092] A cell sample in the carousel may be aspirated via the first pump. For example, as shown in FIG. 12, the sample may be drawn through the probe, toward the selection valve, and into the sample loop with the switching valve 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 may be pushed via the first pump towards the nebulizer, as shown in FIG. 13. A sample capture solution may be used as a carrier to push the sample towards the injector and into the system. An air bubble may be placed between the cell capture solution and the sample to prevent mixing at the interface between them.

[0093] In certain embodiments, the sample loop defines a tightly wound sample loop based on a narrow radius of curvature to achieve a low Dean number, as described in a 2015 Micromachines article. The sample loop may be aligned with a horizontal axis of rotation to minimize settling time for the cell (or particle) flow due to gravity (at the bottom of the loop). An agitator (vibrating rod) may be used as an additional stimulus to prevent cells from settling within the sample loop. However, in contrast to its use in other applications, the vibration does not cause material to settle or clump during transport; it simply adds agitation to the flow to maintain cell suspension within the carrier fluid. This works in the same way as having a summer student periodically tap the sample loop, but without the expense.

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

[0095] The sample introduction device of the present application may include a sample loop, the sample loop winding at least three times. The winding may be around a cylindrical agitator. The sample loop may form a spiral with at least five windings. The spiral defines a cylinder having an axis within 15 degrees of a direction perpendicular to the axis of gravity.

[0096] The sampling system and method of use (e.g., implemented by a computer storage medium) can improve productivity through built-in clog detection, automatic clog declogging (e.g., unclogging), and / or shutdown.

[0097] In certain embodiments, the sample system runs as part of the pellet resuspension workflow and before loading the sample. During mixing, the probe line and fill pump work together and may function like a pipette, aspirating and dispensing a volume (e.g., between 25 μL and 250 μL) multiple times within the tube. The user can select a mixing force value (e.g., between 1 and 10) to define how quickly this volume is dispensed into the sample tube (e.g., 1 represents the least amount of mixing force, and 10 represents the most amount of mixing force). This parameter may be set to a default value (e.g., 5).

[0098] During sample acquisition, the probe line may agitate the sample to prevent settling. The user may select an agitation speed value (e.g., between 0 and 9) to define how fast the probe line rotates (e.g., 0 stops agitation, 9 is the fastest speed). This parameter may be set to a default value (e.g., 5). During acquisition, the inside and / or outside of the probe line may be washed if a pre-wash cycle is selected by the user or executed by the computer-readable medium.

[0099] The default sample loop vibration setting helps to alleviate cells that have settled in the sample loop.

[0100] The device may further include a nebulizer in fluid communication with the sample loop. The nebulizer may be in fluid communication with the nebulizer chamber. The nebulizer chamber may be configured to pass particles through the injector. The device may include a sample holder configured to hold one or more samples. The sample holder may be configured to hold a plurality of tubings, for example, the sample holder may be configured to cool the plurality of tubings. The device may include a plurality of pumps, valves, reservoirs, and / or tubing configured to transfer a suspension of cells from the nebulizer, through the sample loop, and from tubing provided by the sample holder to the probe. The device may optionally further include a pressure sensor that detects a decrease in pressure when the probe becomes clogged with one or more particles, and a computer-readable medium storing a plurality of instructions that, when executed, control a processor to operate the sample guidance system, for example, the instructions may include automatically reversing flow (or adjusting pressure, e.g., increasing pressure) when a clog is detected in the probe, thereby clearing the probe. The processor can operate the sample guidance system to resuspend the cells within the tube prior to transferring the cells.

[0101] The computer-readable media of the present application may include an autosampler module to operate the systems described herein and automate the clog detection and removal workflow. Aspects of the present application include one or more of the following automated method steps and computer-readable media (e.g., of a sample introduction system described herein or a mass cytometry system including such a sample introduction system) configured to perform such steps. There may be at least two pressure sensors within the sample introduction system (such as a push pump and a fill pump as shown in FIGS. 12 and 13). During sample injection, measurements may be taken from the push pump (e.g., at least once per minute, such as once every 5-6 seconds). When the average pressure of the measurements or a predetermined number of consecutive measurements (e.g., two or more measurements, three or more measurements, five or more measurements, such as five measurements) reaches a set threshold, a clog may be detected (e.g., and reported to a user). Sample acquisition (e.g., including sample injection) is then paused, and an automatic clog removal routine is initiated.

[0102] The clog-clearing routine may include switching from injection mode to load mode, allowing the push pump to flush the nebulizer while keeping the sample in the sample loop undisturbed. An initial attempt to clear the clog may be made with a large aspirate and flush pulse (e.g., for about 1-2 minutes). If the clog is not cleared, a clog-identification workflow may be run (where, for example, two pressure sensors are used to determine whether the clog is in the sample loop or downstream of the sample loop, e.g., in the nebulizer line or the nebulizer). The clog-clearing procedure may then focus on the location of the clog. During the clog-clearing procedure, the flow rate level and / or pulse rate may be increased (e.g., in the sample introduction fluidics, such as in the probes described herein). In certain aspects, a clog-clearing pressure (e.g., input by a user) may be applied at any of the steps described above that increase the pressure, flow rate, or pulse rate. For example, the clog removal pressure may be greater than 5 psi, greater than 10 psi, greater than 15 psi, such as between 5 psi and 40 psi, or between 10 psi and 20 psi. In certain embodiments, a predetermined maximum number of clog events (e.g., entered by a user) may trigger the end of sample acquisition.

[0103] If the clog is successfully cleared using the automated workflow, acquisition can be resumed. If it cannot be cleared using the automated workflow, acquisition can be stopped and the sample returned to the sample tube (if a return sample is specified in the acquisition template). The clog may then be manually cleared (e.g., the user may be prompted by the software to manually perform the clog removal). ICP analyzer

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

[0105] In certain embodiments, the mass cytometry systems or sample introduction systems described herein may further include a system monitoring device configured to compile system performance data, such as one or more settings and / or real-time measurements described herein, into a database. The device may operate independently of a computer used to operate the system, or may communicate with such a computer. Alternatively, or in addition, a computer may perform the functions described herein for the device. The device may be a wireless device. The database may include system performance data from another system at a geographically remote location. The device may communicate with the database via a wireless router.

[0106] In certain aspects, the settings may be selected from any parameters or values ​​entered by a user. In certain aspects, the real-time measurements may include measurements internal to the system or measurements made by environmental sensors (e.g., measurements of humidity, pressure, and / or temperature external to the system). The real-time internal measurements may include one or more of the following: one or more clog events (e.g., direct data from a pressure sensor or frequency of clogs), flow rate (e.g., of the sample fluid or one or more gases described herein, such as the nebulizer gas, carrier gas, internal gas, and / or external gas), temperature of one or more components (e.g., in the torch assembly or of the load coil), voltage of a mass detector of the system, current of one or more cones (e.g., the sampling cone and / or skimmer cone of an ICP torch), the frequency or power at which the ICP load coil operates, mass signal from elemental standard beads, mass detector signal (e.g., a detector signal above a predetermined threshold that may indicate detector damage or wear, a cellular event), the amount of pressure of gas in a reservoir (e.g., a gas tank or a dewar supplying gas to the system), system run time, etc.

[0107] When a laser ablation system (e.g., for histology imaging) is configured to transfer a laser ablation plume (e.g., instead of suspended cells or suspended beads) to a mass cytometer of the present application, one or more settings and / or real-time measurements related to the laser ablation system are provided, such as laser ablation settings (e.g., power, operating voltage, operating frequency, operating mode, and / or power dissipation), ablation chamber fluid parameters (e.g., carrier and / or sheath gas flow rates for delivering the ablation plume to the mass), humidity inside the cytometer described herein, the laser ablation fluidics, laser ablation plume width measured by a mass detector, etc.

[0108] In certain embodiments, system performance data from one or more systems running on the same system can be used to identify preventative maintenance needed for that system. At such point, the user may be notified of the need for such maintenance (e.g., prompted to schedule such maintenance). The maintenance may be an instrument adjustment, an instrument cleaning (e.g., cleaning of the sample introduction fluid, the torch body or its tubing, or the sample cone downstream of the ICP torch), or replacement of a particular component (e.g., a laser, a mass detector, a gas reservoir, a torch body or its tubing, a nebulizer, or another element of the sample introduction fluidics described herein, etc.). usefulness

[0109] Aspects of the present systems and methods can extend the life of ICP components, facilitate user operation, and / or reduce maintenance or running costs. For example, aspects of the ICP load coils described herein may have improved lifespans compared to conventional coils. The detachable 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 being replaced). The vortex flow of the torch assembly may reduce the external gas flow required to maintain the plasma. An external ignition electrode can enable a detachable torch and / or have a longer lifespan than an electrode inserted into the torch body. Generally, a longer lifespan may be at least twice the lifespan or at least five times the lifespan before parts break or sustain significant damage (e.g., operating at less than 90% or less than 80% efficiency than when operational, where efficiency is the energy required to operate the device or the strength of the signal detected by an analyzer downstream of the ICP). The sample introduction systems described herein increase user walk-away time and enable automated sample introduction of multiple samples.

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; Equipped with An inductively coupled plasma (ICP) torch box, wherein the load coil is a cylindrical coil and the annular fin extends continuously along the cylindrical coil for at least one-eighth of a turn of the cylindrical coil.

2. (a) 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 that holds a removable inner torch base and a removable outer torch base. 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; an ICP load coil, wherein the annular fin is continuous over at least one-eighth of a turn of the tubular coil, and the ICP load coil is primarily comprised of a metal or alloy that has an oxidation rate during operation lower than that of copper; (c) an external ignition device for igniting a plasma in an inductively coupled plasma (ICP) torch, comprising: the external ignition device includes a circuit; The circuit comprises: an oscillator; a first high voltage transformer coupled to the first electrode; a second high voltage transformer coupled to the second electrode; the external ignition device is configured to ignite the plasma by a dielectric barrier discharge; an external ignition device, the first electrode and the second electrode being positioned on opposite sides of the outer tube and within 5 millimeters of the outer tube; An inductively coupled plasma (ICP) torch box comprising:

3. The removable inductively coupled plasma (ICP) torch holder assembly comprises: 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; 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; (a) the outer tube is separated from the outer tube base; (b) the inner tube is separated from the inner tube base; (c) the torch holder defines an external gas inlet, the external gas inlet in fluid communication with the annular region, the outer tube base defining three or more holes arranged to provide fluid communication between the external gas inlet and the annular region, the three or more holes oriented to generate a vortex flow; (d) the inner tube base and the outer tube base are permanently joined to one another; (e) the torch holder includes a heat-conducting element positioned to heat the injector; (f) metal is the primary material of the torch holder; (g) aluminum alloy is the primary material of the torch holder; (h) the inductively coupled plasma (ICP) torch box of claim 1, wherein the torch holder assembly further comprises a gas supply manifold configured to receive the torch holder, the torch holder including a pin for aligning its position within the gas supply manifold, and wherein twisting the torch holder allows the torch holder to be removed from the gas supply manifold.

4. The inductively coupled plasma (ICP) torch box of claim 3 , wherein the outer tube is detachable from the outer tube base.

5. The inductively coupled plasma (ICP) torch box of claim 3 , wherein the inner tube is detachable from the inner tube base.

6. 4. The inductively coupled plasma (ICP) torch box of claim 3, wherein the torch holder defines an external gas inlet, the external gas inlet in fluid communication with the annular region, the outer tube base defines three or more holes arranged to provide fluid communication between the external gas inlet and the annular region, the three or more holes being oriented to generate a vortex flow.

7. The inductively coupled plasma (ICP) torch box of claim 6 , wherein the outer tube base defines six holes.

8. 7. The inductively coupled plasma (ICP) torch box of claim 6, wherein the hole is within 2.5 cm of the outlet of the inner tube.

9. The inductively coupled plasma (ICP) torch box of claim 3 , wherein the outer tube does not have a taper.

10. The inductively coupled plasma (ICP) torch box of claim 3 , wherein the inner tube base and the outer tube base are each independently removable from the torch holder.

11. The inductively coupled plasma (ICP) torch box of claim 3 , wherein the inner tube base and the outer tube base are permanently joined together.

12. The inductively coupled plasma (ICP) torch box of claim 3 , wherein the inner tube base and the outer tube base are configured to be connected by a fastener.

13. 4. The inductively coupled plasma (ICP) torch box of claim 3, wherein the torch holder comprises a heat conducting element positioned to heat an injector.

14. 4. The inductively coupled plasma (ICP) torch box of claim 3, wherein metal is the primary material of the torch holder.

15. 4. The inductively coupled plasma (ICP) torch box of claim 3, wherein an aluminum alloy is a main material of the torch holder.

16. 4. The inductively coupled plasma (ICP) torch box of claim 3, wherein the outer tube does not have holes for extending electrodes into the outer tube.

17. a gas supply manifold configured to receive the torch holder; 4. The inductively coupled plasma (ICP) torch box of claim 3, wherein the torch holder includes a pin for aligning its position within the gas supply manifold, and wherein twisting the torch holder allows the torch holder to be removed from the gas supply manifold.

18. 20. The inductively coupled plasma (ICP) torch box of claim 17, wherein the torch holder defines an external gas inlet, the external gas inlet in fluid communication with the annular region and an external gas flow region of the gas supply manifold.

19. The removable ICP torch holder assembly includes: 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 for holding a removable inner torch base and a removable outer torch base; The inductively coupled plasma (ICP) torch box of claim 1 , comprising:

20. the outer tube is reversibly detachable from the outer tube base; 3. The inductively coupled plasma (ICP) torch box of claim 2.

21. The annular fin extends and diverges from a central axis.

10. The inductively coupled plasma (ICP) torch box of claim 1.

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