Extensible connecting tube

Abstract

A manifold made of an extensible elastomeric material. The "stretchable" manifold section allows for improved reliability / reproducibility of the connection performance. For example, the stretchability provided may accommodate tolerances between the positions of the imager and the reading device. In one example, the stretchability can accommodate tolerances between the positions of the junction between the rigid portion of the tubing in the IMC unit and the injector to the ionization source. The manifold may also include a reduced inside diameter (ID) compared to standard connection components. A taper in the inside diameter (ID) occurs before the flow enters the extensible manifold. A barbed end may be provided to secure the manifold in place.

Description

[Technical field]

[0001]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 266,550, filed January 7, 2022, and U.S. Provisional Patent Application No. 63 / 437,430, filed January 6, 2023, the entire contents of both of which are incorporated by reference herein in their entirety for all purposes.

[0002]

[0002] The present disclosure relates to manifolds used in conjunction with mass cytometry instruments. The manifolds are designed to secure the instruments together while maintaining laminar flow, reducing the possibility of turbulence, and providing optimal results. In a particular example, the disclosed and improved manifolds transport the ablated plume between a laser ablation source and a mass cytometer, for example, between an Imaging Mass Cytometry™ unit (IMC™) and a CyTOF® unit, including a CyTOF XT™. [Background technology]

[0003]

[0003] Laser ablation combined with inductively coupled plasma mass spectrometry (ICP-MS) or other ionization sources can be used to image biological samples (cells, tissues, etc.) labeled with elemental tags. Each laser pulse produces a plume of material ablated from the sample that can be transported to be ionized for further analysis by a mass analyzer. The information obtained from the laser pulse at each location on the sample can then be used to image the sample based on its analyzed content.

[0004]

[0004] The system may include a laser ablation mass cytometer having (i) a laser ablation source, (ii) an injector adapted to couple the laser ablation source with an ICP (inductively coupled plasma) source or other ionization source, and (iii) a mass analyzer. As shown in Figures 1A and 1B, in one example, pulses of a laser beam are directed at a sample to generate an ablated plume of the sample. The plume of the sample is then transported from the ablation zone to the injector that couples the laser ablation source with the ICP (inductively coupled plasma) of the mass cytometer or other ionization source. The transport may be through a "conduit" that generally includes a rigid tube section, through the injector, and then through a "connection tube" for continuously transporting the plume with the injector to other equipment (all of which may be collectively referred to herein as a "conduit" or "flow path").

[0005]

[0005] Referring now specifically to FIG. 1C, the pixel / sample material starts with the laser ablation and then travels into a conduit, the first part of which is a rigid tube. The term "rigid tube" is used herein to refer to a portion of the transfer conduit between the laser ablation and the injector inlet. That tube is generally rigid in that it is not substantially flexible or stretchable. The rigid tube has an inlet located within the laser ablation source, which captures each separate ablated plume as it is generated. This rigid tube may be a rigid section of a conduit located within an IMC (imaging mass cytometry) unit or other ionization source.

[0006] A manifold may be used to secure the rigid tube section to the injector. The manifold serves to connect the "imager" and "reader" instruments. The injector injects the plume particles dispersed in the gas stream into the ionization source torch. The ionization source ionizes each of the specifically captured, transported, and ablated plumes to generate ions for mass cytometry. An example of such a system is shown and described by co-pending U.S. Patent Application Publication No. 2019 / 0271630, now U.S. Patent No. 10,705,006, the entire contents of which are incorporated herein by reference. The present disclosure provides an improved manifold as part of a transport channel that transports the ablated plume between a laser ablation source and a mass cytometer. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] US Patent Application Publication No. 2019 / 0271630 [Patent Document 2] U.S. Pat. No. 10,705,006 Summary of the Invention

[0008]

[0007] The present disclosure relates to a manifold made of an extensible elastomeric material. The "extensible" manifold section allows for improved reliability / reproducibility of the connection performance. For example, the extensibility provided may accommodate tolerances between the positions of the imager and the reading device. In one example, the extensibility may accommodate tolerances between the positions of the junction between the rigid portion of the tubing in the IMC unit and the injector to the ionization source. In another example, improved laminar flow, reduced potential for turbulence, and / or optimal results may be obtained. The manifold may also have a reduced inner diameter (ID) compared to standard connection components in mass spectrometry and related fields. A taper occurs in the inner diameter (ID) before the flow enters the extensible manifold, and a taper may also occur after the extensible manifold. A barbed end may be provided to secure the manifold in place. [Brief description of the drawings]

[0009] [Figure 1A] FIG. 1 is a prior art schematic diagram of an ablation sampling system. [Figure 1B] FIG. 1B shows a schematic prior art direct injector inlet without the sampling ramp of FIG. 1A. [Figure 1C] FIG. 1B is a schematic diagram of the flow through the system of FIG. 1A. [Diagram 2] FIG. 1 shows an example of a tagged plume and the potential for crosstalk. [Figure 3A] FIG. 13 is a schematic diagram of ablation sampling with a taper in the sampling ramp, according to an embodiment of the present invention. [Figure 3B] FIG. 13 is a schematic diagram of an ablation sampling with a taper in a rigid tube according to an embodiment of the present invention. [Figure 3C] 13 is a schematic diagram of an ablation sampling with a taper at the junction between a rigid tube section and an extendable manifold according to an embodiment of the present invention. FIG. [Figure 3D] 13 is a schematic diagram of an ablation sampling with a taper in the junction between an extendable manifold and an injector according to an embodiment of the present invention. FIG. [Figure 4]FIG. 13 is a side cross-sectional view of a rigid tube portion of an ablation chamber injector that connects to an extendable manifold, according to an embodiment of the present invention. [Diagram 5] FIG. 13 illustrates the injector side of an ablation plume transport channel / conduit according to an embodiment of the present invention. [Figure 6] 1 illustrates a connection end interface according to an embodiment of the present invention. [Figure 7] FIG. 13 shows two joints that function to connect various tube sections together, according to an embodiment of the present invention. [Figure 8] FIG. 1 illustrates an exemplary test setup using an optical post on a linear stage that is driven down into a fixed manifold, in accordance with an embodiment of the present invention. [Figure 9] FIG. 1 is a schematic diagram of a test setup for testing manifold subsidence, according to an embodiment of the present invention. [Figure 10] FIG. 13 is a graphical plot of 800 Hz plume crosstalk versus applied subsidence applied to a manifold in accordance with an embodiment of the present invention. [Figure 11] FIG. 13 is a flow chart diagram of an example process for using a manifold for ablation sampling, according to an embodiment of the present invention. [Figure 12] FIG. 1 is a schematic diagram of an example of a system using a manifold for ablation sampling, in accordance with an embodiment of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010]

[0025] As further background for understanding the present disclosure, Imaging Mass Cytometry™ (IMC™) requires good time fidelity of the arrival of the ablation plume in the plasma in order to construct an ion image from the detected plume sequence. In this context, “time fidelity” refers to how well the arrival time of the plume into the plasma correlates with the time of the corresponding laser shot, as well as how narrowly distributed the arrival times of the individual particles that make up the plume are. These two effects are sometimes described as “arrival time jitter” between individual plumes, and “time spread” of individual plumes during their passage. Without sufficient time fidelity, plumes from successive laser ablation events may blur together, resulting in “smearing” of the final tissue image (reconstructed from the ion signal), or other severe imaging artifacts, which is undesirable.

[0011]

[0026] As shown in Figure 2, each plume is associated with a "tag" input. This is a channel in the acquisition system that identifies the arrival of the laser pulse and allows the system to identify the range of the TOF (time of flight) push that corresponds to that shot. For example, the portion of the signal from one shot that is inside the window of another is the "crosstalk" (CT) of that plume. Typically, several plumes are summed together to produce a clean, representative "cumulative" transient that is used to perform crosstalk measurements.

[0012]

[0027] To ensure low crosstalk, the plume must be transported from the ablation site to the plasma by a laminar flow of gas. Laminar flow ensures predictable and consistent material transfer. Flow turbulence can result in plume spreading and loss of temporal coherence. Turbulence can also lead to aerosol particles in the plume contacting and depositing on the walls of the conduit. This results in sensitivity loss and contamination of the conduit. Therefore, turbulent conditions should be avoided.

[0013]

[0028] As the pixel acquisition rate increases, the requirements to ensure low crosstalk and laminar flow conditions can also become difficult to meet, since a non-zero width of the plume (transient width) will be present. However, since the pixel acquisition rate is currently one of the main limiting parameters that determine the total throughput of an IMC instrument, it is important to ensure that the pixel rate is as high as possible.

[0014]

[0029] One way to limit the spread of transients in the conduit is to reduce the time the plume is in the gas flow. This reduces the time available for the plume aerosol to spread away from the fastest streamline (on the centerline of the flow), which is one of the phenomena that can lead to spreading. Another advantage of reducing the overall travel time is that the arrival of two particles located on streamlines with different velocities (e.g., 100% of the maximum velocity and 90% of the maximum velocity) will be closer to each other in proportion to the overall travel time. However, since the chamber gas flow is determined by the geometry of the ablation sampling ramp and the overall gas flow through the transport channel / conduit is determined by the properties of the plasma in the ionization torch, it is not possible to simply increase the flow to reduce the transit time.

[0015]

[0030] Besides ensuring that laminar flow is maintained, it is also desirable to avoid any abrupt discontinuities in the inner surface of the channel walls, such as those caused by dispersion at the tube section junctions. Such imperfections in the inner surface of the channel can result in localized turbulence, which usually forms in laminar systems, that can disrupt the flow and disturb the plumes as they pass through. It has been found that changes in the inner diameter of the conduits need to be done carefully, as some shapes are more likely to generate turbulence than others. In particular, it may be easier to maintain laminar flow when reducing the conduit ID than when expanding it. Therefore, another attempt to improve the timing of plume migration has been to reduce the inner diameter (ID) of the connecting tube length. However, as the ID was reduced, other problems arose, such as the effects of Dean vortices, and the effects of some mechanical tolerances being misaligned.

[0016]

[0031] Finally, it is desirable for the flow path between the instruments to be kept as straight as possible for best performance, since any curvature of the flow path can result in the development of Dean vortices that cause radial vortices in curved tubes above a certain critical flow rate value. Thus, even with perfect laminar flow, transient performance can be adversely affected if potentially flexible (non-stretchable) parts of the flow path (connecting tube) are significantly curved. However, the use of rigid straight tubes can create alignment tolerance issues. An example of the use of straight, small ID rigid tubes to carry the time-resolved ablation plume from the ablation chamber to the ICP mass analyzer is described in U.S. Pat. No. 10,285,255. Such straight rigid tubes require tight alignment tolerances between the ablation unit and the ICP-MS (or other ionization source). The use of perfectly straight connecting tubes is difficult to achieve using simple alignment tools and risks damaging the tube assembly since any movement of the ablation chamber (free movement on its spring suspension) with respect to the fixed CyTOF XT could destroy the glued seams at the ends of the assembly. A straight tube would be a conduit for vibrations from the CyTOF. Such vibrations would affect the ability of the IMC optics to place ablation shots at equal distances, resulting in image artifacts.

[0017]

[0032] Other analytical techniques use "flexible" and elastic transfer tubes, but these operate under substantially different flow conditions. One example is used to transfer ionized samples to a mass analyzer unit as described in US Patent Application Publication No. 2020 / 0121229. In this publication, the gas sample is partially ionized in the tube. The purpose of the 2020 / 0121229 "flexible" and elastic tubes is not to suppress the spread of the ablation plume, but as a way to connect a sampling area located far away from the inlet of the mass spectrometer. Liquid transfer by the use of flexible / extendable tubes (e.g., via peristaltic pumps) also differs from the present disclosure in that the characteristics that need to be addressed for liquid flows are substantially different from those that need to be addressed for gas flows.

[0018]

[0033] It was therefore desirable to design a coupling tube that would not significantly transfer vibration energy from the CYTOF to the IMC optics, but would be robust enough to withstand the range of motion of the IMC unit, as well as provide improved performance to give good margins on the crosstalk spectrum. It was also desirable to provide a coupling tube that would allow proper alignment using a simple alignment tool. It was also desirable to provide a coupling tube that would allow a strong connection at its ends, rather than involving gluing a semi-flexible plastic tube to a machined hard plastic end piece. The glued seam could slip off the injector, or the injector could be pulled out of the injector holder. It was further desirable to ensure that the IDs were matched across the joint as much as possible to prevent turbulence.

[0019]

[0034] The present disclosure therefore relates to a manifold that addresses many of these problems. The novel manifold comprises an extendable elastomeric tube of any length terminated at each end with a machined barbed joint. The inner diameter (ID) of the manifold is reduced compared to the ID of currently used tube sections near the ablation sampling ramp. In some embodiments, data can be acquired at consistently high speeds, for example in the range of 400-1000 pixels per second or more. To maintain ID continuity, there can be a tapered section inserted between the end of the rigid tube and the manifold. The disclosed features and tapered sections can help avoid alteration of the ablation sampling geometry during plume transport.

[0020]

[0035] stretchable material

[0021]

[0036] In a first aspect, the disclosed coupling tube is made of an extensible elastomeric material. The extensible material of the coupling tube allows for improved reliability / reproducibility of the coupling performance. The stretching shortens / absorbs positional tolerances between the instruments. As used herein, the term "stretchable" is used to mean a material that can stretch for several millimeters or more and return to its original length once released, and has at least some type of elastic rebound. In one example, stretchable means a material that can stretch from about 0.5 mm to about 10 mm. In one example, the stretchable tube section undergoes elastic deformation only up to a maximum of 20% extension and up to about 20 N of applied force. (In contrast, the semi-flexible tube sections of the prior art undergo significant plastic deformation during this extension, requiring substantially higher forces.) By replacing the currently used semi-flexible FEP (fluorinated ethylene propylene) tube sections used in HTI connections with an extensible elastic tube material, the tube section can be kept taut without risking the integrity of the coupling assembly. It can also accommodate variations in the distance between the equipment so that the connection maintains a minimum stretch over a minimum distance. The connecting tube can be slightly shorter than the distance between the machines, so that the distance extends but remains relatively taut. It is also possible to select tubing materials and mounting techniques that ensure that the connecting tube can withstand the stretch corresponding to the maximum distance. Providing an extendable tube allows for a nearly straight connection.

[0022]

[0037] A further advantage of the disclosed extendable tube section is that it simplifies the design of an alignment fixture to place the deuterium unit in the correct position relative to the CyTOF XT unit. The alignment fixture may be used to establish the minimum distance between the two instruments. In that case, it is sufficient to ensure that the length of the tube section is less than the minimum distance between the two machines, so that at the minimum gap, the manifold is still under nominal tension. By keeping the tube section under tension, the tube remains essentially straight between the two connection points. (Gravity may cause the manifold to sink slightly, but its effect is negligible in this application.) Furthermore, imperfect positioning in the vertical or horizontal dimensions may also result in a "non-straight" tube section, although studies have shown that this is tolerable to some extent. The manifold alignment fixture may also serve to laterally align the two machines, which may further minimize any bending of the manifold.

[0023]

[0038] In one embodiment, the connecting tube may be made of a fluoropolymer elastomer. One specific exemplary fluoroelastomer that may be used is Viton™. Products made with Viton™ fluoroelastomer generally maintain their flexibility, shape and seal when exposed to chemicals and high temperatures. Other materials that may be used include, but are not limited to, latex, silicone, flexible and expandable PVC tubing (e.g., Tygon), EPDM, AFLAS, nitrile, neoprene, fluorosilicone, Chemraz (FFKM), or any combination thereof of any of the possible materials listed.

[0024]

[0039] Inner diameter

[0025]

[0040] In another aspect, the inner diameter (ID) of the manifold is reduced compared to conventional or standard manifolds used for this purpose. This reduced ID is expected to have a positive impact on the transient quality (until the diameter reaches a small range where the flow enters turbulent transition, as outlined below). The inner diameter of the manifold may be in the range of 0.1 mm to 10.0 mm. Exemplary inner diameters may be in the range of 0.3 mm to 6.0 mm. The inner diameter of the flow may be about 0.5 mm to about 1.0 mm ID. In a specific example, the ID of the manifold may be about 0.8 mm. The ID range refers to the manifold ID, which may or may not be stretched. That is, the ID may be 0.3 mm to 6.0 mm, whether stretched or not. The reduction in the ID of the manifold provides a reduction in minimum CT (crosstalk) over standard manifold IDs, which are typically about 1.5 to about 2.0 mm. Reducing the manifold ID has been found to reduce transit time. This shortened transit time can in turn reduce transient spreading effects that are exaggerated by long travel distances as well as by timing jitter in transit times from plume to plume. However, it is expected that small diameters (e.g., below the 0.5 mm range) may run into limitations imposed by the laminar to transitional flow transition that is expected to prohibit good transient formation, although small diameters can be tested and are considered to be within the scope of this disclosure.

[0026]

[0041] The ID of the manifold is an important variable that may need to be determined for a given system size. The ablation chamber gas flow is set by the geometry of the sampling ramp, its immediate area, and the total injector flow. The optimum total injector flow rate is determined by the characteristics of the ionization source torch. Changes to either one may require changes to the optimum ID of the tubing.

[0027]

[0042] Taper

[0028]

[0043] With regard to the connecting tube and its associated components, tapers may be provided at various points in the system. In one example, there may be a taper 10 provided on the sampling ramp 26 at the ablation chamber output of the laser ablation source 14. This is shown in FIG. 3A. The sampling ramp 26 is connected to one end of the rigid tubing section 12. By providing a taper 10 on or within the ablation sampling ramp tube (on the output side of the ramp), the taper 10 can be present as soon as possible after the ablation event without substantially changing the gas flow in the vicinity of the ablation site. This approach may have the disadvantage of making the taper location difficult to inspect / clean.

[0029]

[0044] The gas may be provided from a gas source coupled to the laser ablation source 14. The laser ablation source 14 may include a sample holder 23, a laser source, and optics for directing the laser from the laser source to the sample holder 23. The system may further include a mass spectrometer for analyzing plasma products in the inductively coupled plasma chamber.

[0030]

[0045] In another example, there may be a taper 10 on the inside diameter of the rigid tubing section 12 before the flow enters the manifold 20. This is shown in Figure 3B.

[0031]

[0046] In a further example, there may be a taper 10 provided in the junction assembly 28 between the rigid tube section 12 and the manifold 20. This is shown in Figures 3C and 4. As shown in Figure 4, it is also possible to incorporate a barbed joint 18 that is used to secure the manifold 20 in place. (The barbed joint option is described in more detail below.) In this option, the taper 10 may be located within an element of the barbed joint 18 (described in more detail below) on the manifold 20. Providing the barbed joint 18 on the taper 10 inside the junction assembly allows for easy inspection / cleaning of the taper. This approach may have the disadvantage of making the two ends of the manifold assembly dissimilar (e.g., if the output end of the manifold does not incorporate a similar taper).

[0032]

[0047] In another example, there may be a taper 10 at the end of the manifold 20 at the junction assembly 30 that connects to the injector 32. This is shown in Figure 3D. The injector 32 can couple a laser ablation source to an inductively coupled plasma chamber.

[0033]

[0048] These configurations can result in increased transport velocities and short transport times, which are beneficial for reducing the time spread in plume transients. It is generally desirable to provide a low gradual taper that can help suppress turbulence. In one example, the taper may have a maximum angle taper of about 10°. However, it should be understood that the taper angle can be anywhere between about 0.1 degrees to about 180 degrees for the full angle. It is anticipated that 5 to 10 degrees may be a particularly useful taper angle range. The taper between the large and small inner diameter regions can be slow enough to avoid the generation of turbulence. In particular, the taper can be a full angle of 10 degrees or a full angle of 5 degrees. In some configurations, the taper can be 30 degrees or even 60 degrees. The transition between the taper and the inlet to each section of the system may be provided with smooth edges to suppress the generation of turbulence. The transition can be piecewise linear and smoothed by curvature or splines at the junctions on the piecewise elements.

[0034]

[0049] In one particular example, the taper may transition from a channel with an inner diameter of about 1.6 mm to a connecting tube with an inner diameter of about 1.0 mm. The taper distance may be about 0 mm to 50 mm. This distance is related to the angle and change in ID between the input and output of the taper. As an example, an angle of 180 degrees results in a distance of 0 mm where the taper transitions between the large ID and the small ID. Other ranges are of course possible and are considered within the scope of this disclosure. It is also possible to provide a stepped transition with various down step angles, somewhat resembling the smooth nonlinear transition of the internal cross section of an extended telescope, or the slope of a horn (e.g., suspended horn, Bezier horn).

[0035]

[0050] In the example shown, the portion of the large diameter section before the taper remains relatively large (e.g., up to about 2 mm), which can simplify the machining of the ablation sampling area and allows the main part of the tube to be made of a conventional narrow bore tube. The diameter of the narrow bore section (the inner diameter of the tube after the taper) is limited by the diameter that corresponds to the onset of turbulence.

[0036]

[0051] The Reynolds number can be calculated for a circular pipe and a known flow. In general, Reynolds numbers above 4000 indicate turbulent flow in a circular pipe and should be avoided. For a given mass flow rate of gas, the Reynolds number is inversely proportional to the diameter of the conduit. Thus, the diameter of the conduit is limited to small sizes by the Reynolds number and the occurrence of turbulence in the flow. In a typical equipment configuration, the ID of the narrow portion of the conduit can be on the order of about 0.5 mm to about 1.0 mm.

[0037]

[0052] 4, there is shown a taper 10 that reduces the ID of the rigid tube section 12 from the ablation source 14 from 1.58 mm to 1.0 mm. The taper 10 is shown located inside the ablation chamber output plug 16. However, it should be understood that the taper 10 could also be located inside the 1.58 mm ID tube section itself (the current dimensions of the injector tube section from the sampling ramp).

[0038]

[0053] The location of the taper in the system can be varied to any suitable location, with the general goal being to have the taper somewhere in the ablation plume conduit that is located between the region where ablation occurs and the tip of the injector into the plasma.

[0039]

[0054] Additionally, the taper dimensions (angle, ID and length) may vary and may be applied at any location of the taper. It is also possible to have more than one taper located along the length of the flow path. For example, there may be two tapers or three tapers along the length of the conduit. This may incorporate any of the taper locations described herein. For example, the first taper may be 1.6mm to 1.0mm ID and may be located within the rigid tube section 12. The second taper may be 1.0mm to 0.6mm ID and may be located at the junction assembly 30 between the manifold 20 and the injector 32.

[0040]

[0055] Barbed ends for connection

[0041]

[0056] In a further embodiment, one or more machined barbed joints 18, 31 may be provided on the joint assembly 28 and / or the joint assembly 30. For example, a barbed joint 18 may be provided on the joint assembly 28 and a barbed joint 31 may be provided on the joint assembly 30. It is also possible for multiple barbed joints to be connected to the joint assembly 28 or multiple barbed joints to be connected to the joint assembly 30. One or more machined barbed joints 18, 31 may be configured to fit onto the connecting tube 20, examples of which are shown in Figures 4-7. The joint assembly 28, 30 having barbed joints 18, 31 can help ensure a secure attachment of the connecting tube 20 to the joint assembly (28, 30) at its end. The machined barbed joints 18 on the joint assembly 28 and / or the machined barbed joints 31 on the joint assembly 30 provide a significant improvement over previous connection designs. First, they may be made from stainless steel or other metal materials rather than Delrin plastic. This means that the dimensional tolerances of the finished part can be greater, resulting in better centering alignment at the joint between the end of the rigid tube section 12 and the beginning end of the connecting tube 20. The ID of the machined end of the barbed fitting 18 that is received by the connecting tube 20 can be machined into metal (e.g., stainless steel) and therefore can be close to perfectly round. Second, the use of barbed fittings 18 eliminates the need for adhesives in the assembly, removing another potential source of assembly variability. The performance of the system is not as strongly affected by the circular discontinuity of the ID surface that the barbed ends cause. The barbs are relatively small and can be perfectly circular perturbations along the inner surface of the connecting tube 20, so they do not create a significant source of transient spread. The use of barbs simplifies assembly of the system and eliminates the need for adhesives, as was done with older designs, which presented significant manufacturing challenges. The material of the stretchable tube is also suitable for forming a sealed barbed connection that will withstand many connection / disconnection cycles (unlike the hard bendable plastic material used in the original manifold, which uses an FEP material that is bendable but not stretchable).If it were inserted over the barbs, it would not clamp to the barbs with sufficient strength, resulting in potential air and argon leaks into the conduit. Therefore, bendable tubing material is bonded to the joint assembly. Expandable tubing material can be used with the barbed connections providing the advantages discussed above.

[0042]

[0057] Installation

[0043]

[0058] The connecting tube 20 is cut to length, strung through the connecting nut, and pressed onto the barbed joint 18 at both ends. The connecting alignment fixture is designed to ensure that the connecting tube can be held taut by adjusting the separation between the machines for the expected range of tolerance stack-up (e.g., + / - 5mm RMS). A connecting tube pull test may be performed to ensure that the connecting tube ends do not easily pull off when moving between machines. In one example of an exemplary pull test for a 1.0mm PVC connecting pull test, the connecting end did not pull off until a pulling force of about 2kg was applied. The connecting tube showed an elongation of about 46mm (about 20%) before failure. Based on these results, it may be possible to rely entirely on the elongation of the connecting tube to accommodate variations in the axial distance between the machines.

[0044]

[0059] Working Example

[0045]

[0060] During the life test experiments, tests were performed to determine how far the manifold could sink before it began to affect the transients. Figure 8 shows the test setup using a 2 inch optical post on a linear stage driven downward into a fixed manifold 20. The imaging equipment was set up using a manifold alignment fixture to ensure a straight manifold 20. To simulate the effect of a sinking manifold, a device was added to the system that would push the top of the manifold 20 downward. A schematic of the device is shown in Figure 9. The device includes a vertical positioning stage with a 2 inch diameter cylindrical pushing element 36 that contacts the manifold 20. The pushing element 36 was advanced into the manifold 20 in 0.5 mm increments and the transient quality was recorded for each position. The starting position was where the pushing element 36 just contacted the manifold 20. The test was terminated when the system no longer achieved a CT400<12%. For all gas flows, a total subsidence of 6.5 mm was required for the manifold 20 to fail to achieve CT400 (as indicated by the double arrow in FIG. 9). It was determined that a deflection of just 1 mm in the center of the manifold 20 made a measurable difference in transient quality.

[0046]

[0061] Figure 10 shows a graphical plot of 800 Hz plume crosstalk against applied sinkage applied to the manifold. It shows that applying sinkage to the manifold without realigning the gas flow results in a steep degradation in transient quality (blue / upper data points). Although improvement can be achieved by realigning the gas flow after the manifold is bent (orange / lower data points), the manifold never returns to the low crosstalk values ​​obtained with the straight manifold. These results demonstrate the advantage of an extensible manifold that remains substantially straight in normal use due to the elasticity of the material.

[0047]

[0062] Exemplary Methods

[0048]

[0063] 11 is a flow chart diagram of an example process for using a manifold for ablation sampling. In some implementations, one or more process blocks of FIG. 11 may be performed by one or more components of system 1200 of FIG.

[0049]

[0064] In block 1102, a sample is ablated to form an ablation plume. The optical configuration of the laser ablation source 1202 can direct the laser from the laser source to a sample holder that holds the sample. The sample can be a biological sample such as cells, tissue, etc. that are labeled with elemental tags. The laser pulse creates an ablation plume that can be transported to be ionized for further analysis by a mass analyzer.

[0050]

[0065] In block 1104, the ablation plume is laminarly flowed from the sample through a connecting tube 1208 to an injector 1212. The connecting tube 1208 can be made of a stretchable elastomeric material, such as a fluoroelastomer. Additionally, the connecting tube 1208 can have an inner diameter in the range of 0.3 mm to 6.0 mm. Before flowing through the connecting tube 1208, the ablation plume can flow through a rigid tube section 1204 that is coupled to the laser ablation source 1202 and then through a barbed joint of a junction assembly 1206a that couples the rigid tube section 1204 to the connecting tube 1208. The ablation plume can flow from the connecting tube 1208 through another barbed joint of another junction assembly 1206b that couples the connecting tube 1208 to the injector 1212.

[0051]

[0066] In some examples, one or more components may include a taper to increase the transport rate and decrease the transport time of the ablation plume. Increasing the transport rate and decreasing the transport time are beneficial for reducing the time spread in the plume transient. The taper may be in the laser ablation source 1202, the rigid tube section 1204, one or more of the junction assemblies 1206a-b, the manifold 1208, or a combination thereof.

[0052]

[0067] In block 1106, the ablation plume is analyzed using an inductively coupled plasma mass spectrometer, any other ionization source, and any mass analyzer described herein. The inductively coupled plasma mass spectrometer can include an inductively coupled plasma chamber 1216 and a mass spectrometer 1218. The injector 1212 can connect the manifold 1208 to the inductively coupled plasma mass spectrometer, and the mass spectrometer 1218 can analyze the plasma products in the inductively coupled plasma chamber 1216. Information obtained from the laser pulse on the sample can be used to image the sample based on its analyzed content.

[0053]

[0068] Process 1100 may include additional implementations, such as any single implementation or any combination of implementations, in conjunction with one or more other processes described below and / or elsewhere herein.

[0054]

[0069] 11 illustrates example blocks of process 1100, in some implementations, process 1100 may include additional, fewer, different, or differently arranged blocks than those illustrated in FIG 11. Additionally or alternatively, two or more of the blocks of process 1100 may be performed in parallel.

[0055]

[0070] Exemplary System

[0056]

[0071] 12 shows a schematic diagram of an example of a system 1200 using a connecting tube for ablation sampling. The system 1200 includes a laser ablation source 1202, a rigid tube section 1204, a connecting tube 1208, an injector 1212, an inductively coupled plasma chamber 1216, and a mass spectrometer 1218. The injector 1212 can operably connect the laser ablation source 1202 to the inductively coupled plasma chamber 1216, and the connecting tube 1208 can operably connect the laser ablation source 1202 to the injector 1212 to transfer an ablation plume from the laser ablation source 1202 to the injector 1212. Operatively connected may mean that the components are directly connected to each other (e.g., in direct contact) or indirectly connected to each other (e.g., having one or more intermediate components between the connected components).

[0057]

[0072] The laser ablation source 1202 can include a sample holder 1203, a laser source 1205, and an arrangement of optics 1207 for directing the laser from the laser source 1205 to the sample holder 1203. The laser ablation source 1202 can receive a gas flow from a gas source 1201 coupled to the laser ablation source 1202. The laser ablation source 1202 can also include a sampling ramp 1209 operably coupled to the rigid tubing 1204. The sampling ramp 1209 can be tapered to have a smaller diameter proximal to the rigid tubing 1204 compared to distal to the rigid tubing 1204 (e.g., sampling ramp assembly 26 of FIG. 3A).

[0058]

[0073] Connecting tube 1208 can comprise an expandable elastomeric material, such as a fluoroelastomer, and can be characterized by a first inner diameter in the range of 0.3 mm to 6.0 mm, including 0.3 mm to 0.5 mm, 0.5 mm to 1.0 mm, 1.0 mm to 2.0 mm, 2.0 mm to 3.0 mm, 3.0 mm to 4.0 mm, 4.0 mm to 5.0 mm, or 5.0 mm to 6.0 mm. Connecting tube 1208 can be connecting tube 20 or any expandable tube section described herein, laser ablation source 1202 can be laser ablation source 24 or any laser ablation source described herein, rigid tube section 1204 can be rigid tube section 12 or any rigid tube section described herein, and injector 1212 can be injector 32 or any injector described herein.

[0059]

[0074] The rigid tube section 1204 can be operably coupled to a connecting tube 1208. A first end of the rigid tube section 1204 can be operably coupled to the laser ablation source 1202, and a second end of the rigid tube section can be operably coupled to the connecting tube 1208. The rigid tube section 1204 can be characterized by a second inner diameter at the first end that is larger than the first inner diameter of the connecting tube 1208. The second diameter can be up to 2.0 mm (e.g., 0.7 mm, 0.8 mm, 0.9 mm, 1.0 mm, 1.1 mm, 1.2 mm, 1.3 mm, 1.4 mm, 1.5 mm, 1.6 mm, 1.7 mm, 1.8 mm, 1.9 mm, 2.0 mm, or a diameter between and including any two of these numbers). The rigid tube section 1208 may further be characterized by a third inner diameter at the second end, the second inner diameter being greater than the third inner diameter. The rigid tube section 1204 may include a taper (e.g., up to a 10 degree taper) that narrows the third inner diameter at the second end leading to the connecting tube 1208 to within a range of 0.5 mm to 1.0 mm, within a range of 0.5 mm to 0.6 mm, within a range of 0.6 mm to 0.7 mm, within a range of 0.7 mm to 0.8 mm, within a range of 0.8 mm to 0.9 mm, or within a range of 0.9 mm to 1.0 mm. In one particular example, the taper may transition from an inner diameter of about 1.6 mm to an inner diameter of about 1.0 mm with a taper distance between 0 mm and 50 mm.

[0060]

[0075] The system 1200 may also include one or more junction assemblies (e.g., junction assembly 1206a and junction assembly 1206b) that may cooperate with at least one end of the connecting tube 1208. For example, the junction assembly 1206a may couple the rigid tube section 1204 to the connecting tube 1208, and the junction assembly 1206b may couple the connecting tube 1208 to the syringe 1212. The junction assemblies 1206a-b may each include a barbed fitting (e.g., barbed fitting 18) that cooperates with the connecting tube 1208. The barbed fitting may include a taper that reduces a diameter leading to the connecting tube 1208 or the syringe 1212. The junction assemblies 1206a-b may be junction assembly 28 or any junction assembly described herein.

[0061]

[0076] A mass spectrometer 1218 can analyze plasma products in the inductively coupled plasma chamber 1216. The system 1200 may include electrical circuitry and other components for generating a plasma in the inductively coupled plasma chamber 1216. In some embodiments, an ionization chamber may be used in place of the inductively coupled plasma chamber 1216. In embodiments, the system 1200 may also include electrical circuitry and other components for ionizing materials in the ionization chamber.

[0062]

[0077] Although the subject matter of certain embodiments of the present disclosure has been specifically described to meet statutory requirements, this description is not necessarily intended to limit the scope of the claims. The claimed subject matter may be embodied in other ways, may include different elements or steps, and may be used with other existing or future technologies. This description should not be construed as implying a particular order or arrangement between the various steps or elements, unless the order of arrangement of the individual steps or elements is explicitly stated.

[0063]

[0078] It will be understood that different arrangements of the components shown in the drawings or described above, as well as components and steps not shown or described, are possible. Similarly, some features and subcombinations are useful and may be employed without reference to other features and subcombinations. The embodiments of the invention have been described for purposes of illustration and not limitation, and alternative embodiments will become apparent to the reader of this patent. Thus, the invention is not limited to the embodiments described above or shown in the drawings, and various embodiments and modifications are possible without departing from the scope of the following claims.

[0064]

[0079] Any of the methods described herein may be implemented in whole or in part in a computer system including one or more processors that may be configured to perform the steps. For example, the computer system may be in communication with mass flow controllers, power supplies, and other control systems. Thus, the embodiments may be directed to a computer system configured to perform any of the steps of the methods described herein, with potentially different components performing each step or each group of steps. Although presented as numbered steps, steps of the methods herein may be performed simultaneously, at different times, or in different orders that are logically possible. Furthermore, some of the steps may be used with some of the other steps from other methods. Also, all or some of the steps may be optional. Furthermore, any of the steps of any of the methods may be performed using modules, units, circuits, or other means of systems for performing these steps.

[0065]

[0080] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has individual components and features that may be readily separated or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present disclosure.

[0066]

[0081] The above description of exemplary embodiments of the present disclosure has been presented for purposes of illustration and description, and is provided to provide those skilled in the art with a complete disclosure and description of how to make and use the embodiments of the present disclosure. It is not intended to be exhaustive or to limit the present disclosure to the precise forms described, nor are they intended to represent all or the only experiments performed. Although the present disclosure has been described in some detail by way of illustration and example for clarity of understanding, it will be readily apparent to those skilled in the art that, in light of the teachings of the present disclosure, certain changes and modifications can be made without departing from the spirit or scope of the appended claims.

[0067]

[0082] Thus, the foregoing merely illustrates the principles of the present invention. It will be appreciated that those skilled in the art may devise various configurations that embody the principles of the present invention and are within its spirit and scope, although not expressly described or shown herein. Moreover, all examples and conditional language recited herein are intended primarily to aid the reader in understanding the principles of the present disclosure, which are not limited to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the present invention, as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Moreover, such equivalents are intended to include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. Thus, the scope of the present invention is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of the present invention is embodied in the appended claims.

[0068]

[0083] The enumeration of "a," "an," or "the" is intended to mean "one or more," unless specifically indicated to the contrary. The use of "or" is intended to mean "inclusive or," and not "exclusive or," unless specifically indicated to the contrary. A reference to a "first" element does not necessarily require that a second element be provided. Furthermore, a reference to a "first" or "second" element does not limit the referenced element to a particular location unless explicitly stated. The term "based on" is intended to mean "based at least in part on."

[0069]

[0084] The claims may be drafted to exclude any element that may be optional, and thus, this statement is intended to serve as a predicate for use of exclusive terminology, such as "solely," "solely," and the like, in connection with the recitation of claim elements or the use of a "negative" limitation.

[0070]

[0085] Where a range of values ​​is given, it is understood that each intervening value, to the tenth of the unit of the lower limit, between the upper and lower limits of that range is also specifically disclosed, unless the context clearly dictates otherwise. Each smaller range between any stated or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within the embodiments of the disclosure. The upper and lower limits of these smaller ranges may be independently included or excluded in the range, and each range in which either, neither, or both of the limits are included in the smaller range is also encompassed within the disclosure, subject to any specifically excluded limits in the stated range. Where a stated range includes one or both of its limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0071]

[0086] All patents, patent applications, publications, and descriptions mentioned in this specification are incorporated herein by reference in their entireties to disclose and describe the related methods and / or materials in which the publications are cited, as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. None are admitted to be prior art.

Claims

1. a laser ablation source; an injector configured to operably couple the laser ablation source to an inductively coupled plasma chamber; a coupling tube configured to operably couple the laser ablation source to the injector for transporting a plurality of ablation plumes from the laser ablation source to the injector, the connecting tube comprises an expandable elastomeric material; a connecting tube, the connecting tube being characterized by a first inner diameter in a range of 0.3 mm to 6.0 mm; A system comprising:

2. The system of claim 1 , wherein the extensible elastomeric material comprises a fluoroelastomer.

3. The system of claim 1 , further comprising a junction assembly comprising a barbed joint configured to cooperate with at least one end of the manifold.

4. The system of claim 3 , wherein the barbed joint includes a taper that reduces the diameter leading to the manifold.

5. a rigid tube portion configured to operably connect to the connecting tube; the rigid tube having a first end operably coupled to the laser ablation source; the rigid tube portion having a second end operably connected to the connecting tube; the rigid tube portion is characterized by a second inner diameter at the first end that is greater than the first inner diameter of the connecting tube; the rigid tube portion is characterized by a third inner diameter at the second end; The second inner diameter is larger than the third inner diameter. The system of claim 1 .

6. The system of claim 5 , wherein the second inner diameter is in the range of 1.5 mm to 2.0 mm.

7. 6. The system of claim 5, wherein the rigid tube section comprises a 10 degree taper configured to narrow the third inner diameter at the second end that leads to the manifold within a range of 0.5 mm to 1.0 mm.

8. 10. The system of claim 1, further comprising a junction assembly between the manifold and the syringe, the junction assembly emerging at a taper of reducing diameter leading to the syringe.

9. The system of claim 1 further comprising the inductively coupled plasma chamber.

10. The system of claim 1 , further comprising a mass spectrometer configured to analyze plasma products in the inductively coupled plasma chamber.

11. the laser ablation source comprising: A sample holder; A laser source; an optical system for directing a laser from the laser source to the sample holder; The system of claim 1 , comprising:

12. The system of claim 1 , further comprising a gas source operably coupled to the laser ablation source.

13. an inductively coupled plasma chamber; an injector configured to operably connect the inductively coupled plasma chamber to a laser ablation source; a coupling tube configured to operably couple the laser ablation source to the injector for transporting a plurality of ablation plumes from the laser ablation source to the injector, the connecting tube comprises an expandable elastomeric material; a connecting tube, the connecting tube being characterized by a first inner diameter in a range of 0.3 mm to 6.0 mm; An apparatus comprising:

14. a rigid tube portion operably connected to the connecting tube; the rigid tube having a first end operably coupled to the laser ablation source; the rigid tube portion having a second end operably connected to the connecting tube; the rigid tube portion is characterized by a second inner diameter at the first end that is greater than the first inner diameter of the connecting tube; the rigid tube portion is characterized by a third inner diameter at the second end; The second inner diameter is larger than the third inner diameter.

14. The apparatus of claim 13.

15. the laser ablation source comprises a sampling ramp; the sampling ramp is operably coupled to the first end of the rigid tube; the sampling ramp is tapered to have a smaller diameter proximal to the first end of the rigid tubing as compared to distal to the first end of the rigid tubing.

15. The apparatus of claim 14.

16. The apparatus of claim 14, wherein the second inner diameter is in the range of 1.5 mm to 2.0 mm.

17. 15. The apparatus of claim 14, wherein the rigid tube section comprises a 10 degree taper configured to narrow the third inner diameter at the second end that leads to the manifold within a range of 0.5 mm to 1.0 mm.

18. 15. The apparatus of claim 14, further comprising a junction assembly between said rigid tubing section and said manifold, said junction assembly emerging at said junction assembly a taper of reducing diameter leading to said manifold.

19. 20. The apparatus of claim 18, wherein the junction assembly comprises a barbed joint configured to cooperate with a third end of the connecting tube, the barbed joint comprising the taper that reduces the diameter leading to the connecting tube.

20. 14. The apparatus of claim 13, further comprising a junction assembly between the manifold and the syringe, the junction assembly emerging at a taper of reducing diameter leading to the syringe.

21. The apparatus of claim 13 further comprising the laser ablation source.

22. 14. The apparatus of claim 13, further comprising a mass spectrometer configured to analyze plasma products in the inductively coupled plasma chamber.

23. the laser ablation source comprising: A sample holder; A laser source; an optical system for directing a laser from the laser source to the sample holder; The apparatus of claim 13 , comprising:

24. The apparatus of claim 13 further comprising a gas source operably coupled to the laser ablation source.

25. ablating the sample with a laser to form an ablation plume; Flowing the ablation plume in a laminar manner from the sample through a manifold to an injector, the connecting tube comprises an expandable elastomeric material; the connecting tube being characterized by a first inner diameter in a range of 0.3 mm to 6.0 mm; analyzing the ablation plume using an inductively coupled plasma mass spectrometer coupled to the injector; A method comprising:

26. The method of claim 25 , wherein the extensible elastomeric material comprises a fluoroelastomer.

27. 26. The method of claim 25, further comprising the step of channeling the ablation plume through a barbed joint of a joint assembly before or after the step of channeling the ablation plume through the manifold.

28. before the step of channeling the ablation plume through the connecting tube, channeling the ablation plume through a rigid tube; the rigid tube having a first end operably coupled to a laser ablation source including the laser; the rigid tube portion having a second end operably connected to the connecting tube; the rigid tube portion is characterized by a second inner diameter at the first end that is greater than the first inner diameter of the connecting tube; the rigid tube portion is characterized by a third inner diameter at the second end; The second inner diameter is larger than the third inner diameter.

26. The method of claim 25.

29. 29. The method of claim 28, wherein the second inner diameter is in the range of 1.5 mm to 2.0 mm.

30. 30. The method of claim 28, wherein the rigid tube section comprises a 10 degree taper configured to narrow the third inner diameter at the second end that leads to the manifold within a range of 0.5 mm to 1.0 mm.

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