This application is a U.S. national phase filing claiming the benefit of and priority to International Patent Application No. PCT/GB2019/052706, filed Sep. 26, 2019, which claims priority from and the benefit of United Kingdom patent application No. 1815676.0 filed on Sep. 26, 2018. The entire contents of these applications are incorporated herein by reference.
Embodiments described herein relate generally to ionisation sources and in particular to a nozzle for a Matrix Assisted Laser Desorption Ionisation (“MALDI”) ion source.
Mass spectrometers comprising a Matrix Assisted Laser Desorption Ionisation (“MALDI”) ion source are known. MALDI mass spectrometry is a known process which is particularly suited for the analysis of non-volatile mass spectrometry. A suitable matrix material (e.g. an organic solvent) is added to a sample so that the sample becomes embedded in the matrix material. The embedded sample is then positioned on a metal plate and a laser pulse is directed on to the target sample. The laser pulse impinging upon the target sample causes analyte material to be ablated and desorbed from the target sample. Analyte ions are generated by analyte material being protonated and deprotonated in a hot plume of gaseous molecules which is released from the target. The matrix has a strong absorption at the wavelength of the laser pulse and acts as a proton source to encourage ionisation of the analyte. The gaseous plume which is released from the target comprises a mixture of analyte ions together with uncharged material. The mixture of analyte ions and uncharged material is then directed towards the inlet of a mass spectrometer. The ions are directed towards the mass spectrometer by an ion guide using electric fields. The analyte ions are separated from the uncharged material with the analyte ions being onwardly transmitted to a mass analyser of the mass spectrometer in order to be mass analysed.
Although the analyte ions are directed towards the mass spectrometer, the uncharged material may instead disperse into the vacuum chamber. As a result, the matrix may adsorb onto the surrounding surfaces in the vacuum chamber, resulting in a gradual build-up of material.
Any laser optics within the vacuum chamber can be susceptible to this matrix depositing on their surfaces, particularly if they are in a direct line of sight of the ablation. This can cause the transmissivity of the mirror to be reduced (“fogging”), while any material deposited on the optic surface may absorb subsequent laser radiation, causing localized heating at the point that the laser impinges on the surface, resulting in damage to the optic surface. This, in turn, may make the optic surface more absorbing, causing further damage.
Current approaches rely on either creating sufficient distance between the sample and the laser optics to prevent the uncharged material from reaching the laser optics and/or providing a flow of gas across the optic path (“gas curtain”) in an attempt to block the uncharged material from reaching the laser optics. However, this is not always effective and can result in a loss of spatial resolution when the optics are arranged far away from the sample.
Embodiments described herein seek to alleviate or reduce the amount of contaminants reaching the laser optics. Embodiments described herein also seek to alleviate of reduce the amount of damage caused to laser optics during MALDI mass spectrometry. Embodiments described herein also combine the delivery of a cooling gas, with a method of generating a gas shield, to protect the laser optics by reducing the amount of ablated material adsorbed on the optics.
One aspect of the present invention provides a nozzle for an ionisation source comprising:
In at least one embodiment, the gas flow passage is in fluid communication with the light flow passage between the inlet end of the light passage and the outlet end of the light passage.
In at least one embodiment, the nozzle further comprises a window disposed along the light passage.
In at least one embodiment, the window is disposed adjacent the inlet end of the light passage.
In at least one embodiment, the window is received in a recess adjacent the inlet end of the light passage.
In at least one embodiment, the cross-sectional area of the inlet end of the light passage is larger than the cross-sectional area of the outlet end of the light passage.
In at least one embodiment, the light passage is substantially conical.
In at least one embodiment, the nozzle further comprises an aperture configured to receive an ion guide.
In at least one embodiment, the angle between the longitudinal axis of the light passage and the longitudinal axis of the aperture is between 0 and 90 degrees, or between 0 and 45 degrees, or between 0 and 30 degrees.
In at least one embodiment, the nozzle further comprises a viewing passage, wherein the angle between the longitudinal axis of the viewing passage and the longitudinal axis of the aperture is different to the angle between the longitudinal axis of the light passage and the longitudinal axis of the aperture.
In at least one embodiment, the nozzle further comprises a guard which extends in a direction of the longitudinal axis of the aperture.
Another aspect of the present invention provides a nozzle assembly comprising:
In at least one embodiment, the ion guide is a hexapole ion guide.
In at least one embodiment, the extraction electrode is disposed adjacent the outlet end of the light passage.
In at least one embodiment, the extraction electrode comprises a sample inlet configured to receive at least some of an ionised sample in use.
Another aspect of the present invention provides an ionisation assembly comprising:
In at least one embodiment, the nozzle assembly is slidably receivable in the ionisation chamber.
In at least one embodiment, the ionisation chamber is a vacuum chamber.
In at least one embodiment, the ionisation assembly further comprises a gas supply passage having an inlet end outside of the ionisation chamber to receive a flow of gas, and an outlet end, wherein the gas flow passage of the nozzle has an inlet end which is fluidly connected to the outlet end of the gas supply passage when the nozzle is received in the ionisation chamber.
In at least one embodiment, the ionisation assembly further comprises a beam steering arrangement configured to direct a light source through the light passage in use.
In at least one embodiment, the beam steering arrangement comprises at least one mirror disposed inside the ionisation chamber.
In at least one embodiment, the ionisation assembly further comprises a laser light source.
Another aspect of the present invention provides a nozzle for an ionisation source comprising:
Another aspect of the present invention provides a nozzle for an ionisation source comprising:
In
At least an exit end of the gas flow passage 24 is angled with respect to the light passage 21. In at least one embodiment, the angle may be between 1 and 179 degrees. The gas flow passage 24 is not coaxial with the light passage 21. In at least one embodiment, the angle may be 90 degrees. In at least one embodiment, the angle may be between 45 and 89 degrees, such that there is an acute angle between the gas flow passage 24 and the inlet end 22 of the light passage 21. Angling the gas flow passage 24 with respect to the light passage 21, particularly by an acute angle, promotes the flow of gas towards the outlet end 23 of the light passage. If the gas flow passage 24 is angled with respect to the light passage 21 by an angle of substantially 90 degrees, there may be provided a baffle and/or other flow directing means, to promote the flow of gas towards the outlet end 23 of the light passage 21.
In one embodiment, the nozzle is made of an insulting material.
In the embodiment shown in
In one embodiment, the gas provided through the gas flow passage 24 is a collisional cooling gas. Additionally or alternatively, the gas can be used as a carrier gas to transfer reagent molecules into the vicinity of the plume.
The nozzle 20 may also comprise a window 25 disposed along the light passage 21. The window 25 provides a physical barrier that may prevent material (e.g. a gas or ablated material) from exiting through the inlet end 22 of the light passage 21. The window 25 may also help to direct the flow of gas towards the outlet end 23 of the light passage 21 by providing a single flow path for the gas to travel along.
In one embodiment, the window 25 is disposed adjacent the inlet end 22 of the light passage 21. Alternatively or additionally, the window 25 may be received in a recess adjacent the inlet end 22 of the light passage 21. The recess also helps to secure the window 25 in place by constraining movement in at least one direction.
The window 25 may be integrally formed with the nozzle 20 or may be provided as a separate component. Alternatively or additionally, the window 25 may be removable and/or replaceable. This allows the window 25 to be individually removed from the nozzle to be cleaned or replaced, without having to clean or replace the whole nozzle 20.
The light passage 21 shown in
As shown in
The angle between the longitudinal axis of the light passage 21 and the longitudinal axis of the aperture 26 (and thus the longitudinal axis of the ion guide) may be between 0 and 45 degrees. Alternatively, the angle may be between 0 and 30 degrees. It is advantageous to ensure that the angle between the longitudinal axis of the light passage 21 and the longitudinal axis of the aperture 26 is minimised and is as close to zero degrees as possible. This causes a light beam passing through the nozzle 20 to impinge on a sample surface with a less elliptical cross section. In one embodiment, the diameter of the light beam at the sample is about 15 μm. Alternatively, the diameter of the light beam at the sample may be between 5 μm and 20 μm.
The gas flow passage 24 may be substantially perpendicular to the longitudinal axis of the aperture 26 (and thus the longitudinal axis of the ion guide).
It is advantageous for a light beam 32 to impinge on a sample surface 49 along, or close to, the normal of the sample surface 49 (i.e. substantially perpendicular to the sample surface 49) to minimise ellipticity. In the arrangement of
In one embodiment, the nozzle 20 further comprises a viewing passage 28. The viewing passage 28 may be used to visually verify that the arrangement is correctly aligned. The angle between the longitudinal axis of the viewing passage 28 and the longitudinal axis of the aperture 26 may be different to the angle between the longitudinal axis of the light passage 21 and the longitudinal axis of the aperture 26. This may prevent or reduce the amount of light that is reflected directly off of the sample into either the eye of an operator or a camera. The viewing passage 28 may be directly viewed by a user and/or may comprise a camera or other light sensing device to monitor the alignment.
The embodiment shown in
In one embodiment, the ionisation source is a Matrix Assisted Laser Desorption Ionisation source. Additionally or alternatively, the ionisation source may be any other suitable ionisation source.
In one embodiment, a nozzle assembly 30 is provided as shown in
As shown in
In one embodiment, an ionisation assembly 40 is provided, as shown in
In one embodiment, the ionisation assembly 40 comprises a gas supply passage 43. The gas supply passage 43 may have an inlet end 44 outside of the ionisation chamber 41 to receive a flow of gas. The gas supply passage 43 may also comprise an outlet end 45. The gas flow passage 24 of the nozzle 20 has an inlet end 46 which is fluidly connected to the outlet end 45 of the gas supply passage 43 when the nozzle 20 is received in the ionisation chamber 41. Alternatively or additionally, the gas supply passage 43 may comprise a flexible tube. Alternatively or additionally, the gas supply passage 43 may take any other suitable form provided it allows a gas to be delivered into the gas flow passage.
Although
In one embodiment, the ionisation assembly 40 comprises a light source 47. In one embodiment, the light source 47 is a laser light source.
In one embodiment, the ionisation assembly 40 comprises a beam steering arrangement 48. The beam steering arrangement 48 may be configured to direct a light source 47 through the light passage 22. The beam steering arrangement 48 may comprise at least one mirror. The at least one mirror may be disposed inside the ionisation chamber 41.
In one embodiment, the beam steering arrangement 48 additionally comprises at least one lens. The at least one lens may be configured to focus the light source. As discussed previously, it is advantageous to avoid using a long focal length. In one embodiment, the focal length of the light source is 75 mm.
An ion guide 27 used in any of the embodiments discussed herein may comprise a plurality of cylindrical poles arranged, in parallel, in a square, hexagonal or substantially circular pattern. There may be gaps between each of the poles.
Referring to
When used in this specification and claims, numerical ranges are taken to also include the end points of said ranges.
When used in this specification and claims, the terms “comprises” and “comprising” and variations thereof mean that the specified features, steps or integers are included. The terms are not to be interpreted to exclude the presence of other features, steps or components.
The features disclosed in the foregoing description, or the following claims, or the accompanying drawings, expressed in their specific forms or in terms of a means for performing the disclosed function, or a method or process for attaining the disclosed result, as appropriate, may, separately, or in any combination of such features, be utilised for realising the invention in diverse forms thereof.
Number | Date | Country | Kind |
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1815676 | Sep 2018 | GB | national |
Filing Document | Filing Date | Country | Kind |
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PCT/GB2019/052706 | 9/26/2019 | WO |
Publishing Document | Publishing Date | Country | Kind |
---|---|---|---|
WO2020/065312 | 4/2/2020 | WO | A |
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Number | Date | Country |
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2405463 | Jan 2012 | EP |
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Entry |
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Combined Search and Examination Report for GB1815676.0, dated Mar. 27, 2019. |
International Search Report and Written Opinion for International Application PCT/GB/2019/052706, dated Dec. 3, 2019. |
Number | Date | Country | |
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20220005684 A1 | Jan 2022 | US |