Waveguide module and preparation method for MALDI-TOF-MS target plate sample preparation

Abstract

The present invention provides a waveguide module and a preparation method for MALDI-TOF-MS target plate sample preparation. The waveguide module for MALDI-TOF-MS target plate sample preparation includes a frame, a target plate and an excitation element. A first accommodating space is provided inside the frame. The target plate is arranged on the frame. The excitation element is arranged in the first accommodating space, and the excitation element is configured to generate a mechanical vibration wave and the mechanical vibration wave can be transmitted to the target plate. According to the waveguide module and preparation method for MALDI-TOF-MS target plate sample preparation of the present invention, a mechanical vibration wave is generated by an excitation element, and the mechanical vibration wave is transmitted to the target plate, so that the sample in the receiving portion on the target plate generates mechanical vibration, and then in the crystallization process of the co-crystallized film, the unconstrained liquid sample is affected by the mechanical vibration wave, and a homogenization effect similar to "collapse" occurs, thereby improving the resolution and sensitivity of sample detection.

Description

Technical Field

[0001] The invention relates to the technical field of matrix-assisted laser desorption / ionization time of flight mass spectrometry (hereinafter referred to as MALDI-TOF-MS) target plate sample preparation, and in particular to a waveguide module and a preparation method for MALDI-TOF-MS target plate sample preparation. Background Art

[0002] MALDI-TOF-MS is a new soft ionization biomass spectrometry technology developed in recent years. This technology has high sensitivity and high-quality detection range, and can be applied to life sciences, clinical testing, molecular medicine, biosafety, biomedicine and health. The detection application at the molecular level has covered many industries such as clinical diagnosis, clinical molecular diagnosis, epidemic prevention and quarantine, molecular pathology diagnosis, life sciences, bioinformatics, biosafety protection, pharmaceutical research, etc., and is widely used as an important detection method in colleges and universities, scientific research institutes, health and medical institutions, CDC (Centers for Disease Control), animal and plant quarantine, food safety, aquatic animal husbandry, public security, bioterrorism, pharmaceutical research and development and other departments and units.

[0003] The basic principle of using MALDI-TOF-MS for biological sample detection is: the biological sample is mixed with an equal amount of matrix solution or separately applied to the biological sample target plate, and a co-crystallized film of the biological sample and the matrix is ​​formed after the solvent evaporates; the co-crystallized film is irradiated with laser as an energy source, the matrix absorbs energy from the laser to adsorb the biological sample, and charge transfer occurs between the matrix and the biological sample to ionize the biological sample molecules; the biological sample molecules enter the time-of-flight mass spectrometer for mass analysis under the action of the accelerating electric field.

[0004] Unlike conventional physical material testing, the use of MALDI-TOF-MS for biological sample testing usually requires a lot of preliminary processing of the biological sample material and the matrix (usually liquid) to make the spatial distribution morphology of the co-crystallized film as close to the Figure 3 The theoretical state shown is that the biological sample and the matrix form a co-crystallized film as uniformly as possible.

[0005] However, in actual operation, since the preparation amount required for each site of the target plate is usually between dozens of nanoliters and 2 microliters, whether using traditional manual preparation or expensive piezoelectric spray equipment, even professionals who have undergone long-term training cannot guarantee that each prepared site can obtain an ideal test result. In view of this situation, those skilled in the art usually focus on the spray module of the spray equipment, while the present invention provides a waveguide module and preparation method for MALDI-TOF-MS target plate sample preparation, shifting the focus of transformation to the carrier module of the target plate, so as to at least partially solve the above-mentioned defects and deficiencies existing in the prior art. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides a waveguide module for MALDI-TOF-MS target plate sample preparation. The waveguide module for MALDI-TOF-MS target plate sample preparation includes a frame, a target plate and an excitation element. A first accommodating space is arranged inside the frame. The target plate is arranged on the frame. The excitation element is arranged in the first accommodating space, and the excitation element is configured to generate a mechanical vibration wave and the mechanical vibration wave can be transmitted to the target plate.

[0007] Preferably, the frame comprises a base plate, a bracket, a pressure cover and a full-size target plate bracket. The bracket is connected to the base plate. The pressure cover is connected to the bracket, the first accommodation space is arranged between the bracket and the pressure cover, and the pressure cover fixes the excitation element in the first accommodation space. The full-size target plate bracket is connected to the bracket above the bracket.

[0008] Preferably, the target plate is arranged on the full-size target plate holder.

[0009] Preferably, the frame further comprises a replaceable target plate holder, the replaceable target plate holder is detachably connected to the full-size target plate holder, and the target plate is arranged on the replaceable target plate holder.

[0010] Preferably, the target plate is adsorbed onto the frame by permanent magnetic adsorption.

[0011] Preferably, the waveguide module for MALDI-TOF-MS target plate sample preparation further comprises a thermostatic element, and a second accommodating space is provided between the bracket and the full-size target plate bracket, and the thermostatic element is provided in the second accommodating space.

[0012] Preferably, the waveguide module for MALDI-TOF-MS target plate sample preparation further comprises a program-controlled driver and a microprocessor, wherein the program-controlled driver is electrically connected to the excitation element and the constant temperature element, and the program-controlled driver can communicate with the microprocessor.

[0013] The present invention also provides a preparation method for MALDI-TOF-MS target plate sample preparation, wherein the preparation method uses any of the above-mentioned waveguide modules for MALDI-TOF-MS target plate sample preparation.

[0014] According to the waveguide module and preparation method for MALDI-TOF-MS target plate sample preparation of the present invention, a mechanical vibration wave is generated by an excitation element, and the mechanical vibration wave is transmitted to the target plate, so that the sample in the receiving portion on the target plate generates mechanical vibration, and then during the crystallization process of the co-crystallized film, the unconstrained liquid sample is affected by the mechanical vibration wave, and a homogenization effect similar to "collapse" occurs, thereby improving the resolution and sensitivity of sample detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Non-limiting and non-exhaustive embodiments of the present invention are described, by way of example, with reference to the following drawings, in which:

[0016] Figure 1 is a top view schematic diagram of a waveguide module for MALDI-TOF-MS target plate sample preparation according to a preferred embodiment of the present invention;

[0017] Figure 2 yes Figure 1 The waveguide module for MALDI-TOF-MS target plate sample preparation is shown in FIG. Figure 1 A cross-sectional view along line AA; and

[0018] Figure 3 It is a schematic diagram of the effect of the spatial distribution morphology of the co-crystallized film on the analytical test results. DETAILED DESCRIPTION

[0019] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0020] In a first aspect of the present invention, a waveguide module for MALDI-TOF-MS target plate sample preparation is provided. Figure 1 1 is a schematic top view of a waveguide module for MALDI-TOF-MS target plate sample preparation according to a preferred embodiment of the present invention. Figure 2 yes Figure 1The waveguide module for MALDI-TOF-MS target plate sample preparation is shown in FIG. Figure 1 Section view along line AA. Figure 3 This is a schematic diagram of the effect of the spatial distribution of the co-crystallized film on the analysis and test results. Figures 1 to 3 A waveguide module 100 for MALDI-TOF-MS target plate sample preparation according to a preferred embodiment of the present invention is described in detail.

[0021] like Figure 1 to Figure 2 As shown, the waveguide module 100 includes a frame 110 , a target plate 120 disposed on the frame 110 , and an excitation element 1110 .

[0022] like Figure 2 As shown, a first accommodation space 112 is provided inside the frame 110 to accommodate the excitation element 1110 of the waveguide module 100. Preferably, at least a portion of the frame 110 may be made of a metal material having good conduction performance for mechanical vibration waves, which can provide good mechanical vibration wave conduction performance on the one hand, and can provide the mechanical strength required to support the target plate 120 on the other hand.

[0023] like Figure 1 and Figure 2 As shown, the waveguide module 100 includes a target plate 120, which is arranged on the rack 110. The target plate 120 can be a metal target plate, a disposable conductive glass target plate, a disposable disposable plastic target plate or a paper-based target plate. Preferably, the target plate 120 is a metal target plate. On the one hand, the use of a metal target plate is conducive to improving the resolution, sensitivity and other properties of sample detection, so that the reproducibility of sample detection is better; on the other hand, the metal target plate has good conduction performance for mechanical vibration waves and good thermal conductivity; on the other hand, the metal target plate can be reused many times, so that the detection process does not require additional consumables. The target plate 120 includes a plurality of accommodating portions 1201 (i.e., sites) for accommodating a matrix and a sample, and the plurality of accommodating portions 1201 are arranged equidistantly and distributed in a matrix form. Specifically, in the present embodiment, the target plate 120 is set to a rectangular shape, and the target plate 120 includes 96 accommodating portions 1201, and the 96 accommodating portions 1201 are arranged and distributed in a matrix form of 8×12. In addition, the center distance between any two adjacent accommodating portions 1201 can be set as needed, for example, the center distance can be 2.25 mm, 4.5 mm or 9 mm, or larger, so as to meet different detection requirements.

[0024] like Figure 2As shown, the excitation element 1110 is disposed in the first accommodating space 112, and the excitation element 1110 is configured to generate a mechanical vibration wave, and the mechanical vibration wave can be transmitted to the target plate 120. In one embodiment of the present invention, in the working state, the excitation element 1110 can be fixedly disposed in the first accommodating space 112, that is, in the working state, the excitation element 1110 does not move relative to the frame 110. The excitation element is configured to generate a mechanical vibration wave when excited, and the mechanical vibration wave generated by the excitation element 1110 can be transmitted to the target plate 120 through the frame 110, so that the receiving portion 1201 on the target plate 120 generates mechanical vibration, and then the sample (a mixture of the sample and the matrix) in the receiving portion 1201 generates mechanical vibration, so that in the crystallization process of the co-crystallized film, the unconstrained liquid sample is affected by the mechanical vibration wave, and a homogenization effect similar to "collapse" occurs, thereby helping the spatial distribution morphology of the co-crystallized film to be as uniform as possible and tend to be as follows Figure 3 Theoretical state shown to improve the resolution and sensitivity of sample detection.

[0025] Specifically, in the present embodiment, the frame 110 includes a base plate 1106, a bracket 1107, a pressure cover 1109, and a full-size target plate bracket 1104. The bracket 1107 is connected to the base plate 1106. Specifically, the bracket 1107 can be connected to the base plate 1106 by welding connection or threaded fastener connection. The pressure cover 1109 is connected to the bracket 1107, the first accommodating space 112 is provided between the bracket 1107 and the pressure cover 1109, and the pressure cover 1109 fixes the excitation element 1110 in the first accommodating space 112. Specifically, in the present embodiment, the lower part of the bracket 1107 is provided with a first recessed portion concave upward, the pressure cover 1109 is connected to the lower part of the bracket 1107 by screws 1108 and covers the first recess, thereby forming the first accommodating space 112 with the first recess of the bracket 1107, and the pressure cover 1109 fixes the excitation element 1110 in the first recess of the bracket 1107. Figure 1 and Figure 2 As shown, the full-size target plate holder 1104 is connected to the bracket 1107 above the bracket 1107. It should be noted that the full-size target plate holder mentioned here refers to the target plate holder of the maximum size specification, that is, the full-size target plate holder can match the target plate of the maximum size specification. In the working state, the full-size target plate holder 1104 can be fixed to the bracket 1107, that is, in the working state, the full-size target plate holder 1104 does not move relative to the bracket 1107. Specifically, in the present embodiment, the full-size target plate holder 1104 is connected to the base plate 1106 together with the bracket 1107 by screws 1105. Preferably, the full-size target plate holder 1104 and the bracket 1107 are made of a metal material having good conduction performance to mechanical vibration waves and good thermal conductivity.

[0026] In one embodiment of the present invention, the target plate 120 can be directly disposed on the full-size target plate holder 1104. Designed in this manner, a target plate 120 of a maximum size can be used.

[0027] In a preferred embodiment of the present invention, Figure 1 and Figure 2 As shown, the rack 110 further includes a replaceable target plate holder 1103, which is detachably connected to the full-size target plate holder 1104. Specifically, the top of the full-size target plate holder 1104 is provided with a second recessed portion that is concave downward, and the bottom of the replaceable target plate holder 1103 is provided with a protrusion that protrudes downward, and the size and shape of the second recess of the full-size target plate holder 1104 are respectively matched with the size and shape of the protrusion of the replaceable target plate holder 1103, so that the protrusion of the replaceable target plate holder 1103 can be inserted into the second recess of the full-size target plate holder 1104, so that the replaceable target plate holder 1103 is detachably connected to the full-size target plate holder 1104. In addition, the replaceable target plate holder 1103 can be further detachably connected to the full-size target plate holder 1104 by a positioning screw 1102. The target plate 120 is arranged on the replaceable target plate holder 1103. The size of the replaceable target plate holder 1103 is smaller than that of the full-size target plate holder 1104. The multiple replaceable target plate holders 1103 have different sizes. When target plates 120 of different sizes are needed, replaceable target plate holders 103 of corresponding sizes can be selected to meet different detection requirements.

[0028] Preferably, the target plate 120 is adsorbed onto the rack 110 by permanent magnetic adsorption. Specifically, in a preferred embodiment of the present invention, the target plate 120 is adsorbed onto the replaceable target plate holder 1103 by permanent magnetic adsorption. Designed in this way, it is convenient to replace the target plate 120. Of course, in other embodiments not shown in the present invention, the target plate 120 can also be connected to the rack 110 by other detachable connection methods, such as adhesion.

[0029] like Figure 2As shown, the waveguide module 100 further includes a thermostatic element 1111, and a second accommodating space 113 is provided between the bracket 1107 and the full-size target plate holder 1104, and the thermostatic element 1111 is provided in the second accommodating space 113. Specifically, in the present embodiment, a portion of the lower surface of the full-size target plate holder 1104 is concave upward and spaced apart from the bracket 1107, thereby forming a third concave portion at the lower portion of the full-size target plate holder 1104, and the third concave portion forms the above-mentioned second accommodating space 113. Of course, in other embodiments not shown in the present invention, a portion of the upper surface of the bracket 1107 may also be concave downward and spaced apart from the full-size target plate holder 1104, thereby forming a second accommodating space 113 for accommodating the thermostatic element 1111 at the upper portion of the bracket 1107. The constant temperature element 1111 is in thermal contact with the full-size target plate holder 1104 or the replaceable target plate holder 1103, and thus in thermal contact with the target plate 120, so as to control the temperature of the target plate 120 within a predetermined temperature range, thereby accelerating the crystallization speed and improving the time efficiency.

[0030] like Figure 2 As shown, the waveguide module 100 further includes a programmable driver 130, which is electrically connected to the excitation element 1110 and the constant temperature element 1111. The programmable driver 130 can communicate with a microprocessor (MPU) to control the opening and closing of the excitation element 1110 and the constant temperature element 1111.

[0031] In a second aspect of the present invention, a preparation method for MALDI-TOF-MS target plate sample preparation is also provided. The preparation method uses any of the above-mentioned waveguide modules for MALDI-TOF-MS target plate sample preparation.

[0032] The following will refer to Figures 1 to 3 A preparation method for MALDI-TOF-MS target plate sample preparation using the waveguide module 100 of the preferred embodiment of the present invention is described in detail.

[0033] When the target plate sample preparation begins, the excitation component 1110 and the constant temperature module 1111 are put into operation in a timely manner under the scheduling of the MPU. It should be noted that in the working state, all components of the waveguide module 100 are fixed, and no displacement occurs in the plane X and Y directions, and only a small amplitude displacement occurs in the Z direction perpendicular to the XY plane. Figure 2 and Figure 3As shown, the mechanical vibration wave generated by the excitation component 1110 is transmitted to the target plate 120 through the frame 110, so that the receiving portion 1201 on the target plate 120 generates mechanical vibration, and then the liquid sample in the receiving portion 1201 generates mechanical vibration, so that during the crystallization process of the co-crystallized film, the unconstrained liquid sample is affected by the mechanical vibration wave, and a homogenization effect similar to "collapse" occurs, so that the spatial distribution morphology of the co-crystallized film is transformed from an unqualified state before the "collapse" to a qualified state after the "collapse", which significantly improves the resolution and sensitivity of sample detection.

[0034] The waveguide module 100 and preparation method for MALDI-TOF-MS target plate sample preparation according to the present invention can be used not only for mechanical automated sample preparation process, but also for manual sample preparation process. In the waveguide module 100 and preparation method for MALDI-TOF-MS target plate sample preparation according to the present invention, a mechanical vibration wave is generated by the excitation element 1110, and the mechanical vibration wave is transmitted to the target plate 120, so that the sample in the receiving portion 1201 on the target plate 120 generates mechanical vibration, and then in the crystallization process of the co-crystallized film, the unconstrained liquid sample is affected by the mechanical vibration wave, and a homogenization effect similar to "collapse" occurs, thereby improving the resolution and sensitivity of the sample detection. In addition, by replacing the replaceable target plate holder 1103, the waveguide module 100 for MALDI-TOF-MS target plate sample preparation of the present invention can be adapted to target plates of all sizes, such as disposable conductive glass target plates and reusable metal target plates.

[0035] The technical features of the above embodiments may be combined arbitrarily. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0036] Although the present invention has been described in conjunction with the embodiments, it should be understood by those skilled in the art that the above description and the accompanying drawings are only exemplary and non-restrictive, and the present invention is not limited to the disclosed embodiments. Various modifications and variations are possible without departing from the spirit of the present invention.

Claims

1. A waveguide module for MALDI-TOF-MS target plate sample preparation, characterized in that: The waveguide module for MALDI-TOF-MS target plate sample preparation comprises: A frame, wherein a first accommodation space is provided inside the frame; a target plate, wherein the target plate is arranged on the frame; and An excitation element, the excitation element being arranged in the first accommodating space, the excitation element being configured to generate a mechanical vibration wave and the mechanical vibration wave being capable of being transmitted to the target plate, so that the target plate generates a small amplitude displacement only in a Z direction perpendicular to the XY plane; The frame comprises: substrate; a bracket connected to the base plate; a gland connected to the bracket, the first accommodation space being disposed between the bracket and the gland, and the gland fixing the excitation element in the first accommodation space; and A full-size target plate holder is connected to the bracket above the bracket.

2. The waveguide module for MALDI-TOF-MS target plate sample preparation according to claim 1, characterized in that: The target plate is arranged on the full-size target plate holder.

3. The waveguide module for MALDI-TOF-MS target plate sample preparation according to claim 1, characterized in that: The frame further comprises a replaceable target plate holder, which is detachably connected to the full-size target plate holder, and the target plate is arranged on the replaceable target plate holder.

4. The waveguide module for MALDI-TOF-MS target plate sample preparation according to any one of claims 1 to 3, characterized in that: The target plate is adsorbed onto the frame by permanent magnetic adsorption.

5. The waveguide module for MALDI-TOF-MS target plate sample preparation according to any one of claims 2 to 3, characterized in that: The waveguide module for MALDI-TOF-MS target plate sample preparation further comprises a constant temperature element, and a second accommodating space is arranged between the bracket and the full-size target plate bracket, and the constant temperature element is arranged in the second accommodating space.

6. The waveguide module for MALDI-TOF-MS target plate sample preparation according to claim 5, characterized in that: The waveguide module for MALDI-TOF-MS target plate sample preparation further comprises a program-controlled driver and a microprocessor. The program-controlled driver is electrically connected to the excitation element and the constant temperature element, and the program-controlled driver can communicate with the microprocessor.

7. A method for preparing a MALDI-TOF-MS target plate sample, characterized in that: The preparation method uses the waveguide module for MALDI-TOF-MS target plate sample preparation according to any one of claims 1 to 6.

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