A MALDI mass spectrometer matrix spraying instrument

Abstract

This utility model relates to a MALDI mass spectrometer matrix spraying instrument, belonging to the field of mass spectrometry analysis technology. It includes: a matrix spraying assembly and a spraying sealed chamber; the matrix spraying assembly is placed inside the spraying sealed chamber and is capable of spraying mass spectrometry matrix; the matrix spraying assembly includes: a nozzle and a nozzle temperature control assembly; the nozzle temperature control assembly ensures that the nozzle operates within a stable temperature range. This utility model adds an ultrasonic nozzle cooling module, which adjusts and stabilizes the temperature of the ultrasonic nozzle during spraying through a temperature probe and a semiconductor cooling chip, overcoming the defects of unstable spraying and damage to internal nozzle components if excess heat is not dissipated in time.

Description

Technical Field

[0001] This utility model relates to the field of mass spectrometry analysis technology, and in particular to a MALDI mass spectrometry matrix sprayer. Background Technology

[0002] Mass spectrometry is a widely used analytical technique that provides rich information about the mass and structure of molecules. The ion source is a crucial component of a mass spectrometer; it ionizes the analyte molecules into ions, which are then successfully introduced into the subsequent detection apparatus. Therefore, the ionization system is essential for mass spectrometry detection.

[0003] Matrix-assisted laser desorption / ionization (MALDI) is a recently developed soft ionization method for mass spectrometry. MALDI can be used for the ionization analysis of biological macromolecules. Simultaneously, MALDI technology can also be used for mass spectrometric imaging of biological tissue sections to obtain spatial distribution information of molecules on them.

[0004] The characteristic of MALDI technology lies in the fact that when a thin film co-crystallized from a sample and a matrix solution is irradiated with a high-energy pulsed laser, the matrix absorbs the laser energy in this wavelength range, causing the crystals in the irradiated area to instantly desorb, vaporize, and ionize. The ionization mechanism of the analyte molecules is not fully understood, but the two most accepted mechanisms are: analyte ion formation during co-crystallization; and proton transfer in the gas phase within the matrix-analyte pulse stream generated by laser desorption. Regardless of the mechanism, the preparation of the MALDI imaging sample—specifically, how the matrix is ​​deposited on the sample and co-crystallized—is crucial for quality control in MALDI imaging analysis.

[0005] Existing matrix spraying instruments suffer from problems such as heat dissipation difficulties, poor equipment stability, and short service life. The prepared matrix layers are often not uniform and fine enough, which limits the clarity, resolution, and throughput of mass spectrometry imaging analysis. They cannot reflect the specific distribution of biological macromolecules in MALDI imaging analysis. Utility Model Content

[0006] Based on the above analysis, this utility model aims to provide a MALDI mass spectrometer matrix spraying instrument, which solves at least one of the following technical problems of the prior art: (1) poor imaging clarity and low resolution during matrix sample preparation, which cannot reflect the specific distribution of biological macromolecules; (2) poor equipment stability and short service life; (3) difficult heat dissipation, which reduces the accuracy of detection; (4) inability to accurately control the spraying area.

[0007] This utility model provides a MALDI mass spectrometer matrix spraying instrument, comprising:

[0008] A spray-sealed chamber and a matrix spraying assembly disposed inside the spray-sealed chamber;

[0009] The matrix spraying assembly is placed inside the spraying sealed chamber and is capable of spraying mass spectrometry matrix;

[0010] The matrix spraying assembly includes a nozzle temperature control module; the nozzle temperature control module can ensure that the nozzle operates within a stable temperature range.

[0011] Preferably, the nozzle temperature control module includes a nozzle, a semiconductor cooling chip, and a high-conductivity hot water cooling plate; the nozzle, semiconductor cooling chip, and high-conductivity hot water cooling plate are connected in sequence, and the heat generated by the nozzle can be transferred to the high-conductivity hot water cooling plate through the semiconductor cooling chip;

[0012] A water-cooled plate slot is provided between the high-conductivity hot water cooling plate and the Z-axis platform, and the high-conductivity hot water cooling plate is fixed to the Z-axis platform through the water-cooled plate slot.

[0013] Preferably, the nozzle temperature control module further includes a nozzle mounting base;

[0014] The nozzle is fixed to the semiconductor cooling chip via a nozzle mounting base.

[0015] Preferably, the nozzle temperature control module further includes: a heat dissipation water duct, a cooling water tank, and a cooling fan;

[0016] The outlet of the high conductivity hot water cooling plate is connected to the inlet of the heat dissipation water duct, and the outlet of the heat dissipation water duct is connected to the inlet of the cooling water tank; the outlet of the cooling fan faces the heat dissipation surface of the heat dissipation water duct, and uses air flow to cool the coolant in the heat dissipation water duct.

[0017] The coolant at a lower temperature in the cooling water tank circulates to the inlet of the high-conductivity hot water cooling plate to continue cooling the plate.

[0018] Preferably, the matrix spraying assembly further includes a glass slide holder;

[0019] The slide holder is fixed to the base of the mass spectrometer matrix sprayer;

[0020] The projection area of ​​the nozzle is directly opposite the slide holder, and the jet stream from the nozzle covers the slide holder.

[0021] Preferably, the bottom of the nozzle is also provided with a condensate collector to collect condensate and prevent condensate from interfering with the slide holder.

[0022] Preferably, the mass spectrometer matrix sprayer further includes a spraying displacement mechanism. The moving part of the spraying displacement mechanism is fixedly connected to the matrix spraying assembly, and the fixed part is fixedly connected to the base of the mass spectrometer matrix sprayer, which enables the matrix spraying assembly to move in the plane of the base of the mass spectrometer matrix sprayer and in the direction perpendicular to the plane.

[0023] Preferably, the Y-axis displacement assembly includes: a first Y-axis slide rail, a second Y-axis slide rail, a Y-axis platform 4, a first Y-axis slider, a second Y-axis slider, and a Y-axis lead screw nut;

[0024] The first Y-axis slide rail and the second Y-axis slide rail are arranged in parallel and fixedly connected to the base of the mass spectrometer matrix sprayer;

[0025] The first and second Y-axis sliders are arranged parallel to each other at the bottom of the Y-axis platform. The first Y-axis slider is matched with the first Y-axis slide rail, and the second Y-axis slider is matched with the second Y-axis slide rail, so that the first and second Y-axis sliders can drive the Y-axis platform 4 to slide relative to the base of the mass spectrometer matrix sprayer in the Y-axis direction.

[0026] Preferably, the Y-axis displacement assembly further includes: a Y-axis zero-point position sensor, a Y-axis extreme position sensor, and a light-blocking plate for the Y-axis position sensor;

[0027] The light-blocking plate of the Y-axis position sensor generates a signal when it passes the Y-axis zero-point position sensor and the Y-axis extreme position sensor. The signal is used to control the displacement of the Y-axis platform between the zero point and the extreme position.

[0028] Compared with the prior art, the present invention can achieve at least one of the following beneficial effects:

[0029] 1. This utility model adds an ultrasonic nozzle cooling module, which adjusts and stabilizes the temperature of the ultrasonic nozzle during the spraying process through a temperature probe and a semiconductor cooling chip, overcoming the defects of unstable spraying and damage to internal components of the nozzle if excess heat cannot be removed in time.

[0030] 2. This utility model selectively humidifies the sealed spraying chamber using a humidification component, thereby improving the impact of seasonal humidity changes on the uniformity and stability of the spraying, enhancing the stability of the substrate spraying, and is particularly suitable for MALDI detection of macromolecular substances.

[0031] 3. This utility model, by setting up a humidification control board, temperature and humidity sensor, humidification water tank, humidification water trough, ultrasonic humidifier and humidification circulating fan, can not only realize automatic control of the humidification process, but also achieve a better humidification effect by means of ultrasonic atomization.

[0032] 4. Compared with the existing technology that drives the lead screw to rotate via belt drive, thereby driving the four-bar linkage and platform movement, this utility model does not require additional components for belt drive, simplifies the spraying component drive unit, improves power transmission efficiency, enhances system reliability, and achieves precise positioning of the spraying area.

[0033] 5. Existing technology in MALDI matrix sprayers uses only a single central motherboard to control all functions of the machine. This leads to drawbacks such as complex internal wiring, inconvenient maintenance, and low control efficiency during actual use. Here, we divide the entire machine control into four parts: a control motherboard, a motion control board, a humidification control board, and an ultrasonic control motherboard. The control motherboard acts as a host computer, connecting to and controlling the other three motherboards, thus avoiding the influence of the previous complex circuitry and improving control efficiency. This invention avoids the influence of the previous complex circuitry, improves control efficiency, and overcomes the drawbacks of complex internal wiring, inconvenient maintenance, and low control efficiency.

[0034] 6. This utility model, through its partitioned and mutually sealed spraying sealing chamber, motion mechanism chamber, and temperature control equipment chamber, avoids the influence of external humidity on spraying, and also avoids the influence of high humidity environment on internal electrical components; it also avoids the influence of heat generated by the spray head itself on the spraying effect; it overcomes the defects of high humidity environment caused by spraying leading to severe corrosion of electrical components, poor equipment stability, and short service life; it also overcomes the defects of numerous and dense internal components, difficulty in heat dissipation leading to excessively high internal temperature, damage to precision components, denaturation and deactivation of macromolecules in the spray liquid, resulting in distortion and reduced detection accuracy.

[0035] In this invention, the above-described technical solutions can be combined with each other to achieve more preferred combinations. Other features and advantages of this invention will be set forth in the following description, and some advantages will become apparent from the description or be learned by practicing the invention. The objectives and other advantages of this invention can be realized and obtained from the description and accompanying drawings, which are particularly pointed out. Attached Figure Description

[0036] The accompanying drawings are for illustrative purposes only and are not intended to limit the scope of the invention. Throughout the drawings, the same reference numerals denote the same parts.

[0037] Figure 1 A schematic diagram of the external appearance of a multi-module, separately controlled MALDI mass spectrometer matrix sprayer with humidification function;

[0038] Figure 2 A schematic diagram showing the control board setup for a multi-module, separately controlled MALDI mass spectrometer matrix sprayer with humidification function;

[0039] Figure 3 A 3D solid diagram of some components of a multi-module, separately controlled MALDI mass spectrometer matrix sprayer with humidification function;

[0040] Figure 4A 3D solid diagram of the spraying displacement mechanism and matrix spraying component positions and some parts of the MALDI mass spectrometer matrix sprayer with humidification function and multi-module control.

[0041] Figure 5 A 3D solid diagram of the Y-axis displacement assembly and some components of the MALDI mass spectrometer matrix sprayer with humidification function and multi-module control.

[0042] Figure 6 A 3D exploded view of the matrix spraying component of a multi-module, separately controlled MALDI mass spectrometer matrix sprayer with humidification function.

[0043] Figure 7 A 3D solid diagram of the X-axis platform of a multi-module, separately controlled MALDI mass spectrometer matrix sprayer with humidification function;

[0044] Figure 8 A 3D solid diagram of the X-axis displacement component of a multi-module, separately controlled MALDI mass spectrometer matrix sprayer with humidification function;

[0045] Figure 9 Cross-sectional view and detailed enlarged view of the humidification tank of a multi-module, separately controlled MALDI mass spectrometer matrix sprayer with humidification function;

[0046] Figure 10 The mass spectrometry results are for the sample prepared in Example 2.

[0047] Figure 11 The mass spectrometry results are for the sample prepared in Comparative Example 1.

[0048] Figure label:

[0049] 17-Slide holder; 18-First row of fans; 19-Second row of fans;

[0050] 2-Pressure reducing valve;

[0051] 23 - Gas flow meter; 27 - Interactive screen;

[0052] 3-Pressure reducing valve and barometer;

[0053] 45 - Y-axis first slide rail; 46 - Y-axis second slide rail; 47 - Y-axis zero position sensor; 48 - Y-axis extreme position sensor; 49 - Y-axis platform;

[0054] 50-Y-axis displacement assembly;

[0055] 501 - Y-axis first slider; 502 - Y-axis second slider; 52 - Y-axis lead screw nut; 53 - X-axis first slide rail; 54 - X-axis second slide rail; 55 - X-axis limit position sensor; 56 - Nozzle origin position sensor; 57 - X-axis zero position sensor; 58 - X-axis motor mounting bracket; 59 - X-axis stepper motor;

[0056] 60- X-axis displacement assembly;

[0057] 61 - Y-axis position sensor light shield; 62 - X-axis platform; 63 - X-axis lead screw nut; 64 - X-axis second slider; 65 - X-axis first slider; 66 - X-axis position sensor light shield; 67 - X-axis column; 68 - Y-axis stepper motor; 69 - Y-axis motor mounting bracket;

[0058] 70-Z axis displacement assembly;

[0059] 71-Crossbeam; 72-Z-axis base; 73-Z-axis platform; 74-High conductivity hot water cooling plate; 75-Semiconductor cooling chip; 79-Water cooling plate slot; 91-Lower brush strip; 92-Upper brush strip;

[0060] 8-Matrix spraying assembly; 80-Sprayer head; 81-Heat dissipation duct; 82-Cooling water tank; 85-Condensate collector; 86-Hollow sealing partition; 88-Ultrasonic control main board; 89-Humidifying circulating fan; 77-Sprayer head mounting base;

[0061] 9-Humidification component; 90-Humidification water tank; 93-Humidification water bath; 94-Cooling fan; 95-Water outlet valve; 96-Air return valve; 97-Humidification water bath level sensor; 98-Water pump;

[0062] 100 - Drain solenoid valve; 108 - Water tank level sensor;

[0063] 11-Spray coating and sealing chamber; 12-Motion mechanism chamber; 13-Temperature control equipment chamber;

[0064] 415 - Motion control board; 235 - Ultrasonic humidifier; 237 - Power button switch;

[0065] 564-Humidification control board; 565-Water tank pressure protection switch; 566-Temperature and humidity sensor. Detailed Implementation

[0066] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings, which constitute a part of the present invention and are used together with the embodiments of the present invention to illustrate the principles of the present invention, but are not intended to limit the scope of the present invention.

[0067] This utility model discloses a MALDI mass spectrometer matrix spraying instrument, such as Figures 1 to 9 As shown, it includes: a spray sealing chamber 11 and a matrix spraying assembly 8 disposed inside the spray sealing chamber 11;

[0068] The matrix spraying assembly 8 is placed inside the spraying sealed chamber 11 and is capable of spraying the mass spectrometry matrix.

[0069] The matrix spraying assembly 8 includes: a nozzle temperature control module; the nozzle temperature control module is placed inside the temperature control equipment chamber 13, and the nozzle temperature control module can ensure that the nozzle 80 operates within a stable temperature range.

[0070] Compared with the prior art, this utility model selectively humidifies the spraying sealed chamber through a humidification component, thereby improving the impact of seasonal humidity changes on the uniformity and stability of spraying and enhancing the stability of substrate spraying.

[0071] Preferably, the nozzle temperature control module includes a semiconductor cooling chip 75 and a high-conductivity hot water cooling plate 74;

[0072] A water-cooled plate slot 79 is provided between the high-conductivity hot water cooling plate 74 and the Z-axis platform 73, and the high-conductivity hot water cooling plate 74 is fixed to the Z-axis platform 73 through the water-cooled plate slot 79.

[0073] Specifically, the nozzle temperature control module also includes a nozzle mounting base 77;

[0074] The nozzle 80 is fixed to the semiconductor cooling chip 75 via the nozzle mounting base 77.

[0075] During implementation, the high-conductivity hot water cooling plate 74 is clamped in the water cooling plate slot 79; the semiconductor cooling chip 75 is attached to the high-conductivity hot water cooling plate 74 with thermally conductive silicone, and then fixed and clamped by screws through the nozzle fixing seat 77.

[0076] It should be noted that conventional heat dissipation (such as pure air cooling or single water cooling) relies on passive heat conduction due to ambient temperature difference. Compared with existing technologies, this invention achieves active cooling through the Peltier effect of a semiconductor cooling chip. Combined with the high thermal conductivity of the high-conductivity water cooling plate, heat can be quickly transferred to the circulating water circuit, resulting in higher heat dissipation efficiency than purely passive solutions. Simultaneously, the semiconductor cooling chip, in conjunction with a temperature sensor, enables precise temperature control of ±1℃ in the nozzle area, ensuring a stable operating environment for the nozzle.

[0077] When the thermoelectric cooler is working, its cold side faces the ultrasonic nozzle, while the heat from the hot side is carried away by the coolant circulating in the high-conductivity water-cooled plate and further cooled by a cooling fan in the heat sink. A temperature probe is also added to the water-cooled plate slot to monitor the nozzle temperature in real time and send feedback to the control panel. The control panel then adjusts the real-time power of the thermoelectric cooler to maintain a stable low temperature for the ultrasonic nozzle.

[0078] Specifically, the MALDI mass spectrometer matrix sprayer includes: a spraying sealed chamber 11, a motion mechanism chamber 12, a temperature control equipment chamber 13, and heat conduction pipelines;

[0079] The motion mechanism chamber 12 and the temperature control equipment chamber 13 are located on both sides of the spray sealing chamber 11 and are sealed to the spray sealing chamber 11; the motion mechanism chamber 12 and the temperature control equipment chamber 13 are connected by a heat-insulated hollow sealing partition 86.

[0080] The spray sealing chamber 11 is equipped with a spray nozzle 80, and the motion mechanism chamber 12 is equipped with a spray displacement mechanism 6 that controls the movement of the spray nozzle 80.

[0081] The heat conduction pipeline can transfer the heat from the nozzle 80 to the temperature control equipment chamber 13, and exchange heat with the outside through the temperature control equipment chamber 13;

[0082] The electrical components of the spraying displacement mechanism 6 are placed inside the motion mechanism chamber 12, which can prevent the heat and moisture in the spraying sealing chamber 11 and the temperature control equipment chamber 13 from affecting the interior of the motion mechanism chamber 12.

[0083] Compared with existing technologies, this invention avoids the influence of external humidity on spraying by using partitioned and mutually sealed spraying sealing chambers, motion mechanism chambers, and temperature control equipment chambers. It also avoids the influence of high humidity environment on internal electrical components. Furthermore, it avoids the influence of heat generated by the spray nozzle itself on the spraying effect. It overcomes the defects of high humidity environment caused by spraying, which leads to severe corrosion of electrical components, poor equipment stability, and short service life. It also overcomes the defects of numerous and dense internal components, which make heat dissipation difficult, resulting in excessively high internal temperature, damage to precision components, denaturation and deactivation of macromolecules in the spray liquid, and reduced detection accuracy.

[0084] Preferably, the nozzle 80 is an ultrasonic nozzle, which ultrasonically atomizes and sprays the MALDI mass spectrometry matrix solution under the control of the ultrasonic control motherboard 88.

[0085] Preferably, the nozzle temperature control module further includes: a heat dissipation radiator 81, a cooling water tank 82, and a cooling fan 94;

[0086] The outlet of the high-conductivity hot water cooling plate 74 is connected to the inlet of the heat dissipation radiator 81, and the outlet of the heat dissipation radiator 81 is connected to the inlet of the cooling water tank 82; the outlet of the cooling fan 94 is directly facing the heat dissipation surface of the heat dissipation radiator 81, and the air flow is used to cool the coolant in the heat dissipation radiator 81.

[0087] The coolant at a lower temperature in the cooling water tank 82 is circulated to the inlet of the high-conductivity hot water cooling plate 74 to continue cooling the high-conductivity hot water cooling plate 74.

[0088] Specifically, the matrix spraying assembly 8 also includes a glass slide holder 17;

[0089] The slide holder 17 is fixed to the base of the mass spectrometer matrix sprayer;

[0090] The projection area of ​​the nozzle 80 is directly opposite the slide holder 17, and the jet stream from the nozzle 80 covers the slide holder 17.

[0091] Preferably, the nozzle 80 is also provided with a condensate collector 85 at the bottom to collect condensate and prevent condensate from interfering with the slide holder 17.

[0092] Preferably, the mass spectrometer matrix spraying apparatus further includes a humidification component 9, which can adjust the internal humidity of the spraying sealed chamber 11 and regulate the mass spectrometer matrix spraying within a suitable humidity range. Specifically, the humidification component 9 includes: a humidification water tank 90, a humidification water trough 93, an ultrasonic humidifier 235, and a humidification circulating fan 89;

[0093] The humidifying water tank 93 is located inside the spraying sealing chamber 11, and the humidifying water tank 90 is used to fill the humidifying water tank 93 with water;

[0094] The ultrasonic humidifier 235 is located inside the humidification water tank 93 and can throw the water in the humidification water tank to form water mist;

[0095] The outlet of the humidifying circulating fan 89 is positioned close to the ultrasonic humidifier 235, which can bring water mist into the spray-sealed chamber 11 and provide power for the flow and diffusion of water mist.

[0096] During implementation, when the ultrasonic humidifier 235 is working, it throws water mist upwards to form a mist. The humidifying circulating fan 89 located behind the ultrasonic humidifier 235 carries the water mist into the spray-sealed chamber. After the water mist evaporates, it can achieve the humidification effect.

[0097] Preferably, the humidification component 9 further includes: a humidification control board 564 and a temperature and humidity sensor 566;

[0098] The humidification control board 564 is used to receive signals from the temperature and humidity sensor 566 and control whether the ultrasonic humidifier 235 is started.

[0099] The temperature and humidity sensor 566 detects the humidity signal and transmits it to the humidification control board 564. The humidification control board 564 controls the start and stop timing of the ultrasonic humidifier 235, thereby achieving stable humidity control inside the spraying and sealing chamber, keeping it stable within a small range and almost unaffected by ambient humidity.

[0100] As an example, the humidification control board 564 and the temperature and humidity sensor 566 can be humidity sensors and control modules in the prior art.

[0101] Compared with the prior art, this utility model, by setting up a humidification control board 564, a temperature and humidity sensor 566, a humidification water tank 90, a humidification water trough 93, an ultrasonic humidifier 235, and a humidification circulating fan 89, can not only realize automatic control of the humidification process, but also achieve a better humidification effect by means of ultrasonic atomization.

[0102] Preferably, the spraying sealing chamber 11 is also provided with a first row of fans 18 and a second row of fans 19 for dehumidifying when the humidity in the chamber is too high after spraying.

[0103] Specifically, the humidifying water tank 90 is equipped with a water outlet valve 95, a return air valve 96, a water pump 98, and a drain solenoid valve 100; the humidifying water tank 90 and the water pump 98 are located inside the temperature control equipment chamber 13;

[0104] The outlet valve 95 is used to connect the humidification water tank 93 to the humidification water tank 90; the return air valve 96 is used to balance the internal and external air pressure when water flows in or out of the humidification water tank 90; the drain solenoid valve 100 is used for draining.

[0105] Preferably, the humidification component 9 further includes: a water tank level sensor 108 and a water tank pressure protection switch 565;

[0106] During implementation, water pump 98 is turned on, pumping water from an external water source into humidifying water tank 90. ​​When water level sensor 108 detects that the water level in humidifying water tank 90 has reached the full line or the pressure of water tank pressure protection switch 565 has reached the upper limit, the instrument will automatically shut off water pump 98 to prevent the water tank from bursting due to excessive pressure. After that, water is added to the water tank, and air return valve 96 and water outlet valve 95 are opened, allowing water in the water tank to flow to humidifying water tank 93 through water outlet valve 95. When draining water, air return valve 96, water outlet valve 95, and drain solenoid valve 100 are opened, allowing water in humidifying water tank 93 and humidifying water tank 90 to flow out and be drained through water outlet valve 95 and drain solenoid valve 100.

[0107] Preferably, the mass spectrometer matrix sprayer further includes a spraying displacement mechanism 6, wherein the moving part of the spraying displacement mechanism 6 is fixedly connected to the matrix spraying assembly 8, and the fixed part is fixedly connected to the base of the mass spectrometer matrix sprayer, thereby enabling the matrix spraying assembly 8 to move in the plane of the base of the mass spectrometer matrix sprayer and in the direction perpendicular to the plane.

[0108] Compared with the existing technology that drives the lead screw to rotate via belt drive, thereby driving the four-bar linkage and platform movement, this utility model does not require additional components for belt drive, simplifies the spraying component drive unit, improves power transmission efficiency, and enhances the reliability of system operation.

[0109] Specifically, the spraying displacement mechanism 6 includes an X-axis displacement component 60, a Y-axis displacement component 50, and a Z-axis displacement component 70. The X-axis displacement component 60 enables the matrix spraying component 8 to move in the X-axis direction within the plane of the mass spectrometer matrix sprayer's base. The Y-axis displacement component 50 enables the matrix spraying component 8 to move in the Y-axis direction, perpendicular to the X-axis, within the plane of the mass spectrometer matrix sprayer's base. The Z-axis displacement component 70 enables the matrix spraying component 8 to move in the direction perpendicular to the plane of the mass spectrometer matrix sprayer's base. The Y-axis displacement component 50 and the X-axis displacement component 60 are disposed inside the motion mechanism chamber. The Z-axis displacement component is disposed inside the spraying sealing chamber.

[0110] Specifically, the matrix spraying assembly 8 is fixedly connected to the moving part of the Z-axis displacement assembly 70, the fixed part of the Z-axis displacement assembly 70 is fixed to the moving part of the X-axis displacement assembly 60, and the fixed part of the X-axis displacement assembly 60 is fixed to the moving part of the Y-axis displacement assembly 50.

[0111] During implementation, when the X-axis displacement component 60, Y-axis displacement component 50, and Z-axis displacement component 70 move, they can drive the matrix spraying component 8 to move simultaneously or independently in the parallel X-axis, Y-axis, and Z-axis directions relative to the plane of the mass spectrometer matrix sprayer's base.

[0112] Specifically, the Y-axis displacement assembly 50 includes: a first Y-axis slide rail 45, a second Y-axis slide rail 46, a Y-axis platform 49, a first Y-axis slider 501, a second Y-axis slider 502, and a Y-axis lead screw nut 52;

[0113] The first Y-axis slide rail 45 and the second Y-axis slide rail 46 are arranged in parallel and fixedly connected to the base of the mass spectrometer matrix sprayer.

[0114] The first Y-axis slider 501 and the second Y-axis slider 502 are arranged parallel to each other at the bottom of the Y-axis platform 49. The first Y-axis slider 501 is matched with the first Y-axis slide rail 45, and the second Y-axis slider 502 is matched with the second Y-axis slide rail 46, so that the first Y-axis slider 501 and the second Y-axis slider 502 can drive the Y-axis platform 49 to slide relative to the base of the mass spectrometer matrix sprayer in the Y-axis direction.

[0115] Preferably, the Y-axis displacement assembly 50 further includes: a Y-axis zero-point position sensor 47, a Y-axis extreme position sensor 48, and a Y-axis position sensor light-blocking plate 61. The light-blocking plate generates a signal when passing the Y-axis zero-point position sensor 47 and the Y-axis extreme position sensor 48. The signal is used to control the displacement of the Y-axis platform 49 between the zero point and the extreme position.

[0116] Preferably, the Y-axis displacement assembly 50 further includes: a Y-axis stepper motor 68, a Y-axis motor mounting bracket 69, and a Y-axis lead screw nut 52;

[0117] The Y-axis stepper motor 68 is fixed to the base of the mass spectrometer matrix sprayer via the Y-axis motor mounting bracket 69. The moving part of the Y-axis stepper motor 68 is fixedly connected to the moving part of the Y-axis lead screw nut 52. The fixed part of the Y-axis lead screw nut 52 is fixed to the Y-axis platform 49.

[0118] During implementation, the rotation of the Y-axis stepper motor 68 can drive the Y-axis platform 49 to slide relative to the base of the mass spectrometer matrix sprayer in the Y-axis direction; at the same time, due to the high precision of the stepper motor and the control of the Y-axis zero-point position sensor 47 and the Y-axis limit position sensor 48, the sliding of the Y-axis platform 49 in the Y-axis direction is precisely controlled.

[0119] Specifically, the X-axis displacement assembly 60 includes: an X-axis platform 62, an X-axis second slider 64, an X-axis first slider 65, an X-axis first slide rail 53, an X-axis second slide rail 54, and an X-axis position sensor light-blocking plate 66;

[0120] The X-axis first slide rail 53 and the X-axis second slide rail 54 are set in parallel and fixedly connected to the Y-axis platform 49;

[0121] The second X-axis slider 64 and the first X-axis slider 65 are arranged parallel to each other at the bottom of the X-axis platform 62. The second X-axis slider 64 is matched with the second X-axis slide rail 54, and the first X-axis slider 65 is matched with the first X-axis slide rail 53, so that the second X-axis slider 64 and the first X-axis slider 65 can drive the X-axis platform 62 to slide relative to the Y-axis platform 49 in the X-axis direction.

[0122] Preferably, the X-axis displacement assembly 60 further includes: an X-axis extreme position sensor 55, an X-axis position sensor light-blocking plate 66, and an X-axis zero-point position sensor 57, for controlling the displacement of the X-axis platform 62 between the zero point and the extreme position.

[0123] Preferably, the X-axis displacement assembly 60 further includes: an X-axis lead screw nut 63, an X-axis stepper motor 59, and an X-axis motor mounting bracket 58;

[0124] The fixed part of the X-axis stepper motor 59 is fixed to the Y-axis platform 49 by the X-axis motor mounting bracket 58. The moving part of the X-axis stepper motor 59 is fixedly connected to the moving part of the X-axis lead screw nut 63. The fixed part of the X-axis lead screw nut 63 is fixed to the X-axis platform 62.

[0125] During implementation, the rotation of the X-axis stepper motor 59 can drive the X-axis platform 62 to slide relative to the Y-axis platform 49 in the X-axis direction; at the same time, due to the high precision of the stepper motor and the control of the X-axis limit position sensor 55, the X-axis position sensor light-blocking plate 66, and the X-axis zero point position sensor 57, the sliding of the X-axis platform 62 in the X-axis direction is precisely controlled.

[0126] Preferably, the X-axis platform 62 is fixedly provided with an X-axis column 67; the Z-axis displacement assembly includes: a crossbeam 71, a Z-axis base 72, and a Z-axis platform 73;

[0127] One end of the X-axis column 67 is fixedly connected to the X-axis platform 62, and the other end is fixedly connected to the crossbeam 71; the crossbeam 71 has a Z-axis base 72 fixedly connected to the other end away from the X-axis column 67.

[0128] The Z-axis base 72 is connected to the crossbeam 71. One end of the Z-axis base 72 is placed inside the spray sealing chamber 11. The spray sealing chamber 11 is provided with a lower brush strip 91 and an upper brush strip 92 near the contact with the crossbeam 71. The lower brush strip 91 and the upper brush strip 92 work together to achieve the sealing of the area through which the crossbeam 71 passes through the spray sealing chamber 11.

[0129] The Z-axis platform 73 is adjustable relative to the Z-axis base 72 in the Z-axis direction, and this adjustment can be done manually.

[0130] Specifically, the mass spectrometer matrix sprayer is also equipped with a control motherboard and a motion control board 415, which are used to control the spraying displacement mechanism 6 to move in the X-axis and Y-axis directions.

[0131] The motion control board 415, ultrasonic control motherboard 88, and humidification control board 564 are set up separately and control their respective functions independently; the control motherboard acts as a host computer and connects to control the other three motherboards.

[0132] Compared with existing technologies, this utility model avoids the influence of complex circuits, improves control efficiency, and overcomes the shortcomings of complex internal wiring, inconvenient maintenance, and low control efficiency.

[0133] Specifically, the ultrasonic control motherboard 88, the humidification control board 564, and the motion control board 415 can be implemented using existing technologies such as PID control.

[0134] It should be noted that, to avoid the influence of impurities in the air, spraying within the sealed spraying chamber must be carried out under an inert atmosphere such as nitrogen. The mass spectrometry matrix spraying instrument is also equipped with a gas inlet device for the sealed spraying chamber, including: a pressure reducing valve 2, a gas flow meter 23, and a pressure reducing valve pressure gauge 3; external gas is processed by the pressure reducing valve 2 and then input into the sealed spraying chamber at a specific pressure, and the pressure of the input gas after pressure reduction is displayed by the pressure reducing valve pressure gauge 3; the input gas flow rate is adjusted by the gas flow meter 23.

[0135] Specifically, the mass spectrometer matrix sprayer is also equipped with auxiliary devices, such as an interactive screen 27 and a power button switch 237.

[0136] On the other hand, this utility model also discloses a MALDI mass spectrometry matrix spraying method, which uses the above-mentioned mass spectrometry matrix spraying instrument to achieve automatic heat preservation and humidification incubation after spraying of protein hydrolysate. The method includes:

[0137] Mass spectrometer matrix spraying is achieved within a stable humidity range based on humidification components and their control boards;

[0138] The temperature at the nozzle is stabilized by using a nozzle temperature control module and its control board.

[0139] Specifically, the temperature at the nozzle is set to 10℃~20℃, which can be 10℃, 11℃, 12℃, 13℃, 14℃, 15℃, 16℃, 17℃, 18℃, 19℃ or 20℃.

[0140] The applicant's research found that existing ultrasonic nozzles heat up rapidly during use. Temperature changes can lead to potential spraying instability, and if excess heat cannot be dissipated in time, it can damage the internal components of the nozzle.

[0141] Compared with the prior art, this utility model adds a nozzle temperature control module for the ultrasonic nozzle. By measuring the temperature with a temperature probe, the temperature of the ultrasonic nozzle is adjusted and stabilized by a semiconductor cooling chip and a high thermal conductivity cold plate during the spraying process, thus achieving the spraying of liquid within a stable temperature range.

[0142] Specifically, the humidity of the spraying and sealing chamber is set to 85%~95%, which can be 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94% or 95%.

[0143] The applicant's research found that humidity has a significant impact on the preparation of the matrix layer by ultrasonic spraying. Existing spraying methods are affected by changes in external humidity, resulting in reduced clarity and resolution.

[0144] This invention achieves mass spectrometry matrix spraying within a stable humidity range through a humidification component and its control board, thereby improving the clarity and resolution of mass spectrometry imaging.

[0145] Specifically, the spraying methods include:

[0146] S1. Turn on the power of the sprayer main unit, connect the nitrogen gas line, and adjust the pressure value of the pressure gauge 3 of the sprayer to the preset value.

[0147] S2. Adjust the height of the sprayer along the Z-axis so that the distance between the nozzle of the spray head 80 and the plane of the glass slide holder 17 is the target distance;

[0148] S3. Place the glass slide on the glass slide holder 17;

[0149] S4. Set the carrier platform temperature, nozzle temperature, and nozzle carrier gas flow rate;

[0150] S5. Adjust the humidity around the glass slide to the target range, and spray the prepared mass spectrometry matrix to be tested.

[0151] Specifically, in step S1, the pressure value of the pressure reducing valve barometer 3 is 0.01 MPa ~ 0.03 MPa, which can be 0.01 MPa, 0.012 MPa, 0.014 MPa, 0.016 MPa, 0.018 MPa, 0.02 MPa, 0.022 MPa, 0.024 MPa, 0.025 MPa, 0.027 MPa, 0.029 MPa, or 0.03 MPa.

[0152] Specifically, in step S2, the distance between the nozzle of the spray head 80 and the plane of the slide holder 17 is 40mm to 60mm, which can be 40mm, 42mm, 44mm, 45mm, 47mm, 48mm, 49mm, 50mm, 52mm, 54mm, 55mm, 57mm, 58mm, 59mm or 60mm.

[0153] Specifically, in step S4, the carrier gas flow rate of the nozzle is 0.8L / min to 0.9L / min, which can be 0.8L / min, 0.81L / min, 0.82L / min, 0.83L / min, 0.84L / min, 0.85L / min, 0.86L / min, 0.87L / min, 0.88L / min, 0.89L / min, or 0.90L / min.

[0154] The temperature of the carrier platform is 30℃~40℃, which can be 30℃, 31℃, 32℃, 33℃, 34℃, 35℃, 36℃, 37℃, 38℃, 39℃ or 40℃.

[0155] Please supplement the following embodiments and comparative examples to better illustrate this utility model:

[0156] Example 1

[0157] This embodiment discloses a MALDI mass spectrometer matrix spraying instrument, including: a matrix spraying component 8, a humidification component 9, and a spraying sealed chamber 11;

[0158] The matrix spraying assembly 8 is placed inside the spraying sealed chamber 11 and can spray the mass spectrometry matrix; the humidification assembly 9 can adjust the internal humidity of the spraying sealed chamber 11 and adjust the mass spectrometry matrix spraying within a suitable humidity range.

[0159] Humidification component 9 includes: humidification water tank 90, humidification water trough 93, ultrasonic humidifier 235 and humidification circulating fan 89;

[0160] The humidifying water tank 93 is located inside the spraying sealing chamber 11, and the humidifying water tank 90 is used to fill the humidifying water tank 93 with water;

[0161] The ultrasonic humidifier 235 is located inside the humidification water tank 93 and can throw the water in the humidification water tank to form water mist;

[0162] The outlet of the humidifying circulating fan 89 is positioned close to the ultrasonic humidifier 235, which can bring water mist into the spray-sealed chamber 11 and provide power for the flow and diffusion of water mist.

[0163] When the ultrasonic humidifier 235 is working, water mist is thrown upwards and then carried into the spray-sealed chamber by the humidifying circulating fan 89 located behind the ultrasonic humidifier 235. After the water mist evaporates, it can achieve the effect of humidification.

[0164] The humidification component 9 also includes: a humidification control board 564 and a temperature and humidity sensor 566;

[0165] The humidification control board 564 is used to receive signals from the temperature and humidity sensor 566 and control whether the ultrasonic humidifier 235 is started.

[0166] The temperature and humidity sensor 566 detects the humidity signal and transmits it to the humidification control board 564. The humidification control board 564 controls the start and stop timing of the ultrasonic humidifier 235, thereby achieving stable humidity control inside the spraying and sealing chamber, keeping it stable within a small range and almost unaffected by ambient humidity.

[0167] The humidification control board 564 and the temperature and humidity sensor 566 can be humidity sensors and control modules in the prior art.

[0168] The spraying and sealing chamber 11 is also equipped with a first row of fans 18 and a second row of fans 19, which are used to remove moisture when the humidity in the chamber is too high after spraying.

[0169] The humidifying water tank 90 is equipped with a water outlet valve 95, a return air valve 96, a water pump 98, and a drain solenoid valve 100;

[0170] The outlet valve 95 is used to connect the humidification water tank 93 to the humidification water tank 90; the return air valve 96 is used to balance the internal and external air pressure when water flows in or out of the humidification water tank 90; the drain solenoid valve 100 is used for draining.

[0171] The wet assembly 9 also includes: a water tank level sensor 108 and a water tank pressure protection switch 565;

[0172] When water pump 98 is turned on, it pumps water from an external water source into the humidifying water tank 90. ​​When the water level sensor 108 detects that the water level in the humidifying water tank 90 has reached the full line or the pressure of the water tank pressure protection switch 565 has reached the upper limit, the instrument will automatically turn off water pump 98 to prevent the water tank from bursting due to excessive pressure. After that, water is added to the water tank, and the air return valve 96 and the water outlet valve 95 are opened. The water in the water tank will flow to the humidifying water tank 93 through the water outlet valve 95. When draining, the air return valve 96, the water outlet valve 95, and the drain solenoid valve 100 are opened. The water in the humidifying water tank 93 and the humidifying water tank 90 will all flow out through the water outlet valve 95 and the drain solenoid valve 100 to drain completely.

[0173] The mass spectrometer matrix spraying instrument also includes a spraying displacement mechanism 6. The moving part of the spraying displacement mechanism 6 is fixedly connected to the matrix spraying assembly 8, and the fixed part is fixedly connected to the base of the mass spectrometer matrix spraying instrument. It can realize the movement of the matrix spraying assembly 8 in the plane of the base of the mass spectrometer matrix spraying instrument and in the direction of the vertical plane.

[0174] The spraying displacement mechanism 6 includes an X-axis displacement assembly 60, a Y-axis displacement assembly 50, and a Z-axis displacement assembly 70;

[0175] The matrix spraying assembly 8 is fixedly connected to the moving part of the Z-axis displacement assembly 70. The fixed part of the Z-axis displacement assembly 70 is fixed to the moving part of the X-axis displacement assembly 60, and the fixed part of the X-axis displacement assembly 60 is fixed to the moving part of the Y-axis displacement assembly 50.

[0176] The Y-axis displacement assembly 50 includes: a first Y-axis slide rail 45, a second Y-axis slide rail 46, a Y-axis platform 49, a first Y-axis slider 501, a second Y-axis slider 502, and a Y-axis lead screw nut 52;

[0177] The first Y-axis slide rail 45 and the second Y-axis slide rail 46 are arranged in parallel and fixedly connected to the base of the mass spectrometer matrix sprayer.

[0178] The first Y-axis slider 501 and the second Y-axis slider 502 are arranged parallel to each other at the bottom of the Y-axis platform 49. The first Y-axis slider 501 is matched with the first Y-axis slide rail 45, and the second Y-axis slider 502 is matched with the second Y-axis slide rail 46, so that the first Y-axis slider 501 and the second Y-axis slider 502 can drive the Y-axis platform 49 to slide relative to the base of the mass spectrometer matrix sprayer in the Y-axis direction.

[0179] The Y-axis displacement assembly 50 also includes: a Y-axis zero-point position sensor 47, a Y-axis extreme position sensor 48, and a Y-axis position sensor light-blocking plate 61, used to control the displacement of the Y-axis platform 49 between the zero point and the extreme position.

[0180] The Y-axis displacement assembly 50 also includes: a Y-axis stepper motor 68, a Y-axis motor mounting bracket 69, and a Y-axis lead screw nut 52;

[0181] The Y-axis stepper motor 68 is fixed to the base of the mass spectrometer matrix sprayer via the Y-axis motor mounting bracket 69. The moving part of the Y-axis stepper motor 68 is fixedly connected to the moving part of the Y-axis lead screw nut 52. The fixed part of the Y-axis lead screw nut 52 is fixed to the Y-axis platform 49.

[0182] The rotation of the Y-axis stepper motor 68 can drive the Y-axis platform 49 to slide relative to the base of the mass spectrometer matrix sprayer in the Y-axis direction; at the same time, due to the high precision of the stepper motor and the control of the Y-axis zero-point position sensor 47 and the Y-axis limit position sensor 48, the sliding of the Y-axis platform 49 in the Y-axis direction is precisely controlled.

[0183] The X-axis displacement assembly 60 includes: an X-axis platform 62, an X-axis second slider 64, an X-axis first slider 65, an X-axis first slide rail 53, an X-axis second slide rail 54, and an X-axis position sensor light-blocking plate 66;

[0184] The X-axis first slide rail 53 and the X-axis second slide rail 54 are set in parallel and fixedly connected to the Y-axis platform 49;

[0185] The second X-axis slider 64 and the first X-axis slider 65 are arranged parallel to each other at the bottom of the X-axis platform 62. The second X-axis slider 64 is matched with the second X-axis slide rail 54, and the first X-axis slider 65 is matched with the first X-axis slide rail 53, so that the second X-axis slider 64 and the first X-axis slider 65 can drive the X-axis platform 62 to slide relative to the Y-axis platform 49 in the X-axis direction.

[0186] The X-axis displacement assembly 60 also includes: an X-axis extreme position sensor 55, an X-axis position sensor light-blocking plate 66, and an X-axis zero-point position sensor 57, used to control the displacement of the X-axis platform 62 between the zero point and the extreme position.

[0187] The X-axis displacement assembly 60 also includes: an X-axis lead screw nut 63, an X-axis stepper motor 59, and an X-axis motor mounting bracket 58;

[0188] The fixed part of the X-axis stepper motor 59 is fixed to the Y-axis platform 49 by the X-axis motor mounting bracket 58. The moving part of the X-axis stepper motor 59 is fixedly connected to the moving part of the X-axis lead screw nut 63. The fixed part of the X-axis lead screw nut 63 is fixed to the X-axis platform 62.

[0189] The rotation of the X-axis stepper motor 59 can drive the X-axis platform 62 to slide relative to the Y-axis platform 49 in the X-axis direction; at the same time, due to the high precision of the stepper motor and the control of the X-axis limit position sensor 55, the X-axis position sensor light-blocking plate 66, and the X-axis zero point position sensor 57, the sliding of the X-axis platform 62 in the X-axis direction is precisely controlled.

[0190] The X-axis platform 62 is fixedly provided with an X-axis column 67; the Z-axis displacement assembly includes: a crossbeam 71, a Z-axis base 72, and a Z-axis platform 73;

[0191] One end of the X-axis column 67 is fixedly connected to the X-axis platform 62, and the other end is fixedly connected to the crossbeam 71; the crossbeam 71 has a Z-axis base 72 fixedly connected to the other end away from the X-axis column 67.

[0192] The Z-axis base 72 is connected to the crossbeam 71. One end of the Z-axis base 72 is placed inside the spray sealing chamber 11. The spray sealing chamber 11 is provided with a lower brush strip 91 and an upper brush strip 92 near the contact with the crossbeam 71. The lower brush strip 91 and the upper brush strip 92 work together to achieve the sealing of the area through which the crossbeam 71 passes through the spray sealing chamber 11.

[0193] The Z-axis platform 73 is adjustable relative to the Z-axis base 72 in the Z-axis direction, and this adjustment can be done manually.

[0194] The matrix spraying assembly 8 includes a spray nozzle 80 and a glass slide holder 17;

[0195] The slide holder 17 is fixed to the base of the mass spectrometer matrix sprayer;

[0196] The nozzle 80 is fixed to the Z-axis platform 73, and its projection area is directly opposite the slide holder 17. The jet stream of the nozzle 80 covers the slide holder 17.

[0197] A semiconductor cooling chip 75, a high-conductivity hot water cooling plate 74, and a nozzle mounting base 77 are sequentially arranged between the nozzle 80 and the Z-axis platform 73.

[0198] A water-cooled plate slot 79 is provided between the high-conductivity hot water cooling plate 74 and the Z-axis platform 73, and the high-conductivity hot water cooling plate 74 is fixed to the Z-axis platform 73 through the water-cooled plate slot 79;

[0199] The nozzle 80 is fixed to the semiconductor cooling chip 75 via the nozzle mounting base 77.

[0200] The high-conductivity hot water cooling plate 74 is clamped in the water cooling plate slot 79, which is fixed to the Z-axis platform (nozzle cooling system mounting base) 73 by screws. The semiconductor cooling chip 75 is attached to the high-conductivity hot water cooling plate 74 with thermally conductive silicone, and then fixed and clamped by screws through the nozzle mounting base 77.

[0201] Nozzle 80 is an ultrasonic nozzle, which ultrasonically atomizes and sprays the MALDI mass spectrometry matrix solution under the control of the ultrasonic control motherboard 88.

[0202] Preferably, the matrix spraying assembly 8 is further provided with a nozzle temperature control module, which includes: a heat dissipation water drain 81, a cooling water tank 82, and a cooling fan 94;

[0203] The outlet of the high-conductivity hot water cooling plate 74 is connected to the inlet of the heat dissipation radiator 81, and the outlet of the heat dissipation radiator 81 is connected to the inlet of the cooling water tank 82; the outlet of the cooling fan 94 is directly facing the heat dissipation surface of the heat dissipation radiator 81, and the air flow is used to cool the coolant in the heat dissipation radiator 81.

[0204] The coolant at a lower temperature in the cooling water tank 82 is circulated to the inlet of the high-conductivity hot water cooling plate 74 to continue cooling the high-conductivity hot water cooling plate 74.

[0205] Preferably, the nozzle 80 is also provided with a condensate collector 85 at the bottom to collect condensate and prevent condensate from interfering with the slide holder 17.

[0206] Specifically, the mass spectrometer matrix sprayer is also equipped with a control motherboard and a motion control board 415, which are used to control the spraying displacement mechanism 6 to move in the X-axis and Y-axis directions.

[0207] The motion control board 415 is separately configured from the ultrasonic control motherboard 88 and the humidification control board 564, each independently controlling its corresponding function; the control motherboard acts as a host computer, connecting to and controlling the other three motherboards. The ultrasonic control motherboard 88, the humidification control board 564, and the motion control board 415 can be implemented using existing technologies such as PID control.

[0208] This embodiment discloses a control method for achieving good coating with a matrix sprayer, using the aforementioned mass spectrometry matrix sprayer.

[0209] Control methods include:

[0210] S1: Turn on the power of the sprayer main unit, connect the nitrogen gas line, adjust the pressure of the pressure reducing valve and pressure gauge 3 of the sprayer to 0.02MPa, and the gas cylinder pressure to be greater than 0.4MPa;

[0211] S2: Adjust the height of the sprayer in the Z-axis direction, and the position of the X and Y axes so that the nozzle 80 is an ultrasonic nozzle and the distance between the nozzle and the plane of the glass slide holder 17 is 50mm.

[0212] S3: Place the glass slide on the slide holder 17;

[0213] S4: Click on the interactive screen 27 to enter the operation interface. Click "Heating on", "Cooling on", "Blowing on". You can slide to set the platform temperature and nozzle temperature in the lower right corner. Set the platform temperature to 25℃ and the nozzle temperature to 12℃. Adjust the gas flow meter 23 to make the nozzle carrier gas flow rate 0.9L / min.

[0214] S5: Click on Instrument Settings, set the nozzle power to 0.8, click OK to turn on the nozzle. At this time, the nozzle can atomize the liquid pushed into the nozzle by the external peristaltic pump and spray it downwards in a directional manner.

[0215] S6: Click on Method Settings to enter the spraying settings interface. Here you can set parameters such as the area size to be sprayed and the number of sprays. At the same time, the reaction settings on the right can determine whether to turn on the humidification function after spraying is completed. If not needed, do not click on the "Turn on Humidification Reaction" option. After setting the spraying parameters, click "Save Method". After naming it, you can call it at any time in "Open Method".

[0216] S7: Return to the main interface and click "Start Work" to begin the spraying process;

[0217] S8: After the spraying is completed, take out the sprayed sample, clean the liquid phase pipeline, close the gas line and turn off the machine;

[0218] S9: Ultrasonic nozzle 71 needs to be maintained and cleaned after prolonged use.

[0219] The matrix spraying effect obtained using the above control method is shown in the figure below. Figure 10 As shown, the matrix used is α-cyano-4-hydroxycinnamic acid, i.e., CCA; as can be seen from the figure, the coating thickness in each area is relatively uniform, which meets the sample preparation requirements for mass spectrometry.

[0220] Example 2

[0221] The MALDI mass spectrometer matrix sprayer used in this embodiment is the same as that in the previous embodiment.

[0222] This embodiment discloses a control method for achieving good coating with a matrix sprayer, realizing the function of automatic heat preservation and humidification incubation after spraying the protein hydrolysate. Using the matrix sprayer with humidification function of this embodiment, the control method includes:

[0223] S1. Turn on the power of the sprayer main unit, connect the nitrogen gas line, adjust the pressure of the pressure reducing valve and pressure gauge 3 of the sprayer to 0.02MPa, and the gas cylinder pressure to be greater than 0.4MPa;

[0224] S2. Adjust the height of the sprayer in the Z-axis direction so that the distance between the nozzle of the ultrasonic spray head 76 and the plane of the glass slide holder 17 is 50mm.

[0225] S3. Place the glass slide on the glass slide holder 17;

[0226] S4. Click on the interactive screen 27 to enter the operation interface. Click "Heating on", "Cooling on", "Blowing on". You can slide to set the platform temperature and nozzle temperature in the lower right corner. The platform temperature is 25℃ and the nozzle temperature is 12℃. Adjust the gas flow meter 23 to make the nozzle carrier gas flow rate 0.85L / min.

[0227] S5. Click on Instrument Settings, set the nozzle power to 0.8, click OK to turn on the nozzle. At this time, the nozzle can atomize the protease solution pushed into the nozzle by the external peristaltic pump and spray it downwards in a directional manner. At the same time, observe whether the water level diagram of the water tank and water tank above is displayed in blue. If it is still gray, it means that there is no water in the water tank and water tank. You need to click "Turn on Water Tank Add Water" and "Turn on Water Tank Add Water" on the right in sequence. Only after the water tank and water tank are full of humidifying water can you enter the spraying / humidification program settings.

[0228] S6. Click on Method Settings to enter the Spraying / Humidification Settings interface. Here you can set parameters such as the desired spraying area size and number of sprays. The reaction settings on the right determine whether to turn on the humidification function after spraying is complete. Click on "Turn on Humidification Reaction," set the humidification parameters, and then click "Save." The sprayer can then start spraying / humidification at any time. Here, we use the in situ digestion of mouse brain slices with trypsin as an example to illustrate the relevant reaction parameter settings: "Wait Time" is set to 1, "Platform Temperature" is set to 37°C, "Ambient Humidity" is set to 95%, and "Reaction Time" is set to 120.

[0229] S7. Return to the main interface and click "Start Work" to begin the spraying / incubation process;

[0230] S8. After spraying / incubation is complete, remove the completed sample, clean the liquid phase pipeline, shut off the gas line and turn off the machine.

[0231] S9. After prolonged use, the ultrasonic nozzle 71 needs to be maintained and cleaned. If the humidification function is not used for a long time, the humidification liquid in the water tank and water tank needs to be drained. Click "Start Drainage" to do so.

[0232] The image below shows a MALDI mass spectrometry image obtained by in situ digestion of mouse brain slices with trypsin using the above control method. The ion mass-to-charge ratio m / Z shown is 1099.7. The comparison is between mass spectrometry imaging results obtained by incubation using a matrix sprayer under constant temperature and humidity and by incubation using a conventional oven under constant temperature and humidity. The protein digest spraying steps were all performed using the matrix sprayer, only the incubation location was different. The specific conditions were: trypsin digest concentration 10 μg / ml, digest flow rate 10 μL / min, number of spray layers 10, slide temperature 25℃, nitrogen flow rate 0.85 L / min, and reaction conditions: temperature 37℃, humidity 100%RH, reaction time 0.5 hours. Figure 10 The results were obtained from incubation using a matrix sprayer.

[0233] Figure 10 This demonstrates that in situ enzymatic imaging of proteins incubated using a matrix sprayer can more clearly reveal the spatial proteome distribution of tissues and reduce the diffusion of endogenous compounds in tissues during incubation. The figure clearly illustrates the specific distribution of the protein-digested peptide with m / z 1099.7 in the inner and outer capsules of the mouse brain.

[0234] Comparative Example 1

[0235] The difference from Example 2 is that the mass spectrometry matrix spraying instrument used in Example 2 does not include a cavity humidification component unit, and therefore cannot actively control the humidity within the humidification cavity. It is affected by changes in external humidity, which ranges from 30% to 40%. The remaining characteristics of the MALDI mass spectrometry matrix spraying method are the same as in Example 2, and the results are as follows: Figure 11 As shown.

[0236] Figure 11 This reflects the difficulty of clearly revealing the spatial proteome distribution of tissues using existing mass spectrometry matrix spraying instruments. This is due to the diffusion of endogenous compounds on the tissue during incubation, such as... Figure 11 The specific distribution of the protease-digested peptide with m / z 1099.7 in the inner and outer capsules of the mouse brain was far less clear than in Example 2.

[0237] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. A MALDI mass spectrometer matrix spraying instrument, characterized in that, include: A spray-sealed chamber and a matrix spraying assembly disposed inside the spray-sealed chamber; The matrix spraying assembly is placed inside the spraying sealed chamber and is capable of spraying mass spectrometry matrix; The matrix spraying assembly includes: a nozzle temperature control module; the nozzle temperature control module can ensure that the nozzle operates within a stable temperature range; The mass spectrometer matrix spraying instrument also includes a humidification component, which can adjust the internal humidity of the spraying sealed chamber and adjust the mass spectrometer matrix spraying within a suitable humidity range. The humidification components include: a humidification water tank, a humidification water bath, an ultrasonic humidifier, and a humidification circulating fan; The humidifying water tank is located inside the spraying sealed chamber, and the humidifying water container is used to fill the humidifying water tank with water; The ultrasonic humidifier is located inside the humidification tank and can throw the water in the humidification tank to form a water mist; The humidifying circulating fan's outlet is positioned close to the ultrasonic humidifier, enabling it to carry water mist into the spray-sealed chamber and providing power for the flow and diffusion of the water mist.

2. The MALDI mass spectrometer matrix spraying apparatus according to claim 1, characterized in that, The nozzle temperature control module includes a nozzle, a semiconductor cooling chip, and a high-conductivity hot water cooling plate; The nozzle, semiconductor cooling chip, and high-conductivity hot water cooling plate are connected in sequence. A water-cooled plate slot is provided between the high-conductivity hot water cooling plate and the Z-axis platform, and the high-conductivity hot water cooling plate is fixed to the Z-axis platform through the water-cooled plate slot.

3. The MALDI mass spectrometer matrix spraying apparatus according to claim 2, characterized in that, The nozzle temperature control module also includes a nozzle mounting base; The nozzle is fixed to the semiconductor cooling chip via a nozzle mounting base.

4. The MALDI mass spectrometer matrix spraying apparatus according to claim 3, characterized in that, The nozzle temperature control module also includes: a heat dissipation water duct, a cooling water tank, and a cooling fan; The outlet of the high conductivity hot water cooling plate is connected to the inlet of the heat dissipation water duct, and the outlet of the heat dissipation water duct is connected to the inlet of the cooling water tank; the outlet of the cooling fan faces the heat dissipation surface of the heat dissipation water duct, and uses air flow to cool the coolant in the heat dissipation water duct. The coolant at a lower temperature in the cooling water tank circulates to the inlet of the high-conductivity hot water cooling plate to continue cooling the plate.

5. The MALDI mass spectrometer matrix spraying apparatus according to claim 4, characterized in that, The matrix spraying assembly also includes a glass slide holder; The slide holder is fixed to the base of the mass spectrometer matrix sprayer; The projection area of ​​the nozzle is directly opposite the slide holder, and the jet stream from the nozzle covers the slide holder.

6. The MALDI mass spectrometer matrix spraying apparatus according to claim 5, characterized in that, The nozzle is also equipped with a condensate collector at the bottom to collect condensate and prevent it from interfering with the slide holder.

7. The MALDI mass spectrometer matrix spraying apparatus according to claim 6, characterized in that, The mass spectrometer matrix spraying device also includes a spraying displacement mechanism. The moving part of the spraying displacement mechanism is fixedly connected to the matrix spraying assembly, and the fixed part is fixedly connected to the base of the mass spectrometer matrix spraying device. This enables the matrix spraying assembly to move within the plane of the base of the mass spectrometer matrix spraying device and in the direction perpendicular to the plane.

8. The MALDI mass spectrometer matrix spraying apparatus according to claim 7, characterized in that, The matrix spraying assembly is fixedly connected to the moving part of the Z-axis displacement assembly, the fixed part of the Z-axis displacement assembly is fixed to the moving part of the X-axis displacement assembly, and the fixed part of the X-axis displacement assembly is fixed to the moving part of the Y-axis displacement assembly.

9. The MALDI mass spectrometer matrix spraying apparatus according to claim 8, characterized in that, The Y-axis displacement assembly includes: a first Y-axis slide rail, a second Y-axis slide rail, a Y-axis platform, a first Y-axis slider, a second Y-axis slider, and a Y-axis lead screw nut; The first Y-axis slide rail and the second Y-axis slide rail are arranged in parallel and fixedly connected to the base of the mass spectrometer matrix sprayer; The first and second Y-axis sliders are arranged parallel to each other at the bottom of the Y-axis platform. The first Y-axis slider is matched with the first Y-axis slide rail, and the second Y-axis slider is matched with the second Y-axis slide rail, so that the first and second Y-axis sliders can drive the Y-axis platform to slide relative to the base of the mass spectrometer matrix sprayer in the Y-axis direction.

10. The MALDI mass spectrometer matrix spraying apparatus according to claim 9, characterized in that, The Y-axis displacement assembly further includes: a Y-axis zero-point position sensor, a Y-axis extreme position sensor, and a light-blocking plate for the Y-axis position sensor; The light-blocking plate of the Y-axis position sensor generates a signal when it passes the Y-axis zero-point position sensor and the Y-axis extreme position sensor. The signal is used to control the displacement of the Y-axis platform between the zero point and the extreme position.

Images