阅读说明：本技术 试样支承体、试样的离子化方法及质谱分析方法 (Sample support, sample ionization method, and mass spectrometry method ) 是由 泷本未羽 大村孝幸 小谷政弘 于 2019-06-19 设计创作，主要内容包括：一种试样支承体,其用于将试样离子化,其中,具备：基板,其上形成有在第一表面及与第一表面相反侧的第二表面开口的多个贯通孔；导电层,其以不堵塞贯通孔的方式设置于第一表面；以及,加强材料,其配置于多个贯通孔中的一部分贯通孔的内部。(A sample support for ionizing a sample, comprising: a substrate having a plurality of through holes formed therein, the through holes being open at a first surface and a second surface opposite to the first surface; a conductive layer provided on the first surface so as not to block the through hole; and a reinforcing member disposed inside a part of the plurality of through holes.)

1. A sample support body characterized in that,

is a sample support for ionization of a sample,

the disclosed device is provided with:

a substrate having a plurality of through holes formed therein, the through holes being open in a first surface and a second surface opposite to the first surface;

a conductive layer provided on the first surface so as not to block the through hole; and

and a reinforcing member disposed inside a part of the through holes.

2. The sample support according to claim 1,

the width of the through hole is 1nm to 700nm,

the thickness of the substrate is 1-50 μm.

3. Sample support according to claim 1 or 2,

the material of the reinforcing material is resin.

4. Sample support according to claim 1 or 2,

the material of the reinforcing material is metal.

5. The sample support according to any one of claims 1 to 4,

the substrate has: a reinforcing region including a plurality of the through holes, and a measuring region including a plurality of the through holes,

the reinforcing material is disposed inside the plurality of through holes in the reinforcing region,

the reinforcing member is not disposed inside the plurality of through holes in the measurement region.

6. The sample support according to claim 5,

the reinforcing region includes at least a region continuous from one end of the substrate to the other end opposite to the one end, when viewed from the thickness direction of the substrate.

7. Sample support according to claim 5 or 6,

the reinforcing region surrounds each of the plurality of measurement regions when viewed from a thickness direction of the substrate.

8. A sample support body characterized in that,

is a sample support for ionization of a sample,

the disclosed device is provided with:

a substrate having conductivity, and having a plurality of through holes formed therein, the through holes being open on a first surface and a second surface opposite to the first surface; and

and a reinforcing member disposed inside a part of the through holes.

9. A method for ionizing a sample,

the method comprises the following steps:

a first step of preparing a sample and the sample support according to any one of claims 1 to 7;

a second step of disposing the sample support on the sample so that the second surface faces the sample;

a third step of peeling the sample support from the sample; and

a fourth step of ionizing a component of the sample that moves to the first surface side through the through hole, in which the reinforcing member is not disposed, among the plurality of through holes by applying a voltage to the conductive layer and irradiating the first surface with an energy ray.

10. A method for ionizing a sample,

the method comprises the following steps:

a first step of preparing a sample and the sample support according to claim 8;

a second step of disposing the sample support on the sample so that the second surface faces the sample;

a third step of peeling the sample support from the sample; and

a fourth step of ionizing a component of the sample moving to the first surface side through the through hole, in which the reinforcing member is not disposed, among the plurality of through holes by applying a voltage to the substrate and irradiating the first surface with an energy ray.

11. The method of ionizing a sample according to claim 9 or 10,

in the third step, the sample support is peeled from the sample before the component adhering to the substrate is dried.

12. A method of mass spectrometry characterized in that,

the method comprises the following steps:

the steps of the method for ionizing a sample according to any one of claims 9 to 11; and

a fifth step of detecting the component ionized in the fourth step.

Technical Field

The present invention relates to a sample support, a sample ionization method, and a mass spectrometry method.

Background

Conventionally, a sample support for ionizing a sample in mass spectrometry of a sample such as a biological sample is known (for example, see patent document 1). The sample support includes a substrate having a plurality of through holes formed therein, the through holes opening in a first surface and a second surface opposite to the first surface. When the sample support is disposed on the sample so that the second surface faces the sample, the sample can be raised from the second surface side of the substrate toward the first surface side through the through hole by utilizing the capillary phenomenon. When the first surface side is irradiated with an energy ray such as a laser beam, for example, the sample moving toward the first surface side is ionized.

Documents of the prior art

Patent document

Patent document 1: japanese patent No. 6093492

Disclosure of Invention

[ problem to be solved by the invention ]

In the sample support as described above, in order to promote the movement of the sample due to the capillary phenomenon from the second surface side of the substrate to the first surface side via the through hole, the substrate may need to be thinned. However, when the substrate is thinned, for example, it is easily bent and the strength is reduced. As a result, the substrate is easily damaged.

Accordingly, an object of one aspect of the present invention is to provide a sample support, a sample ionization method, and a mass spectrometry method that suppress damage to a substrate.

[ means for solving the problems ]

One aspect of the present invention provides a sample support for ionizing a sample, including: a substrate having a plurality of through holes formed therein, the through holes being open in a first surface and a second surface opposite to the first surface; a conductive layer provided on the first surface so as not to block the through hole; and a reinforcing member disposed inside a part of the plurality of through holes.

In the sample support, a plurality of through holes that are open at a first surface and a second surface opposite to the first surface are formed in a substrate. Therefore, for example, in the case where the sample support is disposed on a sample such as a biological sample so that the second surface of the substrate faces the sample, the sample (component of the sample) can be moved from the second surface side to the first surface side through the through hole by capillary action. Further, when the first surface is irradiated with an energy line such as a laser beam, energy is transmitted to the component of the sample moved to the first surface side through the conductive layer, and therefore, the component of the sample can be ionized. The reinforcing member is disposed inside a part of the plurality of through holes. Thus, the substrate is reinforced by the reinforcing material. This makes the substrate less likely to be bent. Therefore, according to the sample support, the strength of the substrate can be appropriately secured, and the breakage of the substrate can be suppressed.

In the present invention, the following may be used: the width of the through hole is 1nm to 700nm, and the thickness of the substrate is 1 μm to 50 μm. In this case, the above-described movement of the components of the sample due to the capillary phenomenon can be appropriately realized.

In the present invention, the following may be used: the material of the reinforcing material is resin. In this case, the reinforcing material can be easily formed.

In the present invention, the following may be used: the material of the reinforcing material is metal. In this case, when the component of the sample is ionized, the generation of the organic gas can be suppressed, and the noise in the detection result of the ionized sample component can be reduced.

In the present invention, the following may be used: the substrate has a reinforcing region including a plurality of through holes, and a measuring region including a plurality of through holes; a reinforcing material is disposed inside the plurality of through holes in the reinforcing region; the reinforcing member is not disposed inside the plurality of through holes in the measurement region. In this case, since the reinforcing material is disposed inside the plurality of through holes in the reinforcing region, the substrate is less likely to be bent. This can further secure the strength of the substrate, and can further suppress damage to the substrate.

In the present invention, the following may be used: the reinforcing region includes at least a region continuous from one end of the substrate to the other end opposite to the one end, when viewed from the thickness direction of the substrate. In this case, the substrate is less likely to be flexed. This can further secure the strength of the substrate, and can more reliably suppress the breakage of the substrate.

In the present invention, the following may be used: the reinforcing region surrounds each of the plurality of measurement regions when viewed from the thickness direction of the substrate. In this case, the components of the plurality of samples can be ionized in each of the plurality of measurement regions.

A sample support according to another aspect of the present invention is a sample support for ionizing a sample, including: a substrate having conductivity and formed with a plurality of through holes that are open at a first surface and a second surface opposite to the first surface; and a reinforcing member disposed inside a part of the plurality of through holes.

According to this sample support, the conductive layer can be omitted, and the same effects as those of the sample support provided with the conductive layer described above can be obtained.

A method for ionizing a sample according to an aspect of the present invention includes: a first step of preparing a sample and the sample support having the conductive layer; a second step of disposing a sample support on the sample so that the second surface faces the sample; a third step of peeling the sample support from the sample; and a fourth step of ionizing a component of the sample moving to the first surface side through a through hole, in which the reinforcing member is not disposed, among the plurality of through holes by applying a voltage to the conductive layer and irradiating the first surface with an energy ray.

In the above method of ionizing a sample, a plurality of through holes that are open on a first surface and a second surface opposite to the first surface are formed in a substrate. When the sample support is disposed on the sample so that the second surface of the substrate faces the sample, the sample (component of the sample) can be moved from the second surface side to the first surface side through the through hole by capillary action. Further, after the sample support is peeled off from the sample, when a voltage is applied to the conductive layer and the first surface is irradiated with a laser, energy is transferred to the component of the sample moved to the first surface side. This can ionize the components of the sample. In the sample ionization method, the reinforcing member is disposed in a part of the plurality of through-holes. Thus, the substrate is reinforced by the reinforcing material. This makes the substrate less likely to be bent, and therefore, breakage of the substrate can be suppressed when the sample support is peeled from the sample in the third step. Thus, according to this method of ionizing a sample, components of the sample can be ionized while suppressing breakage of the substrate by using a substrate having an appropriately secured strength.

A method for ionizing a sample according to another aspect of the present invention includes: a first step of preparing a sample and a sample support having the substrate having conductivity; a second step of disposing a sample support on the sample so that the second surface faces the sample; a third step of peeling the sample support from the sample; and a fourth step of ionizing a component of the sample moving to the first surface side through a through hole, in which the reinforcing member is not disposed, among the plurality of through holes by applying a voltage to the substrate and irradiating the first surface with an energy ray.

According to this sample ionization method, when a sample support from which the conductive layer is omitted is used, the same effects as those in the case of using a sample support provided with a conductive layer as described above can be obtained.

In the present invention, the following may be used: in the third step, the sample support is peeled off from the sample before the component adhering to the substrate is dried. In this case, the sample support can be more smoothly peeled from the sample before the sample support is adhered to the sample.

A method of mass spectrometry according to an aspect of the present invention comprises: each step of the method for ionizing a sample; and a fifth step of detecting the component ionized in the fourth step.

According to the mass spectrometry, the substrate having an appropriately secured strength is used, whereby mass spectrometry of a sample can be performed while suppressing breakage of the substrate.

[ Effect of the invention ]

According to one aspect of the present invention, a sample support, a sample ionization method, and a mass spectrometry method that suppress damage to a substrate can be provided.

Drawings

Fig. 1 is a plan view of a sample support according to an embodiment.

Fig. 2 is a sectional view of the sample support taken along line II-II shown in fig. 1.

Fig. 3 is a view showing an enlarged image of a measurement region of the substrate shown in fig. 1 as viewed from the thickness direction of the substrate.

Fig. 4 is an enlarged view of the substrate shown in fig. 1 as viewed from the thickness direction of the substrate.

Fig. 5 is a view showing steps of a method for manufacturing a sample support according to an embodiment.

FIG. 6 is a diagram showing steps of a mass spectrometry method according to an embodiment.

Fig. 7 is a diagram showing steps of a mass spectrometry method according to an embodiment.

Fig. 8 is a view showing a sample support according to a modification.

Fig. 9 is a view showing a sample support according to a modification.

Fig. 10 is a view showing a sample support according to a modification.

Fig. 11 is a view showing a procedure of a method for manufacturing a sample support according to a first modification.

Fig. 12 is a view showing steps of a method for manufacturing a sample support according to a second modification.

Detailed Description

Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. In the drawings, the same or corresponding portions are denoted by the same reference numerals, and redundant description thereof is omitted. In order to facilitate understanding of the description, the size or the size ratio of each member (or portion) shown in the drawings may be different from the actual size or size ratio.

[ Structure of sample support ]

Fig. 1 shows a top view of a sample support 1 according to an embodiment. As shown in fig. 1 and 2, the sample support 1 includes a substrate 2, a conductive layer 3, and a reinforcing member 4. The sample support 1 is a sample support for ionizing a sample. The sample support 1 is used to ionize a component of a sample to be measured, for example, in mass spectrometry.

The substrate 2 has a first surface 2a and a second surface 2b opposite to the first surface 2 a. A plurality of through holes 2c are formed in the substrate 2 in the same manner (in a uniformly distributed manner). Each through-hole 2c extends in the thickness direction (hereinafter, simply referred to as "thickness direction") of the sample support 1 (i.e., the substrate 2), and is open at the first surface 2a and the second surface 2 b. The thickness direction is a direction perpendicular to the first surface 2a and the second surface 2 b. The substrate 2 is formed in a rectangular plate shape from an insulating material, for example. The length of one side of the substrate 2 is, for example, about several cm when viewed in the thickness direction. The thickness of the substrate 2 is, for example, about 1 μm to 50 μm. In the present embodiment, the thickness of the substrate 2 is about 5 μm.

The conductive layer 3 is disposed on the first surface 2a of the substrate 2. The conductive layer 3 is provided on the peripheral edge of the through hole 2c on the first surface 2 a. That is, the conductive layer 3 covers a portion of the first surface 2a of the substrate 2 where the through-hole 2c is not formed. That is, the conductive layer 3 is provided so as not to block the through-hole 2 c.

The conductive layer 3 is formed of a conductive material. Among them, as a material of the conductive layer 3, a metal having low affinity (reactivity) with a sample and high conductivity is preferably used for the reason described below.

For example, if the conductive layer 3 is formed of a metal such as Cu (copper) having a high affinity with a sample such as protein, the sample may be ionized in a state where Cu atoms are attached to sample molecules in the process of ionizing the sample, which will be described later, and the detection result may vary depending on the amount of Cu atoms attached in the mass spectrometry, which will be described later. Therefore, as a material of the conductive layer 3, a metal having low affinity with the sample is preferably used.

On the other hand, the more conductive the metal is, the easier and more stable the predetermined voltage is applied. Therefore, when the conductive layer 3 is formed of a metal having high conductivity, a voltage can be uniformly applied to the first surface 2a of the substrate 2. The more electrically conductive the metal, the higher the thermal conductivity tends to be. Therefore, when the conductive layer 3 is formed of a metal having high conductivity, energy of an energy beam such as a laser beam irradiated to the substrate 2 can be efficiently transmitted to the sample through the conductive layer 3. Therefore, as a material of the conductive layer 3, a metal having high conductivity is preferably used.

From the above viewpoint, as a material of the conductive layer 3, for example, Au (gold), Pt (platinum), or the like is preferably used. The conductive Layer 3 is formed to have a thickness of about 1nm to 350nm by, for example, a plating method, an Atomic Layer Deposition (ALD), an evaporation method, a sputtering method, or the like. In the present embodiment, the thickness of the conductive layer 3 is about 10 nm. As a material of the conductive layer 3, for example, Cr (chromium), Ni (nickel), Ti (titanium), or the like can be used.

Fig. 3 is a diagram showing an enlarged image of the substrate 2 viewed from the thickness direction. In fig. 3, the black portions are through holes 2c, and the white portions are partition walls between the through holes 2 c. As shown in fig. 3, a plurality of through holes 2c having a substantially predetermined width are formed in the substrate 2 in the same manner. The through-hole 2c has a substantially circular shape when viewed in the thickness direction, for example. The width of the through-hole 2c is, for example, about 1nm to 700 nm. In the present embodiment, the width of the through-hole 2c is about 200 nm. When the shape of the through-hole 2c viewed from the thickness direction is substantially circular, the width of the through-hole 2c refers to the diameter of the through-hole 2 c; in the case where the shape is other than a substantially circular shape, the width of the through-hole 2c refers to the diameter (effective diameter) of a virtual maximum cylinder accommodated in the through-hole 2 c. The pitch between the through holes 2c is, for example, about 1nm to 1000 nm. When the through-holes 2c are substantially circular in shape as viewed in the thickness direction, the pitch between the through-holes 2c refers to the distance between the centers of the circles; in the case where the shape is other than a substantially circular shape, the pitch between the through holes 2c refers to the distance between the central axes of the imaginary maximum circular columns accommodated in the through holes 2 c. The width of the partition wall between the through holes 2c of the substrate 2 is, for example, about 300 nm.

The aperture ratio of the through-holes 2c (the ratio of all the through-holes 2c to the first surface 2a when viewed in the thickness direction) is 10 to 80%, and particularly preferably 50 to 60%, from the practical viewpoint. The sizes of the plurality of through holes 2c may be different from each other, or a plurality of partial through holes 2c may be connected to each other.

The substrate 2 is an alumina porous film formed by anodizing Al (aluminum), for example. Specifically, the Al substrate is anodized, and the oxidized surface portion is peeled off from the Al substrate, whereby the substrate 2 is obtained. The substrate 2 may be formed by anodizing a valve metal other than Al, such as Ta (tantalum), Nb (niobium), Ti (titanium), Hf (hafnium), Zr (zirconium), Zn (zinc), W (tungsten), Bi (bismuth), and Sb (antimony), or may be formed by anodizing Si (silicon).

Fig. 4 is an enlarged view of the sample support 1 as viewed from the thickness direction. As shown in fig. 1 and 4, the substrate 2 has a reinforcing region 2d and a measuring region 2 e. A mesh-like (in the present embodiment, a honeycomb-like shape, as an example) reinforcing region 2d and a plurality of measuring regions 2e having substantially predetermined widths are formed on the substrate 2 in the same manner. The reinforcing region 2d and the measuring region 2e each include a plurality of through holes 2c (see fig. 2). The plurality of measurement regions 2e are surrounded by the reinforcement region 2d, respectively, when viewed in the thickness direction. That is, the respective reinforced areas 2d of the plurality of measurement areas 2e are separated from each other. The shape of the measurement region 2e viewed from the thickness direction is, for example, substantially hexagonal.

The width w1 of the measurement region 2e is, for example, about 1 μm to 1000. mu.m. In the present embodiment, the width w1 of the measurement region 2e is about 440 μm to 470 μm. The width w1 of the measurement region 2e refers to the diameter (effective diameter) of an imaginary maximum cylinder accommodated in the measurement region 2 e. That is, as shown in the example of fig. 4, when the shape of the measurement region 2e as viewed in the thickness direction is substantially hexagonal, the width w1 of the measurement region 2e is the distance between two sides of the hexagonal shape facing each other. The pitch p between the measurement regions 2e is, for example, about 1 μm to 1100 μm. In the present embodiment, the pitch p between the measurement regions 2e is about 455 to 530 μm. The pitch p between the measurement regions 2e refers to the distance between the central axes of the imaginary maximum cylinders accommodated in the measurement regions 2 e. The width w2 of the reinforcing region 2d is, for example, about 30 μm to 60 μm.

The aperture ratio of the measurement region 2e (the proportion of the entire measurement region 2e to the first surface 2a when viewed in the thickness direction) is 80% or more. The sizes of the plurality of measurement regions 2e may not be the same, or the plurality of measurement regions 2e may be connected to each other in a local portion.

The reinforcing member 4 is disposed inside a part of the through-holes 2c among the plurality of through-holes 2 c. Specifically, the reinforcing material 4 is disposed inside the plurality of through holes 2c in the reinforcing region 2 d. That is, the reinforcing member 4 is disposed inside the through-hole 2c so as to surround each of the plurality of measurement regions 2e separated from each other when viewed in the thickness direction. The reinforcing member 4 is not disposed inside the plurality of through holes 2c in the plurality of measurement regions 2 e. In the reinforcing region 2d, the reinforcing material 4 is disposed inside the plurality of through holes 2c over the first surface 2a and the second surface 2b of the substrate 2. That is, the reinforcing material 4 is filled in the plurality of through holes 2c in the reinforcing region 2 d. The material of the reinforcing material 4 is, for example, resin or the like. The material of the reinforcing material 4 is, for example, photoresist. In the present embodiment, the material of the reinforcing material 4 is a UV curable resin as an example.

[ method for producing sample support ]

Next, a method for manufacturing the sample support 1 will be described. First, as shown in fig. 5 (a), the substrate 2 is set on the printer 20. In the printer 20, a reinforcing material 41(UV curable resin) is provided in a region 41a having the same shape as the reinforcing region 2 d. For example, the substrate 2 is disposed such that the first surface 2a faces the reinforcing member 41. At this time, the reinforcing region 2d of the substrate 2 and the reinforcing material 41 provided in the region 41a face each other. Subsequently, as shown in (b) of fig. 5, the first surface 2a in the reinforcing area 2d is irradiated with the reinforcing material 41 by the printer 20. In the reinforcing region 2d, the reinforcing material 41 is disposed from the first surface 2a to the inside of the through-hole 2 c.

Subsequently, as shown in fig. 5 (c), the substrate 2 is irradiated with UV light L1. The whole of the first surface 2a is irradiated with UV light L1. When the substrate 2 is irradiated with the UV light L1, the reinforcing material 41 disposed inside the through-hole 2c is cured to become the reinforcing material 4 as shown in fig. 5 (d). Subsequently, the conductive layer 3 is provided on the first surface 2 a. The sample support 1 was obtained by the above-described operation.

[ method of ionizing sample ]

Next, a method of ionizing a sample using the sample support 1 will be described with reference to fig. 6 and 7. Here, a laser desorption ionization method using a laser (energy ray) (a part of a mass spectrometry method performed by the mass spectrometer 10) will be described as an example. In fig. 7, the through-hole 2c, the conductive layer 3, and the reinforcing member 4 in the sample support 1 are not shown.

First, as shown in fig. 6 (a), a sample S and the above-described sample support 1 are prepared (first step). The sample S is cut so that its cross section Sa is exposed. Here, the sample S is, for example, a biological sample (water-containing sample). The sample S is fruit such as strawberry. In order to smoothly move the component S1 (see fig. 6 (b)) of the sample S, a solution (e.g., an acetonitrile mixture, acetone, or the like) for reducing the viscosity of the component S1 may be added to the sample S. The sample support 1 may be prepared by manufacturing by a person who performs an ionization method and a mass spectrometry method, or may be prepared by obtaining from a manufacturer or a vendor of the sample support 1.

Next, as shown in fig. 6 (b), the sample support 1 is disposed on the sample S such that the second surface 2b faces the cross section Sa of the sample S (second step). The sample support 1 is disposed on the sample S such that the second surface 2b is in contact with the cross section Sa. The component S1 of the sample S moves from the second surface 2b side of the substrate 2 toward the first surface 2a side of the substrate 2 through the through-hole 2c by capillary action. Specifically, the component S1 of the sample S moves from the second surface 2b side toward the first surface 2a side through the through hole 2c in the measurement region 2 e. More specifically, the component S1 of the sample S moves toward the first surface 2a side through the through-hole 2c, in which the reinforcing member 4 is not disposed, among the plurality of through-holes 2 c. The component S1 moving toward the first surface 2a side of the substrate 2 stays on the first surface 2a side by surface tension. Next, the sample support 1 is peeled off from the sample S (third step). Before the component S1 adhering to the substrate 2 is dried, the sample support 1 is peeled off from the sample S.

Subsequently, as shown in fig. 7, the sample support 1 is placed on the placement surface 6a of the slide (placement portion) 6. The carrier 6 is a glass substrate on which a transparent conductive film such as an ITO (Indium Tin Oxide) film is formed, and the surface of the transparent conductive film is a mounting surface 6 a. The carrier sheet 6 is not limited to the one used as the placement portion, and a member capable of securing conductivity (for example, a substrate made of a metal material such as stainless steel) may be used.

Next, the sample support 1 is fixed to the slide 6. The sample support 1 is fixed to the slide 6 by a conductive tape 7 (e.g., a carbon tape). Tape 7 may be part of sample support 1 or may be prepared separately from sample support 1. When the tape 7 is a part of the sample support 1 (that is, when the tape 7 is provided on the sample support 1), the tape 7 may be fixed to the first surface 2a side in advance at the peripheral edge portion of the substrate 2, for example. More specifically, the tape 7 may be fixed to the conductive layer 3 at the peripheral edge of the substrate 2.

Next, the slide 6 and the sample support 1 are placed on the support portion 12 (e.g., a stage) of the mass spectrometer 10 in a state where the slide 6 and the sample support 1 are fixed to each other. Next, in a state where the component S1 of the sample S adhering to the sample support 1 is dried, a voltage is applied to the conductive layer 3 (see fig. 2) of the sample support 1 via the mounting surface 6a of the carrier 6 and the tape 7 by the voltage applying unit 14 of the mass spectrometer 10 (fourth step). Next, the first surface 2a of the substrate 2 is irradiated with the laser light L by the laser irradiation unit 13 of the mass spectrometer 10 (fourth step). Here, the laser irradiation unit 13 scans the first surface 2a with the laser light L. The scanning of the laser light L with respect to the first surface 2a can be performed by operating at least one of the support unit 12 and the laser irradiation unit 13. In addition, the width of the measurement region 2e is larger than the spot diameter of the laser light L.

By applying a voltage to the conductive layer 3 and irradiating the first surface 2a of the substrate 2 with the laser light L, the component S1 moving toward the first surface 2a of the substrate 2 is ionized, and the sample ion S2 is released (ionized component S1). Specifically, energy is transferred from the conductive layer 3 that absorbs energy of the laser light L to the component S1 that has moved to the first surface 2a side of the substrate 2, and the component S1 that has obtained the energy is vaporized and an electric charge is obtained, becoming the sample ion S2. Each of the above steps corresponds to an ionization method of the sample S using the sample support 1 (here, as an example, a laser desorption ionization method which is a part of a mass spectrometry method).

The released sample ions S2 are accelerated toward a ground electrode (not shown) provided between the sample support 1 and the ion detection unit 15. That is, the sample ion S2 is accelerated toward the ground electrode by the potential difference generated between the conductive layer 3 to which the voltage is applied and the ground electrode. Then, the sample ions S2 are detected by the ion detector 15 of the mass spectrometer 10 (fifth step). Here, the ion detector 15 detects the sample ion S2 so as to correspond to the scanning position of the laser beam L. This makes it possible to visualize the two-dimensional distribution of molecules constituting the sample S. The Mass spectrometer 10 herein is a Mass spectrometer using Time-of-Flight Mass Spectrometry (TOF-MS). Each of the above steps corresponds to a mass spectrometry method using the sample support 1.

As described above, in the sample support 1, the substrate 2 is provided with the plurality of through holes 2c that are open at the first surface 2a and the second surface 2b opposite to the first surface 2 a. Therefore, for example, in the case where the sample support 1 is disposed on the sample S such as a biological sample so that the second surface 2b of the substrate 2 faces the sample S, the component S1 of the sample S can be moved from the second surface 2b side to the first surface 2a side through the through-hole 2c by capillary action. When the first surface 2a is irradiated with the laser light L, energy is transmitted to the component of the sample moved to the first surface 2a side via the conductive layer 3, and therefore the component S1 of the sample S can be ionized. Further, the reinforcing member 4 is disposed inside a part of the through-holes 2c among the plurality of through-holes 2 c. Thus, the substrate 2 is reinforced by the reinforcing material 4. This makes the substrate 2 less likely to be bent. Therefore, according to the sample support 1, the strength of the substrate 2 can be appropriately secured, and the breakage of the substrate 2 can be suppressed.

For example, when the sample support 1 is transported or the sample support 1 is placed on the sample S, the deflection of the substrate 2 is reduced. This suppresses breakage of the substrate 2. In addition, for example, when the sample support 1 is placed on the sample having adhesive force (for example, a cross section of a cut strawberry) S and then peeled off, damage to the substrate 2 due to the adhesive force can be suppressed.

As a method for ionizing a sample, for example, a Surface-Assisted Laser Desorption/Ionization (SALDI) method is known (see, for example, Japanese patent No. 5129628). SALDI is a method of ionizing a sample by dropping the sample on a substrate having a fine uneven structure on the surface thereof and irradiating the sample with laser light. The plurality of recesses formed in the substrate used in the SALDI do not penetrate the substrate. Therefore, when the strength of the substrate is insufficient, the strength of the substrate can be easily increased by increasing the thickness of the portion of the substrate opposite to the recessed portion (the portion where the recessed portion is not formed).

On the other hand, the sample support 1 moves the components of the sample from the second surface 2b side toward the first surface 2a side through the through-holes 2c by capillary action. Therefore, in order to appropriately realize the capillary phenomenon, it may be difficult to increase the thickness of the substrate 2. In this case, it is particularly effective to reinforce the substrate 2 by the reinforcing material 4.

The width of the through-hole 2c is 1nm to 700nm, and the thickness of the substrate 2 is 1 μm to 50 μm. This can appropriately realize the movement of the component S1 of the sample S due to the capillary phenomenon.

The material of the reinforcing material 4 is resin. This enables the reinforcing material 4 to be easily formed.

The substrate 2 has: a reinforcing region 2d including a plurality of through holes 2c, and a measuring region 2e including a plurality of through holes 2 c. The reinforcing material 4 is disposed inside the plurality of through holes 2c in the reinforcing region 2d, and the reinforcing material 4 is not disposed inside the plurality of through holes 2c in the measuring region 2 e. Since the reinforcing material 4 is disposed inside the plurality of through holes 2c in the reinforcing region 2d, the substrate 2 is less likely to be bent. This can further secure the strength of the substrate 2 and further suppress the breakage of the substrate 2.

In the method for ionizing the sample S, the substrate 2 is provided with a plurality of through holes 2c that are open on the first surface 2a and the second surface 2b opposite to the first surface 2 a. When the sample support 1 is disposed on the sample S so that the second surface 2b of the substrate 2 faces the sample S, the component S1 of the sample S can be moved from the second surface 2b side to the first surface 2a side through the through-hole 2c by capillary action. After the sample support 1 is peeled off from the sample S, when a voltage is applied to the conductive layer 3 and the first surface 2a is irradiated with a laser, energy is transferred to the component S1 of the sample S moving to the first surface 2a side. Thereby, the component S1 of the sample S is ionized. In the sample ionization method, the reinforcing member 4 is disposed in a part of the plurality of through-holes 2c inside the through-holes 2 c. Thus, the substrate 2 is reinforced by the reinforcing material 4. This makes the substrate 2 less likely to be bent, and therefore, in the third step, breakage of the substrate 2 can be suppressed when the sample support 1 is peeled off from the sample S. Thus, according to this sample ionization method, by using the substrate 2 having an appropriately secured strength, the component S1 of the sample S can be ionized while suppressing breakage of the substrate 2.

In the third step, the sample support 1 is peeled off from the sample S before the component S1 adhering to the substrate 2 is dried. This allows the sample support 1 to be more smoothly peeled from the sample S before the sample support 1 is adhered to the sample S.

As described above, according to the mass spectrometry, the substrate 2 having an appropriately secured strength is used, whereby the sample S can be mass-analyzed while suppressing breakage of the substrate 2.

[ modified examples ]

As described above, the embodiments of the present invention have been described, but the present invention is not limited to the above embodiments, and various modifications can be made without departing from the scope of the invention.

Although the example in which the reinforcing material 4 is filled in the through-hole 2c is shown, it may not be filled in the through-hole 2 c. The reinforcing material 4 may extend from the first surface 2a of the substrate 2 to the middle of the through hole 2c, for example. That is, the reinforcing material 4 may be disposed only in a part of the through-hole 2 c.

The conductive layer 3 may be provided not on the inner surface of the through-hole 2c as in the above-described embodiment, or may be provided on the inner surface of the through-hole 2 c.

The substrate 2 may have conductivity, and in the mass spectrometry method, a voltage may be applied to the substrate 2 and the laser light L may be irradiated to the first surface 2 a. When the substrate 2 has conductivity, the conductive layer 3 can be omitted from the sample support 1, and the same effects as those in the case of using the sample support 1 including the conductive layer 3 as described above can be obtained. Note that "irradiating the first surface 2a of the substrate 2 with the laser light L" means irradiating the conductive layer 3 with the laser light L when the sample support 1 includes the conductive layer 3; when the substrate 2 has conductivity, the first surface 2a of the substrate 2 is irradiated with the laser light L.

Although an example in which the material of the reinforcing material 4 is a UV curable resin is shown, the material of the reinforcing material 4 may be a thermosetting resin. In addition, the material of the reinforcing material 4 may be metal. The metal is, for example, Ni (nickel) or the like. In the case where the material of the reinforcing material 4 is a metal, when the component S1 of the sample S is ionized, the generation of an organic gas is suppressed, thereby reducing noise in the detection result of the ionized sample ion S2. Further, as described above, the reinforcing material 4 can be made of various materials, and therefore, the reinforcing material 4 can be appropriately provided only in a part of the through-holes 2c by adopting an appropriate processing method depending on the various materials. In the case where the material of the reinforcing member 4 is a thermosetting resin, heat is applied to the substrate 2 instead of the irradiation with the UV light L1 in the above-described method for producing the sample support 1.

Although an example in which the shape of the measurement region 2e is substantially hexagonal when viewed from the thickness direction is shown, the measurement region 2e may take various shapes. For example, as shown in fig. 8 (a), the shape of the measurement region 2e when viewed from the thickness direction may be substantially square. That is, the reinforcing regions 2d may be provided in a lattice shape. As shown in fig. 8 (b), the measurement region 2e may have a substantially rectangular shape extending in one direction when viewed in the thickness direction. That is, the reinforcing region 2d may be formed of a plurality of regions extending in parallel with each other and arranged at substantially equal intervals. The shape of the measurement region 2e when viewed from the thickness direction may be other than the above, and may be, for example, substantially circular or substantially triangular.

Further, although an example in which the reinforcing area 2d is in a mesh shape is shown, the reinforcing area 2d may have various shapes as long as a plurality of through holes 2c in which the reinforcing material 4 is arranged are included. The reinforcing region 2d may include a continuous region from one end of the substrate 2 to the other end opposite to the one end when viewed from the thickness direction. Specifically, the reinforcing region 2d may be an elongated region extending along a diagonal line of the substrate 2, for example, as shown in fig. 9. Here, one end and the other end of the substrate 2 are corner portions facing each other on a diagonal line of the substrate 2, respectively. In this case, the substrate 2 becomes less likely to be flexed. Therefore, the strength of the substrate 2 can be ensured more reliably, and breakage of the substrate 2 can be suppressed more reliably. One end and the other end of the substrate 2 may be side portions of the substrate 2 facing each other. That is, the reinforcing region 2d may be an elongated region extending in a direction perpendicular to the side portion of the substrate 2, instead of the diagonal line of the substrate 2.

Further, although the example in which the width of the measurement regions 2e is about 1 μm to 1000 μm and the pitch between the measurement regions 2e is about 1 μm to 1100 μm is shown, the width of the measurement regions 2e and the pitch between the measurement regions 2e may be about several mm to several cm. In particular, the reinforcing region 2d may also be constituted by one or more elongated regions. That is, as shown in fig. 10 (a), the reinforcing region 2d may be constituted by, for example, an annular region extending along the outer edge of the substrate 2 and one region extending along the diagonal line of the substrate 2. In this case, two measurement regions 2e are formed on the substrate 2. As shown in fig. 10 (b), the reinforcing region 2d may be formed of, for example, an annular region extending along the outer edge of the substrate 2 and two regions extending along two diagonal lines of the substrate 2. In this case, four measurement regions 2e are formed on the substrate 2. As shown in fig. 10 (c), the reinforcing region 2d may be formed of, for example, an annular region extending along the outer edge of the substrate 2, one region extending along one diagonal line of the substrate 2, and a plurality of (three in this case) regions extending along one side portion of the substrate 2. In this case, 8 measurement regions 2e are formed on the substrate 2. As shown in fig. 10 (d), the reinforcing region 2d may be configured by, for example, an annular region extending along the outer edge of the substrate 2, a plurality of (three, here) regions extending along one side portion of the substrate 2, and one region extending along the other side portion of the substrate 2 intersecting the one side portion. In this case, 8 measurement regions 2e are formed on the substrate 2.

In this case, a plurality of samples S may be arranged corresponding to the plurality of measurement regions 2 e. Then, ionization and mass spectrometry of the component S1 of the plurality of samples S can be performed in each of the plurality of measurement regions 2 e. In addition, ionization and mass spectrometry of a plurality of samples S may be performed together.

The sample support 1 may be provided with a frame. In this case, the frame is provided to the first surface 2a of the substrate 2. Specifically, the frame is fixed to the peripheral edge portion of the substrate 2. The frame is fixed to the first surface 2a of the substrate 2 by an adhesive layer. The frame has substantially the same outer shape as the substrate 2 as viewed in the thickness direction. An opening is formed in the frame. The portion of the substrate 2 corresponding to the opening functions as an effective region for moving the component S1 of the sample S toward the first surface 2 a. In this case, the conductive layer 3 may be continuously (integrally) formed on a region corresponding to the opening of the frame (i.e., a region corresponding to the effective region) in the first surface 2a of the substrate 2, an inner surface of the opening, and a surface opposite to the substrate 2 in the frame. By providing such a frame, the strength of the sample support 1 (substrate 2) can be increased, and the handling of the sample support 1 can be improved. In addition, deformation of the substrate 2 due to temperature change or the like can be effectively suppressed.

In the mass spectrometer 10, the following may be used: the laser irradiation unit 13 collectively irradiates the first surface 2a of the substrate 2 with the laser light L, and the ion detection unit 15 detects the sample ions S2 while maintaining the two-dimensional information of the region. That is, the mass spectrometer 10 may be a projection-type mass spectrometer.

Although the example in which the sample support 1 is placed on the slide 6 has been described above, the sample support 1 may be directly placed on the support portion 12 of the mass spectrometer 10.

The application of the sample support 1 is not limited to the ionization of the sample S by the irradiation of the laser light L. The sample support 1 may be used for ionizing the sample S by irradiating the sample with an energy ray other than the laser beam L (for example, an ion beam, an electron beam, or the like).

The sample support 1 may be fixed to the slide 6 by a method other than the tape 7 (for example, a tool using an adhesive, a jig, or the like). Further, a voltage may be applied to the conductive layer 3 without passing through the mounting surface 6a of the carrier 6 and the tape 7. In this case, the slide 6 and the tape 7 may not have conductivity.

The above-described method of ionizing a sample can be used not only for mass spectrometry of molecules constituting the sample S but also for other measurements and experiments such as ion mobility measurement.

The sample S may be a dried sample. In this case, for example, a solvent (for example, an acetonitrile mixture, acetone, or the like) may be added to the sample S in order to move the component S1 of the sample S toward the first surface 2a of the substrate 2 by capillary action.

Next, as a first modification of the method for manufacturing the sample support, a case where the material of the reinforcing member 4 is a negative photoresist will be described. First, as shown in fig. 11 (a), the substrate 2 is set on the printer 20. In the printer 20, a reinforcing material 41 (negative resist) is provided. The substrate 2 is disposed such that the first surface 2a faces the reinforcing material 41, for example. Subsequently, as shown in fig. 11 (b), the reinforcing material 41 is applied to the first surface 2a by the printer 20. The reinforcing material 41 is disposed inside the through-hole 2c from the first surface 2 a. Subsequently, the reinforcing material 41 is cured by heating the substrate 2 provided with the reinforcing material 41.

Subsequently, as shown in fig. 11 (c), a mask 21 is provided on the first surface 2a of the substrate 2. The mask 21 is disposed on the first surface 2a in the measurement region 2 e. Subsequently, the substrate 2 is irradiated with UV light L2, thereby exposing the reinforcing material 41 disposed inside the through-holes 2c in the reinforcing region 2 d. The UV light L2 irradiates the entire surface of the substrate 2 opposite to the second surface 2 b. Since the mask 21 is provided on the first surface 2a in the measurement region 2e, the reinforcing material 41 disposed inside the through-hole 2c in the measurement region 2e is not exposed to light. Subsequently, as shown in (d) of fig. 11, the mask 21 is removed from the first surface 2 a. Subsequently, the reinforcing material 41 disposed inside the through-hole 2c in the measurement region 2e is removed by immersing the substrate 2 on which the reinforcing material 41 is disposed in a developing solution. On the other hand, the reinforcing material 41 disposed in the reinforcing region 2d is not removed and remains because its solubility in the developer is lowered by exposure to light. The reinforcing material 41 thus remaining becomes the reinforcing material 4. Subsequently, the conductive layer 3 is provided on the first surface 2 a. As described above, the sample support 1 was obtained.

Next, as a second modification of the method for manufacturing the sample support, a case will be described in which the material (reinforcing material 41) of the reinforcing material 4 is a metal (for example, Ni). First, as shown in fig. 12 (a), a substrate 2 is prepared. Subsequently, as shown in fig. 12 (b), a mask 22 is provided on the first surface 2a of the substrate 2. The mask 22 is disposed on the first surface 2a in the measurement region 2 e. Subsequently, a metal film is formed by evaporation of the reinforcing material 41 (Ni). A metal film composed of the reinforcing material 41 is formed on the first surface 2a in the reinforcing region 2 d.

Subsequently, as shown in (c) of fig. 12, the mask 22 is removed from the first surface 2 a. Subsequently, as shown in fig. 12 (d), the reinforcing material 41 is disposed in the through-hole 2c by plating. Specifically, first, the substrate 2 on which the metal film made of the reinforcing material 41 is formed is immersed in the electrolytic solution. Subsequently, the reinforcing material 41 is supplied with electric current. When a current is supplied in a state where the reinforcing material 41 is immersed in the electrolytic solution, the metal atoms of the reinforcing material 41 are dissolved in the electrolytic solution. Then, the dissolved metal atoms of the reinforcing material 41 are plated on the inner surface of the through-hole 2c in the reinforcing region 2d inside the through-hole 2 c. The metal atoms of the reinforcing material 41 disposed inside the through-holes 2c function as the reinforcing material 4. Subsequently, the conductive layer 3 is provided on the first surface 2 a. As described above, the sample support 1 was obtained.

[ description of symbols ]

1 sample support

2 base plate

2a first surface

2b second surface

2c through hole

2d reinforced area

2e measurement area

3 conductive layer

4 reinforcing material

L laser (energy line)

S sample

Component S1

S2 sample ions.