JP2023053501A - Specimen support and ionization method - Google Patents

Abstract

To provide a specimen support capable of appropriately implementing processing for transferring a specimen to be ionized on a surface of a substrate, and an ionization method.SOLUTION: A specimen support 1A of an embodiment comprises: a support substrate 2 including a support surface 2a; a sheet-like adhesive layer 3 provided on the support surface 2a; an ionization substrate 4 provided on the support surface 2a via the adhesive layer 3, the ionization substrate 4 including a first surface 4a opposed to the adhesive layer 3, a second surface 4b at an opposite side of the first surface 4a, and a plurality of through holes 4c opening in at least the second surface 4b; and a protective film 6 on a sheet covering a part of the second surface 4b of the ionization substrate 4 and provided removably. The ionization substrate 4 includes a measurement region R1 for transferring a specimen to be measured and a calibration region R2 for calibration. The protective film 6 is provided so as to cover at least the entire calibration region R2.SELECTED DRAWING: Figure 1

Description

The present disclosure relates to sample supports and ionization methods.

Patent Document 1 describes a sample support including a substrate having a measurement area provided with a plurality of through holes, and a frame formed around the measurement area on the surface of the substrate. Patent Document 2 describes a mass spectrometry method in which a sample transferred to a substrate having a fine uneven surface is irradiated with laser light to ionize the sample.

Japanese Patent No. 6093492 Japanese Patent No. 4674875

When a sample to be ionized is transferred onto the surface of a substrate as described in Patent Document 2, if the rigidity of the substrate is insufficient, the substrate may be damaged during transfer. In the sample support described in Patent Document 1, although the substrate is supported by a frame, the measurement area (effective area) of the substrate is not provided with a frame. may be damaged. Further, in order to measure ions with high accuracy, it is preferable to perform calibration using a calibration reagent using the same substrate. For such calibration, it is conceivable to provide a calibration area separate from the measurement area on the substrate, but when the sample is transferred to the measurement area, part of the sample may adhere to the calibration area. .

Accordingly, an object of the present disclosure is to provide a sample support and an ionization method that can suitably perform processing for transferring a sample to be ionized onto the surface of a substrate.

A sample support according to one aspect of the present disclosure includes a support substrate having a support surface, a sheet-like adhesive layer provided on the support surface, and an ionization substrate provided on the support surface via the adhesive layer. an ionization substrate having a first surface facing the adhesive layer, a second surface opposite the first surface, and a plurality of pores opening at least to the second surface; and a second surface of the ionization substrate. and a detachable sheet-like cover member that covers a part of the ionization substrate, and the ionization substrate has a measurement area for transferring the sample to be measured and a calibration area for calibration. , the cover member is provided so as to cover at least the entire calibration area.

In the sample support described above, the first surface of the ionization substrate is fixed to the support substrate via the adhesive layer. Thereby, the rigidity of the ionization substrate can be increased, and the ionization substrate can be prevented from being damaged when the sample is transferred to the measurement area of the ionization substrate. A calibration area for calibration is provided separately from the measurement area, and a cover member that covers the entire calibration area in a detachable manner is provided. This can prevent part of the sample from adhering to the calibration area when the sample is transferred to the measurement area. As described above, according to the sample support, the process of transferring the sample to be ionized onto the surface of the ionization substrate can be preferably carried out.

The adhesive layer may be provided so as to overlap the entire measurement area when viewed in a direction perpendicular to the support surface. According to the above configuration, the entire measurement region of the ionization substrate to which the sample is transferred can be tightly fixed to the support surface by the adhesive layer. This can effectively prevent the portion of the ionization substrate corresponding to the measurement region from being damaged when the sample is transferred.

The ionized substrate may be formed by anodizing a valve metal or silicon, the pores may be through holes opening to the first surface and the second surface, and on the second surface, A conductive layer may be provided so as not to block the pores. According to the above configuration, an ionized substrate having a plurality of through holes (pores) can be easily formed by anodizing the valve metal or silicon. Further, by providing a conductive layer on the second surface of the ionization substrate so as not to block the pores, it is possible to prevent the application of voltage to the second surface of the ionization substrate when ionizing the sample by irradiating the energy beam. It is possible to do it properly.

The pores may be through holes that open to the first surface and the second surface, and the adhesive layer may be provided so as not to overlap the calibration area when viewed in a direction perpendicular to the support surface. good. Calibration using the calibration area involves dropping a calibration reagent (solution) onto the calibration area, drying the calibration reagent, and then irradiating the calibration area with an energy beam to ionize the components of the calibration reagent. , by detecting ionized constituents of the calibration reagent. If the pores provided in the ionization substrate are through holes and the adhesive layer is provided so as to overlap the calibration region, the calibration reagent dropped on the calibration region and the adhesive layer component (adhesive component etc.). As a result, there is a possibility that calibration cannot be performed properly. On the other hand, by providing the adhesive layer so as not to overlap the calibration area as described above, it is possible to prevent the components of the adhesive layer from being mixed into the calibration reagent, and to perform calibration with high accuracy.

The sample support includes a conductive tape that is provided from the second surface of the ionization substrate to a portion of the support surface that does not overlap the ionization substrate, and electrically connects the ionization substrate and the portion of the support surface. Further provided, said portion of the support surface may be electrically conductive. According to the above configuration, by applying a voltage to the portion of the support surface that does not overlap the ionization substrate, it is possible to appropriately apply the voltage to the second surface of the ionization substrate via the conductive tape. .

The conductive tape may be provided with a first opening containing the calibration region therein when viewed in a direction orthogonal to the support surface, and the cover member is conductive so as to cover the first opening. It may be provided on the surface of the tape opposite to the ionization substrate side. According to the above configuration, by interposing the conductive tape between the cover member and the ionization substrate, a part of the cover member (for example, the adhesive surface having slight adhesiveness) is brought into contact with the second surface of the ionization substrate. You can prevent it from slipping. As a result, when removing the cover member, it is possible to appropriately prevent a part of the adhesive surface from adhering to the calibration area. Also, when the cover member is removed, part of the ionized substrate is prevented from peeling off together with the cover member. As described above, the calibration using the calibration region can be performed with higher accuracy.

The pores may be through holes that open to the first surface and the second surface, and the adhesive layer is provided with a second opening corresponding to the calibration area when viewed in a direction perpendicular to the support surface. and the first opening may be larger than the second opening, and the edges of the first opening and the edges of the second opening are In some cases, they may be spaced apart from each other. According to the above configuration, since the edge of the first opening is separated from the edge of the second opening, it is possible to prevent part of the conductive tape from adhering to the calibration area of the ionization substrate. . Furthermore, by using the first opening, which is larger than the second opening (that is, the opening corresponding to the calibration area) as a mark, the operator who carries out calibration can easily grasp the position of the calibration area.

The conductive tape and the cover member may be formed in a frame shape surrounding the measurement area when viewed in a direction perpendicular to the support surface. According to the above configuration, it is possible to suitably apply a voltage to the second surface of the ionization substrate by means of the frame-shaped conductive tape. In addition, since the cover member exists around the measurement area when viewed from the direction orthogonal to the support surface, the handling of the sample support during sample transfer or the like can be improved.

At least a portion of the support substrate that overlaps the measurement area may be transparent, and the adhesive layer may be made of a transparent material. According to the above configuration, since the members (support substrate and adhesive layer) overlapping the measurement area on the first surface side of the ionization substrate are transparent, the light transmission image of the sample transferred to the second surface of the measurement area (that is, It is possible to obtain an observed image by applying illumination light from the surface of the support substrate opposite to the ionization substrate side.

An ionization method according to another aspect of the present disclosure includes a first step of preparing the sample support, a second step of transferring the sample to the second surface of the measurement region of the ionization substrate, and transferring the sample to the second surface. A third step of exposing the calibration region of the ionization substrate by removing the cover member after the transfer is completed; a fourth step of dropping the calibration reagent onto the second surface of the calibration region; a fifth step of ionizing the components of the calibration reagent by irradiating the second surface of the calibration region with energy rays while applying a voltage to the second surface of the ionization substrate after the calibration reagent is dried; and a sixth step of ionizing the constituents of the sample by irradiating the second surface of the measurement region with energy rays while applying a voltage to the second surface of the ionization substrate.

In the above ionization method, since the first surface of the ionization substrate is firmly fixed to the support substrate via the adhesive layer, damage to the ionization substrate in the second step can be suppressed. Further, the ionization substrate 4 is provided with a cover member that covers the entire calibration area in a detachable manner. This can prevent part of the sample from adhering to the calibration area when the sample is transferred to the measurement area in the second step. As described above, according to the ionization method, it is possible to suitably perform the process of transferring the sample to be ionized onto the surface of the ionization substrate.

In the second step, the sample is applied onto the human skin, and the sample support is pressed against the skin with the second surface of the measurement region facing the sample, thereby transferring the sample onto the second surface of the measurement region. You may In the ionization method described above, since the first surface of the ionization substrate is firmly fixed to the support substrate via the adhesive layer, a support member for supporting the ionization substrate on the second surface side of the ionization substrate (for example, Patent Document There is no need to provide a frame with openings for exposing the measurement area as disclosed in 1.). Therefore, the process of pressing the sample support against the human skin and transferring the sample on the skin to the measurement area can be performed smoothly.

After the second step and before the fifth and sixth steps, the ionization method extends from the second surface of the ionization substrate to the portion of the support surface that does not overlap the ionization substrate. and electrically connecting the ionizable substrate and the portion of the support surface such that in each of the fifth and sixth steps, a voltage is applied to the portion of the support surface. Thus, a voltage may be applied to the second surface of the ionizing substrate via the conductive tape. In the above-described ionization method, since part of the second surface of the ionization substrate is not covered with the conductive tape in the second step, the transfer of the sample to the measurement area can be performed more smoothly. Moreover, by providing the conductive tape before the fifth step and the sixth step, it becomes possible to more stably apply a voltage to the second surface of the ionization substrate.

Advantageous Effects of Invention According to the present disclosure, it is possible to provide a sample support and an ionization method that can suitably perform processing for transferring a sample to be ionized onto the surface of a substrate.

FIG. 1 is a plan view of the sample support of the first embodiment. FIG. FIG. 2 is a cross-sectional view of the sample support along line II-II shown in FIG. FIG. 3 is an enlarged cross-sectional view of essential parts showing the schematic configuration of the ionization substrate and the conductive layer of the sample support shown in FIG. 4 is a magnified image of the ionization substrate of the sample support shown in FIG. 1. FIG. FIG. 5 is a diagram showing an example of the manufacturing process of the sample support shown in FIG. FIG. 6 is a diagram showing an example of the manufacturing process of the sample support shown in FIG. FIG. 7 is a diagram showing an example of the manufacturing process of the sample support shown in FIG. FIG. 8 is a diagram showing an example of the manufacturing process of the sample support shown in FIG. FIG. 9 is a diagram showing an example of steps of a mass spectrometry method using the sample support shown in FIG. FIG. 10 is a diagram showing an example of steps of a mass spectrometry method using the sample support shown in FIG. FIG. 11 is a diagram showing an example of steps of a mass spectrometry method using the sample support shown in FIG. FIG. 12 is a diagram showing an example of steps of a mass spectrometry method using the sample support shown in FIG. FIG. 13 is a plan view of the sample support of the second embodiment. 14 is a cross-sectional view of the sample support along line XIV-XIV shown in FIG. 13. FIG. FIG. 15 is a diagram showing the configuration of the sample support of the third embodiment. FIG. 16 is a diagram showing the configuration of the sample support of the fourth embodiment.

Hereinafter, embodiments of the present disclosure will be described with reference to the drawings. In each figure, the same or corresponding parts are denoted by the same reference numerals, and redundant explanations are omitted. In addition, in the drawings, some parts are exaggerated for easy understanding of characteristic parts according to the embodiment, and the dimensions may differ from the actual dimensions.

[First embodiment]
[Configuration of sample support]
A sample support 1A according to the first embodiment will be described with reference to FIGS. 1 to 4. FIG. The sample support 1A is an ionization support substrate used for ionizing a sample. For example, the sample support 1A is used for ionizing a sample applied on human skin (skin surface). More specifically, the sample is transferred to the sample support 1A by pressing the sample support 1A against the sample applied on the human skin. Thereafter, the sample support 1A onto which the sample has been transferred is irradiated with an energy beam such as a laser beam to ionize the sample. Examples of samples include drugs, cosmetics, and the like.

As shown in FIGS. 1 to 3, the sample support 1A includes a support substrate 2, an adhesive layer 3, an ionization substrate 4, a conductive tape 5, a protective film 6 (cover member), and a conductive layer 7. and have. As shown in FIG. 1, the sample support 1A has a substantially rectangular shape in plan view. In the present embodiment, the longitudinal direction along the long sides of the sample support 1A is represented as the X-axis direction, and the short side direction along the short sides of the sample support 1A is represented as the Y-axis direction. The direction orthogonal to the direction (that is, the thickness direction of the sample support 1A) is represented as the Z-axis direction.

The support substrate 2 is a substrate that supports the ionization substrate 4 . The support substrate 2 is formed in, for example, a rectangular plate shape. The support substrate 2 has a support surface 2a. The support surface 2a is a surface on which the ionization substrate 4 is provided with the adhesive layer 3 interposed therebetween. The support surface 2a is a surface parallel to the X-axis direction and the Y-axis direction. That is, the direction perpendicular to the support surface 2a coincides with the Z-axis direction. The support substrate 2 can be formed of, for example, a slide glass. In this embodiment, as an example, the support substrate 2 is a glass substrate (ITO slide glass) on which a transparent conductive film such as an ITO (Indium Tin Oxide) film is formed, and the surface of the transparent conductive film serves as the support surface 2a. there is That is, in this embodiment, the entire support surface 2a is conductive. Moreover, the entire support substrate 2 is made transparent. As an example, the length in the longitudinal direction (X-axis direction) of the support substrate 2 is about 75 mm, the length in the lateral direction (Y-axis direction) is about 25 mm, and the thickness (length in the Z-axis direction) is about 75 mm. is about 1 mm.

The adhesive layer 3 is provided on the support surface 2 a of the support substrate 2 . The adhesive layer 3 is formed in a sheet shape. From the viewpoint of preventing the components of the adhesive layer 3 from penetrating into the through holes 4c of the ionized substrate 4, which will be described later, the adhesive layer 3 is preferably formed of a solid (non-liquid) and non-fluid material. . The adhesive layer 3 is, for example, a double-sided tape (transparent double-sided tape) made of a transparent material. As an example, the adhesive layer 3 is provided in a rectangular shape when viewed from the Z-axis direction, and is arranged in the center of the support surface 2a in the X-axis direction. The thickness of the adhesive layer 3 is, for example, about 50 μm.

The adhesive layer 3 is provided with an opening 3a corresponding to a calibration region R2, which will be described later. As an example, the opening 3a is formed in a circular shape when viewed from the Z-axis direction. In the present embodiment, a plurality of (three as an example) openings 3a are arranged at regular intervals along the Y-axis direction on one side of the adhesive layer 3 in the X-axis direction (the left side in FIG. 1). .

The ionization substrate 4 is provided on the support surface 2a of the support substrate 2 with the adhesive layer 3 interposed therebetween. As an example, the ionization substrate 4 is provided in a rectangular shape having the same size as the adhesive layer 3 when viewed from the Z-axis direction. The ionization substrate 4 is a very thin membrane-like member. The ionized substrate 4 has a first surface 4a and a second surface 4b opposite the first surface 4a. The first surface 4 a is the surface facing the adhesive layer 3 . That is, the first surface 4 a is the surface to be adhered to the adhesive layer 3 . The second surface 4b is a surface onto which the sample is transferred, and is a surface irradiated with energy rays such as laser light in the step of ionizing the components of the sample. The ionization substrate 4 is made of, for example, an insulating material and has a rectangular plate shape. The length of one side of the ionization substrate 4 when viewed in the Z-axis direction is, for example, about several centimeters. The thickness of the ionization substrate 4 is, for example, about 1 μm to 50 μm. As an example, the ionization substrate 4 has a thickness of 5 μm to 15 μm.

The ionization substrate 4 has a measurement region R1 for transferring a sample to be measured and a calibration region R2 for calibration (mass calibration).

The measurement region R1 is a region for transferring the sample and ionizing the components of the sample by irradiation with energy rays. A measurement region R1 is a region of the ionized substrate 4 that is not covered by the conductive tape 5 and the protective film 6 . In the present embodiment, as shown in FIGS. 1 and 2, the measurement area R1 is a rectangular area located in the center of the sample support 1A in the X-axis direction. More specifically, the measurement area R1 includes the protective film 6 formed on one side in the X-axis direction (the left side in FIG. 1) and the conductive film 6 formed on the other side in the X-axis direction (the right side in FIG. 1). This is the area between the conductive tape 5 (conductive tape 5B).

The calibration region R2 is a region where the calibration reagent is dropped, and after the calibration reagent is dried, the components of the calibration reagent are ionized by irradiation with energy rays. The calibration reagent is, for example, a liquid reagent such as a peptide solution. The calibration region R2 is a region that overlaps the opening 3a of the adhesive layer 3. As shown in FIG. That is, the portion of the ionized substrate 4 that does not overlap the adhesive layer 3 when viewed in the Z-axis direction functions as the calibration region R2. In this embodiment, three circular calibration regions R2 are defined by forming three openings 3a in the adhesive layer 3. As shown in FIG. The width of the calibration region R2 (that is, the diameter of the opening 3a) is, for example, about 2.5 mm. As shown in FIGS. 1 and 2, the calibration area R2 is covered with the protective film 6 before the sample is transferred to the measurement area R1.

As shown in FIG. 3, the ionization substrate 4 is formed with a plurality of through holes 4c (pores) uniformly (uniformly distributed). Each through-hole 4c extends along the thickness direction of the ionization substrate 4 (the direction in which the first surface 4a and the second surface 4b face each other and coincides with the Z-axis direction). It is open to the surface 4a and the second surface 4b. The shape of the through-hole 4c when viewed in the Z-axis direction is, for example, substantially circular. The width of the through hole 4c is, for example, about 1 nm to 700 nm.

The width of the through hole 4c is a value obtained as follows. First, an image of each of the first surface 4a and the second surface 4b of the ionized substrate 4 is acquired. FIG. 4 shows an example of an SEM image of a portion of the second surface 4b of the ionization substrate 4. As shown in FIG. In the SEM image, black portions are the through holes 4c, and white portions are partition walls between the through holes 4c. Subsequently, for example, by performing a binarization process on the acquired image of the first surface 4a, a plurality of first openings (openings on the first surface 4a side of the through-holes 4c) in the measurement region R1 correspond to A plurality of pixel groups are extracted, and the diameter of the circle having the average area of the first aperture is obtained according to the size per pixel. Similarly, by performing, for example, binarization processing on the acquired image of the second surface 4b, a plurality of second openings (openings on the second surface 4b side of the through-holes 4c) in the measurement region R1 correspond to A plurality of pixel groups are extracted, and the diameter of the circle having the average area of the second aperture is obtained according to the size per pixel. Then, the average value of the diameter of the circle obtained for the first surface 4a and the diameter of the circle obtained for the second surface 4b is obtained as the width of the through hole 4c.

As shown in FIG. 4, the ionization substrate 4 is uniformly formed with a plurality of through holes 4c having substantially constant widths. The ionized substrate 4 shown in FIG. 4 is an alumina porous film formed by anodizing Al (aluminum). For example, by anodizing the Al substrate, the surface portion of the Al substrate is oxidized and a plurality of pores (portions to become the through holes 4c) are formed in the surface portion of the Al substrate. be done. Subsequently, the oxidized surface portion (anodized film) is peeled off from the Al substrate, and the peeled anodized film is subjected to a pore widening treatment for widening the pores, thereby forming the ionized substrate described above. 4 is obtained. The ionization substrate 4 is Ta (tantalum), Nb (niobium), Ti (titanium), Hf (hafnium), Zr (zirconium), Zn (zinc), W (tungsten), Bi (bismuth), Sb (antimony). ), or may be formed by anodizing Si (silicon).

As described above, since the plurality of through holes 4c are uniformly formed in the ionization substrate 4, both the measurement region R1 and the calibration region R2 are regions including the plurality of through holes 4c. The aperture ratio of the through-holes 4c in the measurement region R1 (the ratio of the through-holes 4c to the measurement region R1 when viewed from the Z-axis direction) is practically 5% to 60%, particularly 10% to 40%. %. The sizes of the plurality of through holes 4c may be uneven, or the plurality of through holes 4c may be partially connected to each other. The calibration region R2 is similar to the measurement region R1.

The conductive tape 5 is provided from the second surface 4b of the ionization substrate 4 to the portion of the support surface 2a that does not overlap the ionization substrate 4, and electrically connects the ionization substrate 4 and the support surface 2a. Connecting. The conductive tape 5 is made of a conductive material. The conductive tape 5 is, for example, aluminum tape, carbon tape, or the like. The thickness of the conductive tape 5 is, for example, 50 μm.

In this embodiment, two conductive tapes 5 extending in the Y-axis direction are provided on each of both side edges of the ionization substrate 4 in the X-axis direction. More specifically, the sample support 1A includes a conductive tape 5A provided along the edge of the ionization substrate 4 on one side in the X-axis direction (the left side in FIG. and a conductive tape 5B provided along the edge on the other side in the direction (the right side in FIG. 1).

The surface of the conductive tape 5 on the support substrate 2 side is an adhesive surface having an adhesive substance. Each conductive tape 5 has a surface at the X-axis direction edge of the ionization substrate 4 (in this embodiment, the surface of the conductive layer 7 on the edge of the ionization substrate 4) and a supporting tape adjacent to the edge of the ionization substrate 4. It is attached to each of the surfaces 2a (in this embodiment, the surface of the conductive layer 7 on the support surface 2a).

The conductive tape 5A is provided so as not to overlap the opening 3a (that is, the calibration region R2) of the adhesive layer 3 when viewed from the Z-axis direction. That is, the end of the conductive tape 5A on the inner side in the X-axis direction (the end on the side of the conductive tape 5B) does not reach the position where the opening 3a of the adhesive layer 3 is formed.

The protective film 6 is a sheet-like member that partially covers the second surface 4b of the ionization substrate 4 and is detachably provided on the second surface 4b. Protective film 6 may be formed of a resin film having a thickness of 100 μm or less, for example. In this embodiment, the protective film 6 is provided so as to straddle the edge of the ionization substrate 4 on one side in the X-axis direction (the left side in FIG. 1). More specifically, the protective film 6 is formed in a rectangular shape larger than the conductive tape 5A when viewed from the Z-axis direction, and is positioned outside the conductive tape 5A (on the side of the conductive tape 5B). It is provided so as to overlap a portion of the support surface 2a on the opposite side), the conductive tape 5A, and a portion of the ionization substrate 4 on the inner side of the conductive tape 5A (on the side of the conductive tape 5B).

The surface of the protective film 6 on the side of the support substrate 2 is an adhesive surface having a slightly adhesive substance having weaker adhesive force than the adhesive substance provided on the conductive tape 5 . The protective film 6 is provided so as to cover at least the entire calibration region R2. That is, the protective film 6 serves to protect the calibration area R2 so that the calibration area R2 is not exposed.

The conductive layer 7 is provided at least on the second surface 4b of the ionization substrate 4 so as not to block the through hole 4c. The conductive layer 7 is made of a conductive material. The conductive layer 7 is made of Pt (platinum) or Au (gold), for example. As the material of the conductive layer 7, it is preferable to use a metal that has a low affinity (reactivity) with the sample and a high conductivity for the reasons described below.

For example, if the conductive layer 7 is formed of a metal such as Cu (copper), which has a high affinity with a sample such as protein, the sample will be exposed to the sample with Cu atoms attached to the sample molecules in the process of ionizing the sample, which will be described later. There is a possibility that detection results in mass spectrometry, which will be described later, will deviate by the amount of Cu atoms that are ionized and attached. Therefore, it is preferable to use a metal having a low affinity for the sample as the material of the conductive layer 7 .

On the other hand, metals with higher conductivity are more easily and stably applied with a constant voltage. Therefore, if the conductive layer 7 is made of a highly conductive metal, it is possible to uniformly apply a voltage to the second surface 4b of the ionization substrate 4 in the measurement region R1. Also, metals with higher conductivity tend to have higher thermal conductivity. Therefore, if the conductive layer 7 is formed of a highly conductive metal, the energy of the laser beam (energy ray) irradiated to the ionization substrate 4 can be efficiently transmitted to the sample via the conductive layer 7. becomes. Therefore, it is preferable to use a highly conductive metal as the material of the conductive layer 7 .

From the above point of view, it is preferable to use, for example, Pt, Au, or the like as the material of the conductive layer 7 . The conductive layer 7 is formed to a thickness of about 1 nm to 350 nm by vapor deposition, sputtering, or the like, for example. As the material of the conductive layer 7, for example, Cr (chromium), Ni (nickel), Ti (titanium), or the like may be used.

As an example, the conductive layer 7 is deposited on the entire surface of the sample support 1A on the support surface 2a side of the support substrate 2 before the conductive tape 5 and protective film 6 are provided. In this case, the conductive layer 7 includes a portion of the support surface 2a where the adhesive layer 3 is not provided, side surfaces (portions exposed to the outside) of the adhesive layer 3 and the ionization substrate 4, and the second surface 4b of the ionization substrate 4. , and is provided so as to straddle the .

[Manufacturing method of sample support]
An example of a method for manufacturing the sample support 1A will be described with reference to FIGS. 5 to 8. FIG. 5 to 8, (A) shows a plan view of the sample support at each manufacturing stage, and (B) shows a cross-sectional view along line BB in (A). .

First, as shown in FIG. 5 , an adhesive layer 3 (transparent double-sided tape in this embodiment) is attached to the support surface 2 a of the support substrate 2 . Subsequently, an ionized substrate 4 is placed on the adhesive layer 3, as shown in FIG. Thereby, the ionization substrate 4 is fixed to the support surface 2 a of the support substrate 2 via the adhesive layer 3 .

Subsequently, a conductive layer 7 is deposited on the support surface 2a side of the support substrate 2, as shown in FIG. As a result, the conductive layer 7 is removed from the portions indicated by the thick lines in FIG. exposed part) as well as on the second surface 4b) of the ionizing substrate 4). The conductive layer 7 may be provided at least on the second surface 4b of the ionization substrate 4, and may not be provided on other portions.

Subsequently, as shown in FIG. 8, conductive tapes 5A and 5B are attached to both side edges of the ionization substrate 4 in the X-axis direction. Finally, as shown in FIG. 1, the sample support 1A is completed by attaching a protective film 6 so as to cover the calibration region R2.

[Mass Spectrometry Method Using Sample Support]
An example of a mass spectrometry method including an ionization method using the sample support 1A will be described with reference to FIGS. 9 to 12. FIG. In FIG. 9, (B) shows a cross-sectional view along line BB in (A).

First, the sample support 1A described above is prepared (first step). Subsequently, as shown in FIG. 9, the sample S1 to be measured is transferred onto the second surface 4b of the measurement region R1 of the ionization substrate 4 of the sample support 1A (second step). In the present embodiment, after the sample S1 is applied to the human skin, the sample support 1A is pressed against the skin with the second surface 4b of the measurement region R1 facing the sample S1, thereby increasing the measurement region R1. A sample S1 is transferred to the second surface 4b.

From the viewpoint of appropriately adhering and holding the sample S1 on the second surface 4b by the transfer, the sample S1 preferably has a relatively small amount of liquid components. For example, sample S1 may have a loss on drying (for example, a value measured by a loss on drying test defined as a general test method) below a predetermined threshold value (for example, 5%). Alternatively, the sample S1 may have no fluidity at body temperature (for example, about 35° C. to 37° C.). The presence or absence of fluidity at human skin temperature is determined, for example, by applying a sample (for example, a cosmetic) on the skin or on a glass plate, and then rubbing it with a finger or a cosmetic applicator. A determination can be made based on whether or not the sample can be moved smoothly. As an example, the presence or absence of fluidity at human skin temperature is determined by applying a sample to the skin or a glass plate so that the amount is 2 mg/cm ² , and applying a constant amount of the sample so that no lumps remain when the sample is rubbed with a finger. It can be judged based on whether it can be spread with a coating thickness of .

Subsequently, as shown in FIG. 10A, after the transfer of the sample S1 to the second surface 4b is completed, the protective film 6 is removed (peeled) from the ionization substrate 4, thereby removing the ionization substrate 4. Exposing the calibration region R2 (third step).

Subsequently, as shown in FIG. 10B, a calibration reagent S2 is dropped onto the second surface 4b of the calibration region R2 (fourth step). Here, since the calibration reagent S2 is a liquid (solution), part of the calibration reagent S2 dropped onto the calibration region R2 moves to the first surface 4a side of the calibration region R2 via the through hole 4c. can. On the other hand, another part of the calibration reagent S2 stays in the through-hole 4c and on the second surface 4b due to surface tension. A portion of the calibration reagent S2 remaining on the second surface 4b and in the through hole 4c near the opening on the second surface 4b side is ionized in the fifth step described later.

Subsequently, after the calibration reagent S2 dropped onto the calibration region R2 is dried, the second surface 4b of the calibration region R2 is irradiated with an energy beam while applying a voltage to the second surface 4b of the ionization substrate 4. to ionize the components of the calibration reagent S2 (fifth step). Further, by applying a voltage to the second surface 4b of the ionization substrate 4 and irradiating the second surface 4b of the measurement region R1 with energy rays, the components of the sample S1 are ionized (sixth step). FIG. 11 is a diagram schematically showing the fifth step, and FIG. 12 is a diagram schematically showing the sixth step.

As shown in FIGS. 11 and 12, the fifth and sixth steps described above can be performed by using the mass spectrometer 10. FIG. The mass spectrometer 10 includes a support section 12 , a sample stage 18 , a camera 16 , an irradiation section 13 , a voltage application section 14 , an ion detection section 15 and a control section 17 . The sample support 1A is placed on the support section 12 . At this time, the sample support 1A is in a state after the protective film 6 has been removed, the sample S1 has been transferred to the measurement region R1, and the calibration reagent S2 has been dropped onto the calibration region R2. The support section 12 is placed on the sample stage 18 . The irradiation unit 13 irradiates the second surface 4b of the sample support 1A with an energy beam such as a laser beam L. As shown in FIG. The voltage applying section 14 applies a voltage to the second surface 4b of the sample support 1A. The ion detection unit 15 detects an ionized component of the sample S1 (ion S1a) or an ionized component of the calibration reagent S2 (ion S2a). The camera 16 is a device that acquires a camera image including the irradiation position of the laser light L by the irradiation unit 13 . The camera 16 is, for example, a small CCD camera attached to the irradiation unit 13 .

The control unit 17 controls operations of the sample stage 18 , the camera 16 , the irradiation unit 13 , the voltage application unit 14 and the ion detection unit 15 . The control unit 17 is, for example, a computer device including a processor (eg, CPU, etc.), memory (eg, ROM, RAM, etc.), and the like.

A voltage is applied to the support surface 2a of the support substrate 2 (the conductive layer 7 formed on the support surface 2a in this embodiment) by the voltage application unit 14 . Thereby, a voltage is applied to the conductive layer 7 on the second surface 4b of the ionization substrate 4 via the supporting surface 2a and the conductive tape 5. FIG. Subsequently, the control unit 17 operates the irradiation unit 13 based on the image acquired by the camera 16 . Specifically, the control unit 17 determines whether the sample S1 or the calibration reagent S2 after drying specified based on the image acquired by the camera 16 in the laser irradiation range (for example, the measurement region R1 or the calibration region R2) is The irradiating section 13 is operated so that the second surface 4b within the existing area) is irradiated with the laser light L. As shown in FIG.

As an example, the control unit 17 moves the sample stage 18 and controls the irradiation operation (such as irradiation timing) of the laser light L by the irradiation unit 13 . That is, the control unit 17 causes the irradiation unit 13 to irradiate the laser light L after confirming that the sample stage 18 has moved by a predetermined distance. For example, the control unit 17 repeats movement (scanning) of the sample stage 18 and irradiation of the laser light L by the irradiation unit 13 so as to perform raster scanning within the laser irradiation range. The irradiation position on the first surface 4a may be changed by moving the irradiation section 13 instead of the sample stage 18, or by moving both the sample stage 18 and the irradiation section 13. good too.

In the fifth step, as shown in FIG. 11, the second surface 4b of the ionization substrate 4 (that is, the conductive layer 7 on the second surface 4b) is dried after the calibration reagent S2 dropped onto the calibration region R2 is dried. By irradiating the second surface 4b of the calibration region R2 with the laser beam L while applying a voltage to the second surface 4b of the calibration region R2, the components of the calibration reagent S2 are ionized and ions S2a are emitted. Specifically, energy is transmitted from the conductive layer 7 that has absorbed the energy of the laser light L to the components of the calibration reagent S2 on the second surface 4b, and the components that have acquired the energy are vaporized and acquire electric charges. , ions S2a. The emitted ions S2a move toward a ground electrode (not shown) provided between the sample support 1A and the ion detection section 15 while being accelerated. That is, the ions S2a move while being accelerated toward the ground electrode due to the potential difference generated between the conductive layer 7 to which the voltage is applied and the ground electrode. Then, the ion detection unit 15 detects the ions S2a. Calibration (mass calibration) is performed based on the detection result of the ions S2a by the ion detection unit 15 .

In the sixth step, as shown in FIG. 12, while applying a voltage to the second surface 4b of the ionization substrate 4 (that is, the conductive layer 7 on the second surface 4b), By irradiating the sample S1 with the laser light L, the components of the sample S1 are ionized and ions S1a are emitted. Specifically, energy is transmitted from the conductive layer 7 that has absorbed the energy of the laser beam L to the component of the sample S1 on the second surface 4b, and the component that has acquired the energy is vaporized and acquires electric charge, resulting in ions. S1a. The emitted ions S1a move toward a ground electrode (not shown) provided between the sample support 1A and the ion detection section 15 while being accelerated. That is, the ions S1a move while being accelerated toward the ground electrode due to the potential difference generated between the conductive layer 7 to which the voltage is applied and the ground electrode. Then, the ion detector 15 detects the ions S1a.

The detection result of the ions S1a by the ion detection unit 15 is associated with the irradiation position of the laser light L. FIG. Specifically, the ion detector 15 detects the ions S1a individually at each position within the laser irradiation range. Thereby, a distribution image (MS mapping data) representing the mass distribution of the sample S1 is obtained. Furthermore, it is possible to image the two-dimensional distribution of the molecules that make up the sample S1. That is, imaging mass spectrometry can be performed. The mass spectrometer 10 here is a mass spectrometer that uses time-of-flight mass spectrometry (TOF-MS).

[Action and effect of the first embodiment]
In the sample support 1A described above, the first surface 4a of the ionization substrate 4 is fixed to the support substrate 2 with the adhesive layer 3 interposed therebetween. As a result, the rigidity of the ionization substrate 4 can be increased, and damage to the ionization substrate 4 when the sample S1 is transferred to the measurement region R1 of the ionization substrate 4 can be suppressed. A calibration region R2 for calibration is provided separately from the measurement region R1, and a protective film 6 is provided to cover the entire calibration region R2 in a detachable manner. This can prevent part of the sample S1 from adhering to the calibration region R2 when the sample S1 is transferred to the measurement region R1 in the second step described above. As described above, according to the sample support 1A, the process of transferring the sample S1 to be ionized onto the surface (second surface 4b) of the ionization substrate 4 can be preferably carried out.

The adhesive layer 3 may be provided so as to overlap the entire measurement region R1 when viewed from the direction (Z-axis direction) perpendicular to the support surface 2a. According to the above configuration, the entire measurement region R1 of the ionization substrate 4 onto which the sample S1 is transferred can be tightly fixed to the support surface 2a by the adhesive layer 3. As shown in FIG. As a result, it is possible to effectively prevent the portion of the ionization substrate 4 corresponding to the measurement region R1 from being damaged when the sample S1 is transferred. More specifically, when the sample support 1A is pressed against the human skin to which the sample S1 is applied and then the sample support 1A is to be pulled away from the skin, the sample S1 left on the skin is In some cases, the portion corresponding to the measurement region R1 of the ionization substrate 4 is pulled toward the skin. In the sample support 1A, the portion corresponding to the measurement region R1 of the ionization substrate 4 is tightly fixed to the support surface 2a by the adhesive layer 3. As shown in FIG. Therefore, even if the tensile force as described above is generated, cracking of the portion corresponding to the measurement region R1 of the ionization substrate 4 can be preferably suppressed.

The ionized substrate 4 may be formed by anodizing a valve metal or silicon. The pores provided in the ionization substrate 4 may be through holes 4c that open to the first surface 4a and the second surface 4b. A conductive layer 7 may be provided on the second surface 4b so as not to block the through hole 4c. According to the above configuration, the ionized substrate 4 having a plurality of through-holes 4c (pores) can be easily formed by anodizing the valve metal or silicon. In addition, by providing the conductive layer 7 on the second surface 4b of the ionization substrate 4 so as not to block the through hole 4c, when ionizing the sample S1 by irradiating the laser beam L (energy ray), the ionization Appropriate application of voltage to the second surface 4b of the substrate 4 becomes possible.

The adhesive layer 3 may be provided so as not to overlap the calibration region R2 when viewed from the direction (Z-axis direction) orthogonal to the support surface 2a. Calibration using the calibration region R2 involves dropping a calibration reagent S2 onto the calibration region R2, and after the calibration reagent S2 has dried, the calibration region R2 is irradiated with a laser beam L (energy beam) for calibration. This is done by ionizing the components of the calibration reagent S2 and detecting the ionized components of the calibration reagent S2 (ie ions S2a). If the pores provided in the ionization substrate 4 are the through holes 4c and the adhesive layer 3 is provided so as to overlap the calibration region R2, the calibration reagent S2 dropped onto the calibration region R2 and the There is a risk of mixing with the components of the layer 3 (adhesive components, etc.). As a result, there is a possibility that calibration cannot be performed properly. More specifically, in the fifth step described above, not only the components of the calibration reagent S2 but also the components of the adhesive layer 3 are ionized, and the background noise contained in the detection result of the ion detector 15 (that is, the adhesive layer 3 (noise caused by the component of ) increases, which may make it difficult to calibrate with high accuracy. On the other hand, as in the present embodiment, by providing the adhesive layer 3 so as not to overlap the calibration region R2 (in other words, a region configured so that the ionized substrate 4 and the adhesive layer 3 do not overlap is set as the calibration region R2. ), it is possible to prevent the components of the adhesive layer 3 from being mixed into the calibration reagent S2, and it is possible to calibrate with high accuracy.

The sample support 1A is provided from the second surface 4b of the ionization substrate 4 to the portion of the support surface 2a that does not overlap the ionization substrate 4, and electrically connects the ionization substrate 4 and the support surface 2a. A connecting conductive tape 5 may be provided. Said part of the support surface 2a may be electrically conductive. In this embodiment, the entire support surface 2a is conductive. According to the above configuration, voltage is applied to the second surface 4b of the ionization substrate 4 via the conductive tape 5 by applying the voltage to the portion of the support surface 2a that does not overlap the ionization substrate 4. becomes possible.

In the present embodiment, as shown in FIG. 7, the conductive layer 7 includes a portion of the support surface 2a where the adhesive layer 3 is not provided, the adhesive layer 3, and the side surfaces of the ionization substrate 4 (portions exposed to the outside). ), and the second surface 4 b of the ionization substrate 4 are continuously formed. In this case, it is also possible to apply a voltage to the second surface 4b of the ionization substrate 4 via the conductive layer 7 by applying a voltage to the supporting surface 2a by the voltage applying section 14. FIG. Therefore, the conductive tape 5 is not necessarily essential in the sample support 1A. However, since the conductive layer 7 is very thin, a step portion (that is, a boundary portion between a portion of the support surface 2a where the adhesive layer 3 is not provided and the side surface of the adhesive layer 3 and the ionization substrate 4, or the adhesive layer 3 and the boundary portion between the side surface of the ionization substrate 4 and the second surface 4b of the ionization substrate 4), the conductive layer 7 may be broken. In this case, there is a possibility that a voltage cannot be appropriately applied from above the support surface 2a to above the second surface 4b of the ionization substrate 4 via the conductive layer 7 . Therefore, by providing the conductive tape 5 so as to cover the boundary portion, conduction from the support surface 2a to the second surface 4b of the ionization substrate 4 can be made more reliable. Further, when the conductive layer 7 is provided only on the second surface 4b of the ionization substrate 4 (that is, when the conductive layer 7 is not provided on the support surface 2a), the conductive tape 5 is provided on the support surface 2a. It plays a role of electrically connecting with the conductive layer 7 on the second surface 4b.

At least a portion of the support substrate 2 that overlaps the measurement region R1 may be transparent. In this embodiment, the entire support substrate 2 is transparent. The adhesive layer 3 may be made of a transparent material. Here, "transparent" means, for example, having transparency to light of a predetermined wavelength (for example, ultraviolet light, visible light, or the like). In this embodiment, the adhesive layer 3 is a transparent double-sided tape. According to the above configuration, since the members (a part of the support substrate 2 and the adhesive layer 3) overlapping the measurement region R1 on the side of the first surface 4a of the ionization substrate 4 are transparent, they are transferred to the second surface 4b of the measurement region R1. It is possible to obtain a light transmission image (image observed by applying illumination light from the surface of the support substrate 2 opposite to the ionization substrate 4 side) of the sample S1.

In addition, in the ionization method using the sample support 1A (first to sixth steps described above), the first surface 4a of the ionization substrate 4 is firmly fixed to the support substrate 2 via the adhesive layer 3. , damage to the ionization substrate 4 (especially the portion corresponding to the measurement region R1) in the second step (during transfer of the sample S1) can be suppressed. The ionization substrate 4 is also provided with a protective film 6 that covers the entire calibration area R2 in a detachable manner. This can prevent part of the sample S1 from adhering to the calibration region R2 when the sample S1 is transferred to the measurement region R1 in the second step. As described above, according to the above-described ionization method, the process of transferring the sample S1 to be ionized onto the surface (second surface 4b) of the ionization substrate 4 can be preferably performed.

In this embodiment, in the second step, the sample S1 is applied onto the human skin, and the sample support 1A is pressed against the skin with the second surface 4b of the measurement region R1 facing the sample S1. A sample S1 is transferred to the second surface 4b of the measurement region R1. Since the first surface 4a of the ionization substrate 4 is firmly fixed to the support substrate 2 via the adhesive layer 3, a supporting member (for example, There is no need to provide a frame or the like provided with an opening for exposing the measurement region R1 as disclosed in Patent Document 1). Therefore, the process of pressing the sample support 1A against the human skin and transferring the sample S1 on the skin to the measurement region R1 can be performed smoothly. More specifically, if the support member (frame) as described above is provided on the second surface 4b side of the ionization substrate 4, during the transfer of the sample S1, the second surface 4b of the measurement region R1 is larger than the second surface 4b. The support member comes into contact with the skin first. Therefore, when the support member as described above is provided, the sample support 1A must be strongly pressed against the skin by the thickness of the support member. Moreover, if the support member is made of a metal material (for example, Ni (nickel), etc.), contact of the support member with the skin may cause a metal allergy. The sample support 1A, in which the ionization substrate 4 is fixed to the support substrate 2 via the adhesive layer 3, avoids the above problems when transferring the sample applied to the human skin. be able to.

[Second embodiment]
A sample support 1B according to the second embodiment will be described with reference to FIGS. 13 and 14. FIG. The sample support 1B differs from the sample support 1A in that a conductive tape 5C is provided instead of the conductive tape 5A. Other configurations of the sample support 1B are the same as those of the sample support 1A.

A conductive tape 5C is provided to cover the calibration region R2. That is, the conductive tape 5C is provided so as to cover the opening 3a (second opening) of the adhesive layer 3. As shown in FIG. Further, the conductive tape 5C is provided with an opening 5a (first opening) including the calibration region R2 (opening 3a) when viewed from the direction (Z-axis direction) perpendicular to the support surface 2a. It is In this embodiment, as shown in FIGS. 13 and 14, the conductive tape 5C is provided with three openings 5a corresponding to each of the three calibration regions R2 (openings 3a). . Each opening 5a has a circular shape that is slightly larger than the opening 3a when viewed in the Z-axis direction. As a result, the edge of the opening 5a and the edge of the opening 3a are separated from each other when viewed from the direction perpendicular to the support surface 2a (the Z-axis direction). More specifically, in a set of openings 5a and 3a corresponding to each other, the center position of the opening 5a and the center position of the opening 3a are aligned, and the width (diameter) of the opening 5a is equal to the opening. It is larger than the width (diameter) of the portion 3a.

In the sample support 1B, the protective film 6 is provided on the surface of the conductive tape 5C opposite to the ionization substrate 4 side so as to cover the opening 5a. That is, the protective film 6 is provided on the ionization substrate 4 via the conductive tape 5C and is not in contact with the ionization substrate 4 (and the conductive layer 7 on the second surface 4b of the ionization substrate 4).

As described above, in the sample support 1B, the conductive tape 5C is provided with the opening 5a containing the calibration region R2 when viewed from the direction perpendicular to the support surface 2a (the Z-axis direction). The protective film 6 is provided on the surface of the conductive tape 5C opposite to the ionization substrate 4 side so as to cover the opening 5a. According to the above configuration, by interposing the conductive tape 5C between the protective film 6 and the ionized substrate 4, a part of the protective film 6 (for example, the adhesive surface having slight adhesiveness) becomes the second ionized substrate of the ionized substrate 4. Contact with the surface 4b (strictly, the conductive layer 7 provided on the second surface 4b) can be prevented. As a result, when the protective film 6 is removed in the third step described above, it is possible to appropriately prevent the components of the adhesive surface of the protective film 6 from adhering to the calibration region R2. Moreover, when the protective film 6 is removed, part of the ionization substrate 4 (including part of the conductive layer 7 on the second surface 4b) is prevented from peeling off together with the protective film 6 . As described above, the calibration using the calibration region R2 can be performed with higher accuracy.

The pores provided in the ionization substrate 4 are through holes 4c that open to the first surface 4a and the second surface 4b. When viewed, there is provided an opening 3a corresponding to the calibration region R2, the opening 5a of the conductive tape 5C being larger than the opening 3a of the adhesive layer 3, and the edge of the opening 5a and the opening. The edges of 3a are separated from each other when viewed in the Z-axis direction. According to the above configuration, since the edge of the opening 5a is separated from the edge of the opening 3a, it is possible to prevent a part of the conductive tape 5C from adhering to the calibration region R2 of the ionization substrate 4. can. Furthermore, by using the opening 5a, which is larger than the opening 3a (that is, the opening corresponding to the calibration region R2) as a mark, the operator who carries out the calibration can easily grasp the position of the calibration region R2.

[Third embodiment]
A sample support 1C according to the third embodiment will be described with reference to FIG. The sample support 1C differs from the sample support 1A in that the conductive tape 5 is not provided. Other configurations of the sample support 1C are the same as those of the sample support 1A. In the sample support 1C, the protective film 6 is provided directly on the second surface 4b of the ionization substrate 4 (in this embodiment, on the conductive layer 7 provided on the second surface 4b) without interposing the conductive tape. It is

When the sample S1 is ionized using the sample support 1C, the above-described ionization method (first to sixth steps) can be modified as follows. That is, in the ionization method using the sample support 1C, the support surface 2a is removed from the second surface 4b of the ionization substrate 4 after the second step and before the fifth and sixth steps. A step of fixing a conductive tape 5 (see FIGS. 11 and 12) so as to cover the portion that does not overlap with the ionization substrate 4, and electrically connecting the ionization substrate 4 and the above-mentioned portion of the support surface 2a. obtain. That is, the sample support 1C does not have the conductive tape 5 in the state before the protective film 6 is removed, and before performing the fifth step and the sixth step (after the protective film 6 is removed) In addition, a conductive tape 5 can be provided on the sample support 1C, if desired. In this case, in each of the fifth step and the sixth step, by applying a voltage to the portion of the support surface 2a (in this embodiment, an arbitrary portion of the support surface 2a that does not overlap with the ionization substrate 4), the conductive A voltage can be applied to the second surface 4 b of the ionizing substrate 4 via the conductive tape 5 and the conductive layer 7 . On the other hand, when the voltage can be stably applied from the support surface 2a to the second surface 4b of the ionization substrate 4 via only the conductive layer 7, the step of providing the conductive tape 5 can be omitted. may

In the ionization method using the sample support 1C described above, since part of the second surface 4b of the ionization substrate 4 is not covered with the conductive tape 5 in the second step, the transfer of the sample S1 onto the measurement region R1 is prevented. can be performed more smoothly. That is, since the conductive tape 5 is not provided, the second surface 4b of the measurement region R1 can be brought into closer contact with the sample applied to the human skin. Further, by providing the conductive tape 5 as necessary before the fifth step and the sixth step, it becomes possible to apply the voltage to the second surface 4b of the ionization substrate 4 more stably.

[Fourth Embodiment]
A sample support 1D according to the fourth embodiment will be described with reference to FIG. The sample support 1D is different from the sample support 1A in that the conductive tape 5D is replaced with the conductive tape 5D and the protective film 6 is replaced with the protective film 6D. Other configurations of the sample support 1D are the same as those of the sample support 1A.

The conductive tape 5D is formed in a frame shape surrounding the measurement area R1 when viewed from the Z-axis direction. As shown in FIG. 16, as an example, the conductive tape 5D is a rectangular portion extending in the X-axis direction between the ends in the Y-axis direction of the conductive tapes 5B and 5C of the second embodiment. It has a shape as if it were connected through The conductive tape 5D is provided with an opening 5b for exposing the measurement region R1. In this embodiment, as an example, the opening 5b is formed in a square shape when viewed from the Z-axis direction. Further, the conductive tape 5D is provided with an opening 5a for exposing the calibration region R2 (opening 3a), similarly to the conductive tape 5C of the second embodiment.

The protective film 6D is formed in a frame shape surrounding the measurement region R1 when viewed from the Z-axis direction. As shown in FIG. 16, as an example, the protective film 6D is formed in a rectangular shape having long sides and short sides approximately the same length as the support substrate 2 when viewed in the Z-axis direction. That is, the long sides and short sides of the protective film 6D are along the long sides and short sides of the support substrate 2 . Further, an opening 6a for exposing the measurement region R1 to the outside is provided at a substantially central portion of the protective film 6D. The opening 6a overlaps the opening 5b in the Z-axis direction. That is, the opening 6a communicates with the opening 5b. In this embodiment, as an example, the opening 6a is formed in the same rectangular shape as the opening 5b.

The surface of the conductive tape 5D on the protective film 6D side is configured as an adhesive surface. The protective film 6D is attached to the protective film 6D side surface (adhesive surface) of the conductive tape 5D.

According to the above configuration, it is possible to suitably apply a voltage to the second surface 4b of the ionization substrate 4 by means of the frame-shaped conductive tape 5D. More specifically, since the conductive tape 5D is provided over the entire circumference of the measurement area R1, electrical continuity from the conductive tape 5D to the measurement area R1 can be made more reliable. In addition, since the protective film 6D exists around the measurement region R1 when viewed from the Z-axis direction, handling of the sample support 1D during sample transfer or the like can be improved. In the example of FIG. 16, the protective film 6D is provided over substantially the entire area of the support surface 2a of the support substrate 2 that does not overlap the measurement region R1. A metal such as aluminum may be vapor-deposited on a portion of the support surface 2a that does not overlap with the measurement region R1 in order to impart conductivity to the support surface 2a. In such a case, the metal-deposited portion can be suitably protected by the protective film 6D. In addition, the operator can work while grasping the portion where the protective film 6D is formed when aligning the sample support 1D at the time of sample transfer or the like. Here, when the protective films 6D are formed on both sides of the measurement region R1 as in the above configuration, both sides of the measurement region R1 (that is, the portions where the protective films 6D are formed) are grasped with both hands to support the sample. Since the body 1D can be held, it is possible to easily perform the work such as the positioning.

[Modification]
Although several embodiments of the present disclosure have been described above, the present disclosure is not limited to the above embodiments. The material and shape of each configuration are not limited to the materials and shapes described above, and various materials and shapes can be adopted. Also, a part of the configuration of one embodiment or modification described above can be arbitrarily applied to the configuration of another embodiment or modification.

The conductive layer 7 may be provided at least on the second surface 4 b of the ionization substrate 4 . That is, as described above, the conductive layer 7 does not have to be provided on the support surface 2a. Also, the conductive layer 7 may be provided on, for example, the first surface 4a in addition to the second surface 4b, or may be provided on all or part of the inner surface of each through hole 4c.

The ionization substrate 4 may have conductivity. For example, the ionization substrate 4 may be made of a conductive material such as a semiconductor. In this case, the conductive layer 7 for applying voltage to the second surface 4b side of the ionization substrate 4 may be omitted. However, even if the ionization substrate 4 has conductivity, the conductive layer 7 may be provided in order to suitably apply a voltage to the second surface 4b side of the ionization substrate 4 .

The pores provided in the ionization substrate 4 are not limited to the through holes 4c described above. The pores provided in the ionization substrate 4 may be non-through holes (recesses) that open at least to the second surface 4b. That is, the ionization substrate 4 may be a substrate having a second surface 4b on which a fine uneven structure is formed. If the ionized substrate 4 does not have the through holes 4c, the components of the adhesive layer 3 arranged on the first surface 4a side of the ionized substrate 4 do not move to the second surface 4b side. Therefore, in this case, the adhesive layer 3 may be provided in a region overlapping the calibration region R2 when viewed from the Z-axis direction. That is, the opening 3a of the adhesive layer 3 in the above embodiment can be omitted.

In the above embodiment, an ITO slide glass that is entirely transparent is used as the support substrate 2, but the support substrate 2 is not limited to the above. For example, in order to observe a light transmission image of the sample S1, at least a portion of the support substrate 2 that overlaps the measurement region R1 should be transparent, and other portions of the support substrate 2 may be non-transparent. . For example, when a non-conductive slide glass having no conductive film such as an ITO film is used as the support substrate 2, the measurement area on the surface of the slide glass is measured in order to make the support surface 2a conductive. A metal such as aluminum may be vapor-deposited on the portion that does not overlap with R1 (for example, the portion that does not overlap with the entire adhesion layer 3 and ionization substrate 4).

When observation of the light transmission image of the sample S1 is unnecessary, the portion of the support substrate 2 that overlaps the measurement region R1 may be non-transparent. In this case, a conductive non-transparent substrate such as a metal plate can be used as the support substrate 2 . Moreover, when observation of a light transmission image is unnecessary, the adhesive layer 3 may also be formed of a non-transparent material.

The number of calibration regions R2 provided on the ionization substrate 4 is not limited to the above embodiment (three). The number of calibration regions R2 may be one, two, or four or more.

A part of the surface of the protective film 6 on the ionization substrate 4 side may be configured as an adhesive surface. For example, in order to prevent part of the ionization substrate 4 (including the conductive layer 7 on the second surface 4b) from peeling off together with the protective film 6 when the protective film 6 is removed, the above-mentioned A portion of the surface that directly faces the second surface 4b of the ionization substrate 4 (or the conductive layer 7 on the second surface 4b) may be configured not to have adhesive properties. For example, only the portion of the surface of the protective film 6 that does not overlap with the ionization substrate 4 may be configured as an adhesive surface. Alternatively, the entire surface of the protective film 6 on the ionization substrate 4 side may be configured as a non-adhesive surface. In this case, at least the surface of the conductive tape 5 that overlaps the protective film 6 and faces the protective film 6 may be configured as an adhesive surface.

In the mass spectrometry method using the sample support, the object to which the voltage is applied by the voltage applying section 14 is not limited to the support surface 2a of the support substrate 2. FIG. For example, the voltage may be applied to a member other than the support substrate 2 (a member electrically connected to the support surface 2 a of the support substrate 2 ), or may be applied directly to the conductive tape 5 .

In the mass spectrometry method using a sample support, the mass spectrometer 10 may be a scanning mass spectrometer or a projection mass spectrometer. In the case of the scanning type, a signal of one pixel having a size corresponding to the spot diameter of the laser light L is obtained for each irradiation of the laser light L by the irradiation unit 13 . That is, scanning (changing the irradiation position) and irradiation of the laser light L are performed for each pixel. On the other hand, in the case of the projection type, a signal of an image (a plurality of pixels) corresponding to the spot diameter of the laser light L is obtained for each irradiation of the laser light L by the irradiation unit 13 . In the case of the projection type, when the spot diameter of the laser light L includes the entire measurement region R1, the imaging mass spectrometry can be performed by irradiating the laser light L once. In the case of the projection type, when the spot diameter of the laser light L does not include the entire measurement region R1, the signal of the entire measurement region R1 is obtained by scanning and irradiating the laser light L in the same manner as in the scanning type. can be obtained. In addition, the ionization method described above can also be used for other measurements and experiments such as ion mobility measurements.

The application of the sample support is not limited to ionization of a sample by irradiation with the laser light L. The sample support can be used for ionizing a sample by irradiation with energy beams such as laser light, electrospray, ion beam, and electron beam. In the ionization method and mass spectrometry method described above, the sample can be ionized by such energy beam irradiation.

1A, 1B, 1C, 1D... sample support, 2... supporting substrate, 2a... supporting surface, 3... adhesive layer, 3a... opening (second opening), 4... ionization substrate, 4a... first surface, 4b Second surface 4c Through hole 5, 5A, 5B, 5C, 5D Conductive tape 5a Opening (first opening) 6, 6D Protective film (cover member) 7 Conductive layer , R1... measurement area, R2... calibration area, S1... sample, S2... calibration reagent.

Claims (12)

a support substrate having a support surface;
a sheet-like adhesive layer provided on the support surface;
An ionizable substrate provided on the support surface via the adhesive layer, the ionized substrate having a first surface facing the adhesive layer, a second surface opposite the first surface, and at least the second surface. a plurality of pores opening into the ionized substrate;
a detachable sheet-like cover member covering a portion of the second surface of the ionization substrate;
with
The ionization substrate has a measurement area for transferring a sample to be measured and a calibration area for calibration,
The cover member is provided so as to cover at least the entire calibration area.
sample support. The adhesive layer is provided so as to overlap the entire measurement area when viewed from a direction orthogonal to the support surface.
A sample support according to claim 1 . The ionized substrate is formed by anodizing a valve metal or silicon,
The pores are through holes that open to the first surface and the second surface,
A conductive layer is provided on the second surface so as not to block the pores,
3. A sample support according to claim 1 or 2. The pores are through holes that open to the first surface and the second surface,
The adhesive layer is provided so as not to overlap the calibration region when viewed in a direction orthogonal to the support surface.
A sample support according to any one of claims 1-3. a conductive tape provided from the second surface of the ionized substrate to a portion of the support surface that does not overlap with the ionized substrate and electrically connecting the ionized substrate and the portion of the support surface; further prepared,
said portion of said support surface being electrically conductive;
A sample support according to any one of claims 1-4. The conductive tape is provided with a first opening containing the calibration area when viewed from a direction orthogonal to the support surface,
The cover member is provided on a surface of the conductive tape opposite to the ionized substrate side so as to cover the first opening.
A sample support according to claim 5 . The pores are through holes that open to the first surface and the second surface,
The adhesive layer is provided with a second opening corresponding to the calibration area when viewed in a direction orthogonal to the support surface,
The first opening is larger than the second opening,
an edge of the first opening and an edge of the second opening are separated from each other when viewed in a direction orthogonal to the support surface;
A sample support according to claim 6 . The conductive tape and the cover member are formed in a frame shape surrounding the measurement area when viewed from a direction orthogonal to the support surface,
A sample support according to any one of claims 5-7. At least a portion of the support substrate that overlaps with the measurement region is transparent,
The adhesive layer is made of a transparent material,
A sample support according to any one of claims 1-8. a first step of providing a sample support according to claim 1;
a second step of transferring a sample onto the second surface of the measurement region of the ionization substrate;
a third step of exposing the calibration region of the ionization substrate by removing the cover member after the transfer of the sample to the second surface is completed;
a fourth step of dropping a calibration reagent onto the second surface of the calibration area;
After the calibration reagent dropped onto the calibration region is dried, the second surface of the calibration region is irradiated with an energy beam while applying a voltage to the second surface of the ionization substrate. a fifth step of ionizing the components of the calibration reagent;
a sixth step of ionizing the components of the sample by irradiating the second surface of the measurement region with an energy beam while applying a voltage to the second surface of the ionization substrate;
A method of ionization, comprising: In the second step, the sample is applied onto the skin of a person, and the sample support is pressed against the skin with the second surface of the measurement region facing the sample. transferring the sample to the second surface of
The ionization method according to claim 10. After the second step and before the fifth step and the sixth step, from the second surface of the ionized substrate to a portion of the support surface that does not overlap the ionized substrate. securing a conductive tape across the substrate to electrically connect the ionizable substrate and the portion of the support surface;
in each of the fifth step and the sixth step, applying a voltage to the second surface of the ionized substrate through the conductive tape by applying a voltage to the portion of the support surface;
The ionization method according to claim 10 or 11.

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