JP6285735B2 - Mass spectrometer - Google Patents

Description

  The present invention relates to a mass spectrometer.

  A time-of-flight mass spectrometer (TOFMS) accelerates and flies ions by applying a certain amount of energy and calculates the mass-to-charge ratio of ions (m / m) from the time required to reach the detector. This is a mass spectrometer for obtaining z).

In TOFMS, accelerating ions at a constant accelerating voltage _{V a.} At this time, the ion velocity v is expressed as follows from the energy conservation law.

  Here, m is the mass of the ion, z is the valence of the ion, and e is the elementary charge.

  Therefore, the flight time T until reaching the detector placed after a certain distance L is expressed by the following equation (3).

  As can be seen from equation (3), the ions can be separated by the m / z value by utilizing the fact that the time of flight T varies depending on the mass m of the ions.

  The mass resolution of TOFMS is defined by the following equation (4), where T is the total flight time and ΔT is the peak width.

  As can be seen from equation (4), if the peak width ΔT is kept constant and the total flight time T can be extended, the mass resolution can be improved.

The simplest ion optical system that performs mass separation is a linear TOFMS that linearly flies ions accelerated by an ion source. A reflective TOFMS that can extend the flight distance by placing a reflection field between the ion source and the detector is also widely used.

  In the linear and reflective time-of-flight mass spectrometers, increasing the total flight time T, that is, extending the total flight distance directly leads to an increase in the size of the apparatus. A multi-circular TOFMS (Non-Patent Document 1) is an apparatus that has been developed to avoid an increase in the size of the apparatus and achieve high mass resolution. In the multi-turn type TOFMS, four toroidal electric fields combining Mazda plates with a cylindrical electric field are used, and the total flight time T is extended by making multiple rounds of the figure-shaped round orbit. Further, in the multi-turn type TOFMS, the spatial spread and the temporal spread on the detection surface due to the initial position, the initial angle, and the initial kinetic energy are successfully converged to the first order term.

  In a time-of-flight mass spectrometer, it is necessary to convert the measured time-of-flight T to m / z. The above equation (3) represents the simplest relationship between the flight time T and m / z, but in practice, it is often converted by a polynomial. The following equation (5) is a case where the relationship between the flight time T and m / z is expressed by a quadratic equation.

  a, b, and c are constants, and can be determined, for example, by measuring three or more compounds with known m / z.

One of the ion sources of TOFMS is a laser desorption / ionization (LDI) method in which a sample applied to a plate or a solid sample itself is irradiated with a laser to ionize a measurement target compound. The laser desorption ionization method may have poor ionization efficiency depending on the measurement target. For this reason, the sample is mixed and dissolved in a matrix (liquid, crystalline compound, metal powder, etc.) that has an absorption band at the wavelength of the laser beam to be used to promote ionization, and the sample is vaporized by laser irradiation. Matrix-assisted laser desorption / ionization (MALDI) for ionization is widely used. In recent years, in order to promote ionization, a surface-assisted laser desorption ionization method (Surface Assisted Laser) in which nanostructures are formed on a plate.
Research on Desorption / Ionization, SALDI) is also underway.

  A technique for obtaining information on the two-dimensional position information and the mass and abundance of a compound contained in each position using such a mass spectrometer is called mass imaging (MSI) (see Non-Patent Document 2). ). In mass imaging, as ionization methods, for example, those using laser desorption ionization centered on MALDI described above, or those using secondary ion mass spectrometry (Secondary Ion Mass Spectrometry, SIMS) using high-speed particles There is. In mass imaging, for example, a stage on which a sample is arranged is scanned two-dimensionally in the X and Y axis directions, and a mass spectrum is measured. The data information is (X, Y, m / z, ion intensity). During or after data acquisition, information on intensity distribution for a specific m / z (that is, a specific compound) and information on a compound localized in a specific region can be obtained.

In mass imaging, the mass axis may drift due to a systematic error due to a relatively long measurement time, a tilt of the sample surface, unevenness of the sample surface (particularly matrix crystal), and the like.
The mass axis drift means that an error occurs when the m / z value is calculated from the flight time T.

  As a method for calibrating the mass axis, for example, in Patent Document 1, a compound (internal standard substance) having a known mass-to-charge ratio is mass-analyzed together with a target sample to obtain an actually-measured mass-to-charge ratio. A method is disclosed in which calibration information is generated by comparing and, and mass calibration is performed based on the calibration information.

JP 2010-205460 A

M.M. Toyoda, D.A. Okumura, M .; Ishihara and I. Katakuse, J. et al. Mass Spectrom, 2003, 38, 1125-1142. R. M.M. Caprioli, T .; B. Farmer, and J.M. Gile, Anal. Chem. , 69, 4751 (1997)

  Here, in mass imaging, since the amount of ions generated in one measurement region (measurement point) cannot be increased so much, it is difficult to increase the SN ratio of each mass spectrum constituting the mass image. For this reason, even if an attempt is made to correct the drift of the mass axis of each mass spectrum constituting the mass imaging, there is a case where the correction cannot be performed or the corrected mass accuracy cannot be improved.

  The present invention has been made in view of the above problems, and one of the objects according to some aspects of the present invention is to provide a mass spectrometer capable of improving mass accuracy. is there.

(1) A mass spectrometer according to the present invention comprises:
A measurement unit that ionizes a sample for each measurement region and performs mass spectrometry for each measurement region;
Based on the measurement result of the measurement unit, a spectrum generation unit that generates a mass spectrum for each measurement region;
At least two of the measurement regions are defined as one block, and the mass spectrum of the measurement region is integrated for each block, and the mass-to-charge ratio of the mass spectrum of the measurement region is calculated for each block based on the integrated mass spectrum. A mass correction unit to be corrected;
Only including,
The measurement unit measures all the measurement areas included in one block, and then measures the measurement area included in the next block .

In such a mass spectrometer, for example, the SN ratio of the spectrum can be improved as compared with the case where the mass-to-charge ratio is corrected based on the mass spectrum of one measurement region.
Therefore, in such a mass spectrometer, the drift of the mass axis of the mass spectrum can be corrected with high accuracy, and the mass accuracy can be improved. Further, in such a mass spectrometer, for example, the influence of systematic errors due to the passage of time can be reduced, and the mass accuracy can be improved.

(2) A mass spectrometer according to the present invention comprises:
A measurement unit that ionizes a sample for each measurement region and performs mass spectrometry for each measurement region;
Based on the measurement result of the measurement unit, a spectrum generation unit that generates a mass spectrum for each measurement region;
At least two of the measurement regions are defined as one block, and the mass spectrum of the measurement region is integrated for each block, and the mass-to-charge ratio of the mass spectrum of the measurement region is calculated for each block based on the integrated mass spectrum. A mass correction unit to be corrected;
Including
In the mass spectrum integrated for each block, when a peak exists in the first range of the mass-to-charge ratio, the mass correction unit calculates the mass-to-charge ratio using the peak that exists in the first range. If there is no peak in the first range, the mass-to-charge ratio is corrected using a peak that exists in a second range having a mass-to-charge ratio different from the first range .

  In such a mass spectrometer, for example, the SN ratio of the spectrum can be improved as compared with the case where the mass-to-charge ratio is corrected based on the mass spectrum of one measurement region. Therefore, in such a mass spectrometer, the drift of the mass axis of the mass spectrum can be corrected with high accuracy, and the mass accuracy can be improved. In such a mass spectrometer, the number of correction blocks that cannot correct the mass-to-charge ratio can be reduced.

(3) In the mass spectrometer according to the present invention,
In the mass spectrum integrated for each block, when a peak exists in the first range of the mass-to-charge ratio, the mass correction unit calculates the mass-to-charge ratio using the peak that exists in the first range. If there is no peak in the first range, the mass-to-charge ratio may be corrected using a peak that exists in a second range having a mass-to-charge ratio different from the first range.

  In such a mass spectrometer, the number of correction blocks that cannot correct the mass-to-charge ratio can be reduced.

(4) In the mass spectrometer according to the present invention,
The mass correcting unit may correct the mass to charge ratio by obtaining a calibration condition in order to convert the time of flight into the mass to charge ratio.

  In such a mass spectrometer, mass accuracy can be improved.

(5) In the mass spectrometer according to the present invention,
For the mass spectrum corrected under the different calibration conditions, including a data processing unit that aligns the data interval of each mass spectrum,
The data processing unit may align the data interval of each mass spectrum with the data interval of any one of the corrected mass spectra.

  In such a mass spectrometer, a mass spectrum with a uniform data interval can be handled in the same manner as a measured mass spectrum. That is, for example, the mass-to-charge ratio axis can be converted to the time-of-flight axis for mass spectra with the same data interval.

(6) In the mass spectrometer according to the present invention,
The measuring unit is
An ion source that ionizes the measurement region by irradiating a laser beam;
A mass analyzer that detects and mass-separates ions generated in the ion source;
May be included.

(7) In the mass spectrometer according to the present invention,
The mass spectrometer may separate ions generated by the ion source based on time of flight.

(8) In the mass spectrometer according to the present invention,
A mass imaging unit that acquires position information of the measurement region and mass information of the compound included in each measurement region from the mass spectrum of each measurement region corrected by the mass correction unit may be included.

  In such a mass spectrometer, for example, it is possible to obtain information on the intensity distribution of a specific compound and information on a compound localized in a specific region.

The figure which shows the structure of the mass spectrometer which concerns on 1st Embodiment. The figure which shows typically the state which divided | segmented the area | region used as the measurement symmetry of a sample for every measurement area | region. The figure which shows typically the ion source of the mass spectrometer which concerns on 1st Embodiment. The figure which shows an example of a structure of the mass spectrometer part of the mass spectrometer which concerns on 1st Embodiment. The figure for demonstrating a correction block. The figure for demonstrating an example of the process which arranges the data interval of a mass spectrum. The figure for demonstrating an example of the process which arranges the data interval of a mass spectrum. The figure for demonstrating the measurement order of the measurement part of the mass spectrometer which concerns on 3rd Embodiment. The figure which shows an example of a structure of the measurement part of the mass spectrometer which concerns on 5th Embodiment.

  DESCRIPTION OF EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the drawings. The embodiments described below do not unduly limit the contents of the present invention described in the claims. In addition, not all of the configurations described below are essential constituent requirements of the present invention.

1. First Embodiment First, a mass spectrometer according to a first embodiment will be described with reference to the drawings. FIG. 1 is a diagram illustrating a configuration of a mass spectrometer (MALDI-TOFMS) 100 according to the first embodiment.

  As shown in FIG. 1, the mass spectrometer 100 includes a measurement unit 10, a processing unit 20, an operation unit 30, a display unit 32, a storage unit 34, and an information storage medium 36.

The measurement unit 10 ionizes the sample S for each measurement region, and performs mass spectrometry for each measurement region. FIG. 2 is a diagram schematically illustrating a state in which a region that is symmetrical to the measurement of the sample S is divided for each measurement region A. FIG. As shown in FIG. 2, the measurement unit 10 divides a region to be measured into a plurality of measurement regions A and performs mass spectrometry. The number of measurement areas A is not particularly limited and can be set arbitrarily. In the illustrated example, the position of the measurement area A is represented as a two-dimensional orthogonal coordinate system (measurement area A (X, Y)). By measuring unit 10 performs measurement for each measurement region A, 1 single mass spectrum M _A for one measurement region A as described later can be obtained.

For example, the measurement unit 10 starts measurement from the measurement region A (0, 0), and 1 in the order of the measurement region A (1, 0), (2, 0), ..., (n _x , 0). When the measurement of the row is performed, the measurement of the second row is performed in the order of the measurement areas A (0, 1), (1, 1), ..., (n _x , 1). The measurement unit 10 performs the same measurement for the second and subsequent rows, and performs measurement up to the measurement region A (n _x , n _y ). Note that _nx and _ny are positive integers.

  As shown in FIG. 1, the measurement unit 10 includes an ion source 10a and a mass analysis unit 10b.

  The ion source 10a ionizes the sample by a predetermined method, and accelerates the generated ions with a constant pulse voltage. In the present embodiment, the ion source 10a ionizes the sample S using a matrix-assisted laser desorption ionization (MALDI) method.

  FIG. 3 is a diagram schematically showing the ion source 10 a of the mass spectrometer 100. As shown in FIG. 3, the ion source 10 a includes a sample stage 12, an ion generation unit 14, and an acceleration unit 16.

A sample plate (target plate) 2 on which a sample S is arranged is placed on the sample stage 12. The sample S is sprayed with a matrix (metal fine particles, organic compounds, etc.) for promoting ionization as necessary. Although not shown, the sample stage 12 is moved in two axial directions XY (in-plane direction of the sample stage 12) by a stage driving unit including a motor and the like.

  The ion generator 14 ionizes the sample S by irradiating the laser beam L. The ion generator 14 includes a laser light source 141 that generates the laser light L, a lens 142 that focuses the laser light L, a mirror 143 that reflects the laser light L, a mirror 144 that reflects an image from the sample S, and a mirror A lens 145 that focuses the image reflected by the lens 144 and a CCD camera 146 that acquires an image incident through the lens 145 are configured.

  In the ion generation unit 14, the laser beam L emitted from the laser light source 141 is irradiated to a predetermined measurement region A of the sample S through the lens 142 and the mirror 143, and the sample S is vaporized or ionized. Since the irradiation position of the laser beam L is fixed, when the sample stage 12 is moved by the stage driving unit, the irradiation region of the laser beam L on the sample S is moved accordingly. The state of the measurement region A of the sample S can be observed by the mirror 144, the lens 145, and the CCD camera 146.

  The acceleration unit 16 includes a first acceleration electrode 162 and a second acceleration electrode 164. By applying a predetermined voltage (pulse voltage) to the acceleration electrodes 162 and 164, the ions I generated by the ion generator 14 are accelerated and introduced into the mass analyzer 10b. In the mass spectrometer 100, a delayed extraction method in which a pulse voltage is applied to the acceleration electrodes 162 and 164 after a predetermined time from the irradiation of the laser light L may be used. By using the delayed extraction method, the initial energy at the time of ion generation can be converged with time, and the mass resolution can be improved.

  The mass analyzer 10b detects and separates the ions I generated by the ion source 10a by mass separation. The mass spectrometer 10b separates the ions I generated by the ion source 10a based on the flight time T. That is, the mass spectrometer 10b performs mass analysis by time-of-flight mass spectrometry.

  FIG. 4 is a diagram illustrating an example of the configuration of the mass spectrometer 10b of the mass spectrometer 100.

  As shown in FIG. 4, the mass spectrometer 10 b is a reflective TOFMS that includes a reflectron 17 and a detector 18.

  The reflectron 17 is disposed in the path of the ions I between the ion source 10a and the detector 18. The ions I generated by the ion source 10a are incident on the reflectron 17. The reflectron 17 can reverse the flight direction of the ions I using an electrostatic field (reflection field). By placing the reflectron 17 between the ion source 10a and the detector 18, the energy convergence can be improved and the flight distance can be extended.

  In the mass analyzer 10b, mass separation is performed by using the fact that the flight time T varies depending on the m / z value according to the above equation (3). The ions I generated by the ion source 10a are mass-separated according to the m / z value until they reach the detector 18 through the reflectron 17.

  The detector 18 detects the ion I and outputs a detection signal corresponding to the amount (intensity) of the incident ion I. The detection signal of the detector 18 is output to the processing unit 20.

The processing unit 20 performs various types of calculation processing according to programs stored in the storage unit 34. The processing unit 20 functions as a spectrum generation unit 220, a mass correction unit 222, a data processing unit 224, and a mass imaging unit 226, which will be described below, by executing a program stored in the storage unit 34. The function of the processing unit 20 includes various processors (CPU, DSP, etc.
Etc.), hardware such as an ASIC (gate array, etc.), and a program. Note that at least a part of the processing unit 20 may be realized by hardware (dedicated circuit).

Spectrum generation unit 220, based on the measurement result of the measuring unit 10, to generate a mass spectrum M _A for each measurement region A. Hereinafter, the processing of the spectrum generation unit 220 will be described in detail.

  First, the spectrum generation unit 220 performs a process of measuring the flight time T based on the detection signal of the detector 18. Specifically, the spectrum generation unit 220 measures the flight time T by detecting a time difference from when a predetermined voltage is applied to the acceleration electrodes 162 and 164 until the ions I reach the detector 18. The spectrum generation unit 220 samples the detection signal of the detector 18, that is, the ion intensity, at a predetermined time interval (equal time interval). And the spectrum production | generation part 220 records the detection signal of the detector 18 in the form at equal intervals in time, integrates it by the designated number of times, and records it as a time-of-flight spectrum.

  The time-of-flight spectrum is a spectrum with the time of flight T on the horizontal axis and the detected intensity on the vertical axis. The time-of-flight spectrum of each measurement region A is recorded in the storage unit 34 together with the position information (X, Y) of the measurement region A.

Next, the spectrum generation unit 220 generates a mass spectrum M _A of each measurement region A from time-of-flight spectrum of each measurement region A. Spectrum generation unit 220 converts the flight time T of the time-of-flight spectrum into a mass-to-charge ratio m / z, to generate a mass spectrum M _A. The calibration condition for converting the time of flight T into the mass-to-charge ratio m / z is expressed by, for example, the following formula (6).

  Three coefficients a, b, and c in Expression (6) are calibration coefficients (initial values), and are stored in the storage unit 34 in advance. The calibration coefficients a, b, and c can be determined, for example, by measuring three or more compounds with known m / z.

A mass spectrum can be generated from the time-of-flight spectrum by converting the time-of-flight T of the time-of-flight spectrum into a mass-to-charge ratio m / z using the above equation (6). Mass spectrum M _A of each measurement region A, the position information (X, Y) of the measurement region A with, is recorded in the storage unit 34.

  Here, the case where the flight time T is converted into the mass-to-charge ratio m / z using the calibration condition represented by the above formula (6) has been described, but the calibration condition is limited to the formula (6). Not. For example, a higher order function of the third order or higher may be used as the calibration condition.

Mass correction unit 222, and one at least two measurement regions A block (also referred to as "correction block"), by integrating the mass spectrum M _A measurement areas A for each said block, based on the integrated by mass spectrum , to correct the mass-to-charge ratio m / z mass spectrum M _a measurement areas a for each corresponding block.

FIG. 5 is a diagram for explaining the correction block B. FIG. In the example shown in FIG. 5, one correction block B includes nine measurement areas A arranged in 3 rows and 3 columns. One correction block B
There are no particular limitations on the number of measurement regions A that constitute two, so long as there are two or more, and a desired number of rows and a desired number of columns can be taken. Further, the number of measurement areas A constituting each correction block B may be different. Moreover, the measurement areas A constituting one correction block B may be close to each other or may be separated from each other. The entire measurement area A measured by the measurement unit 10 is divided into a plurality of correction blocks B (1), B (2), B (3).

Mass correction unit 222, for each correction block B, and corrects the mass-to-charge ratio m / z mass spectrum M _A. Specifically, the mass correction unit 222 obtains the calibration condition for each correction block B, and corrects the mass-to-charge ratio m / z mass spectrum M _A measurement areas A. Hereinafter, the process of the mass correction | amendment part 222 is demonstrated in detail.

First, the mass correction unit 222 integrates the mass spectrum M _A measurement areas A for each correction block B. That is, the weight correcting unit 222 integrates the mass spectrum M _A measurement areas A plurality contained in one correction block B (9 one in the illustrated example). Accordingly, the correction block mass spectrum M _B is obtained. Correction block mass spectrum M _B is obtained by integrating the mass spectrum M _A plurality of measurement regions A included in one correction block B.

In the example of FIG. 5, the mass spectrum MA of the measurement region _A is integrated for each correction block B (1), B (2),..., And each correction block B (1), B (2),. correction block mass spectrum M _B obtained for each ....

Next, the mass correction unit 222, based on the correction block mass spectrum _{M B,} each correction block B (1), B (2 ), obtaining a calibration condition for each ....

  First, the mass correction unit 222 obtains calibration conditions for the correction block B (1).

In the correction block mass spectrum M _B correction blocks B (1), adjust the m / z value of a predetermined peak reference value (theoretical value). The predetermined peak, using the peak observed in common to each correction block mass spectrum M _B (hereinafter also referred to as "common peak").

When correcting the common peak mass-to-charge ratio of the block mass spectrum M _B and (m / z) o, the flight time to To, the formula (6) is expressed by the following equation (7).

  However, | (m / z) o- (m / z) r | <Δm / z. Here, (m / z) r is a reference value (theoretical value) of a common peak, and Δm / z is a threshold value (mass-to-charge ratio range) regarded as a common peak.

  Next, “b” in the above equation (7) is corrected to make (m / z) o and (m / z) r coincide. When the corrected value of “b” is “b ′”, the above equation (6) is expressed by the following equation (8).

  From the equations (7) and (8), “b ′” is expressed by the following equation (9).

  Therefore, the calibration condition for converting the time of flight T into the mass-to-charge ratio m / z is expressed by the following equation (10).

Using the calibration condition of the above equation (10), the measurement areas A (0, 0), (1, 0), (2, 0), (0, 1), ( 1,1), (2,1), (0,2), (1,2), and (2,2), the time of flight T of the flight time spectrum is converted into a mass-to-charge ratio m / z. Thus, for each measurement region A is included in the correction blocks B (1), mass spectrum M _A which axis is corrected mass-to-charge ratio m / z obtained.

The mass correction unit 222 performs the above process on the other correction blocks B (2), B (3),... _A mass spectrum MA with the axis corrected is obtained. Mass spectrum M _A of the corrected each measurement region A has the position information (X, Y) of the measurement region A with, is recorded in the storage unit 34.

Here, since the mass correction unit 222 obtains the calibration condition for each correction block B (1), B (2),..., Each correction block B (1), B (2),. Different calibration conditions are used. Here, when the calibration conditions are the same, the m / z values for the same flight time T are the same, but when the calibration conditions are different, the m / z values are different even for the same flight time. Therefore, for example, as shown in FIG. 5, to select an area C extending over a plurality of measurement areas A, the case of obtaining a mass spectrum of the region C by integrating the mass spectrum M _A measurement areas A included in the region C , it is necessary to align the data interval of the mass spectrum M _a measurement areas a included in the area C.

Thus, the mass spectrometer 100, the data processing unit 224, the mass spectrum M _A corrected in different calibration conditions, performs processing for aligning the corrected data interval of the mass spectrum M _A was.

Figure 6 is a diagram for explaining an example of a process to align the data interval of the corrected mass spectrum M _A. It is assumed that the mass spectrum M _A 1 and the mass spectrum M _A 2 are corrected under different calibration conditions. Therefore, the mass spectrum M _A 1 has data of intensity corresponding to m / z = a1, a2, a3,. In addition, the mass spectrum M _A 2 has data of intensity corresponding to m / z = b1, b2, b3,.

Here, m / z = a1, a2 , a3, ··, and m / z = b 1, b 2, b 3, ·· the detection signals of the detector 18 at predetermined time intervals (time equal interval) Sampling time (flight time T) at the time of sampling is converted into an m / z value using the above equation (10). Since the calibration conditions are different between the mass spectrum M1 and the mass spectrum M2, the interval between m / z = a1, a2, a3,... And the interval between m / z = b1, b2, b3,. ing. Thus, the in mass spectrum M _{A 1} and mass spectrum M _{A 2,} since the data interval is different, it can not be simply multiplying the mass spectrum M _{A 1} and mass spectrum M _{A 2.}

Therefore, the data of the mass spectra M _A 1 and M _A 2 are complemented so that the data interval of the mass spectrum M _A 1 and the data interval of the mass spectrum M _A 2 are equal, and a new mass spectrum M _A 1a is obtained. , M _A 2a is generated. Specifically, from the mass spectrum M _A 1, the intensity at the time of m / z = c1, c2, c3,... Is complemented, and the data interval is set to an arbitrary data interval m / z = c1, c2, c3. _A mass spectrum M _A 1a is obtained. Similarly, from the mass spectrum M _A 2, the intensity at the time of m / z = c1, c2, c3,... Is complemented, and the data interval is set to an arbitrary data interval m / z = c1, c2, c3,. The mass spectrum M _A 2a is obtained. Thereby, the data intervals of the mass spectra M _A 1 and M _A 2 can be made uniform. Therefore, for example, the mass spectrum M _A 1 and the mass spectrum M _A 2 can be integrated.

The data processing unit 224 performs the above-described processing on the corrected mass spectrum M _A of each measurement region A, align the corrected data interval of the mass spectrum M _A was. Information of the mass spectrum M _A data interval is aligned, the position information (X, Y) of the measurement region A with, is recorded in the storage unit 34.

Mass imaging unit 226, from the mass spectrum M _A data interval is aligned with mass spectrum M _A or the data processing unit 224 which is corrected by the weight corrector 222 acquires imaging information. Here, the imaging information is position information of the measurement areas A and information on the mass of the compound included in each measurement area A. That is, the imaging information is recorded in the storage unit 34 in the format of (X, Y, m / z, ion intensity), for example. The processing unit 20 displays, for example, intensity distribution information about a specific m / z (that is, a specific compound), information about a compound localized in a specific region, and the like on the display unit 32 based on imaging information. To perform the process.

  The operation unit 30 performs a process of acquiring an operation signal corresponding to an operation by the user and sending the operation signal to the processing unit 20. The operation unit 30 is, for example, a button, a key, a touch panel display, a microphone, or the like.

  The display unit 32 displays the image generated by the processing unit 20, and its function can be realized by an LCD, a CRT, or the like. The display unit 32 displays, for example, information on intensity distribution for a specific m / z (that is, a specific compound) generated by the processing unit 20 and information on a compound localized in a specific region.

  The storage unit 34 serves as a work area for the processing unit 20, and its function can be realized by a RAM or the like. The storage unit 34 stores programs, data, and the like for the processing unit 20 to perform various calculation processes and control processes. The storage unit 34 is used as a work area of the processing unit 20 and is also used for temporarily storing calculation results and the like executed by the processing unit 20 according to various programs.

The information storage medium 36 (a computer-readable medium) stores programs, data, and the like, and functions as an optical disk (CD, DVD), a magneto-optical disk (MO), a magnetic disk, a hard disk, and a magnetic tape. Alternatively, it can be realized by a memory (ROM). The processing unit 20 performs various processes of the present embodiment based on a program (data) stored in the information storage medium 36. The information storage medium 36 can store a program for causing a computer to function as each unit of the processing unit 20.

  Next, the operation of the mass spectrometer 100 according to the first embodiment will be described.

  The user places the sample S on the sample plate 2 (see FIG. 3), and sprays the sample S with a matrix for promoting ionization as necessary. Then, the sample plate 2 is placed on the sample stage 12.

  Next, the user can measure the area of the sample S in the sample plate 2, the number of measurement areas A (the number of divisions of the area to be measured), and the number of laser irradiations in each measurement area A (eg, several Specify measurement conditions such as 10 to several hundred times. The measurement conditions can be specified via the operation unit 30. When the measurement conditions are specified, the mass spectrometer 100 starts measurement.

  The stage drive unit (not shown) moves the sample stage 12 according to the specified measurement conditions to change the laser irradiation position (measurement region A), and the ion generation unit 14 performs the number of times specified in the measurement region A. Only the laser beam L is irradiated. Each time the laser beam L is irradiated, the ions I pass through the reflectron 17 and are mass separated according to the mass-to-charge ratio (m / z value) before reaching the detector 18, and are detected by the detector 18. Is done.

The spectrum generator 220 records the detection signal of the detector 18 at equal intervals in the flight time T, accumulates the designated number of times, and records it as a flight time spectrum. The time-of-flight spectrum is recorded together with the position information (X, Y) of the measurement area A. Further, spectrum generation unit 220, the flight time axis of time-of-flight spectrum is converted into m / z-axis using the initial calibration conditions (uncorrected) (the formula (6)), mass spectrum M _A obtain. Mass spectrum _{M A} is, position information (X, Y) of the measurement region A is recorded together with. The mass spectrometer 100 performs the above-described processing for all measurement regions A.

Next, the mass correction unit 222 corrects the drift of the mass axis. Specifically, the mass correction unit 222, first, by integrating the respective correction block B mass spectra M _{A (or} time-of-flight spectrum) contained in (see FIG. 5) to obtain the corrected block mass spectrum M _B. Then, the m / z values of peaks observed in common in each correction block mass spectrum M _B (common peak), in accordance with the reference value, to correct the calibration factor. Thereby, a new calibration condition (the above formula (10)) is obtained. Next, using the new calibration condition (the above formula (10)), the flight time axis of the flight time spectrum of the measurement region A included in the correction block B is converted into the m / z axis, and the corrected mass is obtained. _A spectrum MA is obtained. Corrected mass spectrum M _A is, position information (X, Y) of the measurement region A is recorded together with. The mass correction unit 222 performs the above-described processing for all the correction blocks B.

Next, the data processing unit 224, as shown in FIG. 6, equally spaced mass spectrum M _A flight time of each correction block B, performs data interpolation, mass spectrum M which is at equal intervals m / z value Process _A is performed.

Next, the mass imaging unit 226 obtains imaging information for each measurement region A from the mass spectrum M _A corrected by the mass correction unit 222 or the mass spectrum M _A having the m / z values at equal intervals by the data processing unit 224. (X, Y, m / z, ionic strength) is acquired. The imaging information is recorded in the storage unit 34, for example. For example, based on the imaging information, the processing unit 20 causes the display unit 32 to display information on the intensity distribution of a specific m / z (that is, a specific compound) and information on a compound localized in a specific region. Take control.

  The mass spectrometer 100 has the following features, for example.

In the mass spectrometer 100, the mass correction unit 222, at least two measurement regions A and one block B, by integrating the mass spectrum M _A measurement areas A for each the block B, and accumulated the mass spectrum M _B based on, for correcting the mass-to-charge ratio of the mass spectrum M _a measurement areas a for each block B. Therefore, the mass spectrometer 100, for example, as compared with the case of performing the correction of the mass-to-charge ratio based on mass spectrum M _A single measurement region A, to improve the SN ratio of the spectrum. Thus, the drift of the mass axis of the mass spectrum M _A (error in calculating the mass-to-charge ratio from the time-of-flight) can be accurately corrected, it is possible to improve mass accuracy.

In the mass spectrometer 100, the mass imaging unit 226, from the mass spectrum M _A for each measurement region A, which is corrected by the weight corrector 222, and the positional information of the measurement region A, of the mass of the compound contained in each measurement area A Get information. Therefore, the mass spectrometer 100 can obtain information on the intensity distribution of a specific compound and information on a compound localized in a specific region.

2. Second Embodiment Next, a mass spectrometer according to a second embodiment will be described. The configuration of the mass spectrometer according to the second embodiment is the same as that of the mass spectrometer 100 according to the first embodiment shown in FIG. 1 described above. Hereinafter, the difference from the mass spectrometer 100 according to the first embodiment will be described. The point will be described.

In the example of the mass spectrometer 100, the data processing unit 224, as shown in FIG. 6, differs from the mass spectrum M _A corrected calibration conditions, the mass spectrum M an arbitrary data interval data interval _A m / The data intervals were aligned as z = c1, c2, c3,.

Mass contrast, in the mass spectrometer according to the second embodiment, the data processing unit 224, the mass spectrum M _A corrected in different calibration conditions, the data interval of the mass spectrum M _A, the corrected align the data interval of one of the mass spectrum M _a of the spectrum M _a.

Figure 7 is a diagram for explaining an example of a process to align the data interval of the mass spectrum M _A. The mass spectrum M _A 1 and the mass spectrum M _A 2 are corrected under different calibration conditions. In the second embodiment, as shown in FIG. 7, the data interval of the mass spectrum M _{A 2,} to align the data interval of the mass spectrum M _{A 1,} to complement the data of mass spectrum M _{A 2,} a new _A mass spectrum M _A 2a is obtained. Specifically, from the mass spectrum M _A 2, the intensity at the time of m / z = a1, a2, a3,... Is complemented, and the data interval is expressed as m / z = a1, a2, a3,. Mass spectrum M _A 2a is obtained. Thus, the shaft of mass-to-charge ratios of the mass spectrum M _{A 2,} can be aligned to the axis of the mass-to-charge ratio of the mass spectrum M _{A 1.}

The data processing unit 224 performs the above-described processing with respect to the mass spectrum M _A of each measurement region A, align the data interval in each mass spectrum M _A. The reference mass spectrum M _A (mass spectrum M _A 1) can be arbitrarily selected from the entire measurement region A. For example, in the example shown in FIG. 5, a reference data interval first mass spectrum M _A of the measured measurement region A (0,0).

The mass spectrometer according to the second embodiment, the data processing unit 224, a data interval of each mass spectrum M _A, align the data interval of one of the mass spectrum M _A of the corrected mass spectrum M _A . Thus, even data mass spectrum M _A having uniform data interval complement, can be handled in the same manner as measured mass spectrum M _A. That is, for example, be a mass spectrum M _A having uniform data interval complement the data can be converted to the axis of the mass-to-charge ratio to the axis of the flight time T. Thus, for example, it is possible to convert the mass spectrum M _A having uniform data interval to complement the data in the time-of-flight spectrum.

  Further, the mass spectrometer according to the second embodiment can achieve the same effects as the mass spectrometer 100 according to the first embodiment described above.

3. Third Embodiment Next, a mass spectrometer according to a third embodiment will be described. The configuration of the mass spectrometer according to the third embodiment is the same as that of the mass spectrometer 100 according to the first embodiment shown in FIG. 1 described above. Hereinafter, the difference from the mass spectrometer 100 according to the first embodiment will be described. The point will be described.

In the example of the mass spectrometer 100, the measurement unit 10 starts measurement from the measurement region A (0, 0) shown in FIG. 2 and measures the measurement regions A (1, 0), (2, 0),. The first row is measured in the order of (n _x , 0), then the second row is measured, then the third row is measured, and so on, and the measurement area A (n _x , n _y ) is measured. Measurements were made up to.

  On the other hand, in the mass spectrometer according to the third embodiment, the measurement unit 10 measures all the measurement areas A included in one correction block B, and then measures the measurement area A included in the next correction block B. taking measurement.

  FIG. 8 is a diagram for explaining the measurement order of the measurement unit 10 of the mass spectrometer according to the third embodiment. As shown in FIG. 8, the measurement unit 10 first measures all measurement areas A included in the correction block B (1), and then measures all measurement areas A included in the correction block B (2). . Thereafter, the same measurement is performed for the correction block B (3), the correction block B (4),..., And the entire measurement area A is measured.

  The order in which the measurement area A included in one correction block B is measured is not particularly limited. For example, in the illustrated example, the measurement area A (0, 0), (1, 0) in the correction block B (1). , (2,0), (0,1), (1,1), (2,1), (0,2), (1,2), (2,2).

  In the mass spectrometer according to the third embodiment, after all the measurement areas A included in one correction block B are measured, the measurement area A included in the next correction block B is measured. Thereby, the influence of the system | strain error by progress of time can be reduced, and mass accuracy can be improved.

For example, if the measurement unit 10 starts measurement from the measurement area A (0, 0) in FIG. 8, the measurement area A (1, 0), (2, 0),..., (N _x , 0 ) In the order of measurement), and then measurement in the measurement areas A (0, 1), (1, 1), (2, 1)..., (N _x , 1) in the second row. Next, the measurement area A (0, 2), (1, 2), (3, 1)... (N _x , 2) in the third row is measured. In this case, it takes a long time to measure the measurement area A (2, 2) of the correction block B (1) after the measurement area A (0, 0) of the correction block B (1) is measured. That is, it takes a long time to measure all the measurement areas A included in the correction block B (1). Thereby, the influence of the systematic error due to the passage of time increases.

  On the other hand, in the mass spectrometer according to the third embodiment, after measuring all the measurement areas A included in one correction block B, the measurement area A included in the next correction block B is measured. The time from when the measurement area A (0, 0) of the block B (1) is measured to when the measurement area A (2, 2) of the correction block B (1) is measured can be shortened. Therefore, in the mass spectrometer according to the third embodiment, it is possible to reduce the influence of the systematic error due to the passage of time.

  Further, the mass spectrometer according to the third embodiment can achieve the same operational effects as those of the mass spectrometer 100 according to the first embodiment described above.

4). Fourth Embodiment Next, a mass spectrometer according to a fourth embodiment will be described. The configuration of the mass spectrometer according to the fourth embodiment is the same as that of the mass spectrometer 100 according to the first embodiment shown in FIG. 1 described above. Hereinafter, the difference from the mass spectrometer 100 according to the first embodiment will be described. The point will be described.

In the mass correction unit 222 of the mass spectrometer according to the fourth embodiment, a common peak exists in the first range of the mass-to-charge ratio in the mass spectrum M _B (corrected block mass spectrum M _B ) integrated for each correction block B. The mass-to-charge ratio is corrected using the common peak (first peak) existing in the first range, and when there is no common peak in the first range, the first range The mass-to-charge ratio is corrected using a common peak (second peak) that exists in a second range having a different mass-to-charge ratio.

  As described above, the mass correction unit 222 performs correction by specifying another peak for the correction block B in which the first specified common peak is not found. The first range of the mass-to-charge ratio is a range in which a peak existing in the first range can be regarded as the first peak. Further, the second range of the mass-to-charge ratio is a range in which the peak existing in the second range can be regarded as the second peak. The first range and the second range can be arbitrarily set.

The mass spectrometer according to the fourth embodiment, in the correction block mass spectrum M _B, when the common peak is present in the first range of mass to charge ratios, using a common peaks present in the first range When the mass-to-charge ratio is corrected and no common peak exists in the first range, the mass-to-charge ratio is corrected by using the common peak existing in the second range having a mass-to-charge ratio different from the first range. To do. Thereby, the correction block B which cannot be corrected because there is no common peak can be reduced.

  When there is a correction block B that cannot be corrected, for example, the processing unit 20 displays a diagram showing the correction block B shown in FIG. 5 on the display unit 32, and the correction block B that cannot be corrected is corrected. A process of displaying in a color different from that of the completed correction block B is performed.

  Further, the mass spectrometer according to the fourth embodiment can achieve the same operational effects as those of the mass spectrometer 100 according to the first embodiment described above.

5. Fifth Embodiment Next, a mass spectrometer according to a fifth embodiment will be described. The configuration of the mass spectrometer according to the fifth embodiment is the same as that of the mass spectrometer 100 according to the first embodiment shown in FIG. 1 described above. Hereinafter, the difference from the mass spectrometer 100 according to the first embodiment will be described. The point will be described.

In the example of the mass spectrometer 100, the mass analyzer 10b is a reflective TOFMS configured to include a reflectron 17 and a detector 18, as shown in FIG.

  On the other hand, in the mass spectrometer according to the fifth embodiment, the mass analyzer 10b is a multi-circular TOFMS.

  FIG. 9 is a diagram illustrating an example of the configuration of the measurement unit 10 of the mass spectrometer according to the fifth embodiment. As shown in FIG. 9, the mass spectrometer 10b includes sector electrodes 50a, 50b, 50c, and 50d and a detector 18.

  In the mass analyzer 10b, an eight-shaped circular orbit 52 is formed by an electric field generated by the plurality of sector electrodes 50a, 50b, 50c, and 50d. Although not shown, the circular orbit 52 may be circular, elliptical, or the like. Further, the orbit 52 may be a reciprocating orbit in which ions reciprocate along a linear or curved orbit.

  As shown in FIG. 9, the mass analyzer 10 b separates the ions I supplied from the ion source 10 a according to the mass-to-charge ratio m / z by performing multiple rounds along the round orbit 52, and the detector 18 is detected. Thereby, the flight time T can be lengthened and high mass accuracy can be obtained.

In the mass spectrometer according to the fifth embodiment, since the mass spectrometer 10b is a multi-circular TOFMS, high mass accuracy can be obtained. Further, the mass spectrometer according to the fifth embodiment, similarly to the mass spectrometer 100 according to the first embodiment described above, the drift of the mass axis of the mass spectrum M _A can be corrected accurately, the mass accuracy Can be improved. Therefore, in the mass spectrometer according to the fifth embodiment, higher mass accuracy can be obtained.

  The present invention is not limited to the above-described embodiments, and various modifications can be made within the scope of the gist of the present invention.

  For example, in the mass spectrometers according to the first to fifth embodiments described above, the example in which the ion source 10a ionizes the sample S by the matrix-assisted laser desorption ionization method (MALDI) has been described, but the mass spectrometry according to the present invention is described. The ionization method in the ion source of the apparatus is not limited to this. For example, the ion source of the mass spectrometer according to the present invention may ionize a sample by secondary ion mass spectrometry (SIMS). Secondary ion mass spectrometry is a technique in which the surface of the sample S is irradiated with an ion beam (primary ions), and ions (secondary ions) are generated by collision between the ions and a solid surface at the molecular / atomic level. Further, for example, the ion source of the mass spectrometer according to the present invention may ionize the sample by the surface-assisted laser desorption ionization method (SALDI) in which the nanostructure is provided on the sample plate.

  Further, for example, in the mass spectrometers according to the first to fifth embodiments described above, the example in which the mass analyzer 10b is a time-of-flight type has been described. However, the mass analyzer of the mass spectrometer according to the present invention has a time-of-flight. It is not limited to the type. For example, the mass spectrometer of the mass spectrometer according to the present invention may be a quadrupole type, an ion trap type, a magnetic field deflection type, a Fourier transform ion cyclotron resonance type, or the like.

  Further, for example, in the mass spectrometers according to the first to fourth embodiments described above, an example in which the mass analyzer 10b is a reflective TOFMS will be described. In the mass spectrometer according to the fifth embodiment, the mass analyzer An example in which 10b is a multi-circular TOFMS has been described. The mass spectrometer of the mass spectrometer according to the present invention is not limited to these, and may be, for example, a linear TOFMS that is generated by an ion source and linearly flies an ion group.

  The above-described embodiments are examples, and the present invention is not limited to these. For example, the embodiments can be appropriately combined.

  The present invention includes configurations that are substantially the same as the configurations described in the embodiments (for example, configurations that have the same functions, methods, and results, or configurations that have the same objects and effects). In addition, the invention includes a configuration in which a non-essential part of the configuration described in the embodiment is replaced. In addition, the present invention includes a configuration that exhibits the same operational effects as the configuration described in the embodiment or a configuration that can achieve the same object. Further, the invention includes a configuration in which a known technique is added to the configuration described in the embodiment.

DESCRIPTION OF SYMBOLS 2 ... Sample plate, 10 ... Measurement part, 10a ... Ion source, 10b ... Mass analysis part, 12 ... Sample stage, 14 ... Ion production part, 16 ... Acceleration part, 17 ... Reflectron, 18 ... Detector, 20 ... Processing , 30 ... operation part, 32 ... display part, 34 ... storage part, 36 ... information storage medium, 50a, 50b, 50c, 50d ... sector electrode, 52 ... circular orbit, 100 ... mass spectrometer, 141 ... laser light source, 142 ... Lens, 143 ... Mirror, 144 ... Mirror, 145 ... Lens, 146 ... CCD camera, 162 ... First acceleration electrode, 164 ... Second acceleration electrode, 220 ... Spectrum generation unit, 222 ... Mass correction unit, 224 ... Data Processing unit, 226 ... Mass imaging unit

Claims (8)

A measurement unit that ionizes a sample for each measurement region and performs mass spectrometry for each measurement region;
Based on the measurement result of the measurement unit, a spectrum generation unit that generates a mass spectrum for each measurement region;
At least two of the measurement regions are defined as one block, and the mass spectrum of the measurement region is integrated for each block, and the mass-to-charge ratio of the mass spectrum of the measurement region is calculated for each block based on the integrated mass spectrum. A mass correction unit to be corrected;
Only including,
The said measurement part is a mass spectrometer which measures the said measurement area | region contained in the said next block, after measuring all the said measurement area | regions contained in one said block . A measurement unit that ionizes a sample for each measurement region and performs mass spectrometry for each measurement region;
Based on the measurement result of the measurement unit, a spectrum generation unit that generates a mass spectrum for each measurement region;
At least two of the measurement regions are defined as one block, and the mass spectrum of the measurement region is integrated for each block, and the mass-to-charge ratio of the mass spectrum of the measurement region is calculated for each block based on the integrated mass spectrum. A mass correction unit to be corrected;
Only including,
In the mass spectrum integrated for each block, when a peak exists in the first range of the mass-to-charge ratio, the mass correction unit calculates the mass-to-charge ratio using the peak that exists in the first range. A mass spectrometer that corrects and corrects the mass-to-charge ratio using a peak that exists in a second range having a mass-to-charge ratio different from the first range when there is no peak in the first range . In claim 1 ,
In the mass spectrum integrated for each block, when a peak exists in the first range of the mass-to-charge ratio, the mass correction unit calculates the mass-to-charge ratio using the peak that exists in the first range. A mass spectrometer that corrects and corrects the mass-to-charge ratio using a peak that exists in a second range having a mass-to-charge ratio different from the first range when there is no peak in the first range. In any one of Claims 1 thru | or 3,
The mass correction unit corrects the mass-to-charge ratio by obtaining a calibration condition to convert the flight time into the mass-to-charge ratio. In claim 4,
For the mass spectrum corrected under the different calibration conditions, including a data processing unit that aligns the data interval of each mass spectrum,
The data processing unit is a mass spectrometer that aligns the data interval of each mass spectrum with the data interval of any one of the corrected mass spectra. In any one of Claims 1 thru | or 5,
The measuring unit is
An ion source that ionizes the measurement region by irradiating a laser beam;
A mass analyzer that detects and mass-separates ions generated in the ion source;
Including a mass spectrometer. In claim 6,
The mass spectrometer is a mass spectrometer that separates ions generated by the ion source based on time of flight. In any one of Claims 1 thru | or 7,
A mass spectrometer including a mass imaging unit that acquires position information of the measurement region and mass information of a compound included in each measurement region from the mass spectrum of each measurement region corrected by the mass correction unit.

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