Analysis device and analysis method

Abstract

In an analysis method and an analysis device for detecting fluorescence from each light-emitting point in each of a plurality of wavelength bands in order to recognize light emitted by a plurality of types of phosphors from a plurality of light-emitting points, spatial crosstalk and spectral crosstalk exist between signal intensities of the plurality of light-emitting points and the plurality of wavelength bands, and the recognition performance is reduced. By inputting all of the detection signals in the plurality of wavelength bands of each of the plurality of light-emitting points into a predetermined arithmetic expression, spatial crosstalk and spectral crosstalk are eliminated, and the concentration of each light-emitting body at each light-emitting point is derived.

Description

[0001] This application is a divisional application of the Chinese invention patent application "Analysis device and analysis method" with an application date of April 21, 2020 and application number 202080036799.3. Technical Field

[0002] The present disclosure relates to an analysis method and an analysis device for respectively identifying and detecting fluorescence emitted from a plurality of fluorescent substances at a plurality of light-emitting points. Background Art

[0003] In recent years, multi-capillary electrophoresis devices have been widely used, which perform electrophoretic analysis in parallel by filling multiple capillaries with electrolyte solutions or electrophoretic separation media such as polymer gels and polymer electrolyte solutions. The analysis objects range from low molecules to macromolecules such as proteins and nucleic acids. In addition, in the measurement mode, there are a large number of modes such as the following: a mode in which light is irradiated to the absorption point of each capillary and the absorption of light generated when the analysis object passes through the absorption point; or a mode in which laser is irradiated to the luminous point of each capillary and the fluorescence or scattered light generated when the analysis object passes through the luminous point.

[0004] For example, in Patent Document 1, all capillaries around A light-emitting points on A capillaries (A is an integer greater than or equal to 2) are arranged on the same plane, and a laser beam is introduced from the side of the arrangement plane to irradiate the light-emitting points of all capillaries at once, so that the fluorescence generated at each light-emitting point is wavelength-dispersed in a direction perpendicular to the arrangement plane and detected at once. In a detection device, the fluorescence emitted from the A light-emitting points is collimated at once in a condenser lens, transmitted through a transmission-type diffraction grating, and the first-order diffracted light of each fluorescence is imaged at once on a two-dimensional sensor using an imaging lens. Here, by making the arrangement direction of the A light-emitting points and the wavelength dispersion direction based on the diffraction grating perpendicular to each other, the wavelength dispersion images of the light-emitting fluorescence from each capillary do not overlap with each other on the two-dimensional sensor. For the wavelength dispersion image of each capillary, by setting B (B is an integer greater than or equal to 1) arbitrary wavelength band detection areas, B color detection can be performed. The case of B=1 is called single-color detection, and the case of B≥2 is called multi-color detection. In the multi-capillary electrophoresis device of Patent Document 1, for example, DNA sequencing based on the Sanger method can be performed on different DNA samples in each capillary. In the Sanger method, four fluorescent substances are labeled according to the terminal base types A, C, G and T in the DNA fragments included in the DNA sample, and the respective luminescent fluorescence is identified by multi-color detection.

[0005] In Patent Document 2, all capillaries around A light-emitting points on A capillaries (A is an integer greater than or equal to 2) are arranged on the same plane, a laser beam is introduced from the side of the arrangement plane to illuminate the light-emitting points of all capillaries at once, and the fluorescence generated at each light-emitting point in a direction perpendicular to the arrangement plane is split according to the wavelength component and detected at once. In the detection device, the light-emitting fluorescence from the A light-emitting points is collimated by A condensing lenses separately as A light beams, and each light beam is incident in parallel on a group of dichroic mirror arrays in which B (B is an integer greater than or equal to 1) dichroic mirrors are arranged. Each light beam is split into B light beams of different wavelength bands, and the total of A×B light beams generated are incident in parallel on a two-dimensional sensor, and A×B split images are generated on the image. Here, by making the arrangement direction of the A light-emitting points and the B split directions of each light beam based on the dichroic mirror array perpendicular to each other, the A×B split images do not overlap each other on the image, and A×B detection areas can be set. In this way, B color detection of each capillary can be performed. Therefore, in the multicapillary electrophoresis apparatus of Patent Document 2, for example, similarly to the case of Patent Document 1, DNA sequencing based on the Sanger method can be performed on different DNA samples in each capillary.

[0006] However, usually, by only performing multicolor detection, it is impossible to identify the luminescent fluorescence of multiple phosphors. This is because the fluorescence spectra of each phosphor overlap with each other, so the fluorescence of multiple phosphors is mixed in any one band (referred to as spectral crosstalk in the present disclosure). In addition, this is because there are situations where multiple phosphors of different concentrations emit fluorescence at the same time. Therefore, through the next process (referred to as color conversion in the present disclosure), the spectral crosstalk is eliminated, so that the above-mentioned identification can be performed.

[0007] For each of A (A is an integer greater than or equal to 2) light-emitting points, at each time, for the fluorescence emitted by C types (C is an integer greater than or equal to 1) of phosphors, B color detection is performed in a detection area of ​​B types (B is an integer greater than or equal to 1) wavelength bands. Here, it is assumed that B ≥ C. For each light-emitting point and at each time, color conversion is performed on the B color detection result to obtain the concentration of the C type of phosphor. For each light-emitting point P(a) (a=1, 2, ..., A), the fluorescence emitted by the phosphor D(c) (c=1, 2, ..., C) is detected in a detection area W(b) (b=1, 2, ..., B) of a different wavelength band. At any time, the concentration of the phosphor D(c) at the light-emitting point P(a) is set to Z(c), and the signal intensity of the detection area W(b) with respect to the light-emitting point P(a) is set to X(b). Here, assuming that X is a matrix with B rows and 1 column with signal intensity X(b) as an element, Z is a matrix with C rows and 1 column with concentration Z(c) as an element, and Y is a matrix with B rows and C columns with Y(b)(c) as an element, the following equations hold. (Equation 1) to (Equation 4) are equations of relationship between b and c, not a, and are independent at each luminous point P(a). In the case of single-color detection with B=1, since B≥C, C=1, X, Y, and Z are not matrices.

[0008] [Mathematical formula 1]

[0009] X=Y×Z (number 1)

[0010] [Mathematical formula 2]

[0011]

[0012] [Mathematical formula 3]

[0013]

[0014] [Formula 4]

[0015]

[0016] Here, the element Y(b)(c) of the matrix Y of B rows and C columns represents the signal intensity ratio of the luminescent fluorescence of the phosphor D(c) detected in the detection area W(b). By making any one phosphor D(c0) emit fluorescence alone, one column Y(b)(c0) (b=1, 2, ..., B) of the matrix Y can be determined. Here, it is generally difficult to control the concentration of the phosphor D(c0), so it is more convenient to standardize one column Y(b)(c0). For example, preferably, among the B elements, the largest element is 1, and the other elements are represented by the ratio relative to the maximum value. Alternatively, preferably, the ratio of each element is determined so that the total of the B elements is 1. In other words,

[0017] [Mathematical formula 5]

[0018]

[0019] Furthermore, by sequentially performing the above steps for all C types of phosphors D(c), all the columns of the matrix Y can be determined. The matrix Y is determined only by the characteristics of the phosphor D(c) and the detection area W(b) of different wavelengths, and does not change during the ongoing electrophoresis analysis. In addition, as long as the optical system, phosphor D(c), detection area W(b) and other conditions are fixed, the matrix Y remains fixed for different electrophoresis analyses. Therefore, for each luminous point, the concentration Z(c) of the phosphor D(c) at each moment is obtained by the following formula based on the signal intensity X(b) of the detection area W(b) at each moment.

[0020] [Mathematical formula 6]

[0021] Z=Y - ×X (Number 6)

[0022] Here, Y － With C rows and B columns, it is the general inverse matrix of Y, through Y = (Y T ×Y) -1 ×Y T ) is found. When the matrix Y is a square matrix with B = C, Y is equal to the inverse matrix Y －1 .

[0023] (Math 1) is a simultaneous equation that expresses the relationship between the unknown concentration of C-type phosphor and the known intensity of B-color fluorescence, and (Math 6) is equivalent to finding its solution. Therefore, generally, as described above, the condition of B ≥ C is required. Assuming B < C, it is impossible to find a unique solution (in other words, since there may be multiple solutions), it is impossible to perform color conversion as in (Math 6).

[0024] As an example, the case where C=4 in the Sanger method and B=4 in the four-color detection is described in detail. When making copies of DNA fragments of various lengths for the template DNA by the Sanger reaction, four types of fluorescent substances D(1), D(2), D(3), and D(4) are used to label them according to the terminal base types A, C, G, and T, and they are separated by length by electrophoresis and sequentially irradiated with laser beams to emit fluorescence. The four-color fluorescent light is detected in the detection areas W(1), W(2), W(3), and W(4) of the four wavelength bands corresponding to the maximum emission wavelengths of the fluorescent substances D(1), D(2), D(3), and D(4), and the time series data of their signal intensities X(1), X(2), X(3), and X(4) are obtained (referred to as raw data in the present disclosure. However, this is not limited to the case where B=4 and C=4). If the concentrations of the phosphors D(1), D(2), D(3) and D(4) at each moment are set to Z(1), Z(2), Z(3) and Z(4), (Formula 1) becomes the following formula.

[0025] [Formula 7]

[0026]

[0027] Here, the elements Y(b)(c) of the 4-row by 4-column matrix Y represent the intensity ratios of the fluorescence emitted by the phosphor D(c) (c is 1, 2, 3, or 4) detected in the wavelength band W(b) (b is 1, 2, 3, or 4) by spectral crosstalk. In each capillary, the phosphor D(c) (c is 1, 2, 3, or 4) performs electrophoresis analysis on the sample that emits fluorescence alone, thereby determining the elements Y(b)(c) of the matrix Y. For example, the four-color fluorescence intensities X(1), X(2), X(3), and X(4) when the phosphor D(1) emits fluorescence alone are assigned to the elements Y(1)(1), Y(2)(1), Y(3)(1), and Y(4)(1), respectively. In addition, the four-color fluorescence intensities X(1), X(2), X(3), and X(4) when the phosphor D(2) emits fluorescence alone are assigned to the elements Y(1)(2), Y(2)(2), Y(3)(2), and Y(4)(2), respectively. The same is true for the fluorescent substances D(3) and D(4). Y(b)(c) is a fixed value determined only by the characteristics of the fluorescent substance D(c) and the wavelength band W(b), and does not change during electrophoresis. Therefore, for each capillary, the concentration of the fluorescent substances D(1), D(2), D(3), and D(4) at each time is obtained by the following formula which is a concrete form of (Mathematical Formula 6) based on the four-color fluorescence intensities X(1), X(2), X(3), and X(4) at each time.

[0028] [Mathematical formula 8]

[0029]

[0030] Thus, the inverse matrix Y －1 , by multiplying with the four-color fluorescence intensity to eliminate spectral crosstalk, the concentrations of the four fluorescent substances, that is, the time series data of the concentrations of DNA fragments with four bases at the ends are obtained (in this disclosure, referred to as color conversion data. However, it is not limited to the case where B=4, C=4).

[0031] The above color conversion process is as described above, and is performed independently for A capillaries. This is true under the premise that the crosstalk between the capillaries (referred to as spatial crosstalk in the present disclosure) is small enough. In other words, it means that the ratio of the signal intensity from the fluorescence emitted from other capillaries mixed in the signal intensity obtained by detecting the fluorescence emitted from any capillary is small enough. Therefore, the process of determining the matrix Y by sequentially performing the process of making the above-mentioned one kind of phosphor D(c0) emit fluorescence alone on C kinds of phosphors D(c) is substantially performed on A capillaries at the same time, and the matrix Y of each of the A capillaries is obtained in parallel. This is a necessary process to perform the derivation of the matrix Y in a short time without any trouble.

[0032] It is basic to reduce spatial crosstalk based on the design of the optical system, but there are also attempts to reduce it through computational processing. In Patent Document 3, instead of coming from multiple capillaries, the luminescent fluorescence from multiple luminescent points randomly arranged on a plane is imaged on a two-dimensional sensor through a focusing lens. The luminescent fluorescence from each luminescent point is obtained as the signal intensity of the detection area at each imaging position set on the two-dimensional sensor. The ratio of the signal intensity of the luminescent fluorescence from any luminescent point to the spatial crosstalk of the signal intensity of the luminescent fluorescence from other luminescent points can be expressed as a function of the distance between the two luminescent points, or the distance between the corresponding two detection areas, and it is found that it decreases with the above distance. The above function is calculated in advance, and in the fluorescence image of multiple luminescent points randomly arranged on a plane, the mutual spatial crosstalk is calculated based on the distance between any two detection areas and the above function, and the calculated spatial crosstalk is subtracted from the original fluorescence image, thereby achieving the reduction of spatial crosstalk.

[0033] Prior Art Literature

[0034] Patent Literature

[0035] Patent Document 1: Japanese Patent No. 3897277

[0036] Patent Document 2: Japanese Patent No. 6456983

[0037] Patent Document 3: Japanese Patent Application No. 2018-529947 Summary of the invention

[0038] Problems to be solved by the invention

[0039] In either of Patent Document 1 and Patent Document 2, since the fluorescence emitted from A light-emitting points on A capillaries is separated from each other and imaged on the two-dimensional sensor, the spatial crosstalk is essentially suppressed to a low level. Important factors of spatial crosstalk include (1) lens aberrations, (2) multiple reflections between elements of fluorescence generated inside the optical system composed of elements such as polycapillaries, lenses, filters, dichroic mirrors, and two-dimensional sensors, and (3) blooming between pixels of the two-dimensional sensor. In order to reduce spatial crosstalk, it is necessary to construct the optical system in a manner that minimizes the influence of the above-mentioned important factors (1) to (3). For example, in order to suppress the influence of important factor (2), it is considered to select a coating with a lower reflectivity as an anti-reflection coating applied to the lens. However, the spatial crosstalk cannot be completely zero. For spatial crosstalk, in the detection of fluorescence emitted from any capillary, the detection lower limit is effectively increased, so it is possible to reduce the detection dynamic range while reducing the detection sensitivity. Therefore, even a slight reduction in spatial crosstalk is extremely important in alleviating or solving the above-mentioned problems.

[0040] The present inventors tried to reduce the spatial crosstalk by the method of Patent Document 3 for the optical system of Patent Document 1 or Patent Document 2, but the function did not work well. First, the case of single-color detection with B=1 is described. For the signal intensity X(α)(β) of the detection area β of any one of the A light-emitting points, that is, the light-emitting point α, although it is confirmed that the spatial crosstalk given by the signal intensity X(α')(β) of the detection area β of any one of the light-emitting points α' (α≠α') other than the above-mentioned light-emitting points tends to decrease together with the distance between the above-mentioned two detection areas, it is obvious that it cannot be expressed by a function of the above-mentioned distance. For example, even if the distance between the above-mentioned two detection areas is fixed, the ratio of spatial crosstalk is different when the above-mentioned two detection areas are located near the central axis of the optical system and when they are located at a position far from the central axis of the optical system. In addition, the ratio of spatial crosstalk from one of the above-mentioned two detection areas to the other is different from the ratio of spatial crosstalk from the other to one. In the case of multi-color detection with B≥2, the method of Patent Document 3 is even more unable to work. The spatial crosstalk given by the signal intensity X(α)(β) of any one of the A light-emitting points, i.e., the light-emitting point α, and any one of the B detection areas, i.e., the detection area β, to the signal intensity X(α')(β') of any one of the B detection areas, i.e., the detection area β, and the spatial crosstalk given by the signal intensity X(α')(β') of any one of the B detection areas, i.e., the detection area β, with respect to any one of the light-emitting points α' (α≠α') other than the above-mentioned light-emitting points (β=β' or β≠β', β=β' indicates the detection areas of the same wavelength band, and β≠β' indicates the detection areas of different wavelength bands), cannot be expressed as a function of the distance between the above-mentioned two detection areas. In particular, when β≠β', since the spatial crosstalk and the spectral crosstalk exist together, the crosstalk between the above-mentioned two detection areas cannot be essentially expressed as a function of the distance between the two. This subject is described in detail in the "Specific Implementation Method".

[0041] Therefore, in the present disclosure, a method is proposed for reducing spatial crosstalk between multiple light-emitting points generated in an arbitrary optical system that detects light emission from multiple light-emitting points by computational processing, thereby identifying light emission from multiple light-emitting points and detecting the light-emitting points independently. In addition, a method is proposed for reducing spatial crosstalk between multiple light-emitting points generated in an arbitrary optical system that detects fluorescence of multiple phosphors emitted from multiple light-emitting points by computational processing, as well as spectral crosstalk between multiple phosphors for each light-emitting point, thereby identifying fluorescence emission from multiple phosphors at multiple light-emitting points and detecting them independently. Alternatively, the following method is proposed: reducing spatial crosstalk between multiple light-absorbing points generated in an arbitrary optical system that detects absorption at multiple light-absorbing points by computational processing, thereby identifying absorption at multiple light-absorbing points and detecting them independently. In addition, the following method is proposed: by means of computational processing, spatial crosstalk between multiple absorption points and spectral crosstalk between multiple absorbers at each absorption point generated in an arbitrary optical system that separately detects the absorbance of multiple absorbers at multiple absorption points are reduced, thereby identifying the absorbance of multiple absorbers at multiple absorption points and independently detecting the absorbance of multiple types of absorbers at multiple absorption points.

[0042] Means for solving problems

[0043] In the optical system of Patent Document 1 or Patent Document 2, for each of A (A is an integer greater than or equal to 2) light-emitting points, the case of detecting the emission fluorescence of C types (C is an integer greater than or equal to 1) of phosphors in detection areas of B (B is an integer greater than or equal to 1) wavelength bands is described. Here, it is assumed that the detection areas of different wavelength bands detect different wavelength components of fluorescence. In addition, different types of phosphors emit fluorescence having different fluorescence spectra. For each of the light-emitting points P(a) (a=1, 2, ..., A), the emission fluorescence of the phosphor D(a, c) (c=1, 2, ..., C) present in the light-emitting point P(a) is detected in the detection areas W(a, b) (b=1, 2, ..., B) of different wavelength bands. At any time, the concentration of the phosphor D(a, c) at the light-emitting point P(a) is set to Z(a, c), and the signal intensity of the detection area W(a', b) for the light-emitting point P(a') is set to X(a', b). Here, the inventors discovered for the first time that, assuming that X is an A×B row and 1 column matrix with X(a', b) as the element, Z is an A×C row and 1 column matrix with Z(a, c) as the element, and Y is a (A×B) row and (A×C) column matrix with Y(a', b)(a, c) as the element, the following equation holds.

[0044] [Mathematical formula 9]

[0045] X=Y×Z (number 9)

[0046] [Formula 10]

[0047]

[0048] [Mathematical formula 11]

[0049]

[0050] [Mathematical formula 12]

[0051]

[0052] (Math 9) is the same as (Math 1), but if we compare (Math 1) to (Math 4) and (Math 9) to (Math 12), we can see that they are completely different. (Math 9) to (Math 12) are not only the relationship between b and c, but also the relationship between a, and different light-emitting points P(a) are related to each other. That is, in addition to the spectral crosstalk for the same light-emitting point P(a), (Math 9) to (Math 12) can also consider the spatial crosstalk and spectral crosstalk between different light-emitting points P(a), which is essentially different from (Math 1) to (Math 4).

[0053] Here, the element Y(a', b)(a, c) of the matrix Y of (A×B) rows and (A×C) columns indicates: (1) the ratio of the signal intensity of the fluorescence emitted by the phosphor D(a', c) in P(a') detected in any detection area W(a', b) by spectral crosstalk for a'=a, i.e., the same luminous point, and also indicates: (2) the ratio of the signal intensity of the fluorescence emitted by the phosphor D(a, c) at the luminous point P(a) detected in any detection area W(a', b) by spatial crosstalk and spectral crosstalk for a'≠a, i.e., different luminous points. In any luminous point P(a0), by causing any one type of phosphor D(a0, c0) to emit fluorescence alone, one column Y(a, b)(a0, c0) of the matrix Y (a=1, 2, ..., A, and b=1, 2, ..., B) can be determined. Here, it is generally difficult to control the concentration of the fluorescent substance D(a0, c0), so it is convenient to standardize one column Y(a, b)(a0, c0). For example, among A×B elements, the largest element is 1, and the other elements are expressed as the ratio relative to the maximum value. Alternatively, it is preferable to determine the ratio of each element in such a way that the sum of A×B elements is 1. In other words, let

[0054] [Mathematical formula 13]

[0055]

[0056] . Then, by sequentially performing the above steps for all combinations of C types of phosphors D(a, c) in A light-emitting points P(a), all columns of the matrix Y can be determined. The matrix Y is determined only by the characteristics of the light-emitting point P(a), the phosphor D(a, c), and the detection area W(a, b), and does not change while the electrophoresis analysis is in progress. In addition, as long as the conditions such as the optical system, the light-emitting point P(a), the phosphor D(a, c), and the detection area W(a, b) are fixed, the matrix Y remains fixed for different electrophoresis analyses. Therefore, for each light-emitting point, the concentration Z(a, c) of the phosphor D(a, c) at each moment is calculated by the following formula based on the signal intensity X(a, b) of the detection area W(a, b) at each moment.

[0057] [Formula 14]

[0058] Z=Y - ×X (Number 14)

[0059] (Mathematical formula 14) is the same as (Mathematical formula 6), but if (Mathematical formula 2) to (Mathematical formula 4) and (Mathematical formula 6), (Mathematical formula 10) to (Mathematical formula 12) and (Mathematical formula 14) are compared, it can be seen that the two are completely different. Through (Mathematical formula 14), for each moment, by multiplying the general inverse matrix Y- of the pre-calculated matrix Y with A×B signal intensities X(a, b), both spectral crosstalk and spatial crosstalk can be eliminated, and the concentrations Z(a, c) of A×C phosphors can be calculated. Y- is a matrix of (A×C) rows and (A×B) columns. In the present disclosure, eliminating spectral crosstalk is called color conversion, and correspondingly, eliminating spatial crosstalk is called spatial correction. In other words, the (A×C)×(A×B) elements of Y- include both a part that performs color conversion and a part that performs spatial correction. In the present disclosure, the time series data of Z(a, c) obtained by executing (Equation 14) at each time is referred to as color conversion + space correction data.

[0060] Each element of the matrix X may be a value obtained by subtracting background light in advance, or a value subjected to appropriate noise reduction processing. Similarly, each element of the matrix Y may be a value obtained by subtracting background light in advance, or an appropriate change may be added.

[0061] In the case of monochrome detection with B=1 and C=1, the above is simplified as follows. For the luminous point P(a) (a=1, 2, ..., A), the luminescent fluorescence of one kind of fluorescent body is detected in one detection area respectively. At any time, the concentration of the fluorescent body at the luminous point P(a) is set to Z(a), and the signal intensity for the luminous point P(a') is set to X(a'). Here, if X is a matrix with A rows and 1 columns with X(a') as the element, Z is a matrix with A rows and 1 columns with Z(a) as the element, and Y is a matrix with A rows and A columns with Y(a')(a) as the element, then (Formula 10) to (Formula 12) are simplified as follows. (Formula 9) and (Formula 14) are directly established.

[0062] [Mathematical formula 15]

[0063]

[0064] [Formula 16]

[0065]

[0066] [Mathematical formula 17]

[0067]

[0068] (Equation 9) and (Equation 14) to (Equation 17) are the same in form as the general formula used in Patent Document 3, such as [Equation 7] to [Equation 9] in Patent Document 3, but the contents are very different (hereinafter, the formula used in Patent Document 3 is recorded as []). The element αij of the matrix A in [Equation 7] is the luminous point and The function of the distance dij between each detection area is specifically represented by the sum of exponential functions of dij as shown in [Mathematical Formula 10], and αij decays as dij increases. In Patent Document 3, different fluorescence images are obtained for different samples, and in each fluorescence image, the positions of the detection areas of multiple luminous points vary randomly. Therefore, in Patent Document 3, the above function is obtained in advance, and for each fluorescence image, the distance between the detection areas of any two luminous points among the multiple luminous points is obtained for all combinations of two luminous points, and αij is derived by substituting into the above function. In other words, αij, that is, the matrix A, changes for each sample or each fluorescence image. In response to this, in the present disclosure, as described above or as described in [Specific Implementation], it was found that the element Y(a')(a) of the matrix Y of (Mathematical Formula 16) cannot be expressed as a function of the distance between the respective detection areas of the luminous points P(a) and P(a'). In addition, it was found that it is impossible to obtain the element Y(a')(a) by calculation depending on the structure of the optical system, etc. Therefore, in the present disclosure, as a condition that the positions of the detection areas of the plurality of luminous points do not change for different samples, the element Y(a')(a) is obtained by actual measurement under this condition. Specifically, as described above, fluorescence is emitted only at any one luminous point P(a0), thereby determining one column Y(a')(a0) (a'=1, 2, ..., A) of the matrix Y. Then, all the columns of the matrix Y are determined by sequentially performing the above steps on all the A luminous points P(a). The matrix Y is determined only by the characteristics of the luminous point P(a) and does not change during the analysis. In addition, as long as the conditions such as the optical system, the luminous point P(a) and its detection area are fixed, the matrix Y also remains fixed for the analysis of different samples. It is impossible to implement the above steps using the optical system or device structure of Patent Document 3. This is because the positions of the detection areas of the plurality of luminous points cannot be fixed, and even if it is assumed that they can be fixed, each luminous point cannot be made to emit light individually and sequentially. Therefore, the method of the present disclosure cannot be thought of from Patent Document 3.

[0069] The above can be restated as follows. That is, for each of A (A is an integer greater than or equal to 2) luminous points, the luminescence of C (C is an integer greater than or equal to 1) luminous bodies is detected in the detection area of ​​any B (B is an integer greater than or equal to 1) wavelength bands. For each of the luminous points P(a) (a=1, 2, ..., A), the luminescence of the luminous body D(a, c) (c=1, 2, ..., C) present at the luminous point P(a) is detected in the detection area W(a, b) (b=1, 2, ..., B) of different wavelength bands. At any time, the concentration of the luminous body D(a, c) at the luminous point P(a) is set to Z(a, c), and the luminous intensity of W(a', b) for P(a') is set to X(a', b). In this case, (Equation 9) to (Equation 17) hold, and similarly, both spectral crosstalk and spatial crosstalk can be eliminated to obtain the concentration Z(a, c) of A×C luminous bodies. Here, luminescence refers to fluorescence, phosphorescence, scattered light, and the like.

[0070] In addition, the above can also be restated as follows. That is, for each of A (A is an integer greater than or equal to 2) absorption points, the absorption of C types (C is an integer greater than or equal to 1) of absorbers is detected in detection areas of B (B is an integer greater than or equal to 1) arbitrary wavelength bands. For each of the absorption points P(a) (a=1, 2, ..., A), the absorption of the absorber D(a, c) (c=1, 2, ..., C) present at the absorption point P(a) is detected in detection areas W(a, b) (b=1, 2, ..., B) of different wavelength bands. At any time, the concentration of the absorber D(a, c) at the absorption point P(a) is set to Z(a, c), and the absorbance of W(a', b) with respect to the absorption point P(a') is set to X(a', b). In this case, (Equation 9) to (Equation 17) hold true, and similarly, both spectral crosstalk and spatial crosstalk can be eliminated, and the concentrations Z(a, c) of A×C light absorbers can be calculated.

[0071] Alternatively, the above can also be restated as multi-point detection other than optical measurement. That is, for each of A (A is an integer greater than or equal to 2) signal generating points, the signal of C types (C is an integer greater than or equal to 1) of signal generating bodies is detected in the detection area of ​​B (B is an integer greater than or equal to 1) arbitrary frequency bands. For each of the signal generating points P(a) (a=1, 2, ..., A), the signal of the signal generating object D(a, c) (c=1, 2, ..., C) present at the signal generating point P(a) is detected in the detection area W(a, b) (b=1, 2, ..., B) of different frequency bands. At any time, the concentration of the signal generating object D(a, c) at the signal generating point P(a) is set to Z(a, c), and the signal intensity of W(a', b) for the signal generating point P(a') is set to X(a', b). In this case, (Equation 9) to (Equation 17) hold true, and similarly, both spectral crosstalk and spatial crosstalk can be eliminated, and the concentrations Z(a, c) of A×C signal generators can be calculated.

[0072] The above description is mainly based on mathematical formulas, but this is used to facilitate understanding of the contents of the present disclosure. When implementing the technology of the present disclosure, it is sufficient to use a method based on the contents of the present disclosure, and it is not necessary to perform the method as a mathematical formula, and the mathematical formula may be deformed or not used. In addition, in the present disclosure, the description of "the concentration of the luminescent body" can be replaced by "the luminous intensity of the luminescent body", the description of "the concentration of the phosphor" can be replaced by "the fluorescence intensity of the phosphor", and the description of "the concentration of the light absorber" can be replaced by "the absorbance of the light absorber".

[0073] Further features of the present disclosure will become apparent from the description and drawings of this specification. In addition, the present disclosure is achieved and implemented through the combination of elements and various elements and the following detailed description and the appended claims.

[0074] The description in this specification is merely a typical example and does not limit the claims or application examples of the present disclosure in any sense.

[0075] Further features of the present disclosure will become apparent from the description and drawings of this specification. In addition, the present disclosure is achieved and implemented through the combination of elements and various elements and the following detailed description and the appended claims.

[0076] The description in this specification is merely a typical example and does not limit the claims or application examples of the present disclosure in any sense.

[0077] Effects of the Invention

[0078] According to the present disclosure, spatial crosstalk between multiple light-emitting points generated in an arbitrary optical system that detects light emitted from multiple light-emitting points is eliminated or reduced through computational processing, so that light emitted from multiple light-emitting points can be identified and detected independently. In addition, spatial crosstalk between multiple light-emitting points and spectral crosstalk between multiple phosphors about each light-emitting point that are generated in an arbitrary optical system that detects fluorescence emitted from multiple phosphors from multiple light-emitting points are eliminated or reduced through computational processing, so that fluorescence emission from multiple phosphors of multiple light-emitting points can be identified and detected independently.

[0079] Furthermore, by eliminating or reducing spatial crosstalk and spectral crosstalk, it is possible to avoid a reduction in detection sensitivity or a reduction in detection dynamic range that is caused by spatial crosstalk and spectral crosstalk.

[0080] Other problems, structures, and effects than those described above will become apparent from the following description of the embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1 It is a schematic diagram of a simple optical system.

[0082] Figure 2 It is a luminous image obtained by a two-dimensional sensor in a simple optical system.

[0083] Figure 3 is the signal intensity distribution of the luminescence image in the luminescence image.

[0084] Figure 4 It is the relationship between the absolute signal intensity at the center of the luminous image and the absolute signal intensity at a position away from the center of the luminous image to the left.

[0085] Figure 5 It is the relationship between the absolute signal intensity at the center of the luminous image and the absolute signal intensity at a position away from the center of the luminous image to the right.

[0086] Figure 6 It is the relationship between the absolute signal intensity at the center of the luminous image and the absolute signal intensity at a position away from the center of the luminous image to the left.

[0087] Figure 7 It is the relationship between the absolute signal intensity at the center of the luminous image and the absolute signal intensity at a position away from the center of the luminous image to the right.

[0088] Figure 8 is a schematic diagram of the model experimental system.

[0089] Fig. 9 Schematic diagram of spectral crosstalk and spatial crosstalk in the model experimental system.

[0090] Fig.10 is a schematic diagram of the model experimental system.

[0091] Fig.11 It is the raw data obtained through electrophoresis analysis of known samples.

[0092] Fig.12 Yes Fig.11 The original data is the color converted data to which the color conversion is applied.

[0093] Fig.13 Yes Fig.11 The original data has color conversion + space correction implemented on it.

[0094] Fig.14 It is the raw data obtained through electrophoresis analysis of unknown samples.

[0095] Fig.15 Yes Fig.14 The original data is the color converted data to which the color conversion is applied.

[0096] Fig.16 Yes Fig.14 The original data has color conversion + space correction implemented on it.

[0097] Fig.17 It is the raw data obtained by electrophoresis analysis of a sample including an unknown fluorescent substance.

[0098] Fig.18 Yes Fig.17 The original data has color conversion + space correction implemented on it.

[0099] Fig.19 These are raw data obtained by electrophoresis analysis of a sample in which the concentration of a known fluorescent substance is increased stepwise.

[0100] Fig. 20 Yes Fig.19 The original data is the color converted data to which the color conversion is applied.

[0101] Fig.21 Yes Fig.19 The original data has color conversion + space correction implemented on it.

[0102] Fig. 22 This is a schematic diagram of a method of injecting samples into a plurality of capillaries at staggered timings.

[0103] Fig.23 This is a schematic diagram of a method for obtaining fluorescent light emission by staggering the timing in a plurality of capillaries.

[0104] Fig.24 is a schematic diagram of a multi-capillary electrophoresis apparatus.

[0105] Fig.25 This is a wavelength dispersion image of Raman scattered light from four capillaries in the optical system of Patent Document 1.

[0106] Fig.26 Yes means Fig.25 Diagram of the setting of the 20 band detection areas of each capillary.

[0107] Fig. 27 This is a schematic diagram of the optical system of Patent Document 2 in which the emitted fluorescence from four capillaries is divided into four wavelength bands for imaging.

[0108] Fig.28 This is an optical system and schematic diagram that divides the fluorescent light emitted from five light-emitting points arranged on a plane into four bands for imaging.

[0109] Fig.29 is a flow chart of the existing method.

[0110] Fig.30 is a flow chart of the method.

[0111] Fig.31 It is a flowchart for determining the matrix for color conversion + spatial correction of the present method.

[0112] Fig.32 This is a flowchart for a case where a session of analysis is repeated multiple times.

[0113] Fig.33 It is a diagram of the computer structure. DETAILED DESCRIPTION

[0114] [Example 1]

[0115] In order to investigate the characteristics of spatial crosstalk in detail, a Figure 1 A simple optical system is shown. Figure 1 The optical system includes a pinhole plate 1-1, an opening plate 1-2 on the light-emitting point side, a focusing lens 1-3, an opening plate 1-4 on the sensor side, a color glass filter 1-5, a two-dimensional sensor 1-6, and a halogen lamp (light source) for irradiating a halogen light 1-7. Specifically, it is configured as follows Figure 1 Optical system. The pinhole plate 1-1 is illuminated by a halogen light 1-7 from below, thereby forming In addition, there will be The light-emitting point side opening plate 1-2 of the opening is arranged at a position 0.2 mm above the light-emitting point 1-8, and the focusing lens 1-3 with a focal length of f=1.4 mm is arranged at a position 1.54 mm above the light-emitting point 1-8. The sensor side aperture plate 1-4 with an opening is arranged just above the condensing lens 1-3. Furthermore, the two-dimensional sensor 1-6 is arranged 15 mm above the condensing lens 1-3, and the color glass filter 1-5 is arranged just below the two-dimensional sensor 1-6. In the above, the pinhole plate 1-1, the light-emitting point side aperture plate 1-2, the sensor side aperture plate 1-4, the color glass filter 1-5, and the two-dimensional sensor 1-6 are arranged parallel to each other. The light 1-9 emitted from the light-emitting point 1-8 passes through The light is focused by the focusing lenses 1-3 and passes through The opening of the colored glass filter 1-5 forms a Here, the luminous point 1-8 is imaged at a 10-fold magnification while being focused on the two-dimensional sensor 1-6.

[0116] Figure 2 is included in Figure 1 The luminous image of the image 1-10 obtained by the two-dimensional sensor 1-6 in the simple optical system. The sensor size of the two-dimensional sensor 1-6 is 13×13 mm, and the signal range of each pixel is 0 to 65536. Figure 2 (a) and Figure 2 (b) is the same luminescence image, but Figure 2 The signal display scale (gray scale) of (a) is 0 to 50000. Figure 2 The signal display scale of (b) is 0 to 500. The maximum signal value of the luminescent image is about 50000. Figure 2 In (a), the signals of all pixels are almost unsaturated. Figure 2 The luminous image in (b) is saturated. Figure 2 (a), as expected The luminous image of The outer side of the luminous image can be regarded as signal zero. Figure 2 (b) can confirm that The luminous image extends to the outside of the low-intensity hem.

[0117] Figure 3 The α (dotted line) of (a) to (c) indicates Figure 2 The signal intensity distribution in the horizontal direction passing through the center of the luminescence image in the luminescence image. Figure 3 (a) Figure 3 (b) and Figure 3(c) uses the same data, but changes the vertical axis. The horizontal axis is common and represents the distance from the center of the luminescent image. The positive and negative values ​​of the horizontal axis represent the right and left sides of the luminescent image in the luminescent image, respectively. Figure 3 The vertical axis of (a) and (b) represents the absolute signal strength. Figure 3 (a) indicates that the vertical axis scale is 0 to 60000. Figure 3 (b) indicates that the vertical axis scale is 0 to 100. On the other hand, Figure 3 (c) The vertical axis is set to a logarithmic scale, and the vertical axis scale is set to 0.001% to 100% as the relative signal intensity with the maximum signal intensity being 100%. Figure 3 The β (solid line) and γ (dashed line) of (a) to (c) are shown in Figure 2 Under the condition of obtaining the luminous image of , the output intensity of the halogen lamps 1-7 is gradually reduced, and the signal intensity distribution is shown in the same way as α. Figure 3 From (a), we can see that the maximum signal intensities of the luminescent images of α, β, and γ are approximately 50,000, approximately 25,000, and approximately 10,000, respectively. Figure 3 In (b), it can be seen that the signal intensity tends to decrease as it moves away from the center of the luminous image, but it is different from Figure 3 Compared with the size of the luminous image observed in (a) (width of about 0.5mm), it has been expanding with a large underside. In addition, it can be seen that when the maximum signal intensity decreases, the intensity of the underside also decreases. Therefore, when observing Figure 3 When (c) is taken, it is known that α, β, and γ overlap into one. This is a new discovery and brings some important insights. For example, at a distance of ±1 mm from the center of the luminous image, Figure 2 At the position outside the luminescent image shown in (a), a signal intensity of about 0.1% of the maximum signal intensity was observed. Assuming that there is an adjacent luminescent image detection area at a distance of ±1 mm, this means that there is about 0.1% spatial crosstalk. Figure 1 As shown in FIG. 1 , it is surprising that despite the use of an extremely simplified optical system to focus the light-emitting point, there is such a spatial crosstalk of a size that cannot be ignored. Even more surprising is that since α, β, and γ overlap into one, as long as the optical system and the position of the light-emitting point are fixed, regardless of the light intensity of the light-emitting point or the signal intensity of the imaging point, it is determined that the relative signal intensity distribution of the imaging point is constant.

[0118] Figure 4 (a) and Figure 5 (a) is from Figure 3 The exported results. Figure 4 (a) is the position -1, -2, -3, -4, -5, -6 mm away from the center of the imaging point. Figure 5 (a) is a graph in which the absolute signal intensity at the position +1, +2, +3, +4, +5, and +6 mm away from the center of the imaging point is set as the vertical axis, and the absolute signal intensity at the center of the luminous image is set as the horizontal axis in any case. Figure 3 The results show that the three plots at each position (with Figure 3 α, β, γ corresponding to the image point) are all on an approximate straight line passing through the origin. The slope of each approximate straight line represents the spatial crosstalk ratio at the corresponding position. In other words, it can be seen that as the distance from the center of the imaging point increases, the slope of the approximate straight line decreases and the spatial crosstalk ratio decreases. For comparison, straight lines with spatial crosstalk of 0.1% and 0.01% are overlapped and displayed as dotted lines. For example, the spatial crosstalk ratio at a position -1mm away from the center of the imaging point is read as being strong at 0.1%, while the spatial crosstalk ratio at a position -3mm away is about 0.01%. The above results indicate that the absolute signal intensity of the spatial crosstalk at any position away from the center of the imaging point is linear with respect to the absolute signal intensity at the center of the imaging point, and is independent of the absolute signal intensity at the center of the luminous image, and the spatial crosstalk ratio is constant. That is, it has been newly discovered that the spatial crosstalk at any position can be eliminated or reduced by subtracting the value obtained by multiplying the absolute signal intensity at the center of the imaging point by the spatial crosstalk ratio from the absolute signal intensity at the center of the imaging point. This is equivalent to Figure 4 (a) and Figure 5 The absolute signal intensity of each plot in (a) is subtracted from the value of the corresponding approximate straight line at the same position.

[0119] Figure 4 (b) and Figure 5 (b) are respectively Figure 4 (a) and Figure 5 The result of the above subtraction operation is the result of the result of (a). The type of drawing and the horizontal axis representing each position are shown in Figure 4 (a) and Figure 4 (b) Figure 5 (a) and Figure 5 (b) is common. Figure 4 (b) and Figure 5 The vertical axis of (b) represents the absolute signal strength after the above subtraction operation, which is Figure 4 (a) and Figure 5The scale is enlarged compared to (a). For comparison, a straight line with a spatial crosstalk of ±0.01% is overlapped with a dotted line. As a result, it is clear that the absolute signal intensity after subtraction at each position is independent of the absolute signal intensity at the center of the image and is approximately zero, and the spatial crosstalk at each position is less than ±0.01%. If the comparison is made at a position -1mm away from the center of the imaging point, the spatial crosstalk can be reduced by at least one digit by this method. The reason why the absolute signal intensity after subtraction at each position is not completely zero is that there is Figure 4 (a) and Figure 5 The deviation and error of the approximate straight line corresponding to each plot in (a). The coefficient of determination (R 2 ), that is, the higher the linearity of the absolute signal intensity at each position relative to the absolute signal intensity at the center of the imaging point, the smaller the above error, and the closer the absolute signal intensity after the subtraction operation is to zero. Of course, the absolute signal intensity after the subtraction operation sometimes becomes negative. However, as long as the above linearity exists, the magnitude of the absolute signal intensity at each position is at least reduced by the subtraction operation. In addition, the smaller the slope of the approximate straight line, that is, the smaller the spatial crosstalk before the subtraction operation, the smaller the absolute value of the difference between the absolute signal intensity at each position and the approximate straight line, and therefore the smaller the above error.

[0120] Figure 6 as well as Figure 7 is with Figure 4 as well as Figure 5 The same experimental results. Figure 1 In the optical system, temporarily remove the pinhole plate 1-1 and install it again to obtain the same Figure 2 and Figure 3 Same results, derived by the same method Figure 6 as well as Figure 7 The installation position of the pinhole plate 1-1 is roughly the same as the original position, but strictly speaking, it is not exactly the same position. Figure 6 as well as Figure 7 The results and Figure 4 as well as Figure 5 The results are the same, indicating that the method of eliminating or reducing spatial crosstalk at any position through subtraction processing has high reproducibility.

[0121] On the other hand, according to Figures 4 to 7 A comparison of , as shown below, yields another important insight. Figure 4 (a) -1mm, Figure 5 (a) +1mm, Figure 6 (a) -1mm and Figure 7The slopes of these four approximate lines all represent the spatial crosstalk ratio at a position 1 mm away from the center of the imaging point, but they are different from each other. Therefore, for example, it can be seen that if Figure 4 (a) is an approximate straight line of -1 mm. Figure 5 (a) +1mm, Figure 6 (a) -1mm, or Figure 7 If the subtraction operation of each drawing of (a) +1mm is performed, the above error becomes larger and the reduction of spatial crosstalk becomes insufficient. It can be seen that this phenomenon also occurs in other positions. Depending on the situation, it may sometimes lead to the opposite effect of the subtraction operation, that is, increase the spatial crosstalk. The important factors that cause this phenomenon are (1) Figure 1 In such a simple optical system, the point symmetry of the spatial crosstalk caused by imaging is not necessarily high (for example, in Figure 2 (1) The spatial crosstalk ratios in the left and right directions of the imaging point are different), and (2) the spatial crosstalk caused by imaging changes significantly with subtle changes in the optical system such as a slight deviation in the position of the light-emitting point. Therefore, it is clear that the spatial crosstalk ratio generated between the center of the imaging point and an arbitrary position cannot be expressed as a function of the distance between the two. This means that the method shown in Patent Document 3 will not function effectively. In the present disclosure, the difference from Patent Document 3 is that the mutual spatial crosstalk ratios are derived experimentally under the conditions of a fixed optical system and multiple imaging points, that is, multiple detection areas, regardless of the distance between each other, such as Figures 4 to 7 If the conditions change, the spatial crosstalk ratio needs to be re-obtained each time.

[0122] The insights gained from the above experiments were expanded upon, namely, the absolute signal intensity of the spatial crosstalk at any position away from the center of the imaging point is linear with respect to the absolute signal intensity at the center of the imaging point, and the fact that the spatial crosstalk ratio is constant, if generalized, leads to (Equation 9) to (Equation 17).

[0123] [Example 2]

[0124] A model experiment was conducted for the simplest case of an optical system that detects fluorescence emitted from multiple types of phosphors from multiple light-emitting points in detection areas of multiple wavelength bands. In this embodiment and other embodiments using the same model experiment system, as an example, in (Equation 9) to (Equation 12), for A=2 light-emitting points, the case of detecting C=2 types of fluorescence emitted by phosphors in detection areas of B=2 wavelength bands was selected, but it goes without saying that the same effect can be obtained by the same method when A, B, and C are other values.

[0125] Figure 8 Represents the model experimental system. Figure 8 The model experimental system comprises two capillaries Cap (1) and Cap (2) and a light source (not shown) for irradiating a laser beam LB in the arrangement direction of the two capillaries Cap (1) and Cap (2). Light-emitting points P (1) and P (2) are respectively provided on the two capillaries Cap (1) and Cap (2) at the positions irradiated with the laser beam LB. Two substances labeled with fluorescent substances D (1, 1) and D (1, 2) are electrophoresed in the capillary Cap (1), and two substances labeled with fluorescent substances D (2, 1) and D (2, 2) are electrophoresed in the capillary Cap (2). Here, D (1, 1) and D (2, 1) are the same type of fluorescent substances, and D (1, 2) and D (2, 2) are the same type of fluorescent substances. The same type of fluorescent substances means that the fluorescent spectra of the fluorescent substances emitted are equal. The phosphors D(1,1) and D(1,2) emit fluorescence at the light-emitting point P(1) by the irradiation of the laser beam LB, and are detected by the detection areas W(1,1) and W(1,2) provided in the sensor S. Although the detection bands are respectively designed so that the detection area W(1,1) mainly detects the fluorescence emitted by the phosphor D(1,1) and the detection area W(1,2) mainly detects the fluorescence emitted by the phosphor D(1,2), there is a significant spectral crosstalk between them. Similarly, the phosphors D(2,1) and D(2,2) emit fluorescence at the light-emitting point P(2) by the irradiation of the laser beam LB, and are detected by the detection areas W(2,1) and W(2,2) provided in the sensor S. Although the detection bands are respectively designed so that the detection area W(2,1) mainly detects the fluorescence emitted by the phosphor D(2,1) and the detection area W(2,2) mainly detects the fluorescence emitted by the phosphor D(2,2), there is a significant spectral crosstalk between them. In addition, the fluorescence of the fluorescent bodies D(1,1) and D(1,2) at the light-emitting point P(1) is also detected in the detection areas W(2,1) and W(2,2). Similarly, the fluorescence of the fluorescent bodies D(2,1) and D(2,2) at the light-emitting point P(2) is also detected in the detection areas W(1,1) and W(1,2). Spatial crosstalk is significant. If the signal intensities (fluorescence intensities) in the detection areas W(1,1), W(1,2), W(2,1) and W(2,2) are X(1,1), X(1,2), X(2,1) and X(2,2), then Figure 8 As shown on the lower side, for each of the luminescent point P(1) and the luminescent point P(2), time series data of signal intensities X(1,1) and X(1,2), X(2,1) and X(2,2) are obtained along with electrophoresis.

[0126] In addition, Figure 8In the figure, the detection areas W(1, 1), W(1, 2), W(2, 1), and W(2, 2) are depicted as being arranged on one sensor S, but this is not necessary. The detection areas W(1, 1) and W(1, 2) may be arranged on one sensor, the detection areas W(2, 1) and W(2, 2) may be arranged on another sensor, or the detection areas W(1, 1), W(1, 2), W(2, 1), and W(2, 2) may be arranged on four different sensors, respectively. In addition, some imaging unit and spectroscopic unit are required between the light-emitting points P(1) and P(2) and the sensor S, but in the present embodiment, any imaging unit and spectroscopic unit may be used. Figure 8 The imaging unit and the spectroscopic unit are omitted.

[0127] Fig. 9 Yes means Figure 8 Schematic diagram of the relationship between spectral crosstalk and spatial crosstalk. Fig. 9 In (a), the fluorescence emitted by the fluorescent body D(1, 1) at the luminous point P(1) is mainly detected in (i) the detection area W(1, 1), and (ii) is detected as an auxiliary in the detection area W(1, 2). In addition, the above-mentioned luminous fluorescence is also detected in (iii) the detection area W(2, 1) with an intensity that is always smaller than (i) and (ii), and further, it is also detected in (iv) the detection area W(2, 2) with an intensity less than (iii). (i) and (ii) represent spectral crosstalk, and (iii) and (iv) represent spatial crosstalk and spectral crosstalk. In other words, only (i) and (ii) are considered in the existing method, but all of (i) to (iv) are considered in the present disclosure. Fig. 9 In FIG. 1 , the thickness of the arrows, the solid or dotted lines, and the numbers (i) to (iv) are used to show the relationship between the strength of the above detection intensity and the relationship between the crosstalk. Fig. 9 (a) Similarly, Fig. 9 (b) depicts the fluorescence emission of phosphor D(1, 2) at the emission point P(1), Fig. 9 (c) depicts the fluorescence emission of the phosphor D(2, 1) at the emission point P(2), Fig. 9 (d) depicts the fluorescence emission of the phosphor D(2, 2) at the luminous point P(2). In the actual analysis, Fig. 9 (a) to (d) are generated at the same time, and their fluorescence intensities are different.

[0128] Fig.10 Representation and Figure 8 The same model experimental system. However, in Fig.10In the model experimental system, light LL is irradiated from a light source perpendicular to the arrangement direction of two capillaries Cap (1) and Cap (2). Therefore, the luminous fluorescence associated with the irradiation of the laser beam LB is not detected, but the absorption or absorbance associated with the transmission of the light LL is detected. On the two capillaries Cap (1) and Cap (2), absorption points P (1) and P (2) are respectively provided at the positions where the light LL is irradiated. Two types of light absorbers D (1, 1) and D (1, 2) are electrophoresed in the capillary Cap (1), and two types of light absorbers D (2, 1) and D (2, 2) are electrophoresed in the capillary Cap (2). Here, D (1, 1) and D (2, 1) are the same type of light absorbers, and D (1, 2) and D (2, 2) are the same type of light absorbers. The same type of light absorbers means that the light absorption spectra are equal. The light absorbers D(1,1) and D(1,2) absorb light at the light absorption point P(1) by the irradiation of the light source LL, and the light not absorbed is detected by the detection areas W(1,1) and W(1,2) provided in the sensor S. Although the detection wavelength bands are respectively designed so that the detection area W(1,1) mainly detects the light absorption of the light absorber D(1,1) and the detection area W(1,2) mainly detects the light absorption of the light absorber D(1,2), there is a significant spectral crosstalk between the two. Similarly, the light absorbers D(2,1) and D(2,2) absorb light at the light absorption point P(2) by the irradiation of the light source LL, and the light not absorbed is detected by the detection areas W(2,1) and W(2,2) provided in the sensor S. Although the detection bands are designed so that the detection area W(2,1) mainly detects the absorption of the absorber D(2,1) and the detection area W(2,2) mainly detects the absorption of the absorber D(2,2), there is significant spectral crosstalk between them. In addition, the absorption of the absorbers D(1,1) and D(1,2) at the absorption point P(1) is also detected in the detection areas W(2,1) and W(2,2), and similarly, the absorption of the absorbers D(2,1) and D(2,2) at the absorption point P(2) is also detected in the detection areas W(1,1) and W(1,2), and there is significant spatial crosstalk. This is because part of the light transmitted by the lamp LL passing through the light absorption point P(1) is also detected in the detection areas W(2,1) and W(2,2), and part of the light transmitted by the lamp LL passing through the light absorption point P(2) is also detected in the detection areas W(1,1) and W(1,2). If the absorbance in the detection areas W(1,1), W(1,2), W(2,1), and W(2,2) is X(1,1), X(1,2), X(2,1), and X(2,2), then Fig.10 As shown below, for each of the absorption points P(1) and P(2), along with electrophoresis, time series data of absorbance X(1,1), X(1,2), X(2,1), and X(2,2) are obtained. Figure 8 However, when using Fig.10 It goes without saying that the same effect can be achieved in the same situation.

[0129] Fig.11 is used Figure 8 The raw data obtained by injecting known samples into the capillaries Cap (1) and Cap (2) and analyzing them by electrophoresis in the model experimental system. The horizontal axis is set to electrophoresis time (arbitrary unit) and the vertical axis is set to fluorescence intensity (arbitrary unit). Fig.11 (a) and Fig.11 (b) is the same raw data obtained at the light-emitting point P(1), and (b) is a diagram after the vertical axis scale of (a) is enlarged. Fig.11 (c) and Fig.11 (d) is the same raw data obtained at the light emitting point P(2), Fig.11 (d) is to Fig.11 (c) is a graph with the vertical axis scale enlarged. Fig.11 The horizontal axis scales of (a) to (d) represent time 0 to 500. Fig.11 The vertical scale of (a) and (c) represents the fluorescence intensity from 0 to 250. Fig.11 The vertical scale of (b) and (d) represents the fluorescence intensity from -0.2 to 0.8. X(1, 1) and X(2, 1) are represented by solid lines, and X(1, 2) and X(2, 2) are represented by dotted lines. Fig.11 As shown, the sample is prepared so that at time 100, the phosphor D(1, 1) emits fluorescence alone at the light-emitting point P(1) ( Fig. 9 (a) state), at time 200, the fluorescent body D (2, 1) emits fluorescence alone at the light-emitting point P (2) ( Fig. 9 (c) state), at time 300, the fluorescent body D (1, 2) emits fluorescence alone at the light-emitting point P (1) ( Fig. 9 (b) state), at time 400, the fluorescent body D (2, 2) emits fluorescence alone at the light-emitting point P (2) ( Fig. 9 Then, the sample is prepared so that no fluorescence other than these is emitted. Fig.11 (a) and Fig.11 (c), only four large peaks corresponding to the above-mentioned fluorescence emission were observed (with Fig. 9 (i) (ii) corresponds to), but observe Fig.11 (b) and Fig.11(d), four small peaks indicated by arrows, i.e., at time 100, phosphor D(2, 1) is observed at the luminous point P(2), at time 200, phosphor D(1, 1) is observed at the luminous point P(1), at time 300, phosphor D(2, 2) is observed at the luminous point P(2), and at time 400, phosphor D(1, 2) is observed at the luminous point P(1) (same as Fig. 9 (iii) (iv) of FIG. 100). Therefore, these four small peaks can be judged as peaks based on spatial crosstalk and spectral crosstalk. For example, the small peaks of the phosphors D(2, 1) and D(2, 2) at the luminous point P(2) at time 100 are the result of a part of the fluorescent light emission of the phosphor D(1, 1) at the luminous point P(1) at time 100 being detected as X(2, 1) and X(2, 2) through spatial crosstalk and spectral crosstalk (corresponding to FIG. 100). Fig. 9 (iii)(iv) corresponding to (a).

[0130] Without considering the spatial crosstalk, that is, ignoring Fig.11 (c) and Fig.11 (d) of the case of 4 small peaks (in Fig. 9 In the case of only considering (i) and (ii)), as long as the execution Fig.11 (a) and Fig.11 The spectral crosstalk of the four large peaks of (c) can be eliminated, that is, the color conversion based on the existing method can be performed. In this case, (Mathematical formula 6) is expressed as follows.

[0131] [Mathematical formula 18]

[0132]

[0133] [Mathematical formula 19]

[0134]

[0135] Here, Z(1,1), Z(1,2), Z(2,1), and Z(2,2) represent the concentrations of the phosphors D(1,1), D(1,2), D(2,1), and D(2,2) at each time. As shown in (Equation 18) and (Equation 19), in this embodiment, the 2-row 2-column matrix Y and the inverse matrix Y -1 At the light-emitting point P(1) and the light-emitting point P(2), the matrix Y and the inverse matrix Y are the same. However, in general, sometimes at the light-emitting point P(1) and the light-emitting point P(2), the matrix Y and the inverse matrix Y are different. -1Each element of the matrix Y for the light-emitting point P(1) is determined by the intensity ratio of the fluorescence intensities X(1,1) and X(1,2) of the fluorescent body D(1,1) at time 100, and the intensity ratio of the fluorescence intensities X(1,1) and X(1,2) of the fluorescent body D(1,2) at time 300. Each element of the matrix Y for the light-emitting point P(2) is determined by the intensity ratio of the fluorescence intensities X(2,1) and X(2,2) of the fluorescent body D(2,1) at time 200, and the intensity ratio of the fluorescence intensities X(2,1) and X(2,2) of the fluorescent body D(2,2) at time 400.

[0136] Fig.12 It is for Fig.11 The original data is converted into color data by performing the color conversion of (Equation 18) and (Equation 19) at each time. Fig.11 Same. As expected, Fig.11 (a) and Fig.11 (c) The spectral crosstalk of the four large peaks is Fig.12 (a) and Fig.12 The four large peaks of (c) are eliminated. On the other hand, Fig.11 (b) and Fig.11 The spectral crosstalk of the four small peaks in (d) is also reflected by Fig.12 (c) and Fig.12 The four small peaks (similarly indicated by arrows) in (d) are eliminated, but these peaks themselves still remain, which shows that the spatial crosstalk has not been eliminated. This is a problem of the conventional method.

[0137] Therefore, next, for Fig.11 , considering both spectral crosstalk and spatial crosstalk (in Fig. 9 In the above, all (i) to (iv) are considered, and both color conversion and space correction are performed. At this time, (Equation 14) is expressed as follows.

[0138] [Mathematical formula 20]

[0139]

[0140] Matrix Y and inverse matrix Y -1 The 2 rows and 2 columns of (Equation 18) and (Equation 19) are expanded to 4 rows and 4 columns. In (Equation 18) and (Equation 19), the color conversion is performed independently at the light-emitting point P(1) and the light-emitting point P(2), whereas in (Equation 20), the light-emitting point P(1) and the light-emitting point P(2) are mixed and the color conversion + spatial correction is performed at the same time. The elements of the matrix Y of (Equation 20) are calculated according to Fig.11The intensity ratio of the fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) of the fluorescent body D(1,1) at time 100, the intensity ratio of the fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) of the fluorescent body D(2,1) at time 200, the intensity ratio of the fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) of the fluorescent body D(1,2) at time 300, and the intensity ratio of the fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) of the fluorescent body D(2,2) at time 400. The upper left 2 rows and 2 columns and the lower right 2 rows and 2 columns of the matrix Y of (Equation 20) correspond to the matrices Y of (Equation 18) and (Equation 19), and the values ​​of the respective elements are substantially equal to each other. Similarly, the matrix Y of (Equation 20) -1 The upper left 2 rows and 2 columns and the lower right 2 rows and 2 columns and the matrix Y of (Formula 18) and (Formula 19) -1 Correspondingly, the values ​​of each element are approximately equal to each other. In other words, these elements are responsible for color conversion that eliminates spectral crosstalk for the same light-emitting point. Here, the reason why the values ​​of these elements are not completely consistent in (Equation 20) and (Equation 18) and (Equation 19) is the difference in whether or not to consider spatial crosstalk as described below. In contrast, the matrix Y of (Equation 20) and the matrix Y -1 The upper right 2 rows and 2 columns, and the lower left 2 rows and 2 columns do not exist in (Equation 18) and (Equation 19), and are responsible for eliminating the spatial crosstalk between different light-emitting points and the spatial correction of spectral crosstalk and color conversion. (Equation 20) is easier to understand if it is transformed as follows.

[0141] [Mathematical formula 21]

[0142]

[0143] [Mathematical formula 22]

[0144]

[0145] The first term on the right side of (Equation 21) and the second term on the right side of (Equation 22) are responsible for the color conversion of the existing method, and correspond to (Equation 18) and (Equation 19) respectively. In contrast, the second term on the right side of (Equation 21) and the first term on the right side of (Equation 22) are responsible for spatial correction and color conversion that are not processed in the existing method. In this way, color conversion, spatial correction, and color conversion can be performed separately or only one of them can be performed. Or, for example, spatial correction and color conversion can be performed on the light-emitting point P(1), and only color conversion can be performed on the light-emitting point P(2), and the processing can be changed by the light-emitting point.

[0146] Fig.13 It is for Fig.11 The original data is converted into color + space correction data by performing the color conversion + space correction of (Equation 20) at each time. Fig.11 First, with Fig.12 Similarly, it can be seen Fig.11 (a) and Fig.11 (c) The spectral crosstalk of the four large peaks is Fig.13 (a) and Fig.13 The four large peaks of (c) are eliminated. Furthermore, as expected, Fig.11 (b) and Fig.11 The spatial crosstalk and spectral crosstalk of the four small peaks in (d) are also Fig.13 (c) and Fig.13 d), it can be seen that the peak disappears (indicated by the arrow).

[0147] Based on the above, according to the present disclosure, it is shown that spectral crosstalk for the same light-emitting point, as well as spatial crosstalk and spectral crosstalk between multiple light-emitting points, generated in any optical system that separately detects the fluorescence of multiple phosphors emitted from multiple light-emitting points can be eliminated or reduced through computational processing.

[0148] [Example 3]

[0149] Next, based on the experimental results of [Example 2], an analysis of unknown samples was performed. Fig.14 is used Figure 8 The model experimental system is used to inject unknown samples into capillaries Cap (1) and Cap (2) for electrophoresis analysis to obtain the raw data. Since the same model experimental system as in [Example 2] is used, (Equation 18) to (Equation 22) can be used directly. Fig.11 Same. However, Fig.11 The vertical axis scale of (b) and (d) is reduced to fluorescence intensity -0.5 to 2.0. At time 100, 200, 300, and 400, a large peak is observed at the luminescent point P(1), while a small peak (indicated by arrows) is observed at the luminescent point P(2).

[0150] Fig.15 It is for Fig.14 The original data is converted into color data by performing the color conversion of (Equation 18) and (Equation 19) at each time. Fig.14 Same. Fig.15As shown in (a), at the light-emitting point P(1), the light-emitting fluorescence of the phosphor D(1,1) is detected at time 100 and 300, and the light-emitting fluorescence of the phosphor D(1,2) is detected at time 200 and 400. Fig.15 As shown in (d), the original nature of the four small peaks (indicated by arrows) detected at time 100, 200, 300, and 400 at the luminescent point P(2) is unclear. In other words, it is unclear whether they are derived from (1) the fluorescence of the mixture of phosphors D(2,1) and D(2,2) at the luminescent point P(2), (2) the fluorescence of impurities other than phosphors D(2,1) and D(2,2) at the luminescent point P(2), or (3) the spatial crosstalk of the fluorescence of phosphors D(1,1) and D(1,2) at the luminescent point P(1).

[0151] Fig.16 It is for Fig.14 The original data is converted into color + space correction data by performing the color conversion + space correction of (Equation 20) at each time. Fig.14 Same. Fig.16 As shown in (a), Fig.15 (a) Similarly, at the light-emitting point P(1), the light-emitting fluorescence of the phosphor D(1,1) is detected at time 100 and 300, and the light-emitting fluorescence of the phosphor D(1,2) is detected at time 200 and 400. In addition, Fig.16 As shown in (d), it is found that as a result of the elimination of spatial crosstalk and spectral crosstalk of the detection areas W(2,1) and W(2,2) for the fluorescence emission of the phosphors D(1,1) and D(1,2) at the luminescent point P(1), the weak fluorescence emission of the phosphor D(2,2) is detected alone at the time 100 and 300, and the weak fluorescence emission of the phosphor D(2,1) is detected alone at the time 200 and 400 at the luminescent point P(2). The peak intensity of these weak fluorescence emission is weaker than 1% of the peak intensity of the fluorescence emission observed at the luminescent point P(1). On the other hand, Fig.12 The results show that in Figure 8 The spatial crosstalk ratio generated in the model experimental system is also weaker than 1%. Fig.15 In (d), the real weak luminescence fluorescence and the pseudo weak luminescence fluorescence caused by spatial crosstalk coexist, and it is impossible to distinguish between the two.

[0152] The above results show that spatial crosstalk can push up the lower limit of detection in the detection of luminescence from each luminescent point. As in the above example, when the spatial crosstalk ratio is set to 1%, assuming that the lower limit of detection is 0.1% when there is only one luminescent point, it is impossible to distinguish whether the signal below 1% is a real signal or a false signal caused by spatial crosstalk. Therefore, the effective lower limit of detection rises to 1%. In other words, compared with the luminescence detection of one luminescent point, the detection sensitivity and dynamic range are reduced by one digit in the luminescence detection of multiple luminescent points. The present disclosure can solve such a problem and avoid the reduction of detection sensitivity and dynamic range in the luminescence detection of multiple luminescent points.

[0153] [Example 4]

[0154] Next, based on the experimental results of [Example 2], electrophoresis analysis was performed on a substance labeled with an unknown phosphor D(1,3) together with phosphors D(1,1) and D(1,2) at the luminescent point P(1). Fig.17 is used Figure 8 In the model experimental system, only the sample is injected into the capillary Cap (1), and no sample is injected into the capillary Cap (2). The raw data obtained by electrophoresis analysis of both capillaries Cap (1) and Cap (2). Fig.14 The sample is prepared so that at time 100, the phosphor D(1,1) alone emits fluorescence at the light-emitting point P(1), at time 200, the phosphor D(1,2) alone emits fluorescence at the light-emitting point P(1), at time 300, the phosphor D(1,1) alone emits fluorescence at the light-emitting point P(1), and at time 400, the phosphor D(1,3) alone emits fluorescence at the light-emitting point P(1). Then, the sample is prepared so that the other fluorescent substances do not emit. Fig.17 As shown in (a), only four large peaks corresponding to the above-mentioned fluorescence emission are observed. It can be seen that the spectral crosstalk ratio of phosphor D (1, 3) is similar to the spectral crosstalk ratio of phosphor D (1, 2), but slightly different. On the other hand, Fig.17 The four small peaks indicated by arrows in (d) can be judged as Fig.17 (a) The spatial and spectral crosstalk of the luminescent fluorescence from the four large peaks.

[0155] Fig.18 It is for Fig.17 The original data is converted into color + space correction data by performing the color conversion + space correction of (Equation 20) at each time. Fig.14 Same. Fig.18As shown in (a), at the light-emitting point P(1), the fluorescence emitted by the phosphor D(1,1) is detected separately at time 100 and 300, and the fluorescence emitted by the phosphor D(1,2) is detected separately at time 200. However, as shown by the arrow, the fluorescence emitted by the phosphor D(1,3) is detected separately at time 400, but since the spectral crosstalk ratio is different from that of the phosphor D(1,1) and the phosphor D(1,2), the spectral crosstalk is not eliminated. On the other hand, Fig.18 As shown in (d), it is found that although the spatial crosstalk and spectral crosstalk caused by the fluorescence emission of the phosphors D(1,1) and D(1,2) at the light-emitting point P(1) at times 100, 200, and 300 are eliminated, as shown by the arrows, the spatial crosstalk and spectral crosstalk caused by the fluorescence emission of the phosphor D(1,3) at the light-emitting point P(1) at time 400 are not eliminated, and a peak remains. This is because in [Example 2], the fluorescence emission of the phosphors D(1,1) and D(1,2) at the light-emitting point P(1) and the fluorescence emission of the phosphors D(2,1) and D(2,2) at the light-emitting point P(2) are derived by (Formula 20), and the spatial crosstalk and spectral crosstalk characteristics of the fluorescence emission of D(1,3) at the light-emitting point P(1) are different from (Formula 20). Therefore, for the fluorescence emission of the phosphor D(1,3) at the light-emitting point P(1), the above problem can be solved by re-obtaining (Formula 20). The above phenomenon represents one aspect of the present disclosure.

[0156] [Example 5]

[0157] This time, based on the experimental results of [Example 2], an experiment was conducted to perform fluorescence emission by gradually increasing the concentration of a substance labeled with phosphor D(1,1) at the luminescent point P(1). Fig.19 is used Figure 8 The original data obtained by electrophoresis analysis of the capillary Cap (1) and the capillary Cap (2) is shown in the model experimental system. Fig.14 Same. However, Fig.19 The vertical scale of (a) and (c) is reduced to fluorescence intensity 0 to 400. The sample is prepared so that at each time 100, 200, 300, and 400, the fluorescent body D (1, 1) emits fluorescence at the light emitting point P (1) alone, and the concentration and fluorescence intensity of the fluorescent body D (1, 1) increase step by step. In addition, the sample is prepared so that the fluorescent body other than these does not emit. Fig.19 As shown in (a), four large peaks corresponding to the above-mentioned fluorescence emission are observed, but due to Figure 8The sensor S used in the model experimental system is saturated at a fluorescence intensity of 200, so the three peaks at time 200, 300, and 400 are saturated. Fig.19 The four small peaks caused by the spatial crosstalk and spectral crosstalk of the fluorescence emitted by the phosphor D(1, 1) at the light-emitting point P(1) shown in (d) are all observed in an unsaturated state due to their low fluorescence intensity.

[0158] Fig. 20 It is for Fig.19 The original data is converted into color data by performing the color conversion of (Equation 18) and (Equation 19) at each time. Fig.19 Same. Fig. 20 As shown in (a), the spectral crosstalk of the fluorescent light emitted by the phosphor D(1,1) at time 100 in the luminous point P(1) is eliminated and detected separately. However, since the fluorescent light emitted by the phosphor D(1,1) at time 200, 300, and 400 is saturated, the spectral crosstalk is not eliminated. Fig.19 As can be seen from (a), this is because when the fluorescence intensity is saturated, the spectral crosstalk ratio derived from the intensity ratio of X(1,1) and X(1,2) changes and deviates from the spectral crosstalk ratio defined by (Equation 18) and (Equation 19). Fig. 20 As shown in (d), since the fluorescence intensity of the phosphor D(2, 1) is not saturated, the spectral crosstalk is eliminated for each of the four small peaks indicated by arrows. Fig. 20 The intensity ratio of the four peaks shown in (d) represents the intensity ratio of the fluorescence emitted by the phosphor D(1, 1) at the light-emitting point P(1) at time 100, 200, 300, and 400.

[0159] Fig.21 It is for Fig.19 The original data is converted into color + space correction data by performing the color conversion + space correction of (Equation 20) at each time. Fig.19 Same. Fig.21 In (a), according to Fig. 20 The same reason as (a) gives the same result. Fig.21 As shown in (d), both spatial crosstalk and spectral crosstalk are eliminated for the luminescent fluorescence of the phosphor D(2,1) at time 100. However, spatial crosstalk and spectral crosstalk are not eliminated for the luminescent fluorescence of the phosphor D(2,1) at time 200, 300, and 400. This is because at time 200, 300, and 400, Fig.19 The fluorescence intensity of X(1,1) or X(1,2) shown in (a) is saturated. As a result, Fig.19The ratio of the fluorescence intensity of X(2,1) and X(2,2) to the fluorescence intensity of X(1,1) and X(1,2) shown in (d) deviates from the spatial crosstalk ratio defined by (Equation 20). In addition, since the degree of this deviation increases according to the degree of saturation, Fig.21 In (d), the fluorescence intensity at time 300 is larger than that at time 200, and the fluorescence intensity of the fluorescent body D(2, 1) at time 400 is larger than that at time 300.

[0160] [Example 6]

[0161] Here, a method of determining the matrix Y of (Equation 20) by a method different from that of [Example 2] is described. In [Example 2], Fig.11 As shown in (a), the matrix Y is determined by causing the fluorescent bodies D(1, 1), D(2, 1), D(1, 2), and D(2, 2) to emit fluorescence individually in the order of each other, that is, by causing the luminescent point P(1) and the luminescent point P(2) to emit fluorescence alternately. However, if the electrophoresis speed of either of the capillaries Cap(1) and Cap(2) deviates from the assumption, the luminescent point P(1) and the luminescent point P(2) may emit fluorescence simultaneously, and in this case, the matrix Y cannot be determined.

[0162] Fig. 22 represents a more realistic approach to avoid the above problems. Initially, Fig. 22 As shown in (a), the sample is injected into the capillary Cap (1) only, and the capillaries Cap (1) and Cap (2) are used for electrophoresis analysis, so that the fluorescent bodies D (1, 1) and D (1, 2) emit fluorescence light separately, and fluorescence detection is performed in the detection areas W (1, 1), W (1, 2), W (2, 1), and W (2, 2). Next, as Fig. 22 As shown in (b), the sample is injected only into the capillary Cap (2), and electrophoresis analysis is performed using the capillaries Cap (1) and Cap (2), so that the fluorescent bodies D (2, 1) and D (2, 2) emit fluorescence light separately, and fluorescence detection is performed in the detection areas W (1, 1), W (1, 2), W (2, 1), and W (2, 2). By this method, the matrix Y can be determined in the same way as in [Example 2]. In addition, according to this method, the matrix Y can be determined more simply and reliably than in [Example 2]. Fig. 22In order to facilitate understanding, the fluorescent bodies D(1, 1) and D(1, 2) in the capillary Cap (1) and the fluorescent bodies D(2, 1) and D(2, 2) in the capillary Cap (2) are depicted as emitting fluorescence multiple times, but it is sufficient for each to emit fluorescence once. In addition, in the above, after the initial electrophoresis analysis is completed, the next electrophoresis analysis is performed, but the interval between electrophoresis analyses may not be increased. By appropriately staggering the timing of sample injection into the capillary Cap (1) and the timing of sample injection into the capillary Cap (2), the same results as those of Fig. 22 The same effect can be achieved. For example, after injecting the sample into the capillary Cap (1), electrophoresis is performed for a short time, and then the sample is injected into the capillary Cap (2) and then the electrophoresis is restarted. In this way, the time required to determine the matrix Y can be reduced. In addition, according to this method, the sample injected into the capillary Cap (1) and the sample injected into the capillary Cap (2) can be made to have the same composition or to be the same. In this way, the sample to be prepared can be simplified and the cost for it can be reduced.

[0163] Fig.23 Here, although the sample injection into the capillary Cap (1) and the sample injection into the capillary Cap (2) are performed at the same timing, Fig. 22 Similarly, the sample is prepared so that the fluorescent bodies D(1, 1) and D(1, 2) in the capillary Cap (1) and the fluorescent bodies D(2, 1) and D(2, 2) in the capillary Cap (2) emit fluorescent light at different times. In other words, the composition of the sample injected into the capillary Cap (1) and the sample injected into the capillary Cap (2) is changed so that the electrophoretic speeds of the substances labeled with the respective fluorescent bodies differ. For example, the sample injected into the capillary Cap (1) may include a 50-base DNA fragment labeled with the fluorescent body D(1, 1) and a 60-base DNA fragment labeled with the fluorescent body D(1, 2), and the sample injected into the capillary Cap (2) may include a 70-base DNA fragment labeled with the fluorescent body D(2, 1) and a 80-base DNA fragment labeled with the fluorescent body D(2, 2).

[0164] or, Fig.23 This can also be achieved by setting different electrophoresis conditions in the capillaries Cap (1) and Cap (2) even when samples of the same composition are injected into the capillaries Cap (1) and Cap (2) at the same time. For example, during electrophoresis, the applied voltage of Capi (2) is temporarily lowered, or the temperature of Capi (2) is lowered during electrophoresis.

[0165] In the above, it is assumed that a special sample is prepared in advance for determining the matrix Y, but this is not necessarily necessary. In the raw data of the electrophoresis analysis of the actual sample to be analyzed, there is a state where the fluorescence emission of each phosphor in each capillary is generated independently. If this can be determined, the raw data at that time can be used to determine the matrix Y.

[0166] [Example 7]

[0167] Fig.24 FIG. 1 is a structural diagram of a capillary electrophoresis device as an example of an analysis device. The capillary electrophoresis device is widely used as an analysis device for analyzing DNA sequences and DNA fragments. Fig.24As shown, the multi-capillary electrophoresis device includes a capillary 24-1, a cathode 24-4, an anode 24-5, a cathode side buffer 24-6, an anode side buffer 24-7, a pump block 24-9, a syringe 24-11 and a laser light source 24-12. In this embodiment, four capillaries 24-1 are used, and DNA sequencing of different samples is implemented in each capillary 24-1. The sample of the DNA sequence is composed of DNA fragments labeled with four fluorescent substances. One analysis session is performed through the following steps (1) to (6). (1) First, the sample injection ends 24-2 of the four capillaries 24-1 are immersed in the cathode side buffer 24-6, and the sample dissolution ends 24-3 are immersed in the anode side buffer 24-7 via the polymer solution in the polymer block 24-9. (2) Next, close the valve 24-10 of the pump block 24-9, press down the piston of the syringe 24-11 connected to the pump block 24-9 to pressurize the polymer solution inside, and fill the polymer solution inside each capillary 24-1 from the sample dissolution end 24-3 to the sample injection end 24-2. (3) Next, open the valve 24-10, inject different samples into each capillary 24-1 from the sample injection end 24-2, and apply a high voltage between the cathode 24-4 and the anode 24-5 through the power supply 24-8 to start capillary electrophoresis. DNA fragments labeled with four fluorescent substances are electrophoresed from the sample injection end 24-2 toward the sample dissolution end 24-3. (4) In parallel, the position of each capillary 24-1 at a certain distance from the sample injection end 24-2 is used as the light-emitting point 24-14, and the laser beam 24-13 with a wavelength of 505 nm oscillated by the laser light source 24-12 is irradiated to each light-emitting point 24-14. Here, the cover of each capillary 24-1 near the light-emitting point 24-14 is removed in advance, and each capillary 24-1 near the light-emitting point 24-14 is arranged on the same plane. After the laser beam 24-13 is focused, it is introduced from the side of the above-mentioned arrangement plane and along the arrangement plane. (5) Then, the DNA fragments labeled with four kinds of fluorescent substances are electrophoresed in the interior of each capillary 24-1, and when passing through the light-emitting point 24-14, they are excited by the irradiation of the laser beam 24-13 and emit fluorescence. In other words, the four kinds of fluorescent substances are made to emit fluorescence from the four light-emitting points, and the fluorescence intensity of each changes moment by moment with the electrophoresis. (6) Finally, the fluorescence emitted from each luminescent point is detected in multiple colors by a sensor (not shown), and the time series data obtained by a computer (not shown) is analyzed, thereby performing DNA sequencing of the sample injected into each capillary. The analysis session consisting of the above steps (1) to (6) can be repeated multiple times. For example, by analyzing samples (1) to (4) in the first analysis session, analyzing samples (5) to (8) in the second analysis session, ..., a large number of different samples can be analyzed.

[0168] In this embodiment, the multi-color detection (6) is performed using the optical system of Patent Document 1. That is, the fluorescence emitted from A = 4 light-emitting points P(1) to P(4) is collimated by a condenser lens, a transmission-type diffraction grating is passed through, and the first-order diffracted light of each fluorescence is imaged on a two-dimensional sensor through an imaging lens. Fig.25 The two-dimensional sensor image is obtained by irradiating the four capillaries Cap(1) to Cap(4) with a laser beam when the four capillaries are filled with a standard solution, and includes a wavelength dispersion image of the Raman scattered light emitted from the four light-emitting points P(1) to P(4). The horizontal axis is the arrangement direction of the four capillaries, and the vertical axis is the wavelength direction. The four longitudinally extending stripes shown by the arrows are the wavelength dispersion images of the Raman scattered light of the capillaries Cap(1) to Cap(4). Based on this result, the wavelength of the wavelength dispersion image on the two-dimensional sensor image is corrected for each light-emitting point, and the relationship between the pixel position and the wavelength is calculated. Then, based on the result, as shown in FIG. Fig.26 As shown, for each of the luminous points P(1) to P(4) on the two-dimensional sensor image, B=20 detection areas of different wavelength bands are set. The 20 detection areas are respectively set to detect luminescence of 500 to 700 nm in wavelength bands equally divided at 10 nm intervals. For example, the detection areas W(1, 1), W(1, 2), ..., W(1, 20) set for the luminous point P(1) are respectively set to detect luminescence of wavelength bands of 500 to 510 nm, 510 to 520 nm, ..., 690 to 700 nm, and the respective signal intensities are set to X(1, 1), X(1, 2), ..., X(1, 20). The same is true for the luminous points P(2) to P(4). Here, as long as the positions of the luminous points P(1) to P(4) and the optical system are fixed, the relationship between the pixel position and wavelength of each capillary in the two-dimensional sensor image is maintained, so Fig.26 The detection area set in is also valid for multiple different luminescence detections, or multiple different analysis sessions.

[0169] In this embodiment, dR110, dR6G, dTAMRA and dROX are used as C=4 kinds of fluorescent substances, and they are labeled to DNA fragments whose terminal bases are T, C, A and G prepared by Sanger reaction. The maximum fluorescence emission wavelengths of dR110, dR6G, dTAMRA and dROX are 541nm, 568nm, 595nm and 618nm, respectively. Therefore, the fluorescence emission of each is detected with the strongest intensity in the detection area of ​​the wavelength band of 540-550nm, 560-570nm, 590-600nm and 610-620nm. For example, the fluorescence emission of dR110, dR6G, dTAMRA and dROX at the luminous point P(1) is mainly detected in the areas W(1,5), W(1,7), W(1,10) and W(1,12), respectively. Each luminescent fluorescence is also detected in other detection areas of the luminescent point P(1) due to spectral crosstalk, and is also detected at a weaker intensity in the detection areas of the luminescent points P(2) to P(3) due to spatial crosstalk and spectral crosstalk. Hereinafter, dR110, dR6G, dTAMRA, and dROX at the luminescent point P(1) are respectively set to D(1,1), D(1,2), D(1,3), and D(1,4), and their concentrations are respectively set to Z(1,1), Z(1,2), Z(1,3), and Z(1,4). The same is true for the luminescent points P(2) to P(4).

[0170] At this time, (Formula 10) to (Formula 12) in (Formula 9) become the following formulas.

[0171] [Mathematical formula 23]

[0172]

[0173] [Mathematical formula 24]

[0174]

[0175] [Mathematical formula 25]

[0176]

[0177] Here, X is a matrix of 80 rows and 1 column, Y is a matrix of 80 rows and 16 columns, and Z is a matrix of 16 rows and 1 column. By substituting (Formula 23) to (Formula 25) into (Formula 14) to obtain spatial correction and color conversion, both spatial crosstalk and spectral crosstalk can be eliminated, and time series data of the concentrations of the four phosphors in each capillary can be obtained. As a result, DNA sequences of different samples injected into each capillary can be implemented. In addition, the existing method corresponding to this embodiment is to perform color conversion on each of the capillaries (Formula 8). Spatial crosstalk cannot be eliminated in the existing method.

[0178] (Equation 23) to (Equation 25) and (Equation 14) derived from them are effective for multiple different luminescence detections or multiple different analysis sessions as long as the position of each luminescent point, the optical system, and the fluorescent substance used are fixed.

[0179] The method for determining the matrix Y is as described above. However, it is not necessary to set all 80×16 elements. For example, elements with sufficiently small absolute values ​​compared to other elements can be replaced with zero to simplify related calculations. In addition, when the range involved in the spatial crosstalk is limited, (Formula 9) to (Formula 12) can also be defined within the limited range. In this case, as needed, the limited range can also be slid sequentially to perform a wider range analysis. For example, when the range involved in the spatial crosstalk can be limited to two adjacent capillaries, the spatial crosstalk and spectral crosstalk about more than two capillaries can be ignored, and the related calculations can be omitted. Furthermore, in this case, the process of making each light-emitting point emit light separately in order to determine the matrix Y can also be simplified to a process of making multiple light-emitting points that are not affected by spatial crosstalk emit light simultaneously.

[0180] [Example 8]

[0181] In this embodiment, similarly to [Example 7], Fig.24 However, at each time, the optical system of Patent Document 2 detects the fluorescence emission of four kinds of fluorescent substances emitted from four emission points in multiple colors. Fig. 27 The structure of this optical system and the four-divided images of four light-emitting points obtained by this optical system are shown. Fig. 27 The optical system includes a condenser lens array 27-1, a long pass filter 27-3, dichroic mirrors 27-4, 27-5, 27-6 and 27-7, and a two-dimensional sensor 27-8. Fig. 27 (a) and Fig. 27As shown in (b), first, the fluorescence emitted from A = 4 light-emitting points P(1) to P(4) is collimated individually by the four condenser lenses 27-2 constituting the condenser lens array 27-1 to form four light beams 27-9, which are then collectively transmitted through a long-pass filter 27-3, thereby cutting the laser beam light. Next, each light beam 27-9 is collectively incident on a set of dichroic mirror arrays consisting of four dichroic mirrors 27-4, 27-5, 27-6 and 27-7, thereby each light beam 27-9 is respectively split into B = 4 split light beams 27-10, 27-11, 27-12 and 27-13, and then emitted from the dichroic mirror array. Here, by adjusting the spectral characteristics of each dichroic mirror, the split light beams 27-10, 27-11, 27-12 and 27-13 have light in the wavelength bands of 520-550nm, 550-580nm, 580-610nm and 610-640nm, respectively. Finally, the generated A×B=4×4=16 split light beams are incident on the two-dimensional sensor 27-8, as shown in FIG. Fig. 27 As shown in (c), 16 segmented images W(1, 1) to W(4, 4) are obtained. Here, since the arrangement direction of the four capillaries Cap(1) to Cap(4), that is, the arrangement direction of the light-emitting points P(1) to P(4) and the segmentation direction based on the dichroic mirror array, that is, the wavelength direction, are perpendicular to each other, Fig. 27 As shown in (c), in the two-dimensional sensor image 27-14, the segmented images W(1, 1) to W(4, 4) do not overlap with each other and are detected in a queued state. For example, the fluorescence emitted from the light-emitting point P(1) is detected as segmented images of W(1, 1), W(1, 2), W(1, 3), and W(1, 4), which have light components in the wavelength bands of 520 to 550 nm, 550 to 580 nm, 580 to 610 nm, and 610 to 640 nm, respectively. The same is true for the light-emitting points P(2) to P(4). Therefore, the segmented images W(1, 1) to W(4, 4) in the two-dimensional sensor image 27-14 are respectively set as detection areas. In addition, the signal intensity detected in each detection area is set to X(1, 1) to X(4, 4), respectively. Here, as long as the positions of the light-emitting points P(1) to P(4) and the optical system are fixed, the relationship between the pixel position and the wavelength of each capillary in the two-dimensional sensor image 27-14 is maintained. Fig. 27 The detection area set in (c) is also effective for multiple different luminescence detections or multiple different analysis sessions.

[0182] In this example, similarly to [Example 7], dR110, dR6G, dTAMRA, and dROX were used as C=4 kinds of fluorescent substances, and they were labeled with DNA fragments prepared by Sanger reaction with terminal base types of T, C, A, and G, respectively. The maximum fluorescence emission wavelengths of dR110, dR6G, dTAMRA, and dROX are 541 nm, 568 nm, 595 nm, and 618 nm, respectively, so the respective fluorescence emission is detected with the strongest intensity in the detection area of ​​the wavelength band of 520 to 550 nm, 550 to 580 nm, 580 to 610 nm, and 610 to 640 nm. For example, the fluorescence emission of dR110, dR6G, dTAMRA, and dROX at the luminescent point P(1) is mainly detected in the detection areas W(1,1), W(1,2), W(1,3), and W(1,4), respectively. However, each fluorescent light is detected in other detection areas of the light-emitting point P(1) due to spectral crosstalk, and is detected in detection areas of the light-emitting points P(2) to P(3) due to spatial crosstalk and spectral crosstalk. Hereinafter, dR110, dR6G, dTAMRA, and dROX at the light-emitting point P(1) are defined as phosphors D(1,1), D(1,2), D(1,3), and D(1,4), respectively, and their concentrations are defined as Z(1,1), Z(1,2), Z(1,3), and Z(1,4), respectively. The same is true for the light-emitting points P(2) to P(4).

[0183] At this time, (Formula 10) to (Formula 12) in (Formula 9) become the following formulas.

[0184] [Mathematical formula 26]

[0185]

[0186] [Mathematical formula 27]

[0187]

[0188] [Mathematical formula 28]

[0189]

[0190] Here, X is a matrix of 16 rows and 1 column, Y is a matrix of 16 rows and 16 columns, and Z is a matrix of 16 rows and 1 column. By substituting (Equation 26) to (Equation 28) into (Equation 14), both spatial crosstalk and spectral crosstalk can be eliminated, and time series data of the concentrations of the four phosphors in each capillary can be obtained. As a result, DNA sequences of different samples injected into each capillary can be implemented. In addition, the existing method corresponding to this embodiment is to implement color conversion of (Equation 8) for each capillary. Spatial crosstalk cannot be eliminated in the existing method.

[0191] (Equation 26) to (Equation 28) and (Equation 14) derived from them are effective for multiple different luminescence detections or multiple different analysis sessions as long as the position of each luminescent point, the optical system, and the fluorescent substance used are fixed.

[0192] [Example 9]

[0193] Fig.28 A schematic diagram showing a multi-channel elongation reaction device for performing a complementary chain elongation reaction in units of one base by arranging a plurality of reaction tanks on a plane, using different DNA fragments as templates in each channel, and performing multi-color detection of the fluorescence emitted by C=4 kinds of fluorescent bodies labeled with four kinds of bases introduced into the complementary chain during the elongation reaction to perform a DNA sequence of each DNA fragment, and a schematic diagram showing a two-dimensional sensor image obtained by a plurality of two-dimensional sensors. The multi-channel elongation reaction device includes a laser light source 28-1, a dichroic mirror 28-3, a lens 28-4, dichroic mirrors 28-8, 28-9 and 28-10, a lens 28-11, a first two-dimensional sensor 28-12, a lens 28-13, a second two-dimensional sensor 28-14, a lens 28-15, a third two-dimensional sensor 28-16, a lens 28-17 and a fourth two-dimensional sensor 28-18. The maximum fluorescence emission wavelengths of the fluorescent bodies labeled with base types T, C, A and G are 535nm, 565nm, 595nm and 625nm respectively. First, the laser beam 28-2 oscillated by the laser light source 28-1 is passed through the dichroic mirror 28-3, and then focused by the lens 28-4 to irradiate the sample 28-5, i.e., the DNA fragment, in the multi-channel. Next, in each channel, the fluorescence emission 28-6 of each fluorescent body excited by the laser beam irradiation is collimated by the lens 28-4, and the obtained light beam 28-7 is reflected by the dichroic mirror 28-3. The dichroic mirror 28-3 has a spectral characteristic of transmitting the laser beam and reflecting the emitted fluorescence. Next, the light beam 28-7 is divided into four light beams with four different wavelength components by three types of dichroic mirrors 28-8, 28-9 and 28-10. Then, the first split beam is imaged on the first two-dimensional sensor 28-12 through lens 28-11, the second split beam is imaged on the second two-dimensional sensor 28-14 through lens 28-13, the third split beam is imaged on the third two-dimensional sensor 28-16 through lens 28-15, and the fourth split beam is imaged on the fourth two-dimensional sensor 28-18 through lens 28-17.

[0194] Sample 28-19 is a schematic diagram of sample 28-5 observed from a direction perpendicular to the arrangement plane. A=5 channels form luminous points P(1) to P(5) respectively. In fact, there are more channels on the sample, and the luminous fluorescence from each channel is detected together by the above-mentioned optical system. However, here, only the luminous point P(3) of interest and the luminous points P(1), P(2), P(4), and P(5) around the luminous point P(3) that directly cause spatial crosstalk to the luminous point P(3) are taken as the analysis objects, so only the luminous points P(1) to P(5) are briefly depicted in sample 28-19. When the luminous point other than the luminous point P(3) is of interest, its luminous point and the luminous points around it can be analyzed in the same way. When any luminous point is of interest, the same fluorescence detection data can also be analyzed separately after fluorescence detection. The first two-dimensional sensor image 28-20 is a segmented image of the sample 28-19 obtained by the first two-dimensional sensor 28-12. The detection areas W(1, 1) to W(5, 1) are set at the imaging points of the light-emitting points P(1) to P(5), and the respective signal intensities are set to X(1, 1) to X(5, 1). In the first two-dimensional sensor image 28-20, the fluorescence component in the wavelength band of 520 to 550 nm is detected. The second two-dimensional sensor image 28-21 is a segmented image of the sample 28-19 obtained by the second two-dimensional sensor 28-14. The detection areas W(1, 2) to W(5, 2) are set at the imaging points of the light-emitting points P(1) to P(5), and the respective signal intensities are set to X(1, 2) to X(5, 2). In the second two-dimensional sensor image 28-21, the fluorescence component in the wavelength band of 550 to 580 nm is detected. The third two-dimensional sensor image 28-22 is a segmented image of the sample 28-19 obtained by the third two-dimensional sensor 28-16. The detection areas W(1, 3) to W(5, 3) are set at the imaging points of the light-emitting points P(1) to P(5), and the respective signal intensities are set to X(1, 3) to X(5, 3). In the third two-dimensional sensor image 28-22, the fluorescence component in the wavelength band of 580 to 610 nm is detected. The fourth two-dimensional sensor image 28-23 is a segmented image of the sample 28-19 obtained by the fourth two-dimensional sensor 28-18. The detection areas W(1, 4) to W(5, 4) are set at the imaging points of the light-emitting points P(1) to P(5), and the respective signal intensities are set to X(1, 4) to X(5, 4). In the fourth two-dimensional sensor image 28-23, the fluorescence component in the wavelength band of 610 to 640 nm is detected. That is, the four two-dimensional sensors are used to perform B=4-color detection of the luminous fluorescence from each light-emitting point. For example, the fluorescence emitted by the four phosphors in the light-emitting point P(3) is mainly detected in the detection areas W(3,1), W(3,2), W(3,3) and W(3,4) in order of wavelength.However, each fluorescent light is also detected in other detection areas of the light-emitting point P(3) due to spectral crosstalk, and is also detected in the detection areas of the light-emitting points P(1), P(2), P(4), and P(5) due to spatial crosstalk and spectral crosstalk. Hereinafter, the four fluorescent substances in the light-emitting point P(3) are respectively designated as D(3,1), D(3,2), D(3,3), and D(3,4) in order of wavelength, and their concentrations are designated as Z(3,1), Z(3,2), Z(3,3), and Z(3,4). The same is true for the light-emitting points P(1), P(2), P(4), and P(5).

[0195] At this time, (Formula 10) to (Formula 12) in (Formula 9) become the following formulas.

[0196] [Mathematical formula 29]

[0197]

[0198] [Math 30]

[0199]

[0200] [Mathematical formula 31]

[0201]

[0202] Here, X is a matrix of 20 rows and 1 column, Y is a matrix of 20 rows and 20 columns, and Z is a matrix of 20 rows and 1 column. By substituting (Equation 29) to (Equation 31) into (Equation 14), both spatial crosstalk and spectral crosstalk can be eliminated, and time series data of the concentrations of the four phosphors in each channel can be obtained. However, in this embodiment, since only the luminous point P(3) is concerned, only the elements Z(3, 1), Z(3, 2), Z(3, 3), and Z(3, 4) in the matrix Z are extracted. As a result, DNA sequencing can be performed in the channel of the luminous point P(3). The same method can also be used when focusing on other luminous points and performing DNA sequencing in the corresponding channels.

[0203] [Example 10]

[0204] In this embodiment, a flow chart is used to summarize the existing method and the method disclosed in the present invention. Fig.29 is a flow chart showing one analysis session of the conventional method. Fig.29As shown, the analysis session of the conventional method is performed by an analysis system having an analysis device for analyzing a sample 29-1, a computer, a display device, and a database. The analysis device has a sensor (not shown) for incident light from the sample 29-1. First, a sample 29-1 of type A (A is an integer greater than 2) labeled with a fluorescent substance of type C (C is an integer greater than 1) is input into the analysis device. Next, in the analysis device, each sample is analyzed in parallel through step 29-2, and for each analysis, and at each time, the luminescent fluorescence is detected in a band of type B (B is an integer greater than 1), and time series raw data of A×B fluorescence intensities X(a, b) are obtained, and these time series raw data are sent to the computer. Here, a=1, 2, ..., and A, b=1, 2, ..., and B. Next, in the computer, through step 29-3, for analysis (a0), by using a matrix Y with C rows and B columns stored in the database - The color conversion of (a0) is performed to eliminate spectral crosstalk. At each moment, the concentration Z(a0,c) of the C type phosphor is calculated based on the fluorescence intensity X(a0,b). Here, a0=1, 2, ..., A, c=1, 2, ..., C. Finally, in the display device, the time series color conversion data of Z(a0,c) is output through step 29-4. Steps 29-3 and 29-4 are implemented for all a0. Fig.29 As shown, the conventional method is characterized in that A analyses and analyses are performed independently. In (Equation 1) to (Equation 4), the fluorescence intensity X(a, b) is represented as X(b), and the concentration Z(a0, c) is represented as Z(c).

[0205] Fig.30 is a flow chart showing one analysis session of this method. Fig.29 same, Fig.30 The analysis session of the present embodiment shown is performed by an analysis system having an analysis device for analyzing a sample 30-1, a computer, a display device, and a database. The analysis device has a sensor (not shown) for incident light from the sample 30-1. First, a sample 30-1 of type A (A is an integer greater than 2) marked by type C (C is an integer greater than 1) fluorescent bodies is input into the analysis device. Next, in the analysis device, each sample is analyzed in parallel through step 30-2, and for each analysis and at each time, the luminescent fluorescence is detected in B (B is an integer greater than 1) wavelength bands, and time series raw data of A×B fluorescence intensities X (a, b) are obtained, and these time series raw data are sent to the computer. Here, a=1, 2, ..., and A, b=1, 2, ..., and B. However, unlike Fig.29As shown by the dashed arrows in step 30-2, there is significant spatial crosstalk of luminescent fluorescence between different analyses. Next, in the computer, through step 30-3, for all analyses (a), by using the matrix Y with (A×C) rows and (A×B) columns stored in the database - The color conversion + spatial correction is used to eliminate spectral crosstalk and spatial crosstalk. At each moment, the concentration Z(a, c) of the sample of type A or the fluorescent substance of type C to be analyzed is calculated based on the fluorescence intensity X(a, b). Here, a=1, 2, ..., and A, c=1, 2, ..., and C. Finally, in the display device, through step 30-4, for each analysis (a), the time series color conversion + spatial correction data of Z(a, c) is output. Fig.30 As shown, the method is characterized in that the color conversion + spatial correction in step 30-3 is performed on all analyses at the same time. A further characteristic is that the color conversion + spatial correction uses a common matrix Y in all analyses. - .

[0206] Fig.31 It means in Fig.30 The matrix Y previously saved in the database - Flowchart of the method for obtaining . First, a sample 31-1 is input into the analysis device for determining a type A (A is an integer greater than 2) of matrix Y identified by a type C (C is an integer greater than 1) of fluorescent material. The sample is prepared to generate fluorescence light of the fluorescent material (c) in the analysis (a) alone. In other words, a=1, 2, ..., and A, c=1, 2, ..., and C, and fluorescence light is generated for all combinations of these A×C, but fluorescence light of more than 2 combinations is not generated at the same time. Here, the above is achieved by determining the sample 31-1 using the matrix Y, but it can also be achieved by the setting of the analysis device. Next, in the analysis device, through step 31-2, the above A×C individual fluorescence lights are detected in the B type (B is an integer greater than 1) wavelength band, and the time series raw data of A×B fluorescence intensities X (a, b) are obtained, and these time series raw data are sent to the computer. Next, in the computer, through step 31-3, a matrix of (A×B) rows and (A×C) columns is derived from the obtained (A×B)×(A×C) data, and then the general inverse matrix of Y is obtained to obtain the matrix Y of (A×C) rows and (A×B) columns. - Then, the obtained matrix Y - Stored in the database Fig.30 In the following analysis, we will use the matrix Y - .

[0207] Fig.32 Is using Fig.31 The matrix Y obtained in -At the same time, repeat multiple times for Fig.30 Flowchart of the case of an analysis session of steps 30-2 to 30-4 for the type A sample 30-1 shown in FIG. Here, the type A sample 30-1 to be analyzed in each analysis session is different from each other. Fig.32 As shown, for multiple different analysis sessions, the same matrix Y stored in the database is used. - This is a characteristic of this method.

[0208] Fig.33 This shows an example of the structure of a computer. The computer is connected to the analysis device. The computer not only performs data analysis but also controls the analysis device. Figure 29 to Figure 31 In the example, the database and display device are depicted as being external to the computer, but in Fig.33 The computer draws them in the computer. The keyboard as the input unit is used to set the conditions for data analysis and the conditions for analyzing the device. The time series raw data of the fluorescence intensity X(a, b) output from the analyzing device is sequentially stored in the memory. In addition, the matrix Y of (A×C) rows and (A×B) columns stored in the database inside the HDD is - The CPU calculates the fluorescence intensity X(a, b) and Y stored in the memory. - The time series color conversion + space correction data of the phosphor concentration Z(a, c) is derived by product, which is stored in the memory and displayed on the monitor as the display unit. In addition, the analysis results can be compared with the information on the network through the network interface NIF.

[0209] [Modifications]

[0210] The present disclosure is not limited to the above-mentioned embodiments, and includes various modified examples. For example, the above-mentioned embodiments are embodiments described in detail in order to easily explain the present disclosure, and it is not necessary to have all the structures described. In addition, a part of a certain embodiment can be replaced with the structure of other embodiments. In addition, the structure of other embodiments can also be added to the structure of a certain embodiment. In addition, for a part of the structure of each embodiment, a part of the structure of other embodiments can also be added, deleted or replaced.

[0211] Explanation of symbols

[0212] 1-1 Pinhole plate

[0213] 1-2 Lighting point side opening plate

[0214] 1-3 Condenser lens

[0215] 1-4 Sensor side opening plate

[0216] 1-5 Colored Glass Filters

[0217] 1-6 Two-dimensional sensor

[0218] 1-7 Halogen Light

[0219] 1-8 Luminous Points

[0220] 1-9 Light

[0221] 1-10 Luminous Image

[0222] Cap(1) Capillary 1

[0223] Cap(2) Capillary 2

[0224] P(1) The luminous point on Cap(1)

[0225] The luminous point on P(2)Cap(2)

[0226] D(1,1)P(1) Phosphor 1 or light absorber 1 on

[0227] D(1,2)P(1) Phosphor 2 or light absorber 2 on

[0228] D(2,1)P(2) Phosphor 1 or light absorber 1 on

[0229] D(2,2)P(2) Phosphor 2 or light absorber 2 on

[0230] D(1,3)P(1) Phosphor 3 or light absorber 3 on

[0231] W(1,1) is the detection area where D(1,1) is mainly used for detection

[0232] W(1,2) is the detection area where D(1,2) is mainly used for detection

[0233] W(2,1) is the detection area where D(2,1) is mainly used for detection

[0234] W(2,2) is the detection area where D(2,2) is mainly used for detection

[0235] Signal strength of X(1,1)W(1,1)

[0236] Signal strength of X(1,2)W(1,2)

[0237] Signal strength of X(2,1)W(2,1)

[0238] Signal strength of X(2,2)W(2,2)

[0239] Concentration of Z(1,1)D(1,1)

[0240] Concentration of Z(1,2)D(1,2)

[0241] Concentration of Z(2,1)D(2,1)

[0242] Concentration of Z(2,2)D(2,2)

[0243] S Sensor

[0244] LB laser beam

[0245] LL Lighting

[0246] 24-1 Capillary

[0247] 24-2 Sample injection port

[0248] 24-3 Sample dissolution end

[0249] 24-4 Cathode

[0250] 24-5 Anode

[0251] 24-6 Cathode side buffer

[0252] 24-7 Anode side buffer

[0253] 24-8 Power Supply

[0254] 24-9 Polymer Block

[0255] 24-10 Valves

[0256] 24-11 Syringe

[0257] 24-12 Laser Light Source

[0258] 24-13 Laser Beam

[0259] 24-14 Luminous Point

[0260] Cap(a) Capillary a (a=1, 2, 3, and 4)

[0261] P(a) The luminous point on capillary a (a = 1, 2, 3, and 4)

[0262] W(a,b) Detection area of ​​wavelength band b of luminescence from P(a) (a=1, 2, ..., and 4) (b=1, 2, ..., and 20)

[0263] 27-1 Condenser lens array

[0264] 27-2 Condenser lens

[0265] 27-3 Long pass filter

[0266] 27-4, 27-5, 27-6, and 27-7 dichroic mirrors

[0267] 27-8 Two-dimensional sensor

[0268] 27-9 Beam

[0269] 27-10, 27-11, 27-12, and 27-13 Split Beam

[0270] 27-14 2D sensor image

[0271] P(a) luminous point a (a=1, 2, 3, 4, and 5)

[0272] W(a, b) is the detection area of ​​the wavelength band b of the luminescence from P(a) (a=1, 2, 3, 4, and 5) (b=1, 2, 3, and 4)

[0273] 28-1 Laser Light Source

[0274] 28-2 Laser beam

[0275] 28-3, 28-8, 28-9, and 28-10 dichroic mirrors

[0276] 28-4, 28-11, 28-13, 28-15, and 28-17 lenses

[0277] 28-5, and 28-19 samples

[0278] 28-6 Fluorescence

[0279] 28-7 Beam

[0280] 28-12, 28-14, 28-16, and 28-18 2D sensors

[0281] 28-20, 28-21, 28-22, and 28-23 2D sensor images

[0282] 29-1 Sample

[0283] 29-2 Processes in the analytical device

[0284] 29-3 Processes in Computers

[0285] 29-4 Processes in the display device

[0286] 30-1 Sample

[0287] 30-2 Processes in the analytical device

[0288] 30-3 Processes in Computers

[0289] 30-4 Processes in the display device

[0290] 31-1 Sample

[0291] 31-2 Processes in the analytical device

[0292] 31-3 Processes in computers.

Claims

1. An analysis method using a multi-capillary electrophoresis device, wherein the multi-capillary electrophoresis device is configured to use A capillaries Cap(a) to perform electrophoretic analysis on A samples Sam(a) containing a fluorescent substance in parallel, where A is an integer greater than 2, a=1, 2, ..., A, The multi-capillary electrophoresis device comprises: A power supply configured to apply a voltage across each of the A capillaries Cap(a); A luminous points P(a) on the A capillaries Cap(a), wherein the A luminous points are emission sources of fluorescence induced by irradiating the A capillaries Cap(a) with one or more laser beams; an optical system configured to detect A original signals X(a) of fluorescence of the fluorescent body contained in the A samples Sam(a) emitted from the A light-emitting points P(a); A computer is configured to perform a combined processing on the A original signals X(a), To output A processed signals Z(a) corresponding to the concentrations of the phosphor at the A light-emitting points; as well as A display is configured to display the time series of the A processed signals Z(a), the method comprising: preparing a sample comprising a plurality of components labeled with fluorescent bodies of different concentrations, wherein the plurality of components show a plurality of different processed signal peaks in the time series of the A processed signals Z(a); Performing electrophoresis analysis, in which the sample Sam (a=j) injected into one capillary Cap (a=j) among the A capillaries Cap (a) is the one sample, and no sample containing a fluorescent substance is injected into the other capillaries Cap (a≠j), and j is a constant; and The conditions of the polycapillary electrophoresis device are adjusted so that for the luminous point P (a = j) on the one capillary Cap (a = j), in the time series of the processed signal Z (a = j), some intensities of the processed signal peaks are lower than the saturation intensity of the polycapillary electrophoresis device, while other intensities of the processed signal peaks are higher than or equal to the saturation intensity of the polycapillary electrophoresis device.

2. The analysis method according to claim 1, wherein In the combined processing, the polycapillary electrophoresis device is configured to use, in addition to the original signal X(a=j) of the one light-emitting point P(a=j), the original signal X(a≠j) of the light-emitting point P(a≠j) other than the one light-emitting point P(a=j).

3. The analysis method according to claim 1 or 2, wherein: When the processed signal Z(a=j) is below the saturation intensity, the spatial crosstalk between the A processed signals Z(a) is smaller than the spatial crosstalk between the A original signals X(a).

4. The analysis method according to claim 1 or 2, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a ≠ j) for the luminous point P (a ≠ j) on the capillary Cap (a ≠ j) other than the one capillary Cap (a ≠ j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a ≠ j) of the luminous point P (a ≠ j) on the one capillary Cap (a ≠ j), and Each intensity of the one or more crosstalk processed signal peaks increases nonlinearly with the concentration of the corresponding component in the one sample.

5. The analysis method according to claim 3, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a ≠ j) for the luminous point P (a ≠ j) on the capillary Cap (a ≠ j) other than the one capillary Cap (a ≠ j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a ≠ j) of the luminous point P (a ≠ j) on the one capillary Cap (a ≠ j), and Each intensity of the one or more crosstalk processed signal peaks increases nonlinearly with the concentration of the corresponding component in the one sample.

6. The analysis method according to claim 1 or 2, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a ≠ j) for the luminous point P (a ≠ j) on the capillary Cap (a ≠ j) other than the one capillary Cap (a ≠ j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a ≠ j) of the luminous point P (a ≠ j) on the one capillary Cap (a ≠ j), and When the intensity of the corresponding processed signal peak is lower than the saturation intensity, each intensity of the one or more crosstalk processed signal peaks remains low, and when the intensity of the corresponding processed signal peak is higher than or equal to the saturation intensity, each intensity of the one or more crosstalk processed signal peaks increases with the increase of the concentration of the corresponding component in the one sample.

7. The analysis method according to claim 3, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a ≠ j) for the luminous point P (a ≠ j) on the capillary Cap (a ≠ j) other than the one capillary Cap (a ≠ j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a ≠ j) of the luminous point P (a ≠ j) on the one capillary Cap (a ≠ j), and When the intensity of the corresponding processed signal peak is lower than the saturation intensity, each intensity of the one or more crosstalk processed signal peaks remains low, and when the intensity of the corresponding processed signal peak is higher than or equal to the saturation intensity, each intensity of the one or more crosstalk processed signal peaks increases with the increase of the concentration of the corresponding component in the one sample.

8. An analysis method using a multi-capillary electrophoresis device, wherein the multi-capillary electrophoresis device is configured to use A capillaries Cap(a) to perform electrophoretic analysis on A samples Sam(a) containing C types of fluorescent substances in parallel, where A is an integer greater than 2, a=1, 2, ..., A, C are integers greater than 1, The multi-capillary electrophoresis device comprises: A power supply configured to apply a voltage across each of the A capillaries Cap(a); A luminous points P(a) on the A capillaries Cap(a), wherein the A luminous points are emission sources of fluorescence induced by irradiating the A capillaries Cap(a) with one or more laser beams; The optical system is configured to detect C types of phosphors D(a, b) contained in the A samples Sam(a) emitted from the A light-emitting points P(a) in B wavelength bands W(a, b). c) detecting A×B original signals X(a, b) of the fluorescence, where B is an integer greater than 1, b=1, 2, ..., B, c=1, 2, ..., C; A computer configured to perform collective processing on the A×B original signals X(a, b) to output A×C processed signals Z(a, c) corresponding to the concentrations of the C types of fluorescent bodies D(a, c) at the A light-emitting points; as well as a display configured to display a time series of the A×C processed signals Z(a, c), The method comprises: Prepare a sample comprising a plurality of components labeled by a fluorescent substance D (a, c=c0) of C kinds of fluorescent substances with different concentrations, wherein the plurality of components show a plurality of different processed signal peaks in a time series of a processed signal Z (a, c=c0), and c0 is a constant; Performing electrophoresis analysis, in which the sample Sam (a=j) injected into one capillary Cap (a=j) among the A capillaries Cap (a) is the one sample, and no sample containing any fluorescent substance is injected into other capillaries Cap (a≠j) other than the one capillary Cap (a=j), and j is a constant; and The conditions of the polycapillary electrophoresis device are adjusted so that for the luminous point P (a=j) on the one capillary Cap (a=j), in the time series of the processed signal Z (a=j, c=c0), some intensities of the processed signal peaks are lower than the saturation intensity of the polycapillary electrophoresis device, while other intensities of the processed signal peaks are higher than or equal to the saturation intensity of the polycapillary electrophoresis device.

9. The analysis method according to claim 8, wherein: In the combined processing, the polycapillary electrophoresis device is configured to use, in addition to the original signal X (a = j, b = b0) of the one light-emitting point P (a = j), the original signal X (a ≠ j, b = b0) of the light-emitting point P (a ≠ j) other than the one light-emitting point P (a = j), where b0 is a constant.

10. The analysis method according to claim 8 or 9, wherein: When the processed signal Z (a=j, c=c0) is lower than the saturation intensity, the spatial crosstalk and spectral crosstalk between the A×C processed signals Z (a, c) are smaller than the spatial crosstalk and spectral crosstalk between the A×B original signals X (a, b).

11. The analysis method according to claim 8 or 9, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a≠j, c=c0) for the luminous point P (a≠j) on the capillary Cap (a≠j) other than the one capillary Cap (a=j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a=j, c=c0) of the luminous point P (a=j) on the one capillary Cap (a=j), and Each intensity of the one or more crosstalk processed signal peaks increases nonlinearly with the concentration of the corresponding component in the one sample.

12. The analysis method according to claim 10, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a≠j, c=c0) for the luminous point P (a≠j) on the capillary Cap (a≠j) other than the one capillary Cap (a=j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a=j, c=c0) of the luminous point P (a=j) on the one capillary Cap (a=j), and Each intensity of the one or more crosstalk processed signal peaks increases nonlinearly with the concentration of the corresponding component in the one sample.

13. The analysis method according to claim 8 or 9, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a≠j, c=c0) for the luminous point P (a≠j) on the capillary Cap (a≠j) other than the one capillary Cap (a=j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a=j, c=c0) of the luminous point P (a=j) on the one capillary Cap (a=j), and When the intensity of the corresponding processed signal peak is lower than the saturation intensity, each intensity of the one or more crosstalk processed signal peaks remains low, and when the intensity of the corresponding processed signal peak is higher than or equal to the saturation intensity, each intensity of the one or more crosstalk processed signal peaks increases with the increase of the concentration of the corresponding component in the one sample.

14. The analysis method according to claim 10, wherein: The multi-capillary electrophoresis device is configured to: provide one or more crosstalk processed signal peaks in the time series of the processed signal Z (a≠j, c=c0) for the luminous point P (a≠j) on the capillary Cap (a≠j) other than the one capillary Cap (a=j), wherein the one or more crosstalk processed signal peaks are synchronized with a part of the processed signal peaks in the time series of the processed signal Z (a=j, c=c0) of the luminous point P (a=j) on the one capillary Cap (a=j), and When the intensity of the corresponding processed signal peak is lower than the saturation intensity, each intensity of the one or more crosstalk processed signal peaks remains low, and when the intensity of the corresponding processed signal peak is higher than or equal to the saturation intensity, each intensity of the one or more crosstalk processed signal peaks increases with the increase of the concentration of the corresponding component in the one sample.

15. The analysis method according to claim 8 or 9, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

16. The analysis method according to claim 10, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

17. The analysis method according to claim 11, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

18. The analysis method according to claim 13, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

19. The analysis method according to claim 12 or 14, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

20. An analysis method using a multi-capillary electrophoresis device, wherein the multi-capillary electrophoresis device is configured to use A capillaries Cap(a) to perform electrophoretic analysis on A samples Sam(a) containing C types of fluorescent substances in parallel, where A is an integer greater than 2, a=1, 2, ..., A, C are integers greater than 1, The multi-capillary electrophoresis device comprises: A power supply configured to apply a voltage across each of the A capillaries Cap(a); A luminous points P(a) on the A capillaries Cap(a), wherein the A luminous points are emission sources of fluorescence induced by irradiating the A capillaries Cap(a) with one or more laser beams; An optical system configured to detect A×B original signals X(a, b) of fluorescence of C types of fluorescent bodies D(a, c) contained in the A samples Sam(a) emitted from the A light-emitting points P(a) in B bands W(a, b), where B is an integer greater than 1, b=1, 2, ..., B, c=1, 2, ..., C; A computer configured to perform collective processing on the A×B original signals X(a, b) to output A×C processed signals Z(a, c) corresponding to the concentrations of the C types of fluorescent bodies D(a, c) at the A light-emitting points P(a); as well as a display configured to display a time series of the A×C processed signals Z(a, c), The method comprises: Prepare a sample comprising a first component and a second component, wherein the first component is labeled with a first fluorescent body D among the C fluorescent bodies, and the second component is labeled with a second fluorescent body E other than the C fluorescent bodies, and in the time series of the A×C processed signals Z(a, c), the first component shows a first processed signal peak with the largest intensity, and the second component shows a second processed signal peak with the largest intensity; and An electrophoresis analysis is performed, in which the sample Sam (a=j) injected into one capillary Cap (a=j) among A capillaries Cap (a) is the one sample, and no sample containing any fluorescent substance is injected into other capillaries Cap (a≠j) other than the one capillary Cap (a=j), and j is a constant.

21. The analysis method according to claim 20, wherein: In the combined processing, the polycapillary electrophoresis device is configured to use, in addition to the original signal X (a = j, b = b0) of the one light-emitting point P (a = j), the original signal X (a ≠ j, b = b0) of the light-emitting point P (a ≠ j) other than the one light-emitting point P (a = j), where b0 is a constant.

22. The analysis method according to claim 20 or 21, wherein When the fluorescence of the second fluorescent body E is not detected in the time series of the A×C processed signals Z(a, c), the spatial crosstalk and spectral crosstalk between the A×C processed signals Z(a, c) are smaller than the spatial crosstalk and spectral crosstalk between the A×B original signals X(a, b).

23. The analysis method according to claim 20 or 21, wherein: The multi-capillary electrophoresis device is configured to: in electrophoresis analysis, for a luminous point P (a≠j) on a capillary Cap (a≠j) other than the one capillary Cap (a=j), provide a first crosstalk processed signal peak with the largest intensity and a second crosstalk processed signal peak with the largest intensity in a time series of a processed signal Z (a≠j, c), wherein the first crosstalk processed signal peak and the second crosstalk processed signal peak are respectively synchronized with the first processed signal peak and the second processed signal peak in a time series of a processed signal Z (a=j, c) of the luminous point P (a=j) on the one capillary Cap (a=j); and The intensity of the peak value of the second signal after crosstalk processing is greater than the intensity of the peak value of the first signal after crosstalk processing.

24. The analysis method according to claim 22, wherein: The multi-capillary electrophoresis device is configured to: in electrophoresis analysis, for a luminous point P (a≠j) on a capillary Cap (a≠j) other than the one capillary Cap (a=j), provide a first crosstalk processed signal peak with the largest intensity and a second crosstalk processed signal peak with the largest intensity in a time series of a processed signal Z (a≠j, c), wherein the first crosstalk processed signal peak and the second crosstalk processed signal peak are respectively synchronized with the first processed signal peak and the second processed signal peak in a time series of a processed signal Z (a=j, c) of the luminous point P (a=j) on the one capillary Cap (a=j); and The intensity of the peak value of the second signal after crosstalk processing is greater than the intensity of the peak value of the first signal after crosstalk processing.

25. The analysis method according to claim 20 or 21, wherein When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

26. The analytical method according to claim 22, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

27. The analytical method according to claim 23, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

28. The analytical method according to claim 24, wherein: When the (A×B) row and 1 column matrix X with the A×B original signals X(a, b) as elements is represented by the following mathematical formula 1: [Mathematical formula 1] The (A×C)-row-1-column matrix Z with the A×C processed signals Z(a, c) as elements is represented by the following mathematical formula 2: [Mathematical formula 2] and When a matrix Y having (A×B)×(A×C) elements Y(a, b)(a, c) and (A×B) rows and (A×C) columns satisfying the relationship X=Y×Z is represented by the following mathematical formula 3: [Mathematical formula 3] The polycapillary electrophoresis device is configured to obtain in advance a general inverse matrix Y of (A×C) rows and (A×B) columns of the matrix Y. - , and the multi-capillary electrophoresis device is configured to perform Z=Y as the combined processing - Calculation of ×X or processing corresponding to the calculation.

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