Analytical device and analytical method

Abstract

To provide a method for discriminating light emission from multiple light emission points and detecting each independently by computationally reducing spatial crosstalk between the multiple light emission points that occurs in an arbitrary optical system that detects the light emission from the multiple light emission points, respectively.SOLUTION: In an analysis method and an analysis device that detect fluorescence from each light emission point in multiple wavelength bands so as to discriminate light emission of a plurality of types of fluorophores from multiple light emission points, spatial crosstalk and spectral crosstalk exist between signal intensities of the multiple light emission points and the multiple wavelength bands, degrading the performance of the discrimination. By inputting all of detected signals in the multiple wavelength bands for each of the multiple light emission points into a predetermined arithmetic expression, the spatial crosstalk and the spectral crosstalk are eliminated and concentration of each emitter at each light emission point are derived.SELECTED DRAWING: Figure 30

Description

[Technical field]

[0001] The present disclosure relates to an analytical method and an analytical device for detecting fluorescence emitted from a plurality of types of phosphors at a plurality of light-emitting points while identifying each of the fluorescence. [Background technology]

[0002] In recent years, multi-capillary electrophoresis devices have been widely used, in which multiple capillaries are filled with electrophoretic separation media such as electrolyte solutions or electrolyte solutions containing polymer gels or polymers, and electrophoretic analysis is performed in parallel. The analysis targets are wide-ranging, from low molecular weight molecules to high molecular weight molecules such as proteins and nucleic acids. There are also many measurement modes, such as a mode in which lamp light is irradiated onto the absorption point of each capillary and the absorption of the lamp light that occurs when the analysis target passes through the absorption point is detected, or a mode in which laser light is irradiated onto the emission point of each capillary and the fluorescence or scattered light that occurs when the analysis target passes through the emission point is detected.

[0003] For example, in Patent Document 1, all capillaries around A light-emitting points on A capillaries (A is an integer of 2 or more) are arranged on the same plane, a laser beam is introduced from the side of the arrangement plane to irradiate the light-emitting points of all capillaries at once, and the fluorescence generated at each light-emitting point is wavelength-dispersed and detected at once in a direction perpendicular to the arrangement plane. In the detection device, the fluorescence emitted from the A light-emitting points is collimated at once by one focusing lens, transmitted through one transmission type diffraction grating, and the first-order diffracted light of each fluorescence is imaged at once on one two-dimensional sensor by one imaging lens. Here, the arrangement direction of the A light-emitting points and the wavelength dispersion direction by the diffraction grating are made perpendicular to each other, so that the wavelength-dispersed images of the emitted fluorescence from each capillary do not overlap each other on the two-dimensional sensor. For the wavelength-dispersed image of each capillary, B color detection is possible by setting detection areas of B arbitrary wavelength bands (B is an integer of 1 or more). The case of B=1 is called monochromatic detection, and the case of B≧2 is called multicolor detection. In the multi-capillary electrophoresis device of Patent Document 1, for example, DNA sequencing of different DNA samples can be performed by the Sanger method in each capillary. In the Sanger method, DNA fragments contained in a DNA sample are labeled with four types of fluorophores according to the terminal base types A, C, G, and T, and the fluorescence emitted by each is identified by multicolor detection.

[0004] In Patent Document 2, all capillaries around A light-emitting points on A capillaries (A is an integer of 2 or more) are arranged on the same plane, a laser beam is introduced from the side of the arrangement plane to irradiate the light-emitting points of all capillaries at once, and the fluorescence generated at each light-emitting point from the direction perpendicular to the arrangement plane is divided according to wavelength components and detected at once. In the detection device, the emitted fluorescence from the A light-emitting points is individually collimated by A condenser lenses to form A light beams, and each light beam is incident in parallel on a set of dichroic mirror arrays in which B dichroic mirrors (B is an integer of 1 or more) are arranged, and each is divided into B light beams of different wavelength bands, and the total A x B light beams generated are incident in parallel on one two-dimensional sensor, and A x B divided images are generated on the image. Here, by making the arrangement direction of A light emitting points and the B division direction of each light beam by the dichroic mirror array perpendicular to each other, the A x B divided images do not overlap each other on the image, and it becomes possible to set A x B detection areas. This makes it possible to detect B color in each capillary. Therefore, in the multi-capillary electrophoresis device of Patent Document 2, for example, as in Patent Document 1, it is possible to perform DNA sequencing by the Sanger method on different DNA samples in each capillary.

[0005] However, in general, it is not possible to distinguish between the fluorescence emitted by multiple types of phosphors simply by performing multi-color detection. This is because the fluorescence spectra of each phosphor overlap with each other, so the fluorescence of multiple types of phosphors is mixed in any one wavelength band (referred to as spectral crosstalk in this disclosure). Also, multiple types of phosphors may emit fluorescence at different concentrations simultaneously. Therefore, the next step (referred to as color conversion in this disclosure) eliminates the spectral crosstalk, making the above-mentioned distinction possible.

[0006] For each of A (A is an integer of 2 or more) light-emitting points, the fluorescence emitted by C type (C is an integer of 1 or more) phosphor is detected as color B in a detection area of ​​B type (B is an integer of 1 or more) wavelength band at each time, where B is greater than or equal to C. For each light-emitting point and each time, color conversion is performed on the color B detection result to obtain the concentration of the C type phosphor. For each light-emitting point P(a) (a=1, 2, ..., A), the fluorescence emitted by 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 phosphor D(c) at light-emitting point P(a) is Z(c), and the signal intensity of the detection area W(b) for light-emitting point P(a) is X(b). Here, the following equations hold, where X is a B-row, 1-column matrix whose elements are signal intensity X(b), Z is a C-row, 1-column matrix whose elements are concentration Z(c), and Y is a B-row, C-column matrix whose elements are Y(b)(c). (Equation 1) to (Equation 4) are the relational expressions for b and c, but they are not the relational expressions for a and hold independently for each light-emitting point P(a). In the case of monochromatic detection with B=1, C=1 because B≧C, and X, Y, and Z are no longer matrices.

[0007]

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[0008] Here, elements Y(b)(c) of matrix Y with B rows and C columns represent the signal intensity ratios of the fluorescence emitted by phosphor D(c) detected in a detection region W(b) of a different wavelength band due to spectral crosstalk. By making any one type of phosphor D(c0) emit fluorescence alone, one column Y(b)(c0) (b=1, 2, ..., B) of matrix Y can be determined. Here, since it is generally difficult to control the concentration of phosphor D(c0), it is convenient to normalize one column Y(b)(c0). For example, it is good to set the maximum element of the B elements to 1, and express the other elements as a ratio to the maximum value. Alternatively, it is good to determine the ratio of each element so that the sum of the B elements is 1. In other words,

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[0009]

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[0010] (Equation 1) is a system of simultaneous equations showing the relationship between the concentrations of C types of unknown phosphors and the known B-color fluorescence intensity, and (Equation 6) corresponds to obtaining its solution. Therefore, generally, as described above, the condition of B≧C is necessary. If B<C, the solution cannot be uniquely obtained (that is, since multiple solutions may exist), color conversion cannot be performed as in (Equation 6).

[0011] As an example, the case of C = 4 in the Sanger method and B = 4 in four-color detection will be described in detail. When producing copies of DNA fragments of various lengths for the template DNA by the Sanger reaction, they are labeled with four types of phosphors D(1), D(2), D(3), and D(4) according to the terminal base types A, C, G, and T, and while separating them by length by electrophoresis in order, they are irradiated with a laser beam to emit fluorescence. Fluorescent emissions are detected in four colors in four detection regions W(1), W(2), W(3), and W(4) corresponding to the respective maximum emission wavelengths of the phosphors D(1), D(2), D(3), and D(4), and time-series data of their signal intensities X(1), X(2), X(3), and X(4) (in the present disclosure, referred to as raw data. However, it is not limited to the case of B = 4 and C = 4) are acquired. Let the concentrations of the phosphors D(1), D(2), D(3), and D(4) at each time be Z(1), Z(2), Z(3), and Z(4), then (Equation 1) becomes the following equation.

[0012]

Equation

[0013] Here, elements Y(b)(c) of the 4-by-4 matrix Y represent the intensity ratio of the emission fluorescence of fluorophore D(c) (c is 1, 2, 3, or 4) detected in the wavelength band W(b) (b is 1, 2, 3, or 4) due to spectral crosstalk. The elements Y(b)(c) of matrix Y can be determined by electrophoretic analysis of samples in which fluorophore D(c) (c is 1, 2, 3, or 4) emits fluorescence independently in each capillary. For example, the four-color fluorescence intensities X(1), X(2), X(3), and X(4) when fluorophore D(1) emits fluorescence independently give elements Y(1)(1), Y(2)(1), Y(3)(1), and Y(4)(1), respectively. Moreover, the four-color fluorescence intensities X(1), X(2), X(3), and X(4) when phosphor D(2) is emitting fluorescence alone give the elements Y(1)(2), Y(2)(2), Y(3)(2), and Y(4)(2), respectively. The same is true for phosphors D(3) and D(4). Y(b)(c) is a fixed value determined only by the characteristics of phosphor D(c) and wavelength band W(b), and does not change during electrophoresis. Therefore, for each capillary, the concentrations of phosphors D(1), D(2), D(3), and D(4) at each time can be calculated from the four-color fluorescence intensities X(1), X(2), X(3), and X(4) at each time by the following formula, which embodies (Mathematical formula 6).

[0014]

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[0015] In this way, the inverse matrix Y -1 By multiplying this by the four-color fluorescence intensity, spectral crosstalk is eliminated and time series data on the concentrations of the four types of fluorophores, i.e., the concentrations of DNA fragments with four types of bases at their ends, is obtained (in this disclosure, this is referred to as color conversion data; however, this is not limited to the case where B=4 and C=4).

[0016] As already mentioned, the above color conversion process is performed independently for each of the A capillaries. This is based on the premise that the crosstalk between the capillaries (referred to as spatial crosstalk in this disclosure) is sufficiently small. In other words, this means that the ratio of the signal intensity obtained by detecting the fluorescence emitted from any one capillary to the signal intensity derived from the fluorescence emitted from other capillaries is sufficiently small. Therefore, the above-mentioned process of determining the matrix Y by sequentially performing the process of making one type of phosphor D(c0) emit fluorescence independently for C types of phosphor D(c) is performed substantially simultaneously for the A capillaries, and the matrix Y for each of the A capillaries is obtained in parallel. This is an essential process for deriving the matrix Y in a short time and without hassle.

[0017] Although the spatial crosstalk is basically reduced by devising an optical system, there are also attempts to reduce it by computational processing. In Patent Document 3, the emitted fluorescence from multiple light-emitting points randomly arranged on a plane, rather than from multiple capillaries, is imaged collectively on one two-dimensional sensor by one focusing lens. The emitted fluorescence from each light-emitting point is obtained as the signal intensity of a detection area provided at each imaging position on the two-dimensional sensor. It has been found that the ratio of spatial crosstalk that the signal intensity of the emitted fluorescence from any one light-emitting point gives to the signal intensity of the emitted fluorescence from another light-emitting point can be expressed as a function of the distance between those two light-emitting points or the distance between the corresponding two detection areas, and decreases with the distance. The above function is calculated in advance, and the mutual spatial crosstalk is calculated from the distance between any two detection areas and the above function in a fluorescence image of multiple light-emitting points randomly arranged on a plane, and the calculated spatial crosstalk is subtracted from the original fluorescence image, thereby reducing the spatial crosstalk. [Prior art documents] [Patent documents]

[0018] [Patent Document 1] Patent No. 3897277 [Patent Document 2] Patent No. 6456983 [Patent Document 3] Special Publication No. 2018-529947 Summary of the Invention [Problem to be solved by the invention]

[0019] In both Patent Document 1 and Patent Document 2, the fluorescence emitted from A light-emitting points on A capillaries is imaged separately from each other on the two-dimensional sensor, so that spatial crosstalk is essentially suppressed to a low level. Potential causes of spatial crosstalk include (1) lens aberration, (2) multiple reflections between fluorescent elements that occur inside an optical system composed of elements such as multi-capillaries, 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 an optical system so that the effects of the above factors (1) to (3) are minimized. For example, in order to suppress the effect of factor (2), it is possible to select a low-reflection coating for the lens. However, spatial crosstalk cannot be completely reduced to zero. In the detection of fluorescence emitted from any one capillary, spatial crosstalk effectively raises the lower detection limit, which may reduce the detection sensitivity and narrow the detection dynamic range at the same time. Therefore, reducing spatial crosstalk even slightly is extremely important in alleviating or resolving the above issues.

[0020] The present inventors have attempted to reduce spatial crosstalk by the method of Patent Document 3 for the optical system of Patent Document 1 or Patent Document 2, but it did not work well. First, the case of monochromatic detection with B=1 will be described. Although the spatial crosstalk that the signal intensity X(α)(β) of the detection region β for a light-emitting point α, which is an arbitrary light-emitting point among the A light-emitting points, gives to the signal intensity X(α')(β) of the detection region β for an arbitrary light-emitting point α' (α≠α') other than the above tends to decrease with the distance between the two detection regions, it has become clear that it cannot be expressed as a function of the distance. For example, even if the distance between the two detection regions is constant, the ratio of spatial crosstalk differs between the case where the two detection regions are located near the central axis of the optical system and the case where the two detection regions are located far from the central axis of the optical system. Also, the ratio of spatial crosstalk from one of the two detection regions to the other and the ratio of spatial crosstalk from the other to one of the two detection regions differ. In the case of multichromatic detection with B≧2, the method of Patent Document 3 does not work any more. The spatial crosstalk that the signal intensity X(α)(β) of the detection region β, which is any one of the detection regions B, imparts to the signal intensity X(α')(β') of any one of the detection regions β' (β=β' or β≠β', β=β' means a detection region of the same wavelength band, and β≠β' means a detection region of a different wavelength band) of the detection region B, for any one of the light-emitting points α' (α≠α') other than the above, could not be expressed as a function of the distance between the two detection regions. In particular, in the case of β≠β', spatial crosstalk and spectral crosstalk are mixed, so that the crosstalk between the two detection regions essentially could not be expressed as a function of the distance between them. This problem will be described in detail in [Form for carrying out the invention].

[0021] Therefore, in this disclosure, a method is proposed for identifying the emission from multiple emission points and detecting each of them independently by reducing the spatial crosstalk between multiple emission points, which occurs in any optical system that detects the emission from each of the multiple emission points, through computational processing. Also, a method is proposed for identifying the fluorescence emission from multiple types of phosphors from multiple emission points and detecting each of them independently by reducing the spatial crosstalk between multiple emission points and the spectral crosstalk between multiple types of phosphors for each emission point, which occurs in any optical system that detects the fluorescence of multiple types of phosphors emitted from multiple emission points, through computational processing. Alternatively, a method is proposed for identifying the absorption at multiple absorption points and detecting each of them independently by reducing the spatial crosstalk between multiple absorption points, which occurs in any optical system that detects the absorption at multiple absorption points, through computational processing. In addition, we propose a method for identifying the absorption of multiple types of light-absorbing bodies at multiple absorption points and detecting each of them independently by reducing the spatial crosstalk between multiple absorption points and the spectral crosstalk between multiple types of light-absorbing bodies at each absorption point through computational processing, which occurs in any optical system that detects the absorption of multiple types of light-absorbing bodies at multiple absorption points. [Means for solving the problem]

[0022] In the optical system of Patent Document 1 or Patent Document 2, etc., a case will be described where, for each of A (A is an integer of 2 or more) light-emitting points, C types (C is an integer of 1 or more) of fluorescent material are detected in B (B is an integer of 1 or more) wavelength band detection areas. Here, the detection areas of different wavelength bands are assumed to detect different wavelength components of the fluorescent material. Also, each of the different types of fluorescent material is assumed to emit fluorescent material having a different fluorescent spectrum. For each light-emitting point P(a) (a=1, 2, ..., A), the emitted fluorescent material of fluorescent material D(a, c) (c=1, 2, ..., C) present at the light-emitting 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 fluorescent material D(a, c) at the light-emitting point P(a) is Z(a, c), and the signal intensity of detection area W(a', b) for light-emitting point P(a') is X(a', b). Here, the inventors have discovered for the first time that the following equation holds when X is an A×B row and 1 column matrix with X(a',b) as element, Z is an A×C row and 1 column matrix with Z(a,c) as element, and Y is an (A×B) row and (A×C) column matrix with Y(a',b)(a,c) as element.

[0023]

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[0024] Equation 9 is the same as Equation 1, but comparing Equations 1 to 4 with Equations 9 to 12, we can see that they are completely different. Equations 9 to 12 are not just equations for b and c, but also equations for a, in which different light-emitting points P(a) are related to each other. In other words, Equations 9 to 12 are made to take into account the spatial crosstalk and spectral crosstalk between different light-emitting points P(a) all at once, in addition to the spectral crosstalk for the same light-emitting point P(a), and are therefore essentially different from Equations 1 to 4.

[0025] Here, the elements Y(a',b)(a,c) of the matrix Y with (A×B) rows and (A×C) columns represent (1) the signal intensity ratio of the emitted fluorescence of the phosphor D(a',c) at P(a') detected in an arbitrary detection region W(a',b) due to a'=a, i.e., the spectral crosstalk for the same emission point, and (2) the signal intensity ratio of the emitted fluorescence of the phosphor D(a,c) at the emission point P(a) detected in an arbitrary detection region W(a',b) of P(a') due to a'≠a, i.e., the spatial crosstalk and the spectral crosstalk for different emission points. One column Y(a,b)(a0,c0)(a=1,2,...,A, and b=1,2,...,B) of the matrix Y can be determined by making any one type of phosphor D(a0,c0) emit fluorescence independently at any one emission point P(a0). Here, since it is generally difficult to control the concentration of phosphor D(a0, c0), it is convenient to normalize one row Y(a, b)(a0, c0). For example, it is good to set the maximum element of the A×B elements as 1, and express the other elements as a ratio to the maximum value. Alternatively, it is good to determine the ratio of each element so that the sum of the A×B elements is 1. In other words,

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[0026]

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[0027] (14) is the same as (6), but when (2)-(4) and (6) are compared with (10)-(12) and (14), it is clear that they are completely different. (14) uses the generalized inverse matrix Y of the matrix Y previously calculated for each time. - By multiplying A×B signal intensities X(a,b) by Y, it is possible to eliminate both spectral crosstalk and spatial crosstalk and obtain the concentration Z(a,c) of A×C phosphors. - is a matrix with (A×C) rows and (A×B) columns. In this disclosure, the elimination of spatial crosstalk is called spatial correction, in correspondence with the elimination of spectral crosstalk being called color conversion. That is, Y - The (A×C)×(A×B) elements of include both a part that performs color conversion and a part that performs spatial correction. In this disclosure, the time series data of Z(a,c) obtained by executing (Equation 14) for each time is called color conversion + spatial correction data.

[0028] Each element of matrix X may be a value from which background light has been subtracted in advance, or may be a value to which appropriate noise reduction processing has been applied. Similarly, each element of matrix Y may be a value from which background light has been subtracted in advance, or may be appropriately changed.

[0029] In the case of monochromatic detection with B=1 and C=1, the above can be simplified as follows. For each light-emitting point P(a) (a=1, 2, ..., A), the emitted fluorescence of one type of phosphor is detected in one detection area. At any time, the concentration of the phosphor at light-emitting point P(a) is Z(a), and the signal intensity at light-emitting point P(a') is X(a'). Here, if X is an A-row, 1-column matrix with X(a') as its element, Z is an A-row, 1-column matrix with Z(a) as its element, and Y is an A-row, A-column matrix with Y(a')(a) as its element, then (Equation 10) to (Equation 12) can be simplified as follows. (Equation 9) and (Equation 14) hold as they are.

[0030]

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[0031] Although (Equation 9) and (Equation 14) to (Equation 17) are formally the same as the general formulas used in Patent Document 3, for example, [Equation 7] to [Equation 9] in Patent Document 3, the contents are significantly different (hereinafter, the formulas used in Patent Document 3 will be written in [ ]). The element αij of matrix A in [Equation 7] is a function of the distance dij between the detection areas of the light-emitting points φ(meas)i and φ(meas)j, and is expressed as the sum of exponential functions of dij as shown in [Equation 10], and αij attenuates with an increase in dij. In Patent Document 3, different fluorescent images are obtained for different samples, and the positions of the detection areas of multiple light-emitting points change randomly in each fluorescent image. Therefore, in Patent Document 3, the above function is calculated in advance, and the distance between the detection areas of any two light-emitting points among the multiple light-emitting points is calculated for each fluorescent image for all combinations of two light-emitting points, and αij is derived by substituting it into the above function. In other words, αij, i.e., matrix A, changes for each sample or for each fluorescent image. In contrast, in the present disclosure, as described above or as described later in [Form for carrying out the invention], it has been found that the element Y(a')(a) of the matrix Y in (Equation 16) cannot be expressed as a function of the distance between the detection areas of the light-emitting points P(a) and P(a'). It has also been found that it is impossible to calculate the element Y(a')(a) from the configuration of the optical system, etc. Therefore, in the present disclosure, the element Y(a')(a) is actually measured under the condition that the positions of the detection areas of the multiple light-emitting points do not change even for different samples. Specifically, as already described, one column Y(a')(a0) (a'=1, 2, ..., A) of the matrix Y is determined by causing only one of the light-emitting points P(a0) to emit fluorescence alone. Then, all columns of the matrix Y are determined by performing the above process in order for all A light-emitting points P(a). The matrix Y is determined only by the characteristics of the light-emitting point P(a) and does not change during the analysis. Also, as long as the conditions of the optical system, the light-emitting point P(a) and its detection area, etc. are fixed, the matrix Y is kept constant even when analyzing different samples. It is impossible to carry out the above steps with the optical system or device configuration of Patent Document 3.This is because the positions of the detection areas of multiple light-emitting points cannot be fixed, and even if they could be fixed, each light-emitting point cannot be made to emit light independently and in sequence. Therefore, the method of the present disclosure cannot be arrived at from Patent Document 3.

[0032] The above can be rephrased as follows. That is, for each of A (A is an integer of 2 or more) light-emitting points, the light emission of C types (C is an integer of 1 or more) of light-emitting bodies is detected in B (B is an integer of 1 or more) detection areas of arbitrary wavelength bands. For each light-emitting point P(a) (a=1, 2, ..., A), the light emission of light-emitting body D(a, c) (c=1, 2, ..., C) present at light-emitting point P(a) is detected in detection areas W(a, b) (b=1, 2, ..., B) of different wavelength bands. The concentration of light-emitting body D(a, c) at light-emitting point P(a) at an arbitrary time is Z(a, c), and the light emission intensity of W(a', b) for P(a') is 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 light-emitting bodies. Here, luminescence refers to fluorescence, phosphorescence, scattered light, and the like.

[0033] The above may also be interpreted as follows. That is, for each of A absorption points (A is an integer of 2 or more), the absorption of C types (C is an integer of 1 or more) of light absorbers is detected in B detection areas of arbitrary wavelength bands (B is an integer of 1 or more). For each absorption point P(a) (a=1, 2, ..., A), the absorption of light absorber D(a, c) (c=1, 2, ..., C) present at absorption point P(a) is detected in detection areas W(a, b) (b=1, 2, ..., B) of different wavelength bands. The concentration of light absorber D(a, c) at absorption point P(a) at an arbitrary time is Z(a, c), and the absorbance of W(a', b) at absorption point P(a') is X(a', b). In this case as well, equations (9) to (17) hold, and similarly, both the spectral crosstalk and the spatial crosstalk can be eliminated and the concentration Z(a, c) of A × C light absorbers can be found.

[0034] Alternatively, the above can be interpreted as multipoint detection other than optical measurement. That is, for each of A signal generation points (A is an integer of 2 or more), signals of C types (C is an integer of 1 or more) of signal generators are detected in B detection areas of arbitrary frequency bands (B is an integer of 1 or more). For each signal generation point P(a) (a=1, 2, ..., A), signals of signal generators D(a, c) (c=1, 2, ..., C) present at the signal generation point P(a) are detected in detection areas W(a, b) (b=1, 2, ..., B) of different frequency bands. At any time, the concentration of signal generators D(a, c) at signal generation point P(a) is Z(a, c), and the signal intensity of W(a', b) at signal generation point P(a') is X(a', b). In this case as well, equations (9) to (17) hold, and similarly, both the spectral crosstalk and the spatial crosstalk can be eliminated and the concentration Z(a, c) of A × C signal generators can be found.

[0035] The above explanation mainly uses formulas, but this is for the purpose of making the contents of this disclosure easier to understand. When implementing the technology of this disclosure, it is sufficient to use a method based on the contents of this disclosure, and it is not necessary to follow the formulas exactly; the formulas may be modified, or formulas may not be used. Furthermore, in this disclosure, the term "concentration of illuminant" may be read as "luminescence intensity of illuminant," the term "concentration of phosphor" may be read as "fluorescence intensity of phosphor," and the term "concentration of light absorbant" may be read as "absorbance of light absorbant."

[0036] Further features related to the present disclosure will become apparent from the description of the present specification and the accompanying drawings. Also, aspects of the present disclosure may be realized and realized by the elements and combinations of various elements and aspects set forth in the following detailed description and the appended claims. The descriptions in this specification are exemplary and illustrative only and are not intended to limit the scope or application of the present disclosure in any way.

[0037] Further features related to the present disclosure will become apparent from the description of the present specification and the accompanying drawings. Also, aspects of the present disclosure may be realized and realized by the elements and combinations of various elements and aspects set forth in the following detailed description and the appended claims. The descriptions in this specification are exemplary and illustrative only and are not intended to limit the scope or application of the present disclosure in any way. Effect of the Invention

[0038] According to the present disclosure, by eliminating or reducing spatial crosstalk between multiple light-emitting points, which occurs in any optical system that detects light emission from multiple light-emitting points, through computational processing, it becomes possible to identify the light emission from multiple light-emitting points and detect each of them independently.Furthermore, by eliminating or reducing spatial crosstalk between multiple light-emitting points and spectral crosstalk between multiple types of phosphors for each light-emitting point, which occurs in any optical system that detects the fluorescence of multiple types of phosphors emitted from multiple light-emitting points, it becomes possible to identify the fluorescent emission of multiple types of phosphors from multiple light-emitting points and detect each of them independently.

[0039] Furthermore, by eliminating or reducing spatial crosstalk and spectral crosstalk, it is possible to avoid the decrease in detection sensitivity or the decrease in detection dynamic range that occurs due to spatial crosstalk and spectral crosstalk. Problems, configurations and effects other than those described above will become apparent from the following description of the embodiments. [Brief description of the drawings]

[0040] [Figure 1] Schematic diagram of a simple optical system [Diagram 2] Luminescence image captured by a 2D sensor using a simple optical system [Diagram 3] Signal intensity distribution of luminescence image in luminescence image [Figure 4]The relationship between the absolute signal intensity at the center of the luminescence image and the absolute signal intensity at a position far to the left of the center of the luminescence image [Diagram 5] The relationship between the absolute signal intensity at the center of the luminescence image and the absolute signal intensity at a position far to the right of the center of the luminescence image [Figure 6] The relationship between the absolute signal intensity at the center of the luminescence image and the absolute signal intensity at a position far to the left of the center of the luminescence image [Figure 7] The relationship between the absolute signal intensity at the center of the luminescence image and the absolute signal intensity at a position far to the right of the center of the luminescence image [Figure 8] Schematic diagram of the model experiment system [Figure 9] Schematic diagram of spectral crosstalk and spatial crosstalk in a model experimental system [Figure 10] Schematic diagram of the model experiment system [Figure 11] Raw data obtained from electrophoretic analysis of known samples [Figure 12] Color transformation data obtained by performing color transformation on the raw data in Figure 11 [Figure 13] Color conversion + spatial correction data obtained by performing color conversion + spatial correction on the raw data in Figure 11 [Figure 14] Raw data obtained from electrophoretic analysis of unknown samples [Figure 15] Color transformation data obtained by performing color transformation on the raw data in Figure 14 [Figure 16] Color conversion + spatial correction data obtained by performing color conversion + spatial correction on the raw data in Figure 14 [Figure 17] Raw data from electrophoretic analysis of a sample containing an unknown fluorophore [Figure 18] Color conversion + spatial correction data obtained by performing color conversion + spatial correction on the raw data in Figure 17 [Figure 19] Raw data from electrophoretic analysis of samples containing increasing concentrations of known fluorophores. [Figure 20] Color transformation data obtained by performing color transformation on the raw data in Figure 19 [Figure 21] Color conversion + spatial correction data obtained by performing color conversion + spatial correction on the raw data in Figure 19 [Figure 22] Schematic diagram of a method for injecting samples into multiple capillaries at different times [Figure 23] Schematic diagram of a method for obtaining fluorescence emission by shifting the timing using multiple capillaries [Figure 24] Schematic diagram of multi-capillary electrophoresis device [Diagram 25] Wavelength dispersion image of Raman scattered light from four capillaries using the optical system of Patent Document 1 [Figure 26] A diagram showing the setting of detection areas of 20 wavelength bands for each capillary in FIG. [Figure 27] A schematic diagram of the optical system of Patent Document 2 dividing the emitted fluorescence from the four capillaries into four wavelength bands and forming an image. [Figure 28] An optical system that splits the emitted fluorescence from five light-emitting points arranged on a plane into four wavelength bands and forms an image. [Figure 29] Flowchart of conventional method [Diagram 30] Flowchart of this method [Diagram 31] Flowchart for determining the matrix for color conversion and spatial correction of this method [Diagram 32] Flowchart for multiple analysis sessions [Diagram 33] Computer configuration diagram DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0041] [Example 1] In order to investigate the characteristics of spatial crosstalk in detail, a simple optical system shown in Fig. 1 was constructed. The optical system in Fig. 1 includes a pinhole plate 1-1, an aperture plate 1-2 on the light-emitting point side, a condenser lens 1-3, an aperture plate 1-4 on the sensor side, a colored glass filter 1-5, a two-dimensional sensor 1-6, and a halogen lamp (light source) that irradiates halogen lamp light 1-7. Specifically, the optical system in Fig. 1 was constructed as follows. A light-emitting point 1-8 with a diameter of 0.05 mm was formed by irradiating the pinhole plate 1-1 with a pinhole of φ0.05 mm from below with halogen lamp light 1-7. In addition, the aperture plate 1-2 on the light-emitting point side with an aperture of φ0.2 mm was placed 0.2 mm above the light-emitting point 1-8, the condenser lens 1-3 with a focal length of f=1.4 mm was placed 1.54 mm above the light-emitting point 1-8, and the aperture plate 1-4 on the sensor side with an aperture of φ0.7 mm was placed directly above the condenser lens 1-3. Furthermore, the two-dimensional sensor 1-6 was placed 15 mm above the condenser lens 1-3, and the colored glass filter 1-5 was placed directly below the two-dimensional sensor 1-6. Thus, the pinhole plate 1-1, the light-emitting point side aperture plate 1-2, the sensor side aperture plate 1-4, the colored glass filter 1-5, and the two-dimensional sensor 1-6 were arranged parallel to one another. The light 1-9 emitted from the light-emitting point 1-8 passed through an aperture of φ0.2 mm, was collected by the condenser lens 1-3, passed through an aperture of φ0.7 mm, passed through the colored glass filter 1-5, and formed a light-emitting image 1-10 of φ0.5 mm on the two-dimensional sensor 1-6. Here, the light-emitting point 1-8 was focused on the two-dimensional sensor 1-6 and imaged at a magnification of 10 times.

[0042] Figure 2 is a luminescence image including an image 1-10 obtained by a two-dimensional sensor 1-6 in the simple optical system of Figure 1. The sensor size of the two-dimensional sensor 1-6 is 13 x 13 mm, and the signal range of each pixel is 0 to 65536. Figure 2(a) and Figure 2(b) are the same luminescence image, but the signal display scale (gradation) of Figure 2(a) is 0 to 50000, and the signal display scale of Figure 2(b) is 0 to 500. The maximum signal value of the luminescence image is about 50000, and while the signals of all pixels are displayed almost without saturation in Figure 2(a), the luminescence image is displayed saturated in Figure 2(b). According to Figure 2(a), a luminescence image of φ0.5 mm was obtained as expected, and the outside of the φ0.5 mm luminescence image appears to have zero signal. However, according to Figure 2(b), it can be confirmed that a low intensity skirt spreads outside the φ0.5 mm luminescence image.

[0043] The dotted lines α in Figures 3(a) to (c) indicate the horizontal signal intensity distribution passing through the center of the luminescence image in Figure 2. Figures 3(a), 3(b), and 3(c) use the same data, but the vertical axis is changed. The horizontal axis is common and indicates the distance from the center of the luminescence image. The plus and minus signs on the horizontal axis indicate the right and left sides of the luminescence image, respectively. The vertical axes in Figures 3(a) and (b) indicate absolute signal intensity, with the vertical axis scale of 0 to 60,000 in Figure 3(a) and 0 to 100 in Figure 3(b). On the other hand, Figure 3(c) uses a logarithmic vertical axis, with the vertical axis scale of 0.001% to 100% as the relative signal intensity with the maximum signal intensity being 100%. β (solid line) and γ (dashed line) in Figures 3(a)-(c) show the signal intensity distribution similar to α when the output intensity of the halogen lamp light 1-7 is gradually decreased under the conditions for acquiring the luminescence image in Figure 2. As can be seen from Figure 3(a), the maximum signal intensity of the luminescence images of α, β, and γ are about 50,000, about 25,000, and about 10,000, respectively. On the other hand, looking at Figure 3(b), it can be seen that the signal intensity tends to decrease as it moves away from the center of the luminescence image, but compared to the size of the luminescence image seen in Figure 3(a) (about 0.5 mm wide), the tail is much wider. It can also be seen that as the maximum signal intensity decreases, the tail intensity also decreases. Therefore, looking at Figure 3(c), it can be seen that α, β, and γ overlap one another. This is a new discovery and brings several important insights. For example, a signal intensity of about 0.1% of the maximum signal intensity is observed at a position ±1 mm away from the center of the luminescence image, that is, at the outer position of the luminescence image seen in Figure 2(a). If the detection regions of adjacent light emission images exist at positions separated by ±1 mm, this means that there is a spatial crosstalk of approximately 0.1%. As shown in Figure 1, it is surprising that such a non-negligible amount of spatial crosstalk exists even though an extremely simplified optical system is used and the light emission point is just focused. Even more surprising is that, because α, β, and γ overlap, it was found that the relative signal intensity distribution of the imaged point is constant, regardless of the light emission intensity of the light emission point or the signal intensity of the imaged point, as long as the positions of the optical system and the light emission point are fixed.

[0044] Figures 4(a) and 5(a) show the results derived from Figure 3. Figure 4(a) is a graph showing absolute signal intensity at positions -1, -2, -3, -4, -5, and -6 mm away from the center of the imaging point, and Figure 5(a) is a graph showing absolute signal intensity at positions +1, +2, +3, +4, +5, and +6 mm away from the center of the imaging point on the vertical axis, and absolute signal intensity at the center of the emission image on the horizontal axis in both cases. The above absolute signal intensity was taken as the average value of absolute signal intensity in a ±0.1 mm width centered on each position in Figure 3. As a result, it was found that all three plots at each position (corresponding to α, β, and γ in Figure 3) were on an approximated straight line passing through the origin. The slope of each approximated straight line indicates the spatial crosstalk ratio at the corresponding position. In other words, the slope of the approximated straight line becomes smaller as the distance from the center of the imaging point increases, and the spatial crosstalk ratio decreases. For comparison, the lines representing spatial crosstalk of 0.1% and 0.01% are superimposed as dotted lines. For example, it can be seen that the spatial crosstalk ratio at a position -1 mm away from the center of the imaging point is just over 0.1%, while the spatial crosstalk ratio at a position -3 mm away is about 0.01%. The above results indicate that the absolute signal strength of the spatial crosstalk at any position away from the center of the imaging point is linear with respect to the absolute signal strength at the center of the imaging point, and the spatial crosstalk ratio is constant. In other words, it has been newly discovered that it is possible to eliminate or reduce spatial crosstalk at any position by subtracting the value obtained by multiplying the absolute signal strength at the center of the imaging point by the spatial crosstalk ratio from the absolute signal strength at any position away from the center of the imaging point. This is equivalent to subtracting the value of the corresponding approximation line at the same position from the absolute signal strength of each plot in Figures 4(a) and 5(a).

[0045] Figures 4(b) and 5(b) show the results of performing the above subtraction operation on the results of Figures 4(a) and 5(a), respectively. The types of plots showing each position and the horizontal axis are the same for Figures 4(a) and 4(b) and Figures 5(a) and 5(b). The vertical axis of Figures 4(b) and 5(b) shows the absolute signal intensity after the above subtraction operation, and the scale is enlarged compared to Figures 4(a) and 5(a). For comparison, a dotted line showing a spatial crosstalk of ±0.01% is superimposed. As a result, it was found that the absolute signal intensity after the subtraction operation at each position was almost zero regardless of the absolute signal intensity at the center of the image, and the spatial crosstalk at each position was smaller than ±0.01%. When compared at a position -1 mm away from the center of the imaging point, this method was able to reduce the spatial crosstalk by at least one order of magnitude. The reason why the absolute signal intensity after subtraction at each position does not become completely zero is because there is a deviation or error in the approximation line corresponding to each plot in Figure 4(a) and Figure 5(a). The coefficient of determination (R 2 ) is higher (closer to 1), that is, the higher the linearity of the absolute signal strength at each position relative to the absolute signal strength at the center of the imaging point, the smaller the above error will be and the closer the absolute signal strength after the subtraction operation will be to zero. Naturally, the absolute signal strength after the subtraction operation may become negative. However, as long as the above linearity exists, the magnitude of the absolute signal strength at each position will at least be reduced by the subtraction operation. Also, the smaller the slope of the approximation line, that is, the smaller the spatial crosstalk before the subtraction operation, the smaller the absolute value of the difference between the absolute signal strength at each position and the approximation line will be, and therefore the smaller the above error will be.

[0046] Figures 6 and 7 show the same experimental results as Figures 4 and 5. In the optical system of Figure 1, the pinhole plate 1-1 was once removed and then reinstalled, after which the same results as Figures 2 and 3 were obtained, and the results of Figures 6 and 7 were derived using a similar method. The mounting position of the pinhole plate 1-1 was approximately the same as the original position, but not exactly the same position. The results of Figures 6 and 7 are equivalent to the results of Figures 4 and 5, demonstrating the high reproducibility of the method of eliminating or reducing spatial crosstalk at any position by subtraction processing.

[0047] On the other hand, from the comparison of Figures 4 to 7, another important finding was obtained, as shown below. The approximation lines of -1 mm in Figure 4(a), +1 mm in Figure 5(a), -1 mm in Figure 6(a), and +1 mm in Figure 7(a) are compared. It was found that the slopes of these four approximation lines all indicate the spatial crosstalk ratio at a position 1 mm away from the center of the imaging point, but are mutually different. Therefore, for example, if the approximation line of -1 mm in Figure 4(a) is used to perform subtraction processing of each plot of +1 mm in Figure 5(a), -1 mm in Figure 6(a), or +1 mm in Figure 7(a), it became clear that the above error would increase and the reduction of spatial crosstalk would be insufficient. This phenomenon also occurred at other positions, and it was found that in some cases, the subtraction processing would have the opposite effect, that is, increase the spatial crosstalk. The causes of such a phenomenon are considered to be: (1) even in a simple optical system such as that shown in FIG. 1, the point symmetry of the spatial crosstalk caused by the imaging is not necessarily high (for example, in FIG. 2, the spatial crosstalk ratio is different between the left and right directions of the imaging point); and (2) the spatial crosstalk caused by the imaging changes significantly with a subtle change in the optical system, such as a slight shift in the position of the light emitting point. Therefore, it was found that it is impossible to express the spatial crosstalk ratio occurring between the center of the imaging point and an arbitrary position as a function of the distance between them. This means that the method shown in Patent Document 3 does not function effectively. This disclosure differs from Patent Document 3 in that the conditions of the optical system and the multiple imaging points, i.e., the multiple detection areas, are fixed, and the mutual spatial crosstalk ratios are derived by experiment, regardless of the mutual distance, as shown in each of FIGS. 4 to 7. If the conditions are changed, the spatial crosstalk ratio needs to be reacquired each time.

[0048] The findings obtained from the above experiments, namely, that the absolute signal strength of spatial crosstalk at any position away from the center of the imaging point is linear with respect to the absolute signal strength at the center of the imaging point and that the spatial crosstalk ratio is constant, can be expanded and generalized to derive equations (9) to (17).

[0049] [Example 2] A model experiment was carried out for the simplest case of an optical system that detects the fluorescence of multiple types of phosphors emitted from multiple light-emitting points in detection areas of multiple wavelength bands. In this embodiment and other embodiments using the same model experimental system, as an example, in (Equation 9) to (Equation 12), a case was chosen in which the emitted fluorescence of C = 2 types of phosphors is detected in detection areas of B = 2 wavelength bands for each of A = 2 light-emitting points, but it goes without saying that the same effect can be obtained by the same method when A, B, and C are each other numerical values.

[0050] Figure 8 shows a model experimental system. The model experimental system in Figure 8 includes two capillaries Cap(1) and Cap(2) and a light source (not shown) that irradiates a laser beam LB in the arrangement direction of the two capillaries Cap(1) and Cap(2). Emission points P(1) and P(2) are provided on the two capillaries Cap(1) and Cap(2) at positions where the laser beam LB is irradiated. A substance labeled with two types of phosphors D(1,1) and D(1,2) is electrophoresed in the capillary Cap(1), and a substance labeled with two types of phosphors D(2,1) and D(2,2) is electrophoresed in the capillary Cap(2). Here, D(1,1) and D(2,1) are the same type of phosphor, and D(1,2) and D(2,2) are the same type of phosphor. The same type of phosphor means that the fluorescence spectra of the emitted fluorescence are equal. Phosphors D(1,1) and D(1,2) emit fluorescence when irradiated with a laser beam LB at the light-emitting point P(1), and are detected in detection areas W(1,1) and W(1,2) provided in the sensor S. The detection wavelength bands of the detection area W(1,1) and W(1,2) are designed so that the detection area W(1,1) mainly detects the emitted fluorescence of phosphor D(1,1), and the detection area W(1,2) mainly detects the emitted fluorescence of phosphor D(1,2), respectively, but there is significant mutual spectral crosstalk. Similarly, phosphors D(2,1) and D(2,2) emit fluorescence when irradiated with a laser beam LB at the light-emitting point P(2), and are detected in the detection areas W(2,1) and W(2,2) provided in the sensor S. The detection wavelength bands of the detection region W(2,1) and W(2,2) are designed so that the detection region W(2,1) mainly detects the emitted fluorescence of phosphor D(2,1), and the detection region W(2,2) mainly detects the emitted fluorescence of phosphor D(2,2), respectively, but there is significant mutual spectral crosstalk. Furthermore, the fluorescence emission of phosphors D(1,1) and D(1,2) at the emission point P(1) is also detected in the detection regions W(2,1) and W(2,2), and similarly, the fluorescence emission of phosphors D(2,1) and D(2,2) at the emission point P(2) is also detected in the detection regions W(1,1) and W(1,2), and there is significant spatial crosstalk.If the signal intensities (fluorescence intensities) in detection regions 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, as shown in the lower part of FIG. 8, time-series data of signal intensities X(1,1), X(1,2), X(2,1), and X(2,2) are obtained with electrophoresis for light-emitting point P(1) and light-emitting point P(2), respectively.

[0051] In addition, in FIG. 8, the detection areas W(1,1), W(1,2), W(2,1), and W(2,2) are depicted as being provided on one sensor S, but this is not necessarily required. The detection areas W(1,1) and W(1,2) may be provided on one sensor, and the detection areas W(2,1) and W(2,2) may be provided on another sensor, or the detection areas W(1,1), W(1,2), W(2,1), and W(2,2) may be provided on four different sensors. In addition, some kind of imaging means and spectroscopic means are required between the light emitting points P(1) and P(2) and the sensor S, but since the present embodiment does not require any type of imaging means or spectroscopic means, the imaging means and spectroscopic means are omitted in FIG. 8.

[0052] FIG. 9 is a schematic diagram showing the relationship between the spectral crosstalk and the spatial crosstalk in FIG. 8. In FIG. 9(a), the fluorescence emission of the phosphor D(1,1) at the emission point P(1) is (i) mainly detected in the detection region W(1,1) and (ii) secondarily detected in the detection region W(1,2). The above-mentioned emitted fluorescence is also (iii) detected in the detection region W(2,1) with a much smaller intensity than (i) and (ii), and furthermore, (iv) detected in the detection region W(2,2) with an even smaller intensity than (iii). (i) and (ii) indicate the spectral crosstalk, and (iii) and (iv) indicate the spatial crosstalk and the spectral crosstalk. In other words, the conventional method considers only (i) and (ii), but the present disclosure considers all of (i) to (iv). In FIG. 9, the relationship between the strength and weakness of the detection intensity and the relationship between the crosstalk are commonly indicated by the thickness of the arrow, the solid or dotted line, and the numbers (i) to (iv). As with Fig. 9(a), Fig. 9(b) illustrates the fluorescence emission of phosphor D(1,2) at emission point P(1), Fig. 9(c) illustrates the fluorescence emission of phosphor D(2,1) at emission point P(2), and Fig. 9(d) illustrates the fluorescence emission of phosphor D(2,2) at emission point P(2). In an actual analysis, Figs. 9(a)-(d) occur simultaneously at the same time, and each has a different fluorescence intensity.

[0053] Figure 10 shows a model experimental system similar to that of Figure 8. However, in the model experimental system of Figure 10, lamp light LL is irradiated from a light source perpendicular to the arrangement direction of two capillaries Cap(1) and Cap(2). Therefore, the emission fluorescence accompanying the irradiation of the laser beam LB is not detected, but the absorption or absorbance accompanying the transmission of the lamp light LL is detected. Absorption points P(1) and P(2) are provided on the two capillaries Cap(1) and Cap(2), respectively, at the positions where the lamp 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 absorber, and D(1,2) and D(2,2) are the same type of light absorber. The same type of light absorber means that the light absorption spectrum is equal. Light absorbers D(1,1) and D(1,2) absorb light when irradiated with lamp light LL at the absorption point P(1), and the unabsorbed light is detected in the detection areas W(1,1) and W(1,2) provided in the sensor S. The detection wavelength bands of the detection area W(1,1) and W(1,2) are designed so that the absorption of the light absorber D(1,1) and the absorption of the light absorber D(1,2) are mainly detected in the detection area W(1,1), and the absorption of the light absorber D(1,2), respectively, but there is significant mutual spectral crosstalk. Similarly, light absorbers D(2,1) and D(2,2) absorb light when irradiated with lamp light LL at the absorption point P(2), and the unabsorbed light is detected in the detection areas W(2,1) and W(2,2) provided in the sensor S. The detection wavelength bands of the detection region W(2,1) and W(2,2) are designed so that they mainly detect the absorption of the light absorber D(2,1), and the absorption of the light absorber D(2,2), respectively, but there is significant mutual spectral crosstalk. In addition, the absorption of the light absorbers D(1,1) and D(1,2) at the absorption point P(1) is also detected in the detection regions W(2,1) and W(2,2), and similarly, the absorption of the light absorbers D(2,1) and D(2,2) at the absorption point P(2) is also detected in the detection regions W(1,1) and W(1,2), and there is significant spatial crosstalk.This is because a part of the transmitted light of the lamp light LL that passes through the absorption point P(1) is also detected in the detection regions W(2,1) and W(2,2), and a part of the transmitted light of the lamp light LL that passes through the absorption point P(2) is also detected in the detection regions W(1,1) and W(1,2). If the absorbances in the detection regions 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), respectively, time series data of absorbances X(1,1), X(1,2), X(2,1), and X(2,2) are obtained with electrophoresis for the absorption points P(1) and P(2), as shown in the lower part of FIG. 8. In the following, the case of FIG. 8 will be described, but it goes without saying that the same effect can be obtained when FIG. 10 is used.

[0054] Figure 11 shows raw data obtained by electrophoretic analysis of a known sample injected into capillaries Cap(1) and Cap(2) using the model experimental system of Figure 8. In both cases, the horizontal axis indicates electrophoretic time (arbitrary units) and the vertical axis indicates fluorescence intensity (arbitrary units). Figures 11(a) and 11(b) show the same raw data obtained at emission point P(1), and (b) shows an enlarged vertical scale of (a). Similarly, Figures 11(c) and 11(d) show the same raw data obtained at emission point P(2), and (d) shows an enlarged vertical scale of Figure 11(c). The horizontal scale of Figures 11(a) to (d) indicates time 0 to 500, the vertical scale of Figures 11(a) and (c) indicates fluorescence intensity 0 to 250, and the vertical scale of Figures 11(b) and (d) indicates fluorescence intensity -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. As shown in Figure 11, the sample is prepared so that phosphor D(1,1) (state in Figure 9(a)) at light-emitting point P(1) at time 100, phosphor D(2,1) (state in Figure 9(c)) at light-emitting point P(2) at time 200, phosphor D(1,2) (state in Figure 9(b)) at light-emitting point P(1) at time 300, and phosphor D(2,2) (state in Figure 9(d)) at light-emitting point P(2) at time 400 each emit fluorescence independently. The sample is also prepared so that no fluorescence other than these is emitted. 11(a) and 11(c), only four large peaks corresponding to the above-mentioned fluorescence emission are observed (corresponding to (i) and (ii) in Fig. 9), while in Fig. 11(b) and 11(d), four small peaks indicated by arrows are observed, namely, phosphor D(2,1) at emission point P(2) at time 100, phosphor D(1,1) at emission point P(1) at time 200, phosphor D(2,2) at emission point P(2) at time 300, and phosphor D(1,2) at emission point P(1) at time 400 (corresponding to (iii) and (iv) in Fig. 9). Therefore, it can be determined that these four small peaks are due to spatial crosstalk and spectral crosstalk.For example, the small peak of phosphor D(2,1) at emission point P(2) at time 100 is the result of part of the fluorescent emission of phosphor D(1,1) at emission point P(1) at time 100 being detected as X(2,1) and X(2,2) due to spatial crosstalk and spectral crosstalk (corresponding to (iii) and (iv) in Figure 9(a)).

[0055] If spatial crosstalk is not taken into consideration, that is, if the four small peaks in Figures 11(c) and 11(d) are ignored (if only (i) and (ii) in Figure 9 are considered), it is sufficient to eliminate the spectral crosstalk of the four large peaks in Figures 11(a) and 11(c), that is, to perform color conversion using the conventional method. In this case, (Equation 6) is expressed by the following equation.

[0056]

number

number

[0057] Here, Z(1,1), Z(1,2), Z(2,1), and Z(2,2) indicate 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 was the same at the light emitting point P(1) and the light emitting point P(2). However, in general, the matrix Y and the inverse matrix Y -1 Each element of matrix Y for light-emitting point P(1) was determined by the intensity ratio of the fluorescence intensities X(1,1) and X(1,2) due to phosphor D(1,1) at time 100, and the intensity ratio of the fluorescence intensities X(1,1) and X(1,2) due to phosphor D(1,2) at time 300. Each element of matrix Y for light-emitting point P(2) was determined by the intensity ratio of the fluorescence intensities X(2,1) and X(2,2) due to phosphor D(2,1) at time 200, and the intensity ratio of the fluorescence intensities X(2,1) and X(2,2) due to phosphor D(2,2) at time 400.

[0058] FIG. 12 shows color-transformed data obtained by performing color transformations of (Equation 18) and (Equation 19) at each time on the raw data of FIG. 11. The notation method is the same as that of FIG. 11. As expected, it can be seen that the spectral crosstalk of the four large peaks in FIG. 11(a) and FIG. 11(c) is eliminated by the four large peaks in FIG. 12(a) and FIG. 12(c). On the other hand, the spectral crosstalk of the four small peaks in FIG. 11(b) and FIG. 11(d) is also eliminated by the four small peaks in FIG. 12(c) and FIG. 12(d) (also indicated by arrows), but these peaks themselves still remain, and it can be seen that the spatial crosstalk has not been eliminated. This is the problem with the conventional method.

[0059] Next, in FIG. 11, both the spectral crosstalk and the spatial crosstalk are taken into consideration (all of (i) to (iv) in FIG. 9 are taken into consideration), and both the color conversion and the spatial correction are performed. In this case, (Equation 14) is expressed by the following equation.

[0060]

number

[0061] Matrix Y and inverse matrix Y -1has been expanded from 2 rows and 2 columns in (Equation 18) and (Equation 19) to 4 rows and 4 columns. Also, while in (Equation 18) and (Equation 19) color conversion is performed independently at light-emitting point P(1) and light-emitting point P(2), in (Equation 20) light-emitting point P(1) and light-emitting point P(2) are mixed together and color conversion + space correction is performed collectively. Each element of matrix Y in (Equation 20) was determined from the intensity ratio of fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) due to phosphor D(1,1) at time 100, the intensity ratio of fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) due to phosphor D(2,1) at time 200, the intensity ratio of fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) due to phosphor D(1,2) at time 300, and the intensity ratio of fluorescence intensities X(1,1), X(1,2), X(2,1), and X(2,2) due to phosphor D(2,2) at time 400 in FIG. 11. The upper left 2 rows and 2 columns and the lower right 2 rows and 2 columns of matrix Y in (20) correspond to matrix Y in (18) and (19), and the values ​​of each element are almost equal to each other. Similarly, matrix Y in (20) -1 The top two rows and two columns of the upper left and bottom two rows and two columns of the lower right are the matrices Y -1 and the values ​​of the elements are almost equal to each other. In other words, these elements are responsible for color conversion that eliminates spectral crosstalk for the same light-emitting point. The reason why the values ​​of these elements do not completely match between (Equation 20) and (Equation 18) and (Equation 19) is due to the difference in whether or not spatial crosstalk is taken into consideration, as described below. In contrast, the matrices Y and Y -1 The top right two rows and two columns and the bottom left two rows and two columns are responsible for spatial correction and color conversion that eliminates spatial crosstalk and spectral crosstalk between different light-emitting points, which do not exist in (Equation 18) and (Equation 19). The above becomes easier to understand if (Equation 20) is transformed into the following equation.

[0062]

number

number

[0063] The first term on the right-hand side of (Equation 21) and the second term on the right-hand side of (Equation 22) are responsible for the color conversion of the conventional method, and correspond to (Equation 18) and (Equation 19), respectively. In contrast, the second term on the right-hand side of (Equation 21) and the first term on the right-hand side of (Equation 22) are responsible for the spatial correction and color conversion, which are not handled in the conventional method. In this way, color conversion and spatial correction and color conversion may be separated and executed separately, or only one of them may be executed. Alternatively, it is also possible to change the processing depending on the light-emitting point, for example, by applying spatial correction and color conversion to light-emitting point P(1) and only color conversion to light-emitting point P(2).

[0064] FIG. 13 shows the color-converted and spatially corrected data obtained by performing the color conversion and spatial correction of (Equation 20) on the raw data of FIG. 11 at each time. The notation is the same as in FIG. 11. First, as in FIG. 12, it can be seen that the spectral crosstalk of the four large peaks in FIG. 11(a) and FIG. 11(c) is eliminated by the four large peaks in FIG. 13(a) and FIG. 13(c). Furthermore, as expected, it can be seen that the spatial crosstalk and spectral crosstalk of the four small peaks in FIG. 11(b) and FIG. 11(d) are also eliminated in FIG. 13(c) and FIG. 13(d), and the peaks have disappeared (indicated by the arrows).

[0065] As a result of the above, it has been shown that the present disclosure makes it possible to eliminate or reduce, through computational processing, the spectral crosstalk for the same light-emitting point, the spatial crosstalk between multiple light-emitting points, and the spectral crosstalk that occurs in any optical system that detects the fluorescence of multiple types of phosphors emitted from multiple light-emitting points.

[0066] [Example 3] Next, based on the experimental results of [Example 2], an unknown sample was analyzed. Figure 14 shows raw data obtained by electrophoretic analysis of an unknown sample injected into capillaries Cap(1) and Cap(2) using the model experimental system of Figure 8. Since the same model experimental system as in [Example 2] is used, (Equation 18) to (Equation 22) can be used as is. The notation method is the same as in Figure 11. However, the vertical axis scale of Figure 11(b) and (d) is reduced to a fluorescence intensity of -0.5 to 2.0. At times 100, 200, 300, and 400, a large peak was observed at the emission point P(1), while a small peak (indicated by an arrow) was observed at the emission point P(2).

[0067] FIG. 15 shows color-transformed data obtained by performing color conversions of (Equation 18) and (Equation 19) on the raw data in FIG. 14 at each time. The notation method is the same as in FIG. 14. As shown in FIG. 15(a), at the emission point P(1), the emission fluorescence of phosphor D(1,1) was detected alone at times 100 and 300, and the emission fluorescence of phosphor D(1,2) was detected alone at times 200 and 400. On the other hand, as shown in FIG. 15(d), at the emission point P(2), the identity of four small peaks (indicated by arrows) detected at times 100, 200, 300, and 400 was unknown. In other words, it was unclear whether their origin was (1) the fluorescent emission of a mixture of phosphors D(2,1) and D(2,2) at the emission point P(2), (2) the fluorescent emission of impurities other than phosphors D(2,1) and D(2,2) at the emission point P(2), or (3) spatial crosstalk of the fluorescent emission of phosphors D(1,1) and D(1,2) at the emission point P(1).

[0068] Fig. 16 shows color conversion + spatial correction data obtained by performing color conversion + spatial correction (Equation 20) on the raw data in Fig. 14 at each time. The notation method is the same as in Fig. 14. As shown in Fig. 16(a), similar to Fig. 15(a), at the emission point P(1), the emitted fluorescence of phosphor D(1,1) was detected alone at times 100 and 300, and the emitted fluorescence of phosphor D(1,2) was detected alone at times 200 and 400. In addition, as shown in FIG. 16(d), the spatial crosstalk and spectral crosstalk of the fluorescence emission of phosphors D(1,1) and D(1,2) at the emission point P(1) in the detection regions W(2,1) and W(2,2) were eliminated, and it was found that the weak emission fluorescence of phosphor D(2,2) was detected alone at the emission point P(2) at times 100 and 300, and the weak emission fluorescence of phosphor D(2,1) was detected alone at times 200 and 400. The peak intensity of these weak emission fluorescences was just under 1% of the peak intensity of the emission fluorescence observed at the emission point P(1). On the other hand, as can be seen from the results in FIG. 12, the spatial crosstalk ratio occurring in the model experimental system in FIG. 8 was also just under 1%. Therefore, in FIG. 15, the true weak emission fluorescence and the false weak emission fluorescence caused by the spatial crosstalk were mixed, and it became impossible to distinguish between the two.

[0069] The above results show that spatial crosstalk can push up the detection limit in detecting light emission from each light-emitting point. As in the above example, if the spatial crosstalk ratio is 1%, even if the detection limit in the case of one light-emitting point is 0.1%, the effective detection limit rises to 1% because a signal of 1% or less cannot be distinguished as being a true signal or a false signal due to spatial crosstalk. In other words, compared with the detection of light emission from one light-emitting point, the detection sensitivity and dynamic range of light emission from multiple light-emitting points are both one order of magnitude lower. The present disclosure can solve such problems and avoid the decrease in detection sensitivity and dynamic range in the detection of light emission from multiple light-emitting points.

[0070] [Example 4] Next, based on the experimental results of [Example 2], a substance labeled with an unknown phosphor D(1,3) was subjected to electrophoretic analysis at the emission point P(1) together with phosphor D(1,1) and phosphor D(1,2). FIG. 17 shows raw data obtained by electrophoretic analysis of both capillaries Cap(1) and Cap(2) using the model experimental system of FIG. 8, in which a sample was injected only into capillary Cap(1) and no sample was injected into capillary Cap(2). The notation method is the same as FIG. 14. The sample is prepared so that phosphor D(1,1) at the emission point P(1) at time 100, phosphor D(1,2) at the emission point P(1) at time 200, phosphor D(1,1) at the emission point P(1) at time 300, and phosphor D(1,3) at the emission point P(1) at time 400 each emit fluorescence independently. The sample is prepared so that no fluorescence other than these is emitted. As shown in Figure 17(a), only four large peaks corresponding to the above-mentioned fluorescence emissions were observed. The spectral crosstalk ratio of phosphor D(1,3) was found to be similar to that of phosphor D(1,2), but slightly different. Meanwhile, the four small peaks indicated by the arrows in Figure 17(d) can be determined to be the result of spatial and spectral crosstalk of the emitted fluorescence, which is the source of the four large peaks in Figure 17(a), respectively.

[0071] FIG. 18 shows color conversion + spatial correction data obtained by performing color conversion + spatial correction (Equation 20) on the raw data in FIG. 17 at each time. The notation method is the same as in FIG. 14. As shown in FIG. 18(a), at the emission point P(1), the emission fluorescence of phosphor D(1,1) was detected alone at times 100 and 300, and the emission fluorescence of phosphor D(1,2) was detected alone at time 200. However, as shown by the arrow, although the emission fluorescence of phosphor D(1,3) was detected alone at time 400, the spectral crosstalk ratio was different from both phosphor D(1,1) and phosphor D(1,2), so the spectral crosstalk was not eliminated. On the other hand, as shown in Fig. 18(d), although the spatial crosstalk and spectral crosstalk due to the fluorescent emission of phosphors D(1,1) and D(1,2) at the emission point P(1) at times 100, 200, and 300 are eliminated, it was found that the spatial crosstalk and spectral crosstalk due to the fluorescent emission of phosphor D(1,3) at the emission point P(1) at time 400 is not eliminated and a peak remains, as indicated by the arrow. This is because in [Example 2], (Equation 20) was derived for the emitted fluorescence of phosphors D(1,1) and D(1,2) at the emission point P(1) and phosphors D(2,1) and D(2,2) at the emission point P(2), and the emitted fluorescence of D(1,3) at the emission point P(1) has different characteristics of spatial crosstalk and spectral crosstalk from (Equation 20). Therefore, if equation 20 is re-acquired for the emitted fluorescence of phosphor D(1,3) at light-emitting point P(1), the above problem can be solved. The above phenomenon represents one aspect of the present disclosure.

[0072] [Example 5] Next, based on the experimental results of [Example 2], an experiment was conducted in which the concentration of a substance labeled with phosphor D(1,1) was increased stepwise at the emission point P(1) to cause it to emit fluorescence. Figure 19 shows raw data obtained by electrophoretic analysis of both capillaries Cap(1) and Cap(2) using the model experimental system of Figure 8, with a sample injected only into capillary Cap(1) and no sample injected into capillary Cap(2). The notation method is the same as Figure 14. However, the vertical axis scale of Figure 19(a)(c) is reduced to a fluorescence intensity of 0 to 400. The sample is prepared so that phosphor D(1,1) emits fluorescence alone at the emission point P(1) at times 100, 200, 300, and 400, respectively, and the concentration and fluorescence intensity of phosphor D(1,1) increase stepwise. The sample is prepared so that no fluorescence other than these is emitted. As shown in Figure 19(a), four large peaks corresponding to the above-mentioned fluorescence emissions were observed, but because sensor S used in the model experimental system in Figure 8 saturates at a fluorescence intensity of 200, the three peaks at times 200, 300, and 400 were detected as saturated. In contrast, the four small peaks caused by spatial crosstalk and spectral crosstalk of the emitted fluorescence of phosphor D(1,1) at the emission point P(1), shown in Figure 19(d), were observed without being saturated because their fluorescence intensity was low.

[0073] FIG. 20 shows color-converted data obtained by performing color conversion of (Equation 18) and (Equation 19) at each time on the raw data of FIG. 19. The notation method is the same as FIG. 19. As shown in FIG. 20(a), the emission fluorescence of phosphor D(1,1) at the emission point P(1) at time 100 was detected alone, with the spectral crosstalk eliminated. However, the emission fluorescence of phosphor D(1,1) at times 200, 300, and 400 was saturated, and the spectral crosstalk was not eliminated. This is because, as is clear from FIG. 19(a), 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 in (Equation 18) and (Equation 19). In contrast, as shown in FIG. 20(d), the fluorescence intensity of phosphor D(2,1) is not saturated, and the spectral crosstalk is eliminated for each of the four small peaks indicated by the arrows. The intensity ratios of the four peaks shown in FIG. 20(d) represent the intensity ratios of the emitted fluorescence of phosphor D(1,1) at emission point P(1) at times 100, 200, 300, and 400.

[0074] FIG. 21 shows color conversion + spatial correction data obtained by performing color conversion + spatial correction (Equation 20) on the raw data in FIG. 19 at each time. The notation method is the same as in FIG. 19. In FIG. 21(a), the same results are obtained for the same reasons as in FIG. 20(a). In contrast, as shown in FIG. 21(d), for the fluorescence emitted by phosphor D(2,1) at time 100, both spatial crosstalk and spectral crosstalk were eliminated. However, for the fluorescence emitted by phosphor D(2,1) at times 200, 300, and 400, spatial crosstalk and spectral crosstalk were not eliminated. This is because, at times 200, 300, and 400, the fluorescence intensity of X(1,1) or X(1,2) shown in Fig. 19(a) becomes saturated, and as a result, the ratio of the fluorescence intensity of X(2,1) and X(2,2) shown in Fig. 19(d) to the fluorescence intensity of those X(1,1) and X(1,2) deviates from the spatial crosstalk ratio defined in (Equation 20). In addition, since the degree of deviation increases according to the degree of saturation, in Fig. 21(d), the fluorescence intensity of phosphor D(2,1) increases more at time 300 than at time 200, and more than at time 400 than at time 300.

[0075] [Example 6] Here, a method for determining matrix Y of (Equation 20) using a method different from that in [Example 2] is described. In [Example 2], as shown in Fig. 11(a), the matrix Y was determined by causing the phosphors D(1,1), D(2,1), D(1,2), and D(2,2) to emit fluorescence independently in that order, that is, by causing them to emit fluorescence alternately at emission point P(1) and emission point P(2). However, if the electrophoretic speed of either capillary Cap(1) or Cap(2) deviates from the expected speed, there is a possibility that fluorescence will be emitted simultaneously at emission point P(1) and emission point P(2), in which case matrix Y cannot be determined.

[0076] FIG. 22 shows a more practical method for avoiding the above problem. First, as shown in FIG. 22(a), a sample is injected only into the capillary Cap(1), electrophoretic analysis is performed in the capillaries Cap(1) and Cap(2), fluorescent bodies D(1,1) and D(1,2) are caused to emit fluorescence independently, and fluorescence detection is performed in the detection regions W(1,1), W(1,2), W(2,1), and W(2,2). Next, as shown in FIG. 22(b), a sample is injected only into the capillary Cap(2), electrophoretic analysis is performed in the capillaries Cap(1) and Cap(2), fluorescent bodies D(2,1) and D(2,2) are caused to emit fluorescence independently, and fluorescence detection is performed in the detection regions 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]. Furthermore, by this method, the matrix Y can be determined more simply and reliably than in [Example 2]. In FIG. 22, for ease of understanding, the phosphors D(1,1) and D(1,2) in the capillary Cap(1) and the phosphors D(2,1) and D(2,2) in the capillary Cap(2) are depicted as emitting fluorescence multiple times, but it is sufficient to emit fluorescence once each. In addition, in the above, the next electrophoretic analysis is performed after the first electrophoretic analysis is completed, but the interval between electrophoretic analyses does not need to be extended to that extent. By appropriately shifting the timing of sample injection into the capillary Cap(1) and the sample injection into the capillary Cap(2), it is possible to obtain an effect equivalent to that shown in FIG. 22. For example, after injecting a sample into the capillary Cap(1), electrophoresis can be performed for a short time, and then the sample can be injected into the capillary Cap(2) before electrophoresis is resumed. This can reduce the time required to determine the matrix Y. Furthermore, this method allows the sample injected into capillary cap (1) and the sample injected into capillary cap (2) to have the same composition or be the same, which simplifies the sample preparation and reduces the cost.

[0077] Figure 23 shows a method for further simplifying the determination of matrix Y. Here, even though sample injection into capillary Cap(1) and sample injection into capillary Cap(2) are performed at the same timing, the samples are prepared so that fluorophores D(1,1) and D(1,2) in capillary Cap(1) and fluorophores D(2,1) and D(2,2) in capillary Cap(2) emit fluorescence at different times, as in Figure 22. In other words, it is sufficient to change the composition of the sample injected into capillary Cap(1) and the sample injected into capillary Cap(2) so that the electrophoretic speeds of the substances labeled with each fluorophore are different. For example, the sample to be injected into capillary Cap (1) can be prepared to contain a 50-base DNA fragment labeled with fluorophore D (1,1) and a 60-base DNA fragment labeled with fluorophore D (1,2), while the sample to be injected into capillary Cap (2) can be prepared to contain a 70-base DNA fragment labeled with fluorophore D (2,1) and an 80-base DNA fragment labeled with fluorophore D (2,2).

[0078] Alternatively, Fig. 23 can be realized by setting different electrophoresis conditions for capillaries Cap(1) and Cap(2) even if samples of the same composition are injected into capillaries Cap(1) and Cap(2) at the same time. For example, the voltage applied to Cap(2) can be temporarily lowered during electrophoresis, the temperature of Cap(2) can be lowered during electrophoresis, etc.

[0079] In the above, it has been assumed that a dedicated sample for determining matrix Y is prepared in advance, but this is not necessarily required. In the raw data of the electrophoretic analysis of the actual sample to be analyzed, if a state exists in which the fluorescence emission of each fluorophore in each capillary occurs independently and this state can be identified, matrix Y can be determined using the raw data at that time.

[0080] [Example 7] FIG. 24 is a configuration diagram of a multi-capillary electrophoresis device, which is an example of an analytical device. The multi-capillary electrophoresis device is widely used as an analytical device for performing DNA sequence and DNA fragment analysis. As shown in FIG. 24, 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 were used, and DNA sequences of different samples were performed in each capillary 24-1. The DNA sequence sample was composed of DNA fragments labeled with four types of fluorophores. One analytical session was performed by the following steps (1) to (6). (1) First, the sample injection ends 24-2 of the four capillaries 24-1 were immersed in the cathode buffer solution 24-6, and the sample elution ends 24-3 were immersed in the anode buffer solution 24-7 via the polymer block 24-9. (2) Next, the valve 24-10 of the pump block 24-9 was closed, and the piston of the syringe 24-11 connected to the pump block 24-9 was pushed down to pressurize the polymer solution inside, and the polymer solution was filled into each capillary 24-1 from the sample elution end 24-3 toward the sample injection end 24-2. (3) Next, the valve 24-10 was opened, and different samples were injected from the sample injection end 24-2 into each capillary 24-1, and then a high voltage was applied between the cathode 24-4 and the anode 24-5 by the power supply 24-8 to start capillary electrophoresis. The DNA fragments labeled with four kinds of fluorophores were electrophoresed from the sample injection end 24-2 toward the sample elution end 24-3. (4) In parallel, a position of each capillary 24-1 electrophoresed a certain distance from the sample injection end 24-2 was set as the light emission point 24-14, and a laser beam 24-13 with a wavelength of 505 nm emitted from a laser light source 24-12 was irradiated to each light emission point 24-14 at once. Here, the coating of each capillary 24-1 near the light emission point 24-14 was removed in advance, each capillary 24-1 near the light emission point 24-14 was arranged on the same plane, and the laser beam 24-13 was focused and then introduced along the array plane from the side of the above-mentioned array plane.(5) Then, the DNA fragments labeled with the four types of fluorophores electrophoresed inside each capillary 24-1, and were excited by irradiation with the laser beam 24-13 when passing through the light-emitting point 24-14, emitting fluorescence. In other words, the four types of fluorophores emitted fluorescence from the four light-emitting points, and the intensity of each fluorescence changed from moment to moment with electrophoresis. (6) Finally, the fluorescence emitted from each light-emitting point was detected in multiple colors by a sensor (not shown), and the DNA sequence of the sample injected into each capillary was performed by analyzing the time-series data obtained by a computer (not shown). The analysis session consisting of the above steps (1) to (6) can be repeated multiple times. For example, samples (1) to (4) can be analyzed in the first analysis session, samples (5) to (8) can be analyzed in the second analysis session, and so on, allowing many different samples to be analyzed.

[0081] In this embodiment, the multicolor detection of (6) above was 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) was collimated by one condenser lens, passed through one transmission type diffraction grating, and the first-order diffracted light of each fluorescence was imaged on one two-dimensional sensor by one imaging lens. FIG. 25 shows the result of acquiring a two-dimensional sensor image including a wavelength dispersion image of the Raman scattering light emitted from the four light-emitting points P(1) to P(4) by laser beam irradiation when the four capillaries Cap(1) to Cap(4) are filled with a standard solution. The horizontal axis direction is the arrangement direction of the four capillaries, and the vertical axis direction is the wavelength direction. The four streaky images extending vertically, indicated by the arrows, are the wavelength dispersion images of the Raman scattering light of the capillaries Cap(1) to Cap(4), respectively. Based on this result, the wavelength calibration of the wavelength dispersion image on the two-dimensional sensor image was performed for each light-emitting point, and the relationship between the pixel position and the wavelength was obtained. Based on the results, as shown in FIG. 26, detection areas of B=20 different wavelength bands were set for each of the light-emitting points P(1) to P(4) on the two-dimensional sensor image. Each of the 20 detection areas was set to detect light emission 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 light-emitting point P(1) were set to detect light emission in the wavelength bands of 500 to 510 nm, 510 to 520 nm, ..., 690 to 700 nm, respectively, and the respective signal intensities were set as X(1,1), X(1,2), ..., X(1,20). The same is true for the light-emitting points P(2) to P(4). Here, as long as the position of each light-emitting point P(1) to P(4) and the optical system are fixed, the relationship between pixel position and wavelength for each capillary in the two-dimensional sensor image is maintained, so the detection area set in Figure 26 is valid for multiple different light-emitting detections or multiple different analysis sessions.

[0082] In this example, dR110, dR6G, dTAMRA, and dROX were used as C=4 types of fluorophores, and DNA fragments with terminal bases of T, C, A, and G prepared by Sanger reaction were labeled with each of them. Since the maximum fluorescence emission wavelengths of dR110, dR6G, dTAMRA, and dROX are 541 nm, 568 nm, 595 nm, and 618 nm, respectively, the emission fluorescence is detected with the strongest intensity in the detection regions of the wavelength bands of 540-550 nm, 560-570 nm, 590-600 nm, and 610-620 nm. For example, the emission fluorescence of dR110, dR6G, dTAMRA, and dROX at the emission point P(1) is mainly detected in the detection regions W(1,5), W(1,7), W(1,10), and W(1,12), respectively. However, each emitted fluorescence is also detected in other detection areas of the emission point P(1) due to spectral crosstalk, and is also detected at a weak intensity in the detection areas of the emission points P(2) to P(3) due to spatial and spectral crosstalk. Hereafter, dR110, dR6G, dTAMRA, and dROX at the emission point P(1) are referred to as D(1,1), D(1,2), D(1,3), and D(1,4), respectively, and their concentrations are referred to as Z(1,1), Z(1,2), Z(1,3), and Z(1,4). The same applies to the emission points P(2) to P(4).

[0083] In this case, (Equation 10) to (Equation 12) in (Equation 9) become the following equations.

number

number

number

[0084] Here, X is a matrix with 80 rows and 1 column, Y is a matrix with 80 rows and 16 columns, and Z is a matrix with 16 rows and 1 column. By substituting (Equation 23) to (Equation 25) into (Equation 14), spatial correction and color conversion were performed to eliminate both spatial crosstalk and spectral crosstalk, and time series data of the concentrations of four types of fluorophores in each capillary could be obtained. As a result, DNA sequencing of different samples injected into each capillary could be performed. Note that the conventional method corresponding to this embodiment is to perform the color conversion of (Equation 8) for each capillary. The conventional method cannot eliminate spatial crosstalk.

[0085] Equations 23 to 25, and equation 14 derived from them, are valid for multiple different emission detections or multiple different analysis sessions, as long as the position of each emission point, the optical system, and the fluorophores used are fixed.

[0086] The method of determining the matrix Y has already been described. However, it is not necessarily necessary to set all 80 × 16 elements. For example, elements whose absolute values ​​are sufficiently small compared to other elements may be replaced with zero to simplify the related calculations. In addition, if the range to which the spatial crosstalk affects is limited, (Equation 9) to (Equation 12) may be defined in the limited range. In this case, it is also possible to perform a wider range of analysis by sequentially sliding the limited range as necessary. For example, if the range to which the spatial crosstalk affects can be limited to adjacent capillaries on both sides, it is possible to ignore the spatial crosstalk and spectral crosstalk for capillaries that are two or more away, and to omit the related calculations. In this case, it is also possible to simplify the process of causing each light-emitting point to emit light independently in order to determine the matrix Y, into a process of causing multiple light-emitting points that do not affect each other by spatial crosstalk to emit light simultaneously.

[0087] [Example 8] In this embodiment, similarly to [Example 7], DNA sequencing was performed using the multi-capillary electrophoresis device of FIG. 24. However, the fluorescent light emitted from four types of phosphors from four light-emitting points was detected in multiple colors at each time by the optical system of Patent Document 2. FIG. 27 shows the configuration of this optical system and the four-part image of the four light-emitting points acquired by this optical system. The optical system of FIG. 27 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. As shown in FIG. 27(a) and FIG. 27(b), first, the fluorescent light emitted from A=4 light-emitting points P(1) to P(4) was individually collimated by four condenser lenses 27-2 constituting the condenser lens array 27-1 to form four light beams 27-9, and the laser beam light was cut by passing the four light beams 27-9 collectively through one long-pass filter 27-3. 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, and each light beam 27-9 is split into B=4 split light beams 27-10, 27-11, 27-12, and 27-13, which are then emitted from the dichroic mirror array. Here, the spectral characteristics of each dichroic mirror are adjusted so that the split light beams 27-10, 27-11, 27-12, and 27-13 have wavelengths of 520-550 nm, 550-580 nm, 580-610 nm, and 610-640 nm, respectively. Finally, the generated A×B=4×4=16 split light beams were made incident on the two-dimensional sensor 27-8, and 16 split images W(1,1) to W(4,4) were obtained, as shown in Fig. 27(c). Here, the arrangement direction of the four capillaries Cap(1) to Cap(4), i.e., the arrangement direction of the light-emitting points P(1) to P(4), and the splitting direction by the dichroic mirror array, i.e., the wavelength direction, are perpendicular to each other, so that the split images W(1,1) to W(4,4) were detected in an aligned state without overlapping with each other in the two-dimensional sensor image 27-14, as shown in Fig. 27(c).For example, the fluorescence emitted from the light-emitting point P(1) was detected as divided images W(1,1), W(1,2), W(1,3), and W(1,4), which had light components in the wavelength bands of 520-550 nm, 550-580 nm, 580-610 nm, and 610-640 nm, respectively. The same is true for the light-emitting points P(2) to P(4). Therefore, the divided images W(1,1) to W(4,4) in the two-dimensional sensor image 27-14 were set as the detection areas. In addition, the signal intensities detected in each detection area were set as X(1,1) to X(4,4), respectively. Here, as long as the positions and optical systems of the light-emitting points P(1) to P(4) are fixed, the relationship between the pixel position and the wavelength for each capillary in the two-dimensional sensor image 27-14 is maintained, so the detection area set in Figure 27(c) is valid for multiple different light emission detections or multiple different analysis sessions.

[0088] In this example, as in [Example 7], dR110, dR6G, dTAMRA, and dROX were used as C=4 types of fluorophores, and DNA fragments with terminal bases of T, C, A, and G prepared by Sanger reaction were labeled with each of them. Since the maximum fluorescence emission wavelengths of dR110, dR6G, dTAMRA, and dROX are 541 nm, 568 nm, 595 nm, and 618 nm, respectively, the emission fluorescence is detected with the strongest intensity in the detection area of ​​the divided image having light components in the wavelength bands of 520-550 nm, 550-580 nm, 580-610 nm, and 610-640 nm. For example, the emission fluorescence of dR110, dR6G, dTAMRA, and dROX at the emission 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 emitted fluorescence is also detected in other detection regions of the emission point P(1) due to spectral crosstalk, and is also detected in the detection regions of the emission points P(2) to P(3) due to spatial and spectral crosstalk. Hereafter, dR110, dR6G, dTAMRA, and dROX at the emission point P(1) are referred to as fluorophores D(1,1), D(1,2), D(1,3), and D(1,4), respectively, and their concentrations are referred to as Z(1,1), Z(1,2), Z(1,3), and Z(1,4). The same applies to the emission points P(2) to P(4).

[0089] In this case, (Equation 10) to (Equation 12) in (Equation 9) become the following equations.

number

number

number

[0090] Here, X is a matrix with 16 rows and 1 column, Y is a matrix with 16 rows and 16 columns, and Z is a matrix with 16 rows and 1 column. By substituting (Equation 26) to (Equation 28) into (Equation 14), both spatial crosstalk and spectral crosstalk were eliminated, and time series data of the concentrations of four types of fluorophores in each capillary could be obtained. As a result, DNA sequencing of different samples injected into each capillary could be performed. Note that the conventional method corresponding to this embodiment is to perform the color conversion of (Equation 8) for each capillary. The conventional method cannot eliminate spatial crosstalk.

[0091] Equations 26 to 28, and equation 14 derived from them, are valid for multiple different emission detections or multiple different analysis sessions, as long as the position of each emission point, the optical system, and the fluorophores used are fixed.

[0092] [Example 9] 28 shows a configuration diagram of a multichannel extension reaction device in which a complementary strand extension reaction is performed in units of one base in each channel of a multichannel in which a plurality of reaction vessels are arranged on a plane, using a different DNA fragment as a template, and DNA sequencing of each DNA fragment is performed by multicolor detection of the emitted fluorescence of C=4 types of fluorophores labeled with the four types of bases incorporated into the complementary strand in the extension reaction, and a schematic diagram of a two-dimensional sensor image obtained by a plurality of two-dimensional sensors. The multichannel extension 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 fluorophores labeled with the base species T, C, A, and G were set to 535 nm, 565 nm, 595 nm, and 625 nm, respectively. First, the laser beam 28-2 emitted from the laser light source 28-1 was transmitted through the dichroic mirror 28-3, and then collected by the lens 28-4 and irradiated onto the multi-channel sample 28-5. Next, the fluorescence emission 28-6 of each fluorophore excited by the laser beam irradiation in each channel was collimated collectively by the lens 28-4, and the resulting light beam 28-7 was reflected by the dichroic mirror 28-3. The dichroic mirror 28-3 has a spectral characteristic of transmitting the laser beam light and reflecting the emitted fluorescence. Next, the light beam 28-7 was divided into four light beams having four different wavelength components by three types of dichroic mirrors 28-8, 28-9, and 28-10. Then, the first split light beam is imaged by lens 28-11 onto a first two-dimensional sensor 28-12, the second split light beam is imaged by lens 28-13 onto a second two-dimensional sensor 28-14, the third split light beam is imaged by lens 28-15 onto a third two-dimensional sensor 28-16, and the fourth split light beam is imaged by lens 28-17 onto a fourth two-dimensional sensor 28-18.

[0093] Sample 28-19 is a schematic diagram of sample 28-5 observed from the front. A=5 channels each formed light-emitting points P(1) to P(5). In reality, many more channels exist on the sample, and the emitted fluorescence from each channel was detected collectively by the above optical system. However, here, only the light-emitting point P(3) of interest and the light-emitting points P(1), P(2), P(4), and P(5) around the light-emitting point P(3), which directly affect the light-emitting point P(3) by spatial crosstalk, are analyzed, so only light-emitting points P(1) to P(5) are drawn in sample 28-19 for simplicity. When focusing on a light-emitting point other than light-emitting point P(3), it is sufficient to analyze that light-emitting point and the light-emitting points around it in the same way. Regardless of which light-emitting point is focused on, it can be analyzed individually for the same fluorescence detection data after fluorescence detection. The first two-dimensional sensor image 28-20 is a divided image of the sample 28-19 acquired by the first two-dimensional sensor 28-12, and detection areas W(1,1) to W(5,1) are set at the respective image points of the light-emitting points P(1) to P(5), and the respective signal intensities are X(1,1) to X(5,1). In the first two-dimensional sensor image 28-20, fluorescent components in the wavelength band of 520 to 550 nm are detected. The second two-dimensional sensor image 28-21 is a divided image of the sample 28-19 acquired by the second two-dimensional sensor 28-14, and detection areas W(1,2) to W(5,2) are set at the respective image points of the light-emitting points P(1) to P(5). In the second two-dimensional sensor image 28-21, fluorescent components in the wavelength band of 550 to 580 nm are detected. The third two-dimensional sensor image 28-22 is a divided image of the sample 28-19 acquired by the third two-dimensional sensor 28-16, and detection areas W(1,3) to W(5,3) were set at the respective image points of the light-emitting points P(1) to P(5). In the third two-dimensional sensor image 28-22, fluorescent components in the wavelength band of 580 to 610 nm were detected. The fourth two-dimensional sensor image 28-23 is a divided image of the sample 28-19 acquired by the fourth two-dimensional sensor 28-18, and detection areas W(1,4) to W(5,4) were set at the respective image points of the light-emitting points P(1) to P(5). In the first two-dimensional sensor image 28-23, fluorescent components in the wavelength band of 610 to 640 nm were detected.That is, the emitted fluorescence from each light-emitting point was detected in B = 4 colors using four two-dimensional sensors. For example, the emitted fluorescence of the four types of phosphors at 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, respectively. However, each emitted fluorescence 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 and spectral crosstalk. Hereafter, the four types of phosphors at the light-emitting point P(3) are referred to as D(3,1), D(3,2), D(3,3), and D(3,4), respectively, in order of wavelength, and their concentrations are referred to as Z(3,1), Z(3,2), Z(3,3), and Z(3,4). The same applies to the light-emitting points P(1), P(2), P(4), and P(5).

[0094] In this case, (Equation 10) to (Equation 12) in (Equation 9) become the following equations.

number

number

number

[0095] Here, X is a matrix with 20 rows and 1 column, Y is a matrix with 20 rows and 20 columns, and Z is a matrix with 20 rows and 1 column. By substituting (Equation 29) to (Equation 31) into (Equation 14), both spatial crosstalk and spectral crosstalk were eliminated, and time series data of the concentration of four types of fluorophores in each channel could be obtained. However, in this embodiment, since only the light-emitting point P(3) is focused on, only the elements Z(3,1), Z(3,2), Z(3,3), and Z(3,4) were extracted from the matrix Z. As a result, DNA sequencing could be performed in the channel of the light-emitting point P(3). A similar method can be used when focusing on other light-emitting points and performing DNA sequencing in the corresponding channels.

[0096] [Example 10] In this embodiment, the conventional method and the method disclosed herein are summarized in a flowchart. FIG. 29 is a flowchart showing one analysis session of the conventional method. As shown in FIG. 29, the analysis session of the conventional method is executed by an analysis system having an analysis device that analyzes a sample 29-1, a computer, a display device, and a database. The analysis device has a sensor (not shown) into which light from the sample 29-1 is incident. First, A type (A is an integer of 2 or more) sample 29-1 labeled with C type (C is an integer of 1 or more) fluorophores is input to the analysis device. Next, in the analysis device, each sample is analyzed in parallel by step 29-2, and for each analysis and at each time, emission fluorescence is detected in B type (B is an integer of 1 or more) wavelength bands, and A×B pieces of time-series raw data of fluorescence intensity X(a, b) are obtained, and these time-series raw data are sent to the computer. Here, a=1, 2, ..., and A, and b=1, 2, ..., and B. Next, in step 29-3, the computer calculates the matrix Y - The spectral crosstalk is eliminated by color conversion (a0) using (a0), and the concentration Z(a0,c) of C types of phosphors is obtained from the fluorescence intensity X(a0,b) at each time. Here, a0=1, 2, ..., or A, c=1, 2, ..., and C. Finally, the display device outputs the time-series color conversion data of Z(a0,c) in step 29-4. Steps 29-3 and 29-4 are performed for all a0. As shown in FIG. 29, the conventional method is characterized in that the A analyses and analysis are performed independently. In addition, in (Equation 1) to (Equation 4), the fluorescence intensity X(a,b) is expressed as X(b), and the concentration Z(a0,c) is expressed as Z(c).

[0097] FIG. 30 is a flow chart showing one analysis session of this method. As in FIG. 29, the analysis session of this embodiment shown in FIG. 30 is executed by an analysis system having an analysis device that analyzes sample 29-1, a computer, a display device, and a database. The analysis device has a sensor (not shown) into which light from sample 29-1 is incident. First, A type (A is an integer of 2 or more) sample 30-1 labeled with C types (C is an integer of 1 or more) of fluorophores is input to the analysis device. Next, in the analysis device, each sample is analyzed in parallel by step 30-2, and emission fluorescence is detected in B types (B is an integer of 1 or more) of wavelength bands for each analysis at each time, and A×B pieces of time-series raw data of fluorescence intensity X(a, b) are obtained, and these time-series raw data are sent to the computer. Here, a=1, 2, ..., and A, and b=1, 2, ..., and B. However, unlike FIG. 29, there is significant spatial crosstalk of emission fluorescence between different analyses, as indicated by the dotted arrows in step 30-2. Next, in step 30-3, the computer calculates the matrix Y - The spectral crosstalk and spatial crosstalk are eliminated by color conversion + spatial correction using the matrix Y, and the concentration Z(a,c) of type C of fluorophore for type A of sample or analysis is calculated collectively at each time from the fluorescence intensity X(a,b). Here, a=1, 2, ..., and A, and c=1, 2, ..., and C. Finally, in step 30-4, the display device outputs the time-series color conversion + spatial correction data Z(a,c) for each analysis (a). As shown in Figure 30, this method is characterized in that the color conversion + spatial correction in step 30-3 is performed collectively for all analyses. Furthermore, the color conversion + spatial correction is performed using a matrix Y common to all analyses. - Another feature is the use of

[0098] Figure 31 shows the matrix Y to be stored in the database, which is performed before Figure 30. -This is a flowchart showing how to obtain the matrix Y determination sample 31-1. First, a type A (A is an integer of 2 or more) labeled with a type C (C is an integer of 1 or more) of fluorophores is input to the analysis device. This sample is prepared so that the fluorescent emission of the fluorophor (c) in the analysis (a) occurs alone. That is, a=1, 2, ..., and A, c=1, 2, ..., and C, and fluorescence is emitted for all combinations of A x C of these, but the fluorescent emission of two or more combinations does not occur simultaneously. Here, the above is realized by the matrix Y determination sample 31-1, but the above may be realized by the setting of the analysis device. Next, in the analysis device, in step 31-2, the above A x C single fluorescent emissions are detected in a wavelength band of type B (B is an integer of 1 or more), and time-series raw data of the fluorescent intensity X (a, b) of A x B is obtained, and these time-series raw data are sent to a computer. Next, in step 31-3, a matrix with (A×B) rows and (A×C) columns is derived from the (A×B) × (A×C) pieces of data obtained by the computer, and the matrix Y with (A×C) rows and (A×B) columns, which is the generalized inverse matrix of Y, is derived. - Then, the matrix Y - is stored in the database, and the matrix Y - Utilize the following.

[0099] Figure 32 shows the matrix Y obtained in Figure 31. - 32 is a flowchart for repeating the analysis session of steps 30-2 to 30-4 for the type A sample 30-1 shown in FIG. 30 multiple times while utilizing the matrix Y - A distinctive feature of this law is that it makes use of the following:

[0100] FIG. 33 shows an example of the computer configuration. The computer is connected to an analytical device. The computer not only performs data analysis, but also controls the analytical device. In FIGS. 29 to 31, the database and display device are depicted outside the computer, but in FIG. 33, they are depicted inside the computer. Conditions for data analysis and conditions for controlling the analytical device are set via the keyboard, which serves as the input section. Time-series raw data of fluorescence intensity X(a,b) output from the analytical device is stored in memory sequentially. Also, a matrix Y with (A×C) rows and (A×B) columns stored in a database inside the HDD is - The CPU calculates the fluorescence intensity X(a, b) and Y - The product of these is calculated, and the time series color conversion + space correction data for the phosphor concentration Z(a,c) is derived, which is then stored in memory one by one and simultaneously displayed on the monitor, which is the display unit. In addition, the analysis results can be collated with information on the network via the network interface NIF.

[0101] [Variations] The present disclosure is not limited to the above-described embodiments, and includes various modified examples. For example, the above-described embodiments have been described in detail to clearly explain the present disclosure, and it is not necessary to include all of the configurations described. In addition, a part of an embodiment can be replaced with a configuration of another embodiment. In addition, a configuration of another embodiment can be added to a configuration of an embodiment. In addition, a part of the configuration of each embodiment can be added to, deleted from, or replaced with a part of the configuration of another embodiment. [Explanation of symbols]

[0102] 1-1 Pinhole plate 1-2 Light-emitting point side aperture plate 1-3 Condenser lens 1-4 Sensor side opening plate 1-5 Colored Glass Filters 1-6 2D Sensor 1-7 Halogen lamp light 1-8 Light emitting point 1-9 light 1-10 Luminous image Cap(1) Capillary 1 Cap(2) Capillary 2 P(1) Emission point on Cap(1) P(2) Emission point on Cap(2) D(1,1) Fluorescent or absorbent 1 on P(1) D(1,2) Fluorescent or absorbent 2 on P(1) D(2,1) Fluorescent or absorbent 1 on P(2) D(2,2) Fluorescent or absorbent 2 on P(2) D(1,3) Fluorescent or absorbent 3 on P(1) W(1,1) Detection area that mainly detects D(1,1) W(1,2) Detection area that mainly detects D(1,2) W(2,1) Detection area that mainly detects D(2,1) W(2,2) Detection area that mainly detects D(2,2) Signal strength of X(1,1) W(1,1) Signal strength of X(1,2) W(1,2) Signal strength of X(2,1) W(2,1) Signal strength of X(2,2) W(2,2) Cardinality of Z(1,1) D(1,1) Concentration of Z(1,2) D(1,2) Cardinality of Z(2,1) D(2,1) Cardinality of Z(2,2) D(2,2) S Sensor LB Laser Beam LL Lamp Light 24-1 Capillary 24-2 Sample injection port 24-3 Sample elution end 24-4 Cathode 24-5 Anode 24-6 Cathode buffer solution 24-7 Anode side buffer solution 24-8 Power supply 24-9 Polymer Block 24-10 Valve 24-11 Syringe 24-12 Laser light source 24-13 Laser Beam 24-14 Light Point Cap(a) Capillary a (a=1, 2, 3, and 4) P(a) Emission point on capillary a (a=1, 2, 3, and 4) W(a,b) is the detection area of ​​the emission from P(a) in wavelength band b (a=1, 2, …, and 4) (b=1, 2, …, and 20). 27-1 Condenser lens array 27-2 Condenser lens 27-3 Long pass filter 27-4, 27-5, 27-6, and 27-7 Dichroic Mirrors 27-8 2D Sensor 27-9 Luminous flux 27-10, 27-11, 27-12, and 27-13 Split beam 27-14 2D Sensor Image P(a) Emission point a (a=1, 2, 3, 4, and 5) W(a,b) is the detection area of ​​the emission from P(a) in wavelength band b (a=1, 2, 3, 4, and 5) (b=1, 2, 3, and 4). 28-1 Laser light source 28-2 Laser beam 28-3, 28-8, 28-9, and 28-10 Dichroic Mirrors 28-4, 28-11, 28-13, 28-15, and 28-17 Lenses 28-5, and 28-19 Samples 28-6 Fluorescence 28-7 Luminous flux 28-12, 28-14, 28-16, and 28-18 2D Sensors 28-20, 28-21, 28-22, and 28-23 2D Sensor Images 29-1 Sample 29-2 Processes in analytical equipment 29-3 Computer-based processes 29-4 Display device processes 30-1 Sample 30-2 Processes in analytical equipment 30-3 Computer-based processes 30-4 Display device processes 31-1 Sample 31-2 Processes in analytical equipment 31-3 Computer-based processes

Claims

1. An analyzer for measuring raw signals of a plurality of samples in parallel; a database in which common information relating to mutual crosstalk of the raw signals of the plurality of samples is stored in advance before the measurement; a computer that processes raw signals of the plurality of samples collectively using the common information; and a display device for displaying processed signals of the plurality of samples obtained by the batch processing, An analytical system characterized in that the mutual crosstalk existing between the processed signals of the multiple samples is reduced by the batch processing compared to the mutual crosstalk existing between the raw signals of the multiple samples.

2. 2. The analytical system according to claim 1, the raw signals of the plurality of samples are time-varying; The computer executes the batch processing at each time using the common information; the display device displays a time variation of the processed signal of the plurality of samples; An analysis system characterized in that the mutual crosstalk existing between processed signals of the multiple samples at each time is reduced by the batch processing compared to the mutual crosstalk existing between raw signals of the multiple samples at each time.

3. In the analysis system according to claim 1 or 2, Each of the plurality of samples contains a component labeled with a fluorophore; The raw signals of the plurality of samples each include a fluorescence intensity of a fluorophore labeled to the component, An analytical system, wherein the processed signals of the plurality of samples each include a concentration of a fluorophore labeled to the component.

4. In the analysis system according to claim 1 or 2, Each of the plurality of samples contains components labeled with a plurality of types of fluorophores; The raw signals of the plurality of samples each include fluorescence intensities of the plurality of types of fluorophores in a plurality of wavelength bands, 11. An analytical system, wherein the processed signals of the plurality of samples each include a concentration of the plurality of types of fluorophores.

5. The analytical system according to claim 4, An analytical system characterized in that the mutual crosstalk existing between the concentrations of the multiple types of phosphors in each of the multiple samples is reduced by the batch processing, compared to the mutual crosstalk existing between the fluorescence intensities in multiple wavelength bands of the multiple types of phosphors in each of the multiple samples.

6. In the analysis system according to claim 1 or 2, An analytical system characterized in that the same common information is used for the batch processing of each of the raw signals of the multiple samples in different combinations obtained by multiple measurements performed at different times for the multiple samples in different combinations.

7. An analyzer for measuring raw signals of a plurality of samples in parallel; A computer and In an analysis system including a database, An analytical system characterized in that the analytical device includes a step of measuring each of the raw signals of the multiple samples at different times to obtain common information related to mutual crosstalk of the raw signals of the multiple samples.

8. The analytical system according to claim 7, The raw signals of the plurality of samples obtained by the process are converted into the common information by the computer; The analysis system is characterized in that the common information is stored in the database.

9. The analytical system according to claim 8, The computer processes the raw signals of the plurality of samples collectively using the common information; The analysis system includes a display device that displays processed signals of the plurality of samples obtained by the batch processing, An analytical system characterized in that the mutual crosstalk existing between the processed signals of the multiple samples is reduced by the batch processing compared to the mutual crosstalk existing between the raw signals of the multiple samples.

10. an analyzer for measuring raw signals of A samples in parallel, A being an integer equal to or greater than 2; A database in which common information relating to mutual crosstalk of the raw signals of the A samples is stored in advance before the measurement; a computer for processing the raw signals of the A samples collectively using the common information; and a display device for displaying processed signals of the A samples obtained by the batch processing, An analysis system characterized in that the mutual crosstalk existing between the processed signals of the A samples is reduced by the batch processing compared to the mutual crosstalk existing between the raw signals of the A samples.

11. The analytical system according to claim 10, The raw signal of the A samples is time-varying, The computer executes the batch processing at each time using the common information; the display device displays the time variation of the processed signal of the A samples; An analysis system characterized in that the mutual crosstalk existing between the processed signals of the A samples at each time is reduced by the batch processing compared to the mutual crosstalk existing between the raw signals of the A samples at each time.

12. In the analysis system according to claim 10 or 11, Each of the A samples contains components labeled with C types of fluorophores, where C is an integer of 1 or more, The raw signals of the A samples each include fluorescence intensities of B wavelength bands of C types of fluorophores labeled on the components, where B is an integer of 1 or more, The fluorescence intensity is A×B at each time, The processed signals of the A samples each include a concentration of the C type of fluorophores labeled to the component, The analysis system is characterized in that the concentrations are present in A×C quantities at each time.

13. The analytical system according to claim 12, An analytical system characterized in that the mutual crosstalk existing between the A x C concentrations is reduced by the batch processing compared to the mutual crosstalk existing between the A x B fluorescence intensities.

14. The analytical system according to claim 12, The analysis system according to claim 1, wherein the common information is information corresponding to a generalized inverse matrix of a matrix of (A×B) rows and (A×C) columns.

15. The analytical system according to claim 13, The analysis system according to claim 1, wherein the common information is information corresponding to a generalized inverse matrix of a matrix of (A×B) rows and (A×C) columns.

16. In the analysis system according to claim 10 or 11, An analysis system characterized in that the same common information is used for the batch processing of each of the raw signals of the A samples in D different combinations obtained by the measurements performed at the D different timings for the A samples in D different combinations, where D is an integer greater than or equal to 2.

Images