JP2013221839A - Data analysis device, program, recording medium and data analysis method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To reduce mistakes occurring when correlating corresponding points with each other in a plurality of measurement data and to highly accurately correlate the points with each other.SOLUTION: A data analysis device correlates points with each other which are associated with a measuring object as to the points showing the measuring object detected from measurement data. The data analysis device includes: input means inputting a plurality of measurement data; point information showing a plurality of appearance positions of the points in the measurement data and measurement conditions when measuring the measurement data; data correction means correcting the measurement data and the point information on the basis of correction amount calculated from the measurement conditions in the measurement data to be correlated; correlating point detection means preparing point correlating information correlating the points with each other on the basis of the measurement data and the point information corrected between the measurement data having the different positions of the points in the measurement data.

Description

  The present invention relates to a data analysis device, a program, a recording medium, and a data analysis method for detecting a point indicating the presence of a measurement object in measurement data obtained from the measurement object and associating corresponding points in a plurality of measurement data About.

After the completion of the Human Genome Project in 2003, the relationship between various diseases and biopolymers is becoming clear to date. In particular, a protein that is one of biopolymers is directly involved in the functions of living cells, organs, and organs, and is different in amino acid sequence or three-dimensional structure of the protein, or chemically such as sugar chain or phosphorylation. It is beginning to become clear that many diseases can be caused by the presence or absence of modification.
In such a situation, proteome analysis is actively performed. Proteome means a specific cell, organ, and the entire protein that is translated and produced in the organ. Examples of the analysis (proteome analysis) include protein profiling or functional analysis. It is known that proteins synthesized in vivo after protein translation have their functions controlled by post-translational modifications such as phosphorylation. Therefore, obtaining information on chemical modification of proteins can be one of the important matters in future proteome analysis. Therefore, a method for separating and detecting a sample in which a plurality of proteins are mixed with high accuracy is regarded as important, and development of an apparatus for that purpose is being promoted.

Currently, gel electrophoresis or the like is used as a method for separating and detecting proteins.
In gel electrophoresis, individual proteins are separated by taking advantage of the unique properties they have in surface charge and molecular weight. For example, there are one-dimensional electrophoresis in which proteins are separated depending only on charge or molecular weight, and two-dimensional electrophoresis in which both are separated in combination. Among them, two-dimensional electrophoresis is widely used in proteome analysis because it can separate many proteins at once and analyze them comprehensively.

Proteins separated by two-dimensional electrophoresis are observed as fluorescent regions or stained regions (hereinafter referred to as spots) on a digital image. The amount of protein corresponds to the volume that is the integrated value of the spot, and in order to quantify the amount of the separated protein, the position of the spot is detected and the volume is calculated.
By comparing spot volumes between two-dimensional electrophoresis images taken by two-dimensional electrophoresis of samples from normal cells and samples from cancer cells, for example, proteins related to cancer, ie Discovery of marker proteins has also been conducted.

In this case, it is necessary to associate spots derived from the same protein between a plurality of two-dimensional electrophoresis images. In addition, the spot which shows the characteristic derived from the component of the measurement object which appears in measurement data, such as a two-dimensional electrophoresis image, is hereafter called a point.
In a two-dimensional electrophoresis image, the points derived from the same protein should ideally be placed at the same position for each two-dimensional electrophoresis. However, in reality, the points derived from the same protein are not arranged at the same position among a plurality of images due to the flexibility of the gel and the variation from experiment to experiment. Therefore, it is necessary to visually associate points derived from the same protein among a plurality of images, and it takes enormous time and labor.

In addition, in order to shorten the time and labor required for point association, a method of performing association between points derived from the same protein using a computer has been attempted.
For example, Patent Literature 1 discloses a method of associating a new image point using a master composite image generated by averaging selected images. Specifically, the operator selects a set of points to be anchor points from the master composite image. Then, the operator visually searches for an anchor point existing in the new image, and checks whether there is an anchor point. Next, the operator associates the anchor point in the master composite image with the anchor point in the new image corresponding to the anchor point. Thereafter, the computer associates points other than the anchor point between the master composite image and the new image based on the position of the anchor point selected by the operator.

Special Table 2001-500614

In gel electrophoresis, in which electrophoresis separates proteins depending on molecular weight, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) separates proteins by molecular weight using polyacrylamide gel containing sodium dodecyl sulfate (SDS). Is done. SDS is a molecule with a strong negative charge and forms a complex with the polypeptide chain of a protein. In SDS-PAGE, each protein is separated according to its molecular weight by the moving speed at which the SDS-protein complex moves through the gel when a voltage is applied to a porous polyacrylamide gel (PAGE). The moving speed at which the SDS-protein complex moves in the gel is slower as the protein molecular weight is higher, and the moving speed is higher in the gel as the protein molecular weight is lower.
In SDS-PAGE, the position of the point changes depending on the migration time or the voltage applied during electrophoresis. The longer the migration time and the higher the voltage applied during electrophoresis, the longer the protein moves.
In reality, the electrophoresis time and voltage vary for each electrophoresis experiment. Therefore, the points derived from the same protein are not arranged at the same position among a plurality of images.

 However, in the method disclosed in Patent Document 1, when aligning a master composite image and a new image, an operator needs to manually select a point of the master composite image and a point of a new image corresponding thereto. . For this reason, time and labor are required, and if the person who performs the association is different, there is a possibility of selecting points that do not originate from the same protein. As a result, there is a possibility that the accuracy of associating points derived from the same protein is lowered.

  Therefore, the present invention has been made in view of the above-mentioned drawbacks of the prior art, and reduces time and labor when matching corresponding points in a plurality of measurement data, and also supports a plurality of measurement data. An object of the present invention is to provide a data analysis device, a program, a recording medium, and a data analysis method capable of reducing the time required for associating points with each other and preventing association between incorrect points.

  (1) The present invention has been made to solve the above-described problems, and one aspect of the present invention relates to a point representing a measurement object detected from measurement data, between points related to the measurement object. A data analysis device that performs association, and inputs a plurality of the measurement data, point information representing the appearance positions of the plurality of points in the measurement data, and measurement conditions when the measurement data is measured A correction amount of the measurement data by comparing the measurement conditions in the measurement data to be associated with the data, a data correction unit for correcting the measurement data and the point information based on the correction amount, and the measurement data The measurement data corrected by the data correction means is corrected between the measurement data having different appearance positions of the points. A corresponding point detecting means for creating a point correspondence information that associates the point to each other on the basis of the point information, a data analyzer, characterized in that it comprises a.

  (2) In another aspect of the present invention, the measurement data is data in which a plurality of the points appear in two-dimensional data, and the appearance position of the point is indicated by the position of the point in the two-dimensional data. The data analyzing apparatus according to (1), wherein

  (3) In another aspect of the present invention, the measurement data is a two-dimensional electrophoretic image, and the portion of the two-dimensional electrophoretic image having a high density value indicating the color of the image or the density of brightness for each pixel. Appears as the point, the data analysis apparatus according to (2).

  (4) Another aspect of the present invention is characterized in that the data correction means calculates a correction amount of measurement data based on an applied voltage of electrophoresis and a migration time when a two-dimensional electrophoresis image is measured. The data analysis apparatus according to (3).

  (5) In another aspect of the present invention, the correction amount is calculated based on a total applied voltage that is a product of the applied voltage and the running time of electrophoresis when measuring a two-dimensional electrophoresis image. The data analysis apparatus according to (4), wherein

  (6) In another aspect of the present invention, the correction amount is calculated by using a ratio of the total applied voltage when measuring a two-dimensional electrophoresis image between two-dimensional electrophoresis images to be applied as a correction amount. The data analyzing apparatus according to (5), characterized in that:

  (7) In another aspect of the present invention, the correction of the measurement data and the point information is performed by enlarging or reducing the two-dimensional electrophoresis based on the ratio of the total applied voltages, and appears in the two-dimensional electrophoresis image The data analysis device according to (6), wherein the appearance position of the point being converted is converted.

  (8) In another aspect of the present invention, two-dimensional electrophoresis expanded or reduced based on the ratio of the total applied voltages, and an appearance position of a point converted based on the ratio of the total applied voltages. The data analysis apparatus according to (7), further including an output unit that outputs the data.

  (9) Another aspect of the present invention includes any one of (1) to (8), characterized in that it includes output means for outputting the measurement data corrected by the data correction means and the corrected point information. A data analysis apparatus according to claim 1.

  (10) Another aspect of the present invention is a program for causing a computer to function as the input unit, the data correction unit, or the corresponding point detection unit according to any one of (1) to (9). is there.

  (11) Another aspect of the present invention is a computer-readable recording medium on which the program according to (10) is recorded.

  (12) Another aspect of the present invention is a data analysis method for associating points related to the measurement object using measurement data in which points indicating the measurement object appear, and a plurality of the measurement A step of inputting data, point information representing the appearance positions of the plurality of points in the measurement data, and a measurement condition when the measurement data is measured is compared with the measurement condition in the measurement data to be associated A correction amount of the measurement data is calculated, and the correction data is corrected by the correction step between the step of correcting the measurement data and the point information based on the correction amount, and the measurement data in which the appearance position of the point in the measurement data is different. Point association information in which the points are associated with each other based on the measured data and point information. A data analysis method characterized by comprising the steps of forming a.

  According to the present invention, when a point indicating the presence of a measurement object is detected in measurement data obtained from the measurement object, and the corresponding points are associated with each other in a plurality of measurement data, occurrence of an incorrect point-to-point association is generated. Can be prevented, and points corresponding to each other with high accuracy can be associated with each other. In addition, it is possible to reduce the time and labor required for associating corresponding points in a plurality of measurement data, and to reduce the time required for associating corresponding points in a plurality of measurement data.

It is a block diagram which shows an example of a structure of the data analyzer which concerns on one Embodiment of this invention. It is a figure which shows an example of the two-dimensional electrophoresis image which is the electrophoresis result obtained by image | photographing what carried out the two-dimensional electrophoresis of the sample containing a some protein. It is a figure for demonstrating an example which detects point information. It is a figure for demonstrating an example which detects point information. It is a figure for demonstrating an example which detects point information. It is sectional drawing which shows the principal part structure of an electrophoresis apparatus. It is a control block diagram of an electrophoresis apparatus. It is a flowchart which shows an example of the data analysis method by the data analyzer which concerns on one Embodiment of this invention. It is a flowchart which shows an example of the expansion / contraction process of the image by the data analyzer which concerns on one Embodiment of this invention, and the conversion process of point position information. It is a figure for demonstrating the position of the point in a two-dimensional electrophoresis image. It is a flowchart which shows an example of the point matching process by the data analyzer which concerns on one Embodiment of this invention. It is a figure for demonstrating the position of the point in a two-dimensional electrophoresis image. It is a figure which shows the example of an output of the analysis result by the data analyzer which concerns on one Embodiment of this invention. It is a figure which shows the other output example of the analysis result by the data analyzer which concerns on one Embodiment of this invention. It is a figure for demonstrating the electrophoresis time and the measured secondary electrophoresis image of electrophoresis. It is a figure for demonstrating the applied voltage of electrophoresis and the measured secondary electrophoresis image. It is a figure for demonstrating the relationship between the migration time of electrophoresis, and migration distance. It is a figure for demonstrating the relationship between the applied voltage and the migration distance of electrophoresis. Two-dimensional electrophoresis image when two-dimensional electrophoresis is performed using a sample containing a plurality of proteins, and reduced two-dimensional electrophoresis when analysis is performed by the data analysis apparatus according to one embodiment of the present invention It is a figure which shows an example of an image.

Hereinafter, an embodiment of the present invention will be described in detail with reference to the drawings. With reference to FIG. 1, the outline of the data analysis apparatus according to the present embodiment will be described. FIG. 1 is a diagram illustrating an example of a configuration of a data analysis system including a data analysis apparatus 100 according to the present embodiment.
As shown in FIG. 1, the data analysis system includes a data analysis device 100, an external detection device 200, and an electrophoresis device 300. The data analysis device 100 and the external detection device 200 are connected. The external detection device 200 outputs measurement data and the like that are analyzed by the data analysis device 100 to the data analysis device 100. Further, the data analysis apparatus 100 and the electrophoresis apparatus 300 are connected. The electrophoresis apparatus 300 outputs the electrophoresis time and applied voltage in SDS-PAGE to the data analysis apparatus 100.
The data analysis device 100 includes a control device 101, an input device 102, a data processing device 103, a storage device 104, an external storage device 105, and an output device 106.
The data processing apparatus 103 includes a reference image selection unit 131, a data correction unit 132, and a corresponding point detection unit 133.
The storage device 104 includes a data storage unit 141, a reference image information storage unit 142, and a correction data storage unit 143.

The data analysis apparatus 100 according to the present embodiment detects points indicating the presence of a measurement target in measurement data obtained by measuring a substance including the measurement target, and associates corresponding points in a plurality of measurement data. . The point indicating the presence of the measurement target is a point indicating a characteristic derived from the component of the measurement target that appears in the measurement data in the present embodiment.
Moreover, in this embodiment, the measurement object detected from measurement data is protein. Note that the present invention is not limited to a protein whose measurement target is a protein, and may be, for example, an arbitrary component for identifying a component constituting a certain substance.
The data analysis apparatus 100 associates corresponding points between the plurality of measurement data based on the plurality of measurement data.
Here, the “corresponding point” is a portion representing in the measurement data the presence of a measurement target (here, protein) derived from the same component. The point is a concept indicating a certain region including a peak value in an image which is two-dimensional measurement data.

Hereinafter, an example in which the data analysis apparatus 100 associates corresponding points among a plurality of two-dimensional data using two-dimensional data having points will be described. Here, a case where measurement data of a migration result of two-dimensional electrophoresis (hereinafter referred to as a two-dimensional electrophoresis image) is used as the two-dimensional data will be described as an example.
This two-dimensional electrophoresis is a technique for separating in two dimensions using the electrical properties of proteins. Two-dimensional electrophoresis is based on two types of electrophoresis: isoelectric focusing, which separates proteins depending on charge, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which separates depending on molecular weight. Become.
First, in the first dimension, isoelectric focusing is performed in which proteins are separated at isoelectric points using an isoelectric focusing gel. This isoelectric focusing is separation in the first dimension, and an isoelectric focusing gel having a constant pH gradient is used. In this isoelectric focusing, using the property that when a voltage is applied to both ends of the gel, the net charge of each protein moves to a pH of 0 (isoelectric point), isoelectric focusing of each protein. Identifying points.

In the second dimension, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) is performed in which a protein is separated by molecular weight using a polyacrylamide gel containing sodium dodecyl sulfate (SDS). This SDS-PAGE is separation in the second dimension and uses sodium dodecyl sulfate (SDS) which is a kind of anionic surfactant. SDS is a molecule with a strong negative charge and forms a complex with the polypeptide chain of a protein. In SDS-PAGE, when a voltage is applied to a porous polyacrylamide gel (PAGE), the molecular weight of each protein is identified by the moving speed at which the SDS-protein complex moves in the gel.
The first-dimensional separation direction and the second-dimensional separation direction are orthogonal to each other. As a result, a protein separation pattern separated in the first-dimensional direction and the second-dimensional direction is obtained.
The protein separated on the electrophoresis gel is visualized by staining with Coomassie Brilliant Blue (CBB) or a fluorescent dye. A two-dimensional electrophoretic image is obtained by photographing the visualized image with an image reading apparatus such as a camera or a scanner. FIG. 2 shows an example of a two-dimensional electrophoresis image. FIG. 2 shows measurement data (two-dimensional electrophoresis image) of a migration result obtained by two-dimensional electrophoresis of a sample containing a plurality of proteins.

As shown in FIG. 2, in the obtained two-dimensional electrophoresis image, proteins are separated in the first dimension (X-axis direction in the figure) by the difference in isoelectric point, and the second dimension (in the figure) In the Y-axis direction) by the difference in molecular weight. A plurality of points represented by black spots on the two-dimensional electrophoresis image represent different types of proteins. In the two-dimensional electrophoresis image shown in FIG. 2, the isoelectric point increases as it goes from 0 to the positive direction of the X axis, and the molecular weight increases as it goes from 0 to the positive direction of the Y axis.
As described above, when there are a plurality of measurement data having points, the data analysis device 100 is a device that associates corresponding points among the plurality of measurement data. Specifically, the data analysis apparatus 100 includes a two-dimensional electrophoresis image that is measurement data, information indicating the position of a point included in each two-dimensional electrophoresis image, an electrophoresis time at the time of electrophoresis, and an applied voltage. Using the information, corresponding points are associated with each other between a plurality of two-dimensional electrophoresis images. The processing contents will be described in detail later.

Returning to FIG. 1, each component of the data analysis apparatus 100 according to the present embodiment will be described.
The control device 101 controls the data analysis device 100 in an integrated manner.
The input device 102 is a device such as a keyboard or a pointing device for inputting an instruction from the outside (user). The user inputs instruction information to the input device 102 using a keyboard, a pointing device, or the like. The input device 102 is connected to the external detection device 200 and inputs information from the external detection device 200. For example, when the control device 101 instructs the input device 102 to read information from the external detection device 200, the input device 102 reads information from the external detection device 200. Further, the input device 102 is connected to the electrophoresis device 300 and inputs information from the electrophoresis device 300. For example, when the control device 101 instructs the input device 102 to read information from the electrophoretic device 300, the input device 102 reads information from the electrophoretic device 300.
The data processing device 103 operates under program control and performs each process based on a two-dimensional electrophoresis image obtained for each measurement by electrophoresis. For example, the data processing apparatus 103 performs a reference image selection process for selecting a reference image that is a reference for correspondence from a plurality of two-dimensional electrophoresis images. In addition, the data processing device 103 performs a process of enlarging or reducing the two-dimensional electrophoretic image and a process of converting the position information of the points using information on the electrophoretic time during electrophoresis and the applied voltage. Furthermore, the data processing device 103 performs a point association process for associating corresponding points between two-dimensional electrophoresis images that are a plurality of measurement data.
The storage device 104 is, for example, a random access memory (RAM) that temporarily stores information.

The external storage device 105 is, for example, a hard disk drive (HDD) that stores information for a long time. The external storage device 105 stores information input from the outside. In the present embodiment, a two-dimensional electrophoresis image that is a result of electrophoresis obtained by performing two-dimensional electrophoresis on a sample containing a plurality of proteins is stored in the external storage device 105. In this case, there are a plurality of (1, 2,..., N) two-dimensional electrophoresis images stored in the external storage device 105.
The external storage device 105 stores, for example, a two-dimensional electrophoresis image that is a migration result obtained by performing two-dimensional electrophoresis on each sample collected from a plurality of specimens.
Furthermore, the external storage device 105 stores point information (point information) regarding points included in each two-dimensional electrophoresis image in association with each two-dimensional electrophoresis image. This point information includes point identification information for identifying each point detected from the two-dimensional electrophoresis image and point position information indicating the position of each point in the two-dimensional electrophoresis image. Furthermore, the external storage device 105 stores information on SDS-PAGE electrophoresis time and applied voltage when two-dimensional electrophoresis is performed in association with each two-dimensional electrophoresis image. The information on the migration time and applied voltage includes the time from the start to the end of SDS-PAGE and the applied voltage value for each unit time during electrophoresis.
For example, when the control device 101 instructs the input device 102 to read measurement data from the external detection device 200, the input device 102 reads measurement data from the external detection device 200. Measurement data read from the input device 102 is stored in the external storage device 105. For example, when the control device 101 instructs the input device 102 to read measurement data from the electrophoretic device 300, the input device 102 provides information on SDS-PAGE migration time and applied voltage to the electrophoretic device 300. Read from. Information on the migration time and applied voltage of SDS-PAGE read from the input device 102 is stored in the external storage device 105. In this embodiment, an example in which the external storage device 105 is built in the data analysis device 100 will be described. However, the present invention is not limited to this, and is provided as an external device. 100 may be connected.
The output device 106 is a display device or a printing device that displays or prints the processing result of the data processing device 103 to the outside.

[Example of point information detection method]
Here, an example of the point information detection method will be described with reference to FIGS.
A two-dimensional electrophoretic image is data having a pixel as a constituent unit, and is information indicated by a density value (that is, a pixel value (or luminance value)) indicating a density of an image color or brightness for each pixel. In the present embodiment, the point information is detected by the external detection device 200 and input to the data analysis device 100.
The external detection device 200 detects, for example, a pixel having a density value larger than the density values of all adjacent pixels as a center position of a point in a two-dimensional electrophoresis image. Further, in the present embodiment, the external detection device 200 detects the pixel having the highest density value at each detected point as the center position of the point.
For example, the relationship between the density values of each pixel in the two-dimensional electrophoresis image is shown in FIG. FIG. 3 is a graph showing an example of the relationship between the density values of each pixel in the two-dimensional electrophoresis image. In FIG. 3, the horizontal axis indicates the pixels in the same row or the same column, and the vertical axis indicates the density value of each pixel. Note that the actual two-dimensional electrophoresis image is a two-dimensional image in which a plurality of pixels spread in a matrix in the X (row) direction and the Y (column) direction. Only one row or column of pixels is shown on the axis.

The external detection device 200 detects the position of the peak value as shown in FIG. 3 based on the density value of each pixel. The position of this peak value can be represented by coordinates P (Px, Py) in the two-dimensional electrophoresis image. In the example illustrated in FIG. 3, the position of the peak value 1 (the center position of the point) is, for example, the point position P1 (Px ₁ , Py ₁ ). The position of the peak value 2 (the center position of the point) is, for example, the point position P2 (Px ₂ , Py ₂ ).

Next, the external detection device 200 performs fitting by the least square method between the Gaussian function and the density value of the two-dimensional electrophoresis image for each position of the detected peak value, and determines the parameter of the Gaussian function. At this time, the external detection apparatus 200 performs fitting by matching the center position of the Gaussian function and the peak position of the two-dimensional electrophoresis image. For example, the fitting is shown in FIG. FIG. 4 is a graph showing an example of fitting performed by matching the center position of the Gaussian function and the peak position of the two-dimensional electrophoresis image. In the graph shown in FIG. 4, a Gaussian function corresponding to each peak value is displayed superimposed on the value shown in FIG.
The external detection device 200 determines an image area corresponding to a point using a parameter value representing the spread of the Gaussian function among the determined parameters of the Gaussian function. Note that the spread of the Gaussian function represents the spread of the parameter in the pixel direction around the peak value. In the present embodiment, the external detection device 200 determines an image area corresponding to a point that is an ellipse centered at the point position with a radius obtained by multiplying the value of the parameter representing the spread of the Gaussian function by j. The image area corresponding to this point is shown in FIG. FIG. 5 is a diagram illustrating an example of an image area corresponding to a point.

Here, an example of determining the spread of the Gaussian function and the image area corresponding to the point according to the present embodiment will be specifically described.
The Gaussian function can be expressed by the following equation.

  In Equation (1), c is a value representing the spread of the Gaussian function. a represents the peak height (density value), and b represents the coordinates (pixel) of the peak position. These ac are shown in the figure as shown in FIG. As shown in the figure, the value of the Gaussian function spread c is the distance between the x coordinate and the peak coordinate of the inflection point of the Gaussian function.

  Furthermore, the value of y when the spread of the Gaussian function becomes the value of c (that is, the value of y when x = b + c) can be calculated as follows.

Therefore, for example, the width of the Gaussian function at a height of about 0.6065 times the peak height of the Gaussian function is the value of c. In this case, the x coordinate value [pixel] of the inflection point is (b + c), and the y coordinate value [density value] of the inflection point is the Gaussian function peak value a × about 0.6065.
In Expressions (1) and (2), x indicates the position (pixel) of the two-dimensional electrophoresis image, which is the horizontal axis in FIGS. y shows the density value in each pixel of the two-dimensional electrophoresis image which is the vertical axis of FIGS.

In the present embodiment, the external detection device 200 has an image area corresponding to a point in the ellipse centered at the point position with a radius obtained by multiplying the value c of the parameter representing the spread of the Gaussian function calculated in this way by j. It has been decided. The value of j is determined empirically so that the size of the point area is such that the point area is not so narrow and the adjacent points are not included so much.
If the value of j is 0 or less, the point area cannot be taken. In addition, the value of the Gaussian function decreases as the Gaussian function moves away from the peak position. For this reason, even if the value of j is too large, there is a possibility that the point area becomes enormous and the next point is included. Therefore, it is preferable that the value of j is a value that is larger than 0 and is not included up to the adjacent point. In the present embodiment, by setting the value of j to about 1.665, it is possible to determine an appropriate point area size that does not include the adjacent points.

  In this way, the external detection device 200 identifies each point based on point position information indicating, for example, the coordinate P (Px, Py) of the point position, based on the two-dimensional electrophoresis image that is measurement data. Point information that is associated with point identification information, which is unique information assigned for the purpose, is detected. For example, one or more point information corresponds to one two-dimensional electrophoresis image.

[An example of a method for detecting information of SDS-PAGE migration time and applied voltage]
Next, an example of a method for detecting information of SDS-PAGE migration time and applied voltage will be described with reference to FIGS.
FIG. 6 is a cross-sectional view showing a configuration of an electrophoresis apparatus 600 that performs SDS-PAGE by two-dimensional electrophoresis.
The electrophoresis apparatus 600 includes a gel 601 sandwiched between a first insulator 610 and a second insulator 611, and the gel 601 can come into contact with the outside through the first opening 620 and the second opening 621. .
The first opening 620 and the second opening 621 face the first buffer tank 630 and the second buffer tank 631, respectively. In order to execute electrophoresis, the first buffer solution tank 630 and the second buffer solution tank 631 are filled with the first buffer solution and the second buffer solution, respectively. The first buffer tank 630 and the second buffer tank 631 are provided with a first electrode 640 and a second electrode 641, and the first electrode 640 and the second electrode 641 are connected to a power source 650 by a wiring 660. . When a voltage is applied to the gel 601 by the power source 650 through the first electrode 640 and the second electrode 641, the sample is developed / separated from the first opening 620 toward the second opening 621.
A first-dimensional gel (not shown) including a sample separated by isoelectric focusing that is separation in the first-dimensional direction is separated by the gel 601 after being adhered to the first opening 620 of the gel 601. Is done. Further, the electrophoresis apparatus 600 includes a voltmeter 670 for measuring a voltage value applied during electrophoresis.

FIG. 7 shows a control block diagram of the electrophoresis apparatus 600. The electrophoresis apparatus 600 includes a control unit 700 for appropriately controlling electrophoresis. The control unit 700 outputs a control signal to the voltage application unit 701 and inputs information from the voltmeter 702 in order to perform electrophoresis successfully.
The control means 700 calculates the migration time from the voltage application start time and the voltage application end time, and outputs it to the data analysis apparatus 100. Further, the control means 700 measures the applied voltage per unit time with the voltmeter 702 and outputs it to the data analysis apparatus 100.

[Example of point matching method]
Next, an example of a processing flow of a point association process for the data analysis apparatus 100 to associate points corresponding to each other between a plurality of two-dimensional electrophoresis images will be described with reference to FIG. explain.
FIG. 8 is a flowchart showing a flow until the data analysis apparatus 100 according to the present embodiment associates corresponding points between two-dimensional electrophoresis images that are a plurality of measurement data.
(Step ST201)
When the user inputs an execution instruction for the process of associating each point in the two-dimensional electrophoresis image, which is measurement data (point association process), to the input apparatus 102, the input apparatus 102 Is output to the control device 101, the data processing device 103, and the external storage device 105.
Receiving the information indicating the execution instruction from the input device 102, the external storage device 105 stores the two-dimensional electrophoretic image, the point information, and the SDS-PAGE migration time and applied voltage information stored therein. Output to the unit 141. The data storage unit 141 stores the two-dimensional electrophoresis image, point information, and SDS-PAGE migration time and applied voltage information.

(Step ST202)
As described above, the input device 102 also outputs information indicating an execution instruction to the reference image selection unit 131 of the data processing device 103. The reference image selection unit 131 reads a two-dimensional electrophoresis image from the data storage unit 141 when information indicating an execution instruction is input from the input device 102.

(Step ST203)
The reference image selection unit 131 selects a reference image from the read two-dimensional electrophoresis image. In the present embodiment, the reference image selection unit 131 selects, as a reference image, an image having the largest number of point identification information included in the associated point information from among the n two-dimensional electrophoresis images. That is, the reference image selection unit 131 selects a two-dimensional electrophoresis image (measurement data) including the most points as a reference image. When there are a plurality of two-dimensional electrophoretic images having the most points, the reference image selection unit 131 selects an arbitrary image from these two-dimensional electrophoretic images.
The reference image selection unit 131 stores information indicating that the two-dimensional electrophoresis image selected as the reference image is an image selected as the reference image in the reference image information storage unit 142. For example, the reference image selection unit 131 stores image identification information (image ID) for identifying the two-dimensional electrophoresis image selected as the reference image in the reference image information storage unit 142.
The reference image selection unit 131 may select a two-dimensional electrophoresis image designated by the user as a reference image in accordance with an instruction from the user input via the input device 102. Further, the reference image selection unit 131 may be controlled such that the control device 101 selects a two-dimensional electrophoresis image that satisfies a predetermined condition as a reference image.

  Further, the reference image selection unit 131 outputs information indicating that the selection of the reference image has been completed to the data correction unit 132. The data correction unit 132 that has received the information indicating that the selection of the reference image has been completed from the reference image selection unit 131, the two-dimensional electrophoresis image, the point information, and the SDS-PAGE migration time and applied voltage from the data storage unit 141. The image identification information indicating the reference image is read from the reference image information storage unit 142.

(Step ST204)
The data correction unit 132 performs an enlargement or reduction process (hereinafter referred to as an enlargement / reduction process) on the two-dimensional electrophoresis image based on the information of the SDS-PAGE migration time and the applied voltage. Note that the enlargement / reduction processing of the two-dimensional electrophoresis image by the data correction unit 132 will be described later with reference to FIG.
Briefly described here, the data correction unit 132 obtains information on the SDS-PAGE migration time and applied voltage when the reference image is measured, and a two-dimensional electrophoresis image other than the reference image (hereinafter referred to as a slave image). The slave image is enlarged / reduced by comparing the migration time of the SDS-PAGE when measured and the information of the applied voltage. Further, the point position information included in the point information of the slave image is converted so as to coincide with the point position in the enlarged or reduced image (hereinafter referred to as an enlarged / reduced image) of the slave image. In SDS-PAGE, the longer the migration time or the higher the applied voltage, the longer the migration distance of the points. Therefore, for example, when the migration time of SDS-PAGE when measuring a slave image is long compared to the migration time of SDS-PAGE when measuring a reference image, the migration direction in SDS-PAGE in the slave image, That is, the migration distance of the points becomes longer in the molecular weight direction (Y-axis direction in FIG. 2). As a result, the slave image becomes an image in which the image extends in the molecular weight direction compared to the reference image. On the other hand, for example, when the migration time of SDS-PAGE when measuring a slave image is shorter than the migration time of SDS-PAGE when measuring a reference image, the migration of points in the molecular weight direction in the slave image The distance becomes shorter. Thereby, in the slave image, the image is contracted in the molecular weight direction as compared with the reference image. Further, for example, when the applied voltage of SDS-PAGE when measuring the slave image is higher than the applied voltage of SDS-PAGE when measuring the reference image, the migration distance of points in the molecular weight direction in the slave image Becomes longer. As a result, the slave image becomes an image in which the image extends in the molecular weight direction compared to the reference image. Conversely, for example, when the applied voltage of SDS-PAGE when the slave image is measured is lower than the applied voltage of SDS-PAGE when the reference image is measured, in the slave image, the migration of points in the molecular weight direction is performed. The distance becomes shorter. Thereby, in the slave image, the image is contracted in the molecular weight direction as compared with the reference image.

  As described above, when the migration time or applied voltage of SDS-PAGE when measuring the reference image and the migration time or applied voltage of SDS-PAGE when measuring the slave image are different, the reference image and the slave image are different. The position of the corresponding point will be different. In order to correct the difference in the point position, the SDS-PAGE migration time and applied voltage information when the reference image is measured, and the SDS-PAGE migration time and applied voltage information when the slave image is measured. In comparison, slave image enlargement / reduction processing is performed. For example, when the SDS-PAGE migration time when the slave image is measured is longer than the SDS-PAGE migration time when the reference image is measured, the slave image is reduced in the molecular weight direction. On the other hand, for example, when the SDS-PAGE migration time when the slave image is measured is shorter than the SDS-PAGE migration time when the reference image is measured, the slave image is enlarged in the molecular weight direction. For example, when the applied voltage of SDS-PAGE when measuring a slave image is higher than the applied voltage of SDS-PAGE when measuring a reference image, the slave image is reduced in the molecular weight direction. Conversely, for example, when the applied voltage of SDS-PAGE when the slave image is measured is lower than the applied voltage of SDS-PAGE when the reference image is measured, the slave image is enlarged in the molecular weight direction.

Further, the position in the molecular weight direction in the point position information included in the point information of the slave image, that is, the Y coordinate is converted in accordance with the enlargement / reduction ratio of the slave image. The data correction unit 132 stores the created enlarged / reduced image of the slave image and the converted point information in the correction data storage unit 143.
Further, the data correction unit 132 outputs information indicating that the slave image enlargement / reduction processing has been completed to the corresponding point detection unit 133. Receiving information indicating that the slave image enlargement / reduction processing has been completed from the data correction unit 132, the corresponding point detection unit 133 reads the reference image and the point information from the data storage unit 141, and reads the reference image information storage unit 142 from the reference image information storage unit 142. And the enlarged / reduced image of the slave image and the converted point information are read from the correction data storage unit 143.

(Step ST205)
The corresponding point detection unit 133 detects corresponding points between the two-dimensional electrophoresis images and associates them. Details of the correspondence between points by the corresponding point detection unit 133 will be described later.
Briefly described here, the corresponding point detection unit 133 compares the reference image stored in the data storage unit 141 with the enlarged / reduced image of the slave image stored in the correction data storage unit 143, and determines the slave image. The point which exists in the expansion / contraction image of an image is matched with the point in the reference | standard image corresponding to this. For example, the corresponding point detection unit 133 uses the distance between the point appearing in the reference image and the point appearing in the enlarged / reduced image of the slave image, in the enlarged / reduced image of the slave image in the reference image. The information (henceforth point correlation information) which matched the point in the appearance position corresponding to this point is produced. This point association information is information in which point identification information of points in the enlarged / reduced image of the corresponding slave image is associated with point identification information of points in the reference image. In the present embodiment, the corresponding point detection unit 133 calculates the inter-point distance based on the point information between the measurement data having different point appearance positions in the measurement data, and associates the points with each other based on the inter-point distance. Creating information.
Then, the corresponding point detection unit 133 outputs the created point association information to the output device 106.

  The output device 106 outputs a result of point association between final two-dimensional electrophoresis images based on the point association information input from the corresponding point detection unit 133. Thereby, the user can confirm the point association result between the final two-dimensional electrophoresis images.

  In the above-described embodiment, an example has been described in which the data analysis apparatus 100 outputs an association result of points between final two-dimensional electrophoresis images to the output apparatus 106. However, the present invention particularly relates to this. It is not limited. For example, the result of the reference image selection process by the reference image selection unit 131, the result of the enlargement / reduction process of the slave image by the data correction unit 132, and the point association process between the two-dimensional electrophoresis images by the corresponding point detection unit 133 The results may be output to the user via the output unit 106, respectively.

Next, details of each process performed by each component of the data analysis apparatus 100 will be described.
[Reference Image Selection Processing by Reference Image Selection Unit 131]
Hereinafter, an example of a reference image selection process in which only one reference image is selected from a plurality of two-dimensional electrophoresis images by the reference image selection unit 131 will be described in detail. This reference image selection process is a process corresponding to step ST203 in FIG.
As described above in steps ST201 and ST202, the reference image selection unit 131 receives a two-dimensional data storage unit 141 from the data storage unit 141 upon receiving an instruction to execute the process of associating points in the two-dimensional electrophoresis image from the input device 102. Read the electrophoresis image. The reference image selection unit 131 outputs all the read two-dimensional electrophoresis images to the output device 106. The output device 106 outputs all the two-dimensional electrophoresis images and presents them to the user. The user selects a reference image via the input device 102 based on the presented two-dimensional electrophoresis image, and inputs information indicating that the two-dimensional electrophoresis image selected by the user is the reference image to the input device 102. To do. For example, the user selects a two-dimensional electrophoresis image using a mouse or a pointing device that is the input device 102. In the present embodiment, the reference image is a two-dimensional electrophoresis image having the largest number of points, and the user selects a two-dimensional electrophoresis image corresponding to the reference image.
At this time, only one image can be selected as the reference image. The image identification information of the reference image selected in this way is stored in the reference image information storage unit 142 by the reference image selection unit 131.

[Image Enlargement / Reduction Processing by Data Correction Unit 132]
Next, the image enlargement / reduction processing in the two-dimensional electrophoresis image by the data correction unit 132 will be described in detail with reference to FIG. FIG. 9 is a flowchart illustrating an example of a flow in which the data correction unit 132 performs the enlargement / reduction processing of the two-dimensional electrophoresis image. This image enlargement / reduction process is a process corresponding to step ST204 in FIG.
(Step ST301)
First, when the data correction unit 132 receives information indicating that the selection of the reference image has been completed from the reference image selection unit 131, the two-dimensional electrophoresis image, point information, and SDS-PAGE migration time are received from the data storage unit 141. And the applied voltage information, and the image identification information indicating the reference image is read from the reference image information storage unit 142.
Based on the image identification information of the read reference image, the data correction unit 132 converts the two-dimensional electrophoresis image, point information, and SDS-PAGE migration time and applied voltage information into the reference image, point information, and SDS-PAGE migration. Information is divided into time and applied voltage information, slave images other than the reference image, point information, SDS-PAGE migration time and applied voltage information. Then, the data correction unit 132 enlarges / reduces the slave image other than the reference image. Further, the data correction unit 132 converts the point position information included in the point information of the slave image so as to coincide with the point position in the enlarged / reduced image of the slave image.

(Step ST302)
In the image enlargement / reduction process, the data correction unit 132 first calculates an image enlargement rate or reduction rate (hereinafter referred to as an enlargement / reduction rate) based on information on the migration time and applied voltage of SDS-PAGE. More specifically, the data correction unit 132 calculates the total applied voltage of the SDS-PAGE when the reference image is measured from the information on the migration time and applied voltage of the SDS-PAGE when the reference image is measured. Further, the total applied voltage of SDS-PAGE when the slave image is measured is calculated from the information of the migration time and applied voltage of SDS-PAGE when the slave image is measured. Then, a ratio between the total applied voltage of the SDS-PAGE when the reference image is measured and the total applied voltage of the SDS-PAGE when the slave image is measured is calculated, and the ratio of the total applied voltage is enlarged for the slave image. The reduction rate.
The total applied voltage of SDS-PAGE can be calculated according to the following equation (3).

Here, t shown in Expression (3) is time. Also, t _end is the time at the _end of SDS-PAGE. v _t is the applied voltage value at time t. Therefore, the total applied voltage of SDS-PAGE is the total sum of applied voltages from the start to the end of SDS-PAGE. The ratio of the total applied voltage can be calculated according to the following formula (4).

For example, FIG. 10 shows an example of the reference image and the slave image. As shown in FIG. 10, the reference image 1001 has three points, and the slave image 1002 has three points. Note that the point information corresponding to the two-dimensional electrophoresis image of the reference image 1001 includes point identification information ID1011 to ID1013 and point position information indicating a position in the reference image 1001. Point identification information is assigned to each point position information.
The point information corresponding to the two-dimensional electrophoresis image of the slave image 1002 includes point identification information ID1021 to ID1023 and point position information indicating a position in the slave image 1002, respectively. Point identification information is assigned to each point position information.
Here, it is assumed that the point identification information indicating the points 1011 to 1013 and the points 1021 to 1023 and the point position information indicating the positions of the points are as shown in Table 1.

  In addition, the SDS-PAGE migration time when measuring the reference image 1001 is 1 hour, the applied voltage is 100 V, the SDS-PAGE migration time when measuring the slave image 1002 is 1.25 hours, the applied voltage is 120 V, Suppose that The total applied voltage at this time can be calculated as 1 hour × 100 V = 100 Vh for the reference image and 1.25 hours × 120 V = 150 Vh for the slave image. Therefore, the ratio of the total applied voltage can be calculated as 100 Vh ÷ 150 Vh = 2/3. Therefore, the enlargement / reduction ratio of the slave image is 2/3.

(Step ST303)
Next, the data correction unit 132 enlarges / reduces the slave image based on the enlargement / reduction ratio of the slave image. Further, the point position information included in the point information of the slave image is converted so as to match the point position in the enlarged / reduced image of the slave image.
In the slave image enlargement / reduction process, the enlargement / reduction process is performed in the migration direction in SDS-PAGE, that is, in the molecular weight direction (Y-axis direction in FIG. 2) in the slave image. The enlargement / reduction processing of the slave image can be performed by a general method such as a two-arrest neighbor method, a bilinear method, or a bicubic method. For example, in the two-arrest neighbor method, the enlarged / reduced image of the slave image can be calculated by the following equation (5).

Here, f (x, y) shown in Expression (5) is a pixel value at coordinates (x, y) in the enlarged / reduced image of the slave image, and g (x, y) is the slave image before enlargement / reduction. The pixel value at the coordinates (x, y). Further, a is an enlargement / reduction ratio. The symbol [] represents truncation of the decimal part.
Regarding the method of converting the point position information included in the point information of the slave image so as to match the point position in the enlarged / reduced image of the slave image, the y coordinate of the point position information of the slave image is expressed by the following formula ( Convert according to 6).

Here, y ₁ represented by the formula (6) is the y-coordinate after conversion, y ₂ is the y coordinate before conversion. Further, a is an enlargement / reduction ratio. The symbol [] represents truncation of the decimal part. That is, the [(a × y ₂ ) +0.5] portion in the formula (6) means that the value of (a × y ₂ ) is rounded off.
A specific example will be described with reference to FIG. In the case of the image of FIG. 10, the enlargement / reduction ratio of the slave image is calculated as 2/3 in step ST302. Therefore, the enlargement / reduction of the slave image is performed according to the equation (5). For example, the pixel value f (100, 100) of the coordinate (100, 100) in the enlarged / reduced image of the slave image is expressed by the following equation (7). Become.

That is, for example, the pixel value f (100, 100) of the coordinate (100, 100) in the enlarged / reduced image of the slave image is the same as the pixel value of the coordinate (100, 150) in the slave image before the enlarged / reduced image. Become.
The conversion of the point position information included in the point information of the slave image is as follows. Point position information of points existing in the slave image 1002 shown in FIG. 10 is as shown in Table 1. In this case, according to the equation (6), the point position information of the slave image is converted as shown in Table 2 below.

In this way, the data correction unit 132 performs slave image enlargement / reduction processing based on the enlargement / reduction rate, and further converts the point position information included in the point information of the slave image.
The enlarged / reduced image of the slave image and the converted point information are stored in the correction data storage unit 143 by the data correction unit 132. Here, when the enlargement / reduction processing and conversion of the point position information of one slave image are completed, the data correction unit 132 still performs the enlargement / reduction processing and conversion of the point position information from the slave images other than the reference image. One non-slave slave image is selected, and enlargement / reduction processing and conversion of the point position information are started in the new slave image.
At this time, for example, the data correction unit 132 counts the number of slave images other than the reference image so that the data correction unit 132 finishes the enlargement / reduction processing and conversion of the point position information for all the slave images. May be provided. Specifically, the reference image selection unit 131 assigns numbers sequentially from 1 to slave images other than the reference image in the read two-dimensional electrophoresis image. When the selection of the reference image by the reference image selection unit 131 is completed, the data correction unit 132 initializes the value counted by the counter to 1 and initializes the two-dimensional electrophoresis image of the reference image, its point information, SDS-PAGE. The information of the migration time and applied voltage of the first time, the two-dimensional electrophoretic image of the first slave image among the slave images other than the reference image, the point information thereof, and the information of the migration time and applied voltage of the SDS-PAGE Read from the storage unit 141. Then, when the enlargement / reduction processing of the first slave image and the conversion of the point position information among the slave images other than the reference image are completed, the data correction unit 132 increments the value counted by the counter by one, and the other than the reference image Among the slave images, the two-dimensional electrophoretic image of the second slave image, its point information, and the SDS-PAGE migration time and applied voltage information are read from the data storage unit 141. As described above, the counter sequentially increases the count value according to the number of slave images for which the point association processing has been completed, and when the count value reaches a predetermined number n−1, the data correction unit 132 It is preferable that the work is completed.

[Corresponding Point Detection Processing by Corresponding Point Detection Unit 133]
Finally, the process of associating each point by the corresponding point detection unit 133 will be described in detail with reference to FIG. FIG. 11 is a flowchart showing a flow until the corresponding point detection unit 133 associates each point. This association process is a process corresponding to step ST205 in FIG.
(Step ST401)
First, when the corresponding point detection unit 133 receives information indicating that the slave image enlargement / reduction processing has been completed from the data correction unit 132, the corresponding point detection unit 133 reads image identification information indicating the reference image from the reference image information storage unit 142. Based on the image identification information indicating the image, the reference image and the point information are read from the data storage unit 141, and the enlarged / reduced image of the slave image and the converted point information are read from the correction data storage unit 143. Then, the corresponding point detection unit 133 selects one slave image enlargement / reduction image from among the enlargement / reduction images of the slave images other than the reference image, and enlarges the slave image other than the reference image with respect to the point of the reference image. The point of the enlarged / reduced image of one slave image selected from the reduced images is associated with the point corresponding to the appearance position included in the reference image.

(Step ST402)
In the point association, the corresponding point detection unit 133 first calculates the inter-point distance between the point of the reference image and the point of the enlarged / reduced image of the slave image. More specifically, the corresponding point detection unit 133 generates correspondence table data indicating all combinations between the points of the reference image and the enlarged / reduced image of the slave image. Then, the corresponding point detection unit 133 calculates the distance between each point in each combination.
For example, FIG. 12 shows an example of the enlarged and reduced images of the reference image and the slave image. As shown in FIG. 12, it is assumed that there are three points in the reference image 1201 and three points in the enlarged / reduced image 1202 of the slave image. Note that the point information corresponding to the two-dimensional electrophoresis image of the reference image 1201 includes point identification information ID 1211 to ID 1213 and point position information indicating the position in the reference image 1201. Point identification information is assigned to each point position information.
The point information corresponding to the two-dimensional electrophoretic image of the enlarged / reduced image 1202 of the slave image includes point identification information ID1221 to ID1223 and point position information indicating the position of the slave image in the enlarged / reduced image 1202, respectively. ing. Point identification information is assigned to each point position information.
Here, as an example of the point position information, it is assumed that the points 1211 to 1213 and 1221 to 1223 are as shown in Table 3 below.

  In this case, the corresponding point detection unit 133 as correspondence table data indicating all combinations between the points 1211 to 1213 of the reference image 1201 and the points 1221 to 1223 of the enlarged / reduced image 1202 of the slave image, as shown in Table 4 below. To generate a data indicating a correspondence table. Then, the corresponding point detection unit 133 calculates an inter-point distance as shown in Table 4 for each combination of points.

  Note that the corresponding point detection unit 133 calculates the distance between the points according to the calculation formula (8) shown below.

Here, l shown in Expression (8) is a distance between points. Also, x ₁ and y ₁ are the x and y coordinates of the point of the reference image, and x ₂ and y ₂ are the x and y coordinates of the point of the enlarged / reduced image of the slave image.

(Step ST403)
Next, the corresponding point detection unit 133 selects one point of the reference image that minimizes the inter-point distance for each point of the enlarged / reduced image of the slave image based on the inter-point distance, and associates each of them with each other. Create point association information.
In the case of Table 4, the point distance between the points 1221 of the enlarged / reduced image 1202 of the slave image and the points 1211 to 1213 of the reference image 1201 is calculated by the corresponding point detection unit 133. The distance between the points is the shortest in the case of the combination of the points 1221 and 1211. Therefore, the corresponding point detection unit 133 associates the point identification information of the point 1221 of the enlarged / reduced image 1202 of the slave image with the point identification information of the point 1211 of the reference image 1201 and writes it in the point association information. Similarly, the corresponding point detection unit 133 also associates the points 1222 and 1223 of the enlarged / reduced image 1201 of the slave image. In the example shown in Table 4, the corresponding point detection unit 133 associates the point 1222 of the enlarged / reduced image 1202 of the slave image with the point 1212 of the reference image 1201. Also, the corresponding point detection unit 133 associates the point 1223 of the enlarged / reduced image 1202 of the slave image with the point 1213 of the reference image 1201.

In this way, the corresponding point detection unit 133 corresponds to any one of the points 1211 to 1213 of the reference image 1201 that minimizes the distance between points for all the points 1221 to 1223 of the enlarged / reduced image 1202 of the slave image. In addition, point association information indicating these correspondences is generated.
As described above, the correspondence point detection unit 133 associates points between the reference image and the enlarged / reduced image of the slave image. The association result is output to the output device 106 by the corresponding point detection unit 133.

Here, when the correspondence point detection unit 133 finishes associating the points between the reference image and the enlarged / reduced image of one slave image, the corresponding point detection unit 133 still associates the correspondence among the enlarged / reduced images of the slave images other than the reference image. Using the enlarged / reduced image of another slave image that has not been performed as an enlarged / reduced image of a new slave image, point association processing is started between the reference image and the enlarged / reduced image of the new slave image.
At this time, in order for the corresponding point detection unit 133 to finish associating the points for all the images, for example, the corresponding point detection unit 133 includes a counter function for counting the number of enlarged / reduced images of slave images other than the reference image. May be. Specifically, the corresponding point detection unit 133 assigns numbers sequentially from 1 to the enlarged / reduced images of the slave images stored in the correction data storage unit 143 by the data correction unit 132. When the corresponding point detection unit 133 receives information indicating that the slave image enlargement / reduction processing has been completed from the data correction unit 132, the corresponding point detection unit 133 sets the counter value to 1 and initializes it. The image identification information indicating the reference image is read from the reference image information storage unit 142, the reference image and the point information are read from the data storage unit 141 based on the image identification information indicating the reference image, and the slave image is enlarged from the correction data storage unit 143. A reduced image and converted point information may be read.
Then, when the point association with the enlarged / reduced image of the first slave image among the enlarged / reduced images of the slave images other than the reference image is completed, the value counted by the counter is incremented by one. Then, the corresponding point detection unit 133 reads, from the correction data storage unit 143, the enlarged / reduced image of the second slave image among the enlarged / reduced images of the slave images other than the reference image and the converted point information. In this manner, the corresponding point detection unit 133 increases the value counted by the counter in the order in which the point association processing is performed. A configuration may be adopted in which the corresponding point detection unit 133 ends the work when the value counted by the counter reaches a predetermined number n-1.

As described above, the data analysis apparatus 100 according to the present embodiment performs enlargement or reduction processing on slave images other than the reference image based on the information on the SDS-PAGE migration time and applied voltage. Further, the point position information included in the point information of the slave image is converted so as to match the point position in the enlarged / reduced image of the slave image. Thereafter, a combination of points of the reference image and the enlarged / reduced images of all the slave images is generated between a plurality of two-dimensional electrophoretic images, and between the reference image and the enlarged / reduced image of the slave image between the combined points. Calculate the distance. Then, the corresponding point detection unit 133 of the data analysis apparatus 100 associates the point of the reference image with the point of the enlarged / reduced image of the slave image based on the calculated inter-point distance. Thereby, the data analysis apparatus 100 according to the present embodiment can associate all the points included in the enlarged / reduced image of the slave image with the points in the reference image without relying on human hands.
As described above, the data analysis apparatus 100 according to the present embodiment performs the enlargement / reduction process on the slave image based on the information of the SDS-PAGE migration time and the applied voltage, and further, points in the enlarged / reduced image of the slave image. By converting the point position information included in the point information of the slave image so as to match the position, the point position in the molecular weight direction in the two-dimensional electrophoretic image due to the variation in SDS-PAGE migration time and applied voltage Variations can be corrected. Furthermore, by detecting the corresponding points between the two-dimensional electrophoretic images using the enlarged / reduced image of the slave image and the converted point information, and associating these points, the accuracy of the point correspondence is improved. be able to.

[Display Screen Example of Output Device 106 of Data Analysis Device 100]
Next, an example of a display image displayed on the output device 106 of the data analysis device 100 is shown in FIGS. 13 and 14.
For example, FIG. 13 shows an example of an image displayed to the user when the data correction unit 132 is configured to output the result of image enlargement / reduction processing of the two-dimensional electrophoresis image to the output device 106. FIG. 13 shows the reference image and the total applied voltage of SDS-PAGE in the reference image and the point information of the reference image, the total applied voltage of SDS-PAGE in the slave image and the slave image, the point information of the slave image, and the enlarged / reduced image of the slave image In this example, point information after conversion is displayed.
In the example shown in FIG. 13, there are three points detected in the reference image, and IDs 001 to 003 are assigned to these points. For example, as shown in the figure, three points assigned IDs 001 to 003 are displayed superimposed on the reference image. The three points assigned IDs 001 to 003 are each represented by a lead line drawn from the pixel corresponding to the peak position of the point.
Further, in the example shown in FIG. 13, there are three points detected in the slave image, and IDs 101 to 103 are attached to these points. For example, as shown, three points assigned IDs 101 to 103 are displayed superimposed on the reference image. The three points to which IDs 101 to 103 are assigned are represented by lead lines drawn from pixels corresponding to the peak positions of the points. The same is displayed in the enlarged / reduced image of the slave image, and the same ID is given to the same point between the slave image and the enlarged / reduced image of the slave image.
The user clicks, for example, an icon in the screen shown in FIG. 13 in accordance with the point association result button 1301, and selects this point association result button 1301. As a result, the output device 106 can display an image showing the result of point association as shown in FIG. 14 instead of the image shown in FIG.

For example, FIG. 14 illustrates an example of a screen displayed to the user when the data analysis apparatus 100 adopts a configuration in which the corresponding point detection unit 133 detects the corresponding point detection result and outputs the result to the output device 106. FIG. 14 is an example in which the reference image, the enlarged / reduced image of the slave image, and the inter-point distance are displayed for each associated point.
In the example shown in FIG. 14, there are three associated points, and the same ID is assigned to each associated point in the enlarged image of the reference image and the slave image. For example, as shown in the figure, three points assigned IDs M001 to M003 are displayed superimposed on the enlarged and reduced images of the reference image and the slave image, respectively. The three points to which the IDs of M001 to M003 are assigned are represented by lead lines drawn from the pixels corresponding to the peak positions of the points.
Here, when the user selects a button 1401 described on the next screen illustrated in FIG. 14, the data analysis apparatus 100 can associate points between the next slave image and the reference image.

By the way, although the example which matched the point using the two-dimensional electrophoresis image of protein was mentioned above, you may match | combine a point using the two-dimensional electrophoresis image of biopolymers, such as DNA or RNA. .
The present invention is not limited to the above-described embodiments, and various modifications can be made within the scope shown in the claims. That is, embodiments obtained by combining technical means appropriately modified within the scope of the claims are also included in the technical scope of the present invention.

[Program and recording medium]
Finally, each unit included in the data analysis apparatus 100 may be configured by hardware logic. Alternatively, it may be realized by software using a CPU as follows.
That is, the data analysis apparatus 100 includes a CPU that executes instructions of a program that realizes each function, a RAM that stores the program, and that expands the program into an executable format, and a memory that stores the program and various data And other storage devices (recording media). With this configuration, the object of the present invention can also be achieved by a predetermined recording medium.
This recording medium only needs to record the program code (execution format program, intermediate code program, source program) of the program of the data analysis apparatus 100, which is software that implements the functions described above, so that it can be read by a computer. This recording medium is supplied to the data analysis apparatus 100. Thus, the data analysis apparatus 100 (or CPU or MPU) as a computer may read and execute the program code recorded on the supplied recording medium.

  The recording medium that supplies the program code to the data analysis apparatus 100 is not limited to a specific structure or type. That is, this recording medium is, for example, a tape system such as a magnetic tape or a cassette tape, a magnetic disk such as a floppy (registered trademark) disk / hard disk, or an optical disk such as CD-ROM / MO / MD / DVD / BD / CD-R. A disk system including IC, a card system such as an IC card (including a memory card) / optical card, or a semiconductor memory system such as mask ROM / EPROM / EEPROM / flash ROM.

  The object of the present invention can be achieved even if the data analysis device 100 is configured to be connectable to a communication network. In this case, the program code is supplied to the data analysis apparatus 100 via the communication network. The communication network may be any network that can supply program codes to the data analysis apparatus 100, and is not limited to a specific type or form. For example, the Internet, intranet, extranet, LAN, ISDN, VAN, CATV communication network, virtual private network, telephone line network, mobile communication network, or satellite communication network may be used.

  The transmission medium constituting the communication network may be any medium that can transmit the program code, and is not limited to a specific configuration or type. For example, infrared rays such as IrDA or a remote control, Bluetooth (registered trademark), 802.11 wireless, HDR even with a wired line such as IEEE1394, USB, power line carrier, cable TV line, telephone line, or ADSL (Asymmetric Digital Subscriber Line) line It can also be used wirelessly, such as a mobile phone network, a satellite line, or a terrestrial digital network. The present invention can also be realized in the form of a computer data signal embedded in a carrier wave in which the program code is embodied by electronic transmission.

  The embodiment of the present invention has been described in detail with reference to the drawings. However, the specific configuration is not limited to this embodiment, and includes design changes and the like within a scope not departing from the gist of the present invention.

The present invention will be further described below using examples.
In the examples, a soluble fraction of mouse liver that was fluorescently labeled on the sample was used. Using this sample, two-dimensional electrophoresis was performed while changing the SDS-PAGE electrophoresis time or applied voltage in two-dimensional electrophoresis. Using the two-dimensional electrophoresis image that is the result of the two-dimensional electrophoresis, the corresponding points are associated using the method described in the embodiment of the present invention.

(First dimension electrophoresis)
Isoelectric focusing, which is first-dimensional electrophoresis, was performed as follows.
A pH gradient immobilized gel having a pH of 3-10 was used as the first dimension gel. After introducing the sample into the first-dimensional gel, voltage was applied and electrophoresis was performed. The voltage was applied at 200 V for 30 minutes, then boosted to 1000 V over 30 minutes, further applied at 1000 V for 10 minutes, then boosted to 5000 V over 30 hours, and applied at 5000 V for 10 minutes.
(Second dimension electrophoresis)
SDS-PAGE, which is second-dimensional electrophoresis, was performed as follows.
An acrylamide gel having a concentration of 10% was used as the second dimension gel. The voltage application conditions were two types: one in which the electrophoresis time was changed while the voltage was fixed, and one in which the voltage was changed with the migration time fixed. In the case where the voltage was fixed and the migration time was changed, the applied voltage was fixed at 150 V, and the migration time was performed under four conditions of 10 minutes, 20 minutes, 30 minutes, and 40 minutes. In the case where the voltage was changed by fixing the migration time, the migration time was fixed for 30 minutes, and the applied voltage was 4 conditions of 50V, 100V, 150V, and 200V.

(Imaging)
After SDS-PAGE, the gel was taken out and observed with a fluorescent scanner. Since two-dimensional electrophoresis is performed using a sample stained with a fluorescent dye, the result of protein separation by electrophoresis can be observed with a fluorescence scanner. Each two-dimensional electrophoresis image when the applied voltage is fixed and the electrophoresis time is changed is shown in FIG. 15, and each two-dimensional electrophoresis image when the electrophoresis time is fixed and the applied voltage is changed is shown in FIG. Shown in

(Relationship between SDS-PAGE applied voltage or migration time and migration distance)
In the images shown in FIG. 15, the Y coordinates of the positions of the points 1511 to 1514 corresponding to the images are acquired. The distance between the Y coordinate of the migration start point and the Y coordinate of the position of each point was calculated as the migration distance. A graph was prepared with the migration time on the horizontal axis and the migration distance on the vertical axis (FIG. 17). FIG. 17 revealed that there is a linear relationship between the migration time and the migration distance.
Further, in the images shown in FIG. 16, the Y coordinates of the positions of the points 1611 to 1614 corresponding to the images are acquired. The distance between the Y coordinate of the migration start point and the Y coordinate of the position of each point was calculated as the migration distance. A graph was created with the applied voltage on the horizontal axis and the migration distance on the vertical axis (FIG. 18). From FIG. 18, it was revealed that there is a linear relationship between the applied voltage and the migration distance.

(Associating points)
Corresponding points are associated using the method described in the embodiment of the present invention. The two-dimensional electrophoresis image used for the association is shown in FIG. The applied voltage of SDS-PAGE when measuring the reference image is 150 V, the migration time is 20 minutes, that is, the total applied voltage is 50 Vh, the applied voltage of SDS-PAGE when measuring the slave image is 200 V, and the migration time is 30 minutes. That is, the total applied voltage was 100 Vh. An enlarged / reduced image of the slave image is shown in FIG. As a result of associating corresponding points between the enlarged and reduced images of the reference image and the slave image, 93 points out of 100 detected points were correctly associated.
On the other hand, for comparison, the slave image was not enlarged or reduced, and the points were associated between the slave image and the reference image. As a result, 15 points were correctly associated among the detected 100 points.
From the above, it has been shown that by using the point association method according to the present invention, points can be associated with high accuracy.

When there are a plurality of measurement data having points such as two-dimensional electrophoresis images such as protein, DNA, or RNA, the present invention is suitably used when the points are associated between the measurement data. The data analysis apparatus 100 according to the present invention uses, for example, SDS-PAGE migration time and applied voltage information in two-dimensional electrophoresis, compared to a general image analysis apparatus, and thus SDS-PAGE migration time and The shift of the point position between the two-dimensional electrophoretic images resulting from the variation in the applied voltage is corrected, and then correspondence is performed. This makes it possible to associate points with high accuracy.
In a general point matching technique, points are correlated using the two-dimensional electrophoresis image obtained as a result of the two-dimensional electrophoresis as it is, and therefore, between the images due to variations in SDS-PAGE migration time and applied voltage. If the corresponding point position is greatly shifted in step 1, such a point cannot be associated. According to the present invention, even when the corresponding point position is greatly deviated between images, by using the information of the SDS-PAGE migration time and applied voltage, it is derived from the variation in the SDS-PAGE migration time and applied voltage. Since the shift of the point position between the two-dimensional electrophoretic images is corrected, the corresponding points between the images appear in approximately the same position in the corrected image. Therefore, points can be associated with high accuracy.

As described above, the present invention calculates the correction amount based on the SDS-PAGE migration time and applied voltage in the two-dimensional electrophoresis in matching the corresponding points in the two-dimensional electrophoresis image. Based on the amount, the two-dimensional electrophoresis image is enlarged or reduced, and the position of the point is converted, and then the corresponding point is detected.
Thereby, since the corresponding point detection is performed after correcting the shift of the point position between the two-dimensional electrophoretic images, it is possible to accurately associate the points.
The present invention can be applied to, for example, a protein analysis device or a biopolymer analysis device such as DNA or RNA.

  DESCRIPTION OF SYMBOLS 100 ... Data analysis device, 101 ... Control device, 102 ... Input device, 103 ... Data processing device, 104 ... Storage device, 105 ... External storage device, 106 ... Output Device 131... Reference image selection section 132 132 Data correction section 133. Corresponding point detection section 141... Data storage section 142. Reference image information storage section 143. Correction data storage unit, 200 ... external detection device, 300 ... electrophoresis device, 600 ... electrophoresis device, 601 ... gel, 610 ... first insulator, 611 ... second Insulator, 620 ... first opening, 621 ... second opening, 630 ... first buffer tank, 631 ... second buffer tank, 640 ... first electrode, 641 ... Second electrode, 650 ... Power supply, 660 ... Wiring 670 ... Voltmeter, 700 ... Control means, 701 ... Voltage application means, 702 ... Voltmeter, 1001 ... Reference image, 1002 ... Slave image, 1011-1023 ... Point , 1201... Reference image, 1202... Enlarged / reduced image of slave image, 1211 to 1223... Point, 1301... Button, 1401. ···point

Claims (12)

For a point representing a measurement object detected from within measurement data, a data analysis device for associating points related to the measurement object,
Input means for inputting a plurality of the measurement data, point information representing the appearance positions of the plurality of points in the measurement data, and measurement conditions when the measurement data is measured;
Data correction means for calculating a correction amount of the measurement data by comparing the measurement conditions in the measurement data to be associated, and correcting the measurement data and the point information based on the correction amount;
Correspondence for creating point association information in which the points are associated with each other based on the measurement data corrected by the data correction unit and the corrected point information between the measurement data having different appearance positions of the points in the measurement data Point detection means;
A data analysis apparatus comprising:   The measurement data is data in which a plurality of the points appear in two-dimensional data, and the appearance position of the point is indicated by the position of the point in the two-dimensional data. Data analysis equipment.   The measurement data is a two-dimensional electrophoresis image, and a portion having a high density value indicating a color density or brightness density of an image appears for each pixel in the two-dimensional electrophoresis image as the point. The data analysis apparatus according to claim 2. The data correction means includes
4. The data analysis apparatus according to claim 3, wherein a correction amount of the measurement data is calculated based on an applied voltage of electrophoresis and a migration time when a two-dimensional electrophoresis image is measured. The calculation of the correction amount is as follows:
The data analysis apparatus according to claim 4, wherein the correction amount of the measurement data is calculated based on a total applied voltage that is a product of the applied voltage of electrophoresis and the migration time when a two-dimensional electrophoresis image is measured. The calculation of the correction amount is as follows:
6. The data analysis apparatus according to claim 5, wherein a ratio of total applied voltages when measuring two-dimensional electrophoresis images is set as a correction amount between two-dimensional electrophoresis images to be associated. Correction of the measurement data and point information is as follows:
Expanding or reducing the two-dimensional electrophoresis based on the ratio of the total applied voltages;
7. The data analysis apparatus according to claim 6, wherein the appearance position of a point appearing in the two-dimensional electrophoresis image is converted.   Output means for outputting two-dimensional electrophoresis expanded or reduced based on the ratio of the total applied voltages and the appearance position of the point converted based on the ratio of the total applied voltages. The data analysis apparatus according to claim 7.   The data analysis apparatus according to any one of claims 1 to 8, further comprising an output unit that outputs measurement data corrected by the data correction unit and corrected point information.   The program for functioning a computer as an input means as described in any one of Claims 1-9, a data correction means, or a corresponding point detection means.   The computer-readable recording medium which recorded the program of Claim 10. A data analysis method for associating points related to the measurement object using measurement data in which a point indicating the measurement object appears,
Inputting a plurality of the measurement data, point information representing the appearance positions of the plurality of points in the measurement data, and measurement conditions when measuring the measurement data;
Calculating a correction amount of the measurement data by comparing the measurement conditions in the measurement data to be correlated, and correcting the measurement data and point information based on the correction amount;
Creating point association information in which the points are associated with each other based on the measurement data and point information corrected by the correction step, between the measurement data having different appearance positions of the points in the measurement data;
A data analysis method comprising: