JP3401040B2 - How to display densitograms - Google Patents

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for displaying a densitogram in an electrophoresis apparatus. 2. Description of the Related Art In an electrophoresis apparatus, a sample such as serum is applied to a support such as a cellulose acetate membrane by an applicator, and is electrophoresed in an electrophoresis tank for a predetermined time.
After dyeing, decoloring, and drying in the dyeing tank, photometry is performed using a densitometer. Based on the sampling data, albumin (Alb), α ₁ -globulin (α _1-
G), α ₂ -globulin (α ₂ -G), β-globulin (β
-G) and each fraction% of γ-globulin (γ-G), α
_The A / G ratio, which is the fractional ratio of Alb to the total globulin of _1- G, α ₂ -G, β-G, and γ-G, is calculated and written together with the electrophoresis pattern (densitogram) in a given report. It is displayed on a display device such as a CRT. [0003] However, in the conventional display method, the densitogram displayed on the report or the display device is only that of the measurement sample, and the densitogram has the highest concentration. Autospan is performed so that the peak of Alb is always constant so that the concentration of each fractionated protein is relatively expressed. For this reason, there is a problem that a change in the absolute amount of each fraction cannot be determined from the densitogram, a difference in electric mobility and the presence of M protein cannot be found, and it is difficult to visually analyze a disease state. In order to solve such a problem, the applicant of the present invention disclosed, for example, in Japanese Patent Application Laid-Open No. 62-42033, normalized the electrophoresis length of a densitogram of a measurement sample together with a standard normal serum densitogram. We have already proposed a method of displaying densitograms that are duplicated. According to this display method, since the densitogram of the measurement sample and the reference densitogram are displayed in an overlapping manner, there is an advantage that the pathological condition can be easily analyzed by visual inspection based on the comparison. . However, this method has a problem in that it is not possible to immediately understand the course of the disease, even if the disease state at the time of measurement can be understood. Actually, for example, in a hospitalized patient or the like, it is known in advance that the abnormality is abnormal. In addition, there is a problem in that the analysis conditions may change depending on different date and time or the apparatus, for example, because different types of electrophoresis apparatuses are used in the hospital or the operator is changed, so that the progress cannot be directly compared. . The present invention has been made in view of such conventional problems, and it is easy and accurate to visually determine the change in the concentration of each fraction even if the amount of sample applied or the migration length varies. It is an object of the present invention to provide a method for displaying a densitogram, which can be performed easily and accurately while visually observing the progress of a disease state. In order to achieve the above object, according to the present invention, a measurement sample is measured together with a reference sample, and their measurement data is stored in a storage means. The measurement data of the desired subject and the measurement data of the subject at another date and time or under different analysis conditions are respectively extracted, and these extracted measurement data are converted into the measurement data of the reference sample measured at the same time. Then, the normalized measurement data is overlapped and displayed. In the present invention, the measurement sample is measured each time together with the reference sample, and the measurement data is stored in the storage means. By specifying a desired subject from the measurement data stored in the storage means, measurement data of the subject at different dates and times or under different analysis conditions are extracted. The extracted measurement data is normalized based on the measurement data of the reference sample measured at the same time, and the normalized measurement data is overlapped and displayed. FIG. 1 is a diagrammatic cross-sectional view showing a configuration of a main part of an example of a densitometer in an electrophoresis apparatus embodying the present invention. The support 1 dyed, decolorized and dried in the dyeing tank is conveyed by a feed roller 2 to a photometric unit 4 containing decalin 3 and is subjected to photometric scanning by a photometric device 5 at a constant speed, for example, 8 mm / sec. rear,
The paper is discharged by the paper discharge roller 6. The photometric device 5 is configured to receive the light from the light source 5a passing through the support 1 with the light receiving element 5b, and the photometric device 5 having the light source 5a and the light receiving element 5b is connected to the support 1 as shown in FIG. By moving in the scanning direction b orthogonal to the transport direction a,
The electrophoretic image 7 formed on the support 1 is subjected to photometric scanning. FIG. 3 is a block diagram showing a configuration of a main part of an example of a data processing device for processing an output of the photometric device 5. As shown in FIG. The data processing device includes a logarithmic amplifier 12, an A / D converter 13, a CPU 14, a memory 15, a keyboard 16,
CRT 17, floppy disk 18, and printer 1
9 is provided. The photoelectric conversion output obtained from the light receiving element 5b by photometric scanning is logarithmically amplified by the logarithmic amplifier 12, converted into absorbance, and then sampled at the A / D converter 13 by a sampling rate (for example, 12 msec) determined by analysis conditions such as migration time. ) Is sampled and converted into a digital signal in synchronization with the clock corresponding to
The data is stored in the memory 15 under the control of No. 4. In one embodiment of the present invention, as shown in FIG. 4, a sample A is compared with a sample B which is the same subject as this sample A but is collected on another day. For this purpose, first, normal serum and sample A, which are the standards for normalization, are applied to the support, respectively, and electrophoresed. The densitogram obtained by photometry of the electrophoresis image is transferred to a floppy disk 18.
Respectively. Further, a normal serum and a sample B, which are the standards for normalization, are applied to another support, respectively, and electrophoresed. The densitogram obtained by photometry of the electrophoresis image is stored in the floppy disk 18. next,
The photometric data of each of the measurement samples A and B is normalized based on the photometry data of normal serum migrated on the same support.
In addition to the display on the report 20 as a display member by the printer 19, the difference and the ratio of the densitograms of the measurement samples A and B are obtained, and the increase / decrease of each fraction is similarly displayed on the CRT 17 and the report 20. FIG. 5 shows a flowchart of an example of the normalization processing. In this embodiment, the number of data points related to a predetermined migration length in the normalization is set to 350, and the peak position of Alb which is stable as a prominent peak in the migration image at 100 data points is set to 200 data points in the migration image. To match the peak position of β-G whose appearance position is stable. That is, using the respective peak positions of Alb and β-G as reference points, these reference points are used as 100 data points and 200 data points at 350 data positions in normalization.
Match each data point. Note that the number of points of the sampling data in the scanning range of the photometric device 5 is more than 350 points. In this normalization process, first, for each electrophoresis image, a reference point which is a peak position of each of Alb and β-G related to electrophoresis length is detected from the sampling data stored in the memory 15. The method of detecting this reference point will be described below. First Method : As shown in FIG. 6, data exceeding a predetermined threshold value is extracted from the sampling data of the electrophoresis image so that both end points are both ends of the electrophoresis image. Detecting the presence or absence of a peak in predetermined ranges l ₁ and l ₂ ,
The highest concentration in the detected peak was determined as the Al
The reference point is detected as the peak position of b and the peak position of β-G. Second Method The sampling data of the electrophoresis image is sequentially integrated from both end points,
Positions where those values are respectively a predetermined value or a predetermined ratio with respect to the total integrated value are defined as both end points of the electrophoretic image, and peaks in a predetermined range are respectively determined from the both end points in the same manner as the first method. The reference points are detected by detecting the peak positions and using the peak positions as the respective peak positions of Alb and β-G. For example, if the integrated value of the migration polarity from the positive electrode side to the Alb direction is set to 2% of the total integrated value, and the integrated value from the negative electrode side to the γ-G direction is set to 1% of the total integrated value, the migration length obtained by visual observation is approximately obtained. Matches. Third Method First, the peak position of Alb is detected for the reference normal serum, and then the third peak position in the negative direction of the migration polarity from Alb is detected as the β-G peak position.
Thereafter, the peak position of Alb is detected for the measurement sample, and is equal to the number of data points related to the migration length between the Alb peak position and the β-G peak position of the reference normal serum previously determined from the Alb peak position. A peak near the position of the number of data points is detected, and each reference point is detected using the peak position as a β-G peak position. In detecting the Alb peak position from the sampling data of the electrophoresis image, a relatively high threshold value is set because Alb is stable as a conspicuous peak, and the data having the maximum concentration of the data that exceeds this value is regarded as the Alb peak. It may be detected as a position.
81,943 No. As disclosed in Japanese detects the maximum value D _M from all data sampled, then detects the 1/16 or more peaks of the maximum value as a peak position P _M of the Alb from the positive electrode side of the electrophoresis polarity May be. Here, 1/1
6 to remove the peaks of prealbumin, as D _M is P _M be other components not Alb is detected as a peak position of Alb, which was empirically determined by various conditions, It is not particularly fixed. In this manner, the peak position of Alb, which is stable as a conspicuous peak position, and the peak position of β-G, whose appearance position is stable, are detected as reference points. Next, the detected Alb peak position and β-G peak position are respectively set at 100 data points and 200 data points on the X-axis having 350 data points related to a predetermined migration length. The X axis is normalized so that they match. For example, if the peak position of Alb is 120 data points and the peak position of β-G is 230 data points, those data positions are matched with 100 data points and 200 data points on the X-axis, and the other sampling data is The migration length between the reference points is 110 data points (23
0-120), while it corresponds to 100 data points (200-100) on the X-axis, and is shifted to data points on the X-axis according to the ratio. Here, when there is no sampling data corresponding to the data point on the X-axis, an interpolation process or an extrapolation process, that is, a process for aligning with both end data is performed. As described above, the normalization of the X-axis with respect to the migration length is completed. Next, the value of the sampled data at 350 points on the X-axis is converted to the number of data points between the reference points so that the integrated value becomes substantially equal to the integrated value between the corresponding original data positions. The normalization of the Y-axis is performed based on the ratio of the number of data (100 in this example) while these reference points are located on the X-axis. For example, in the above example, the Alb peak position was 120 data points, the β-G peak position was 230 data points, and the number of 110 data points between them was normalized to the number of 100 data points. The value of the sampling data at the data point is (110/10
0) to perform normalization on the Y axis. The above processing is performed by reading out the sampling data stored in the memory 15 under the control of the CPU 14, and storing the processed data on the floppy disk 18. Next, density normalization is performed to make the integrated value and the absolute amount of density correspond to each fraction. In normalizing the concentration, the total protein value or the Alb concentration value of the measurement sample is measured in advance by a biochemical analyzer or the like, and the value is connected via the keyboard 16 or from or to the biochemical analyzer. The data is input online or off-line via the computer system for testing and stored in the floppy disk 18. For the reference normal serum, the total protein value or the concentration value of Alb or the like is input via the keyboard 16 for the floppy disk. School 18
To memorize it. In addition, the reference integrated value of the input absolute amount with respect to the unit concentration (1 g / dl) is also stored. For example, when an Alb concentration value is input, A
lb is a reference integrated value for 1 g / dl, for example, 150
00 (by A / D conversion value). Where Alb
Assuming that the density value of is input as 4 g / dl,
First, the integrated value of the Alb fraction of the normalized data is determined, and then the ratio of the integrated value to the reference integrated value corresponding to the input Alb concentration value is determined. For example, normalized Alb
Assuming that the integrated value of the fraction is 80000 (in A / D conversion value), the input Alb concentration value is 4 g / dl, and the reference integrated value is 4 (g / dl) × 15000 = 6000.
Since it is 0, the ratio of 80,000 to 60,000 is
(60000/80000) = 0.75. next,
This ratio is multiplied by the data value of each point to complete the density normalization. In this density normalization processing, as disclosed in Japanese Patent Application Laid-Open No. S61-196154 proposed by the present applicant, differences in the staining properties of each fraction can be corrected simultaneously. When the total protein value is input, the same processing can be performed by calculating the ratio between the reference integrated value corresponding to the input value and the integrated value of all the fractions. As described above, when the normalization process is completed for each of the sample A and the sample B, the densitogram of each of the samples A and B together with the densitogram of the reference normal serum is subjected to CR.
T17 and the predetermined area of the report 20 are overlapped and displayed, and each fraction%, A / G ratio, and the like of the measurement sample are calculated, and the result is similarly displayed in the predetermined area of the CRT 17 and the report 20. indicate. Hereinafter, an example of overlapping display of the densitogram of the sample A and the densitogram of the sample B will be described. FIG.
7A shows the densitogram I of the measurement sample B indicated by a wavy line, and the densitogram II of the measurement sample A indicated by a solid line.
B shows both densitograms I and II as solid lines, and the line of densitogram II of measurement sample A is thicker or darker than densitogram I of measurement sample B. FIG. 8 shows a portion where the densitogram II of the measurement sample A is higher than that of the densitogram I of the measurement sample B by red hatching, and a portion below the densitogram II by blue mesh hatching. Of course, the type can be set arbitrarily as long as it is a hatching type and a color that clearly distinguishes the two. FIG. 9 shows the densitogram III of the test sample C.
8 and the densitogram IV of the same subject one month later is displayed in the same manner as in FIG. 8, and the densitogram v of the normal serum is additionally displayed. As described above, the number of densitograms to be compared is not limited to two, and can be set to any number. FIG. 10 additionally shows arrows in addition to the display method shown in FIG. 8 to increase or decrease each fraction. This arrow can also be replaced with another symbol as long as the increase or decrease can be clearly understood. In the above, an example in which a plurality of waveforms are completely overlapped is shown. However, if the specimen is normalized, the size of the densitogram can be absolutely compared. Alternatively, each densitogram can be displayed shifted in the Y-axis direction. According to this embodiment, the concentration of the measurement sample and the control measurement sample of the same subject are normalized based on the X-axis and Y-axis and the concentration value or the total protein value of the input single fraction. As a result, the densitogram of the sample to be measured is displayed in duplicate with the pattern related to the densitogram of the reference normal serum. This has the same effect as creating a migration image with the same migration length. Therefore, it is possible to obtain a densitogram corresponding to the concentration change of each fraction, that is, the span is proportional, and it is possible to represent the difference in the mobility of each fraction, the presence of the M protein, and the presence of the waveform band. The disease state can be easily analyzed from the densitogram. Also, when comparing samples,
Since the position of each data point can be used as an absolute reference, it can be done easily and accurately. Furthermore, by displaying the densitogram of the control measurement sample together with the pattern related to the densitogram of the reference measurement sample,
The change in increase / decrease of each fraction can be easily compared, and if the reference measurement sample and the control sample are of the same patient and the measurement is performed on another day, the follow-up of the patient can be visually observed. FIG. 12 is a diagram for explaining a reference example of a method for displaying a densitogram developed with the present invention. In this reference example, in order to compare and display the sample A and the sample B which is the same subject as the sample A but collected on another day, a normal serum and a sample serving as a standard for normalization are provided on the support. A
Are applied and electrophoresed, their electrophoretic images are photometrically measured, and the densitogram of the specimen A is normalized based on photometric data of normal serum which has been simultaneously photometrically stored, and stored on the floppy disk 18. In addition, normal serum as a standard for normalization and sample B are applied to another support, and each sample is electrophoresed. Photometry of the electrophoresis image is performed, and the densitogram of sample B is measured at the same time. The data is normalized on the basis of the data and stored in the floppy disk 18. The normalization is performed in the same manner as in the previous embodiment. Thereafter, when it is desired to duplicately display, the densitograms of the measurement samples A and B are called from the floppy disk 18 and displayed on the CRT 17 or the report 20. In this reference example, the densitograms of samples A and B are normalized, respectively,
8, there is no need to perform normalization each time the content is overlapped and displayed. Therefore, the overlapping display can be quickly performed. It should be noted that the present invention is not limited to the above-described embodiment, and that various changes and modifications are possible. For example, in the normalization processing, the same effect can be obtained by using only the X axis or using only the X axis and the Y axis. In the above embodiment, the density normalization processing is performed last, but this processing may be performed before the X-axis normalization processing. Further, the reference point in the electrophoretic image is not limited to the Alb peak position and the β-G peak position, but may be a peak position of another fraction, an end point of the electrophoresis length, or the like.
The reference sample used for the normalization process is not limited to a normal sample, but may be a 5-fraction sample electrophoresed on the same support.
It can also be used as a reference sample. In addition, a one-dimensional array sensor or a two-dimensional array sensor can be used as a light receiving element included in the photometric device. Further, the data of the densitogram can be stored not only in a floppy disk but also in any storage means such as a hard disk, a magneto-optical disk, and a flash memory. Also,
The data stored in the storage means may be measurement data under different analysis conditions. As described above, according to the present invention,
Densitograms of the same subject obtained at different times or under different analysis conditions were normalized with the densitograms of the reference samples measured at the same time, and displayed as duplicates. Data can be compared even if the analysis conditions are different due to differences in equipment manufacturers, models, operators, etc., and it is possible to easily and accurately determine changes in the concentration of each fraction visually, and to visually analyze the pathological progress of the disease. Can also be performed easily and accurately.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagrammatic sectional view showing a configuration of a main part of an example of a densitometer in an electrophoresis apparatus embodying the present invention. FIG. 2 is a diagram illustrating a scanning direction of an electrophoretic image. FIG. 3 is a block diagram illustrating a configuration of a main part of an example of a data processing device. FIG. 4 is a diagram for explaining an embodiment of the present invention. FIG. 5 is a flowchart illustrating an example of a normalization process. FIG. 6 is a diagram for explaining an example of reference point detection. FIG. 7 is a diagram illustrating an example of overlapping display. FIG. 8 is a diagram showing an example of overlapping display. FIG. 9 is a diagram showing an example of the overlap display. FIG. 10 is a diagram showing an example of overlapping display. FIG. 11 is a diagram showing an example of overlapping display. FIG. 12 is a diagram illustrating a reference example of the present invention. [Description of Signs] 1 support 2 feed roller 3 decalin 4 photometer 5 photometer 5a light source 5b light receiving element 6 paper discharge roller 7 electrophoretic image 12 logarithmic amplifier 13 A / D converter 14 CPU 15 memory 16 keyboard 17 CRT 18 Floppy disk 19 Printer 20 Report

Continuation of the front page (56) References JP-A-62-174631 (JP, A) JP-A-2-103460 (JP, A) JP-A-2-103459 (JP, A) JP-A-1-100436 (JP) JP-A-62-42033 (JP, A) JP-A-61-182552 (JP, A) JP-A-61-181942 (JP, A) JP-A-63-85349 (JP, A) 62-278442 (JP, A) JP-A-62-251651 (JP, A) JP-A-62-47534 (JP, A) JP-A-62-42034 (JP, A) (58) Fields investigated (Int. Cl ^7, DB name) G01N 21/00 -. 21/01 G01N 21/17 - 21/61 G01N 27/26 - 27/40 G01N 27/42 - 27/56 PATOLIS

Claims (1)

(57) [Claims] [Claim 1] A measurement sample is measured together with a reference sample,
The measurement data is stored in the storage means, and the measurement data of the desired subject stored in the storage means and the measurement data of the subject at another date and time or under different analysis conditions are respectively extracted. A method for normalizing the extracted measurement data based on the measurement data of the reference sample measured simultaneously, and comparing and displaying the normalized measurement data in an overlapping manner.