JPS58120153A - Measuring method for density in densitometer - Google Patents

Abstract

PURPOSE:To measure again a component density automatically, by switching a fraction sensitivity corresponding to the abnormality of fraction of an abnormal substance to be inspected, in a densitometer for measuring the fraction density of a sample wherein a detecting unit of an optical system is moved in the directions of longitudinal and lateral axes in relation to the sample. CONSTITUTION:The density is measured for all substances a1-an to be inspected, numbering n, which are made to migrate on one migration film 1, with the fraction sensitivity selected beforehand. Thereafter, a component density is measured again for an abnormal substance a2 to be detected. Based on a memory value stored in CPU, a sample conveying table 11 is controlled to move at a distance l1 in the direction of an X axis from the position of the substance a1 to be inspected as a datum point, and thereby a detecting unit 12 of an optical system is positioned at the position of the abnormal substance a2. The fraction sensitivity is switched from a low level to high automatically in a prescribed width according to the number of fractions of the substance a2 and is set.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for measuring bait content in a densitometer, and more specifically, when an abnormality in fractionation occurs in either electrophoresed or electrophoresed specimen, re-measurement improves the fractionation sensitivity. The present invention relates to a method for measuring the degree of protection that can be performed automatically in sequence while making switching adjustments.

As is well known, a densitometer uses electrophoresis to fractionate a sample on a support such as a cellulose acetate membrane, and automatically measures the concentration of each component such as serum protein from the obtained sample (electrophoresis pattern). It is a device for recording, and in recent years, with the remarkable increase in the number of specimens per day, facilities such as hospitals, medical care, or testing centers have exclusively used devices for multiple specimens.

In such a densitometer, as shown in Fig. 1, a plurality of specimens (approximately 10 to 20 specimens) are placed at regular intervals of m1 in the lateral direction (so-called X-axis direction) on the same flat support. A sample in which a plurality of components of each sample are electrophoresed in a straight line in the vertical direction (the so-called YI1411 direction) is held in a frame such as a cassette, and the frame is held in a plane along the vertical and horizontal axes. By moving the sensor in the direction, the fractional concentrations of multiple samples are automatically measured, and the measured values are automatically recorded together with a densitometry map. Furthermore, many of these densitometers for multiple samples are configured to sequentially and continuously measure a large number of samples, for example, several dozen or more, by installing a plurality of frames holding the above-mentioned samples.

However, in such conventional performance sub-determination, the following problems related to the electrophoresis method have been pointed out.

During this electrophoresis, sample samples such as serum proteins are usually separated into 50% of each globulin in the order of (albumin, α1, α2, β, γ).
The electrophoresis pattern is obtained by separating and electrophoresing the fractions.
Fluctuations in the current pressure applied to the support membrane, ambient temperature,
Due to electrophoresis conditions such as humidity or sample coating conditions, the electrophoresis pattern that should normally consist of 5 fractions changes to, for example, 4 fractions, 6 fractions, etc., resulting in some samples not being separated into the normal 5 fractions. As a result, the measurement results obtained may be inaccurate and unreliable, reducing the reliability of clinical trials and causing problems that may lead to misjudgments.

Conventionally, the concentration of all samples on the support membrane was measured once, and then the operator manually separated each sample with a fractional abnormality into five regular fractions and re-measured the sample. I was doing it.

However, this method requires manual operation, which makes the work extremely cumbersome and requires time and effort.Furthermore, constant monitoring by the operator is required for re-measurement, and large numbers of samples must be monitored. This has been an obstacle when automating concentration measurements.

The present invention was made in view of such conventional drawbacks, and an object of the present invention is to enable the above-mentioned re-measurement operation to be performed automatically and accurately.

An embodiment of the present invention will be described in detail below with reference to the drawings.

First, a concentration measurement operation in a concentration meter to which the present invention is applied will be explained.

This operation is performed by the procedure shown below.

(a) Scanning the center line of the first sample A of the electrophoretic membrane 1 in which a large number of samples, for example 20 samples, as shown in FIG. ．． , stored in the densitometer's built-in memory.

From these stored values, the center point P of albumin, which is identified as the point with the highest concentration (see Figures 2 and 3), is determined, and the starting point 0 of the movement (this position will be described later) is determined. 5- Optical system detection Determine the distance f from the position (°) determined by the positioning of the part 12, determine U't to that extent, and store it in the CPU built in the densitometer.
It is easily calculated by pu.

2 f=--XF N.

Here, F is the total length of the sample from the starting point O to the ending point Q, N,
, N2 are the sampling numbers between the starting point 0 and points Q and 2, respectively.

Next, when aligning to the next sample B, the sample transport table is again moved in the vertical axis direction to the position of the center point P of the albumin and stopped there.

(b) Next, without moving the sample carrier in the horizontal axis (X) direction, the optical system detecting section detects the voltage change as described in (a), and based on the amount of change in voltage, the next sample B Continuing to identify albumin as the highest concentration point, the CPU
Let me remember it. In this case, the locus of point P in the horizontal axis direction is Y'-Y'.

(c) Further, the sample carrier continues to move in the horizontal axis direction until it reaches the midpoint R with sample B (at the transparent part between samples A and B = 6- as shown in Figure 4). Stop at the peak, i.e. the point at which the concentration is the lowest.

(d) In order to determine the albumin center point S of the next sample B, the CPU is caused to perform the following editing operation.

The albumin fractionation range 81 to S2 is specified from the stored values described in items (b) and jc) above (see FIG. 4).

The voltage level in this range 81 to S2 is actually the fifth
As shown in the figure, it fluctuates by a considerable amount and is not uniform. Therefore, the center point S cannot be determined based only on changes in density. For this purpose, the above voltage fluctuation (maximum point and minimum point)
By setting certain conditions for detection, such as the difference (amount of change) between the orange dog value and the local minimum value, and canceling extreme level fluctuations, data fluctuations can be reduced.
Data smaller than the amount of change in the set variation range is treated as the same data. By performing the above editing operations, 8
The values of 1 to S2 are the same data, and the center point S of sample B
can be determined as the midpoint of the range S1-S2 (FIG. 6).

(e) Next, the sample carrier is returned to the albumin center point S-4 of sample B, and the sample carrier is further moved in the direction of the original axis to return this carrier to the movement start point 0. It becomes possible to determine the concentration fi11 of all fraction components from albumin hW to each globulin r in the sample.

Thereafter, by repeating the above method, the concentration of the next sample C and subsequent samples is measured.

In addition, -1m'i'+1-. 'The slits in the optical system detection section are, for example, slits 2 and 3 located at right angles to each other, as shown in FIGS. 7(a) and 7(b).
A slit plate 4 provided with a slit plate 4 is mounted on a support frame 5 so as to be slidable in the directions of arrows 6 and 7. This slit 2 is arranged perpendicular to the vertical axis direction, that is, the direction of fractionation of the sample, and is used exclusively as a slit for concentration measurement. Further, the slit 3 is perpendicular to the horizontal axis direction, that is, the arrangement direction of each specimen, and is used as a slit 1 for detecting sample spacing in the vertical and horizontal axis directions.

Thus, the slit 3 allows the fractionation sensitivity of each specimen sample to be continuously measured while automatically detecting the sample spacing in the vertical and horizontal directions.

In addition, the stiff slit is not limited to this, and as previously proposed by the applicant, other various configurations may be used.
It is Noh.

In addition, in this example, when measuring the concentration, the optical system detection section is fixed and the sample transport table is moved in the vertical and horizontal directions relative to it. The transport table may be fixed and the optical system detection unit may be moved in the direction of the yarn loss/horizontal axis relative to the conveyance table. In this case as well, it goes without saying that the gravel level measurement can be performed without any problem in the same manner as described above.

In this embodiment, as shown in FIG.
1, and is sequentially transferred to the optical system detection unit 12 in the horizontal axis (X-axis) direction and the vertical axis (Y@) direction at a fixed timing υ. This transfer, for example,
This is performed by a rack and pinion mechanism in the axial direction and a timing belt in the Y-axis direction, each driven by a stepping motor. In this case, the electrophoretic film 1 in the cassette 10 on which the electrophoretic backbone has been formed in the electrophoresis process is subjected to a transparent treatment, and then the cassette 10 is placed on the sample carrier 11.

Next, the concentration measuring method according to the present invention will be explained.

(In this example, abnormal fractionation means that the electrophoresis pattern of the sample is not separated into the normal 5 fractions (albumin, α1, α2, β, and γ globulins), and the number of fractions is 5 fractions. First, all of the n samples a, ~an migrated on one electrophoretic membrane 1 shown in FIG.
Concentration measurements are then carried out with a pre-selected fractional sensitivity.
A fractional concentration map as shown in FIG. 2 is created for each sample, and at the same time, the measurement results are printed out on report paper by a recording device installed in the densitometer.

This concentration measurement is performed according to the measurement procedure described above.

In this example, the above fractional sensitivities are HH (highest), H (high), M
..., L low, and LL (lowest).

When a fractional abnormality occurs in any of the 10- and 10-an, it is automatically switched according to the instructions from the CPU built in the liA meter. This switching is necessary when the number of fractions of the sample is more than 5 fractions, the level of fractionation sensitivity (
It is carried out by lowering as in L→LL, M−+L, H→M), and increasing as in (L−+M, M→H, f(−→HH)) if it is less than 5 fractions.

To start concentration measurement, the operator operates a sensitivity switching key (not shown) to select H@,
An arbitrary level of fractionation sensitivity is selected and set in advance from three levels: M..., L low. Thereafter, under this selected sensitivity, all the specimens a1 to anK (8) are subjected to immersion measurement. At this time, it is assumed that a fractionation abnormality has occurred in any of these specimens a1 to an. For example, As shown in FIG. 10, sample a2 is divided into 4 fractions, and sample a is divided into 6 fractions.

It is also assumed that sample a7 is fractionated into seven fractions.

In this case, these abnormal specimens a2+a5+a? The measurement results are recorded along with the fraction a diagram for all other analytes.

=11= On the other hand, these abnormal samples a2+a5+a? For, the position from the first sample a1 and its sample number, that is, which sample from the first sample a1 has a fractionation abnormality, and what is the distance between them is the agricultural degree. It is stored in the built-in CPU of the meter, and at the same time, the fraction number of each abnormal sample is determined and stored by this CPU.

Thereafter, when the concentration is measured for all the samples a1 to an and the sample transport table 11 reaches the measurement end position, the transport table 11 moves back to the measurement start position as shown by the arrow 13 in FIG. As shown by the blank line in FIG. 10, the optical system detection section 12 is positioned at the position of the first specimen a1.

After that, each abnormal specimen a2+a5+a? The component concentration will be remeasured. This re-measurement is performed as follows.

First, the sample transport table 11 is controlled to move over a distance t in the X-axis direction with the position of the sample a1 as a reference based on the memory value stored in the old CPU, and the optical system detection unit 12 detects the abnormal sample a.
It is located at position 2.

12- At the same time, the fractionation sensitivity is automatically switched from a low level to a high level in a predetermined range according to the number of fractions 4 of the sample a2. This setting is performed based on the stored value according to a command from the CPU. Thereafter, sample a2 is remeasured using this fractional sensitivity according to the above measurement procedure.

As a result, during the first measurement, for example, albumin At and α2
α1, in which there was a fractional abnormality, that is, no fraction appeared in between.
As shown in Fig. 11(b), the part of sample a is clearly fractionated by increasing the level of fractionation sensitivity.
A degree diagram is drawn on the normal # divided into 5 regular fractions per 2.

Next, the sample transport table 11 moves a distance Z2+ in the X-axis direction based on the position of the sample a8, that is, a distance (t
2-41), and the optical system detection unit 12 is positioned at the measurement start position of the next abnormal sample a. In this case, the number of fractions for sample a is 6, and therefore, at the same time as this positioning, the fractionation sensitivity is automatically changed from ``high'' to ``low'' in accordance with the instruction from the CPU. ’” level at a predetermined width, and then, under this fractionation sensitivity, sample a
, is automatically remeasured according to the measurement procedure described above. Then, as shown by the arrow (b) in FIG. 12(a), the abnormal fractionation that occurred in the β-globulin portion during the first measurement, for example, causes the level of fractionation sensitivity to be lowered by a predetermined width. As shown in FIG. 12(b), a normal fractional concentration map is drawn for this sample a, which is divided into five normal fractions.

Thereafter, the sample carrier 11 is further controlled to move by a distance t3 in the X-axis direction based on the position of the sample a, that is, a distance (z, -22) from the sample a, and the optical system detection unit 12 After being positioned at the measurement position of a7 and then adjusting the fractionation sensitivity by the same procedure as above,
What are the components of this sample a7? ! IF is remeasured.

In addition, the operation control of each part, such as the movement control of the sample transport table 11 or the positioning control of each sample with respect to the optical system detection part 12, is performed using the built-in densitometer based on the memorized value of 14- at the time of the first measurement. This is performed by driving and controlling a pulse motor (not shown) that drives the multi-sample transport platform 11 in the X-axis and Y-axis directions in accordance with instructions from the CPU.

In this way, by appropriately adjusting the level of fractionation sensitivity by switching up or down within a predetermined range, the component concentrations of all of the abnormal samples a2+a5+a7 are remeasured, and these sample a
2~a? A normal concentration map divided into five regular fractions is drawn for all of the samples, and the measurement results are recorded on report paper using a recording device and printed out.

According to the method of the present invention, the switching and adjustment of fractionation sensitivity and a series of remeasurement operations are all performed under automatic control without human intervention, so there is no need to rely on manual labor as in the past. Therefore, it is possible to save labor in measurement work.

Furthermore, since the operations of the above-mentioned parts are controlled based on commands from the CPU built into the densitometer, each part can be operated accurately without errors.

In addition, in the above example, among the samples a, ~a, the wave body a2+ +a? Although the case has been explained above, the present invention is not limited to this, and re-measurement can be performed using the same procedure as above regardless of the number of abnormal samples. . Further, in the above embodiment, after the initial concentration measurement is completed, the optical system detection section 12 is once positioned at the measurement start position of the first sample a, and then the angle measurement of the sample in which the abnormal fraction has occurred is performed. Although the optical system detecting section 12 is described as being performed, it is also possible to configure the optical system detecting section 12 to be directly positioned at the position of the first abnormal specimen a2.

Next, FIG. 13 is a basic processing flow diagram when implementing the method of the present invention, and FIG. 14 is a block diagram centered on the CPU 1 showing the overall configuration of the device used when implementing the method of the present invention. be.

In FIGS. 13 and 14, the start/stop switch provided on the operation panel 101 is operated to drive the device, and the data input key provided on the keyboard section (not shown) of the same panel 101 is operated. CP the initial data by operating
When input to U100, a control signal is sent from the CPU 100 to the mechanism section 102 consisting of a pulse motor, etc., and the optical system detection section 12 is activated by the command from CI) (Jloo). Each specimen a8 ~.

Concentration measurements of fractionated components for each an are performed. At this time, the detection signal received and detected by the optical system detection section 12 is transmitted to the amplifier 10.
After being amplified by 3, the A/D converter 104 converts the signal into a mathematical signal, which is then taken into the CPU 100. At the same time, a command signal is sent from the CPU 100 to the function generator 105, which performs various operations on the converted numerical signal. Processed data, the result is CPU 1
00 and is printed out from the printer 106. At the same time, a fractional concentration map is created. And the above i1 I! If a fractionation abnormality occurs in any of the samples during the measurement process, the number of fractions at that position or the sample distribution number is stored in the CPU 100, so it is counted as 1. After that, all samples a1 to a
When # degree measurement is completed for n, the re-measurement of the sample in which the fractionation abnormality has occurred is automatically performed according to the above-mentioned procedure based on a command from the CPU 100. 17- Then, the pan of the abnormal sample is re-measured, and once measurement data is obtained for these, all operations are completed and the apparatus is stopped.

In this example, messages accompanying the various operations mentioned above or their operating procedures are displayed on the display F of the single display n107, and at the same time are printed out from the printer 108 to detect errors caused by erroneous operations or perform debugging thereof. This allows for reliable operation. In this case, the switching and adjustment of the fractional sensitivity level, etc., are automatically performed based on the commands of the CPU 100, as described above.

As explained hereafter, in the #concubine measurement method according to the present invention, when an abnormality in fractionation occurs in any of the specimens when measuring the concentration of the specimen, fractionation is performed in accordance with the number of fractions of the abnormal specimen. The image sensitivity level is automatically switched up and down within a predetermined range, and each abnormal sample can be remeasured automatically in sequence while adjusting the fractional sensitivity level up and down. This method has the advantage that re-measuring can be performed much more quickly, reliably, and much more easily than conventional manual operations. Therefore, the method of the present invention has the effect of significantly improving the work efficiency of this remeasurement compared to the conventional method.

Moreover, in this invention, 5 of the correct inkstones are used for each abnormal specimen.
Since re-measurement can be reliably performed in a state separated into fractions, accurate measurement data can be obtained for all specimens at all times, which also has the effect of improving the reliability of determination.

In addition, in this invention, all of the re-measurement operations described above can be performed automatically, making it possible to completely automate the measurement operations in the densitometer, which allows for significant labor savings, and allows rapid processing of multiple samples. The effect of contributing to the field of clinical testing, which is in demand, is significant.

[Brief explanation of drawings]

Figure 1 shows an example of the electrophoresis pattern for multiple samples;
Figures 8 to 8 are diagrams explaining the concentration measurement method according to the present invention, in which Figure 2 is a fractional concentration diagram, and Figure 3 is an explanation of electrophoresis patterns and sample interval detection operations when sample intervals are different. FIGS. 4 to 6 are voltage level diagrams for the case shown in FIG. 3, and FIG. Before switching,
(b) shows the state after switching. Fig. 8 is an explanatory diagram of the main parts of a densitometer to which the method of the present invention is applied, Figs. 9 to 12 (a),
(b) is a diagram explaining the order of re-measurement of abnormal samples in the method of the present invention, Figure 13 is a basic processing flow diagram when implementing the method of the present invention, and Figure 14 is a CPU showing the overall configuration of the present densitometer. (This is a block diagram centered on i7. 1... Electrophoretic membrane (support) 11... Sample transport stage 12... Optical system detection sections a1 to an. ...Sample a2 +aB +a7... Abnormal specimen Fig. 1 -> Figure 12 (0) (b)

Claims (3)

[Claims] (1) A plurality of specimens are arranged at regular intervals in the horizontal direction on the same flat support, and multiple components of each specimen are migrated onto a vertical straight ball. The fractional concentration of the electrophoresed sample is measured by moving the vertical axis and the horizontal axis orthogonal to the vertical axis, or by moving the autopsy detection unit in the vertical and horizontal directions with respect to the sample. 1. A method for measuring concentration in a densitometer, comprising automatically re-measuring the component concentration of an abnormal specimen using the method described in (a) to <b) below. (a) After measuring the agricultural yield of all the samples on support J with the fractionation sensitivity selected and set in advance, and detecting and memorizing the position of the abnormal sample where the fractionation abnormality occurred and its sample number. , move the sample carrier or optical system detection unit back to the measurement start position. (b) Then, based on the above-mentioned memory, the sample transport table or the optical system detection section is controlled to move to the position of the above-mentioned abnormal specimen, and at the same time, the J: period fractionation sensitivity is increased or decreased in response to the abnormality in the fractionation of the abnormal specimen. After automatically switching and adjusting with a predetermined width, the concentration of all components of this abnormal sample is measured. (2) The method according to claim 1, characterized in that, after the concentration measurements of all the samples are completed, the sample transport table or the optical system detection unit is controlled to move directly to the measurement start position of the abnormal sample. Concentration measurement method using a densitometer. (3) The concentration in the densitometer according to claim 1, characterized in that the fractionation sensitivity is configured to be switchable and adjustable in multiple stages up and down in response to the fractionation of the abnormal specimen. Measuring method.