JPS58195139A - Fractionation processing method in densitometer - Google Patents

Abstract

PURPOSE:To obtain perfect data with all abnormal specimens, by the constitution wherein the operation for remeasurement of the specimens that cause something abnormal in fractionation is accomplished automatically. CONSTITUTION:When initial data is inputted to a CPU100 by operating an input key, an optical detection part 12 measures the density in the fractionated components for each of specimens a1-an on a migration membrane 1 in accordance with the command of the CPU100. The detection signal thereof is loaded through an amplifier 103 and an AD converter 104 into the CPU. At the same time, a command is inputted from the CPU100 to a function generator 105 and after the arithmetic processing, the results thereof are printed out from a printer 106 and a divided density chart is plotted. If abnormality in division arises in any of the specimens during the course of such density measurement, the position thereof, the number of divisions, the number of the specimen, etc. are stored in the CPU100, and the remeasurement of the specimen showing abnormality is accomplished automatically and satisfactorily upon ending of the density measurement with all the specimens a1-an.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for measuring concentration using a densitometer, and more specifically, when a fractional abnormality occurs in any of the electrophoresed specimen samples, re-measurement improves the fractional sensitivity. The present invention relates to a fractionation processing method that can be sequentially and automatically performed while switching and adjusting.

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 arranged at regular intervals in the horizontal direction j (the so-called X-axis direction) on the same flat support. , and a sample in which multiple components of each sample are migrated on a straight line in the vertical direction (the so-called Y-axis direction) is held in a frame such as a cassette, and the frame is aligned in a plane along the vertical, horizontal, and vertical axes. By moving the sensor in this direction, the fractional concentrations of multiple samples are automatically measured, and these measured values are automatically recorded along with a fractional concentration diagram. In addition, such a □concentration meter for multiple samples is one that is configured to sequentially and continuously process a large number of samples, for example, several dozen or more, by installing several frames holding the above-mentioned samples:::□11- There are many.

By the way, the electrophoresis performed in such concentration measurement is usually carried out on sample samples such as serum proteins (albumin, α1, etc.).
．． α3. Although each globulin is subjected to wtK separation for 5 minutes in the order of β, r) to obtain a migration pattern, in reality, changes in the current and voltage applied to the support membrane, ambient temperature, and pH
, due to electrophoresis conditions such as ionic strength or sample application conditions, an electrophoresis pattern that should normally consist of 5 fractions may become, for example, 4 fractions.
There may be fluctuations in fractions or 6 fractions, etc., and the value of the 5 fractions cannot be determined, resulting in inaccurate and unreliable measurement results. A problem arises in that the necessary clinical data for everything is not available.

Therefore, in order to re-measure the abnormal sample in which such a fractionation abnormality has occurred, and to obtain the normal 5 fractions, we have removed the peaks that occurred outside the peak of the basic electrophoresis pattern, i.e., the yellow fraction point ( Albumin, α1.α,.

β, γ) Noi Hajime: Calculated by including it in Reka's globulin: 1
゜71su + ttmuv applied 0 ga 6 hi 6 gabe Previously, the work of fractionation processing in step 1 was carried out manually by the operator, who looked at the electrophoresis pattern and the electrophoresis image visually, making the work extremely difficult. This required time and effort and was inefficient.

This invention was developed in view of the above-mentioned drawbacks of the conventional art, and allows re-measurement of samples with abnormal fractionation to be automatically and reliably corrected and separated into the normal 5-fraction state. The purpose is to obtain complete measurement data for abnormal specimens.

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

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

This operation is performed according to the procedure shown below.

(a) The center line to the first sample of the electrophoretic membrane 1 in which multiple samples, for example 20 samples, are fractionated as shown in the m1 diagram is scanned over its entire length, and the voltage change due to this scanning is simulated. The center point P of albumin, which is determined as the point with the highest concentration from these stored values, is stored in the memory built into the densitometer.
(see Figures 2 and 8), calculate the distance f from the starting point 0 of movement (this position is determined by the positioning of the optical system detection unit 12, which will be described later), and after determining the position, the above-mentioned concentration Store it in the built-in CPU. This f is easily calculated by the CPU using the following equation.

Here, F is the total length of the sample from the starting point O to the ending point Q, N,
, N are the numbers of samples from the starting point O to the points Q and P, respectively.

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

(i)) Next, while moving the sample carrier in the horizontal axis (X-axis) direction, the optical system detecting section detects the voltage change in the manner described in (a), and based on the amount of change in voltage, the next sample B Continuing to identify albumin as the area with the highest concentration, this is determined as CP.
Let U remember it. In this case, the trajectory of point P in the horizontal axis direction is Y'-Y'.

(C) Further, continue moving the sample transport table in the horizontal axis direction until it reaches the midpoint R with sample B (the 4th point in the transparent area between samples A and B).
As shown in the figure, the voltage value is stopped at the peak (ie, the point where 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.

From the stored values described in (b) and (C) above, the albumin fractionation range 81 to S is specified (see Figure 4).7 The voltage level in this range 81 to S is actually shown in Figure 5. As shown in the figure, it fluctuates considerably and is not uniform. Therefore, the center point S cannot be determined based only on changes in density. For this purpose, certain conditions must be met to detect the voltage fluctuations (maximum and minimum points), such as the difference (amount of change) between the maximum and minimum values.
By setting and canceling extreme level fluctuations, data fluctuations are minimized (at the same time, data that is smaller than the amount of change in the set fluctuation range is treated as the same data. S by doing
, ~S2 become the same small 1.1 nita, and the center point S of the B sample can be determined as the midpoint of the range 5j-S (FIG. 6).

(e) Next, the sample carrier is returned to the center point S of the albumin of the sample B, and further the sample carrier is moved in the vertical axis direction to return this carrier to the movement start point O. It becomes possible to measure the concentration of all fraction components from At to each γ globulin.

Thereafter, by repeating the above method, the concentration measurement in the following document C is performed.

As a slit in the optical system detection section, for example, as shown in FIGS. 7(a) and 7(b), a slit plate 4 is provided with slits 2.3 located at right angles to each other.
The support frame 5 is equipped with one slide in the directions of arrows 6 and 7. This slit 2 is perpendicular to the vertical axis direction, that is, the fractionation direction of the sample, and is used exclusively as a slit for concentration measurement. It is perpendicular to the i-column direction of the specimen, vertical, 1 car, o eka - 6, 1 yo.-"!77) &
L”Cut 11 I want to do it.
In this way, the fractional concentration of each specimen sample is continuously measured by the slit 8 while automatically detecting the sample interval in the vertical and horizontal axis directions.

Similarly, these slits are not limited to this, but as previously proposed by the present applicant, it is possible to adopt various other configurations.

In addition, in this example, the optical system detecting section is fixed and the sample transport table is moved in the vertical and horizontal axes directions in the above concentration measurement. The table may be fixed and the optical system detection unit may be moved in the vertical and horizontal directions with respect to the table. In this case as well, the concentration can be measured in the same manner as described above (of course, it is possible to do so without any problem).

In this embodiment, as shown in FIG.
1, and is arranged to be sequentially transferred in the horizontal axis (X-axis) direction and the vertical axis (Y-axis) direction with respect to the optical system detection unit 12 at a predetermined timing. This transfer is performed, for example, by a rack and pinion mechanism in the X-axis direction and by a timing belt in the Y-axis direction, which are moved by stepping motors. In this case, after the electrophoretic membrane 1 in the cassette 10 on which the electrophoretic pattern has been formed in the electrophoretic treatment process is subjected to a transparent treatment, the cassette 10 is placed on the sample carrier 11.

Next, the fractionation processing method in the present invention will be explained.

(In this example, abnormal fractionation refers to the 5 fractions in which the electrophoresis pattern of the specimen is normal (albumin alpha).

α2. β, r globulin) is not separated, and the number of fractions increases or decreases from 5 fractions to 4 fractions or less or 6 fractions or more. ) First, the concentration of all n samples al to an electrophoresed on a single electrophoresis membrane 1 shown in FIG. 9 was measured using a pre-selected fractional sensitivity, and At the same time that a fractional concentration map is created for each sample, the measurement results are printed out on report paper using a recording device attached to the densitometer.

This concentration measurement was carried out according to the measurement procedure described above.
Ru.

In this example, the above fractional sensitivity is Hfl (＠＾), H(＾)
It can be switched and adjusted to 5 levels: M (medium), L (low), and LL (lowest), and as described later, when a fractionation abnormality occurs in either sample a or ~a, the abnormality can be adjusted. It is designed to be automatically switched according to a command from the CPU built into the densitometer. This switching changes the fraction sensitivity level (L→LL, M-
, L, H, M), or if it is less than 5 fractions, increase it as (L→M, M, H, H→HH).

At the start of concentration measurement, the operator selects an arbitrary level of fractionation sensitivity from among eight levels: H (high), M (medium), and L (low) by operating a sensitivity switching key (not shown). is selected and set in advance. Then, under this selected sensitivity, all samples a, ~an
degree measurements are taken.

At this time, suppose that a fractional abnormality occurs in one of these specimens ~an (・). a, 6 fractions,
Sample a again? Suppose that it is fractionated into 7 fractions.

In this case, these abnormal samples al+allea? On the other hand, for these abnormal samples at + 36 + "I,
The position of the first specimen a1 and its specimen number, that is,
The number of fraction abnormalities in the first sample al and the distance between them are stored in the built-in CPU of the densitometer, and at the same time, the number of fractions in each abnormal sample is stored in the CPU. It is determined and stored by the CPU.

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

After that, each abnormal specimen a! h a6 r a? What is the 111 constant of the component concentration? , be caught. This re-measurement is carried out as follows - First, based on the memory value stored in the CPU, the sample transport table 11 is controlled to move by a distance in the X-axis direction with respect to the position of the specimen a1, and the optical The system detection unit 12 is positioned at the position of the abnormal sample a.

At the same time, the fractionation sensitivity is automatically switched and set in a predetermined range from a "low" level to a "high" level in accordance with the number of fractions 4 of the sample a. This setting is performed based on the stored value according to a command from the CPU. Thereafter, sample a is remeasured using this fractional sensitivity according to the measurement procedure described above, and the measurement results for this abnormal sample a are recorded together with the fractional concentration diagram.

Based on this measurement result, it is automatically determined whether or not the fraction has been corrected to the regular 5 fractions.

As a result, during the first measurement, Fig. 11(a)
As shown by the arrow (a), for example, albumin A4 and α
α that no fractionation abnormality, that is, no fractionation point appeared during 1
When part 1 is clearly corrected and separated as shown in Figure 11(b) and measurement data as 5 fractions are obtained, sample a,
remeasurement is completed.

On the other hand, if the corrected and separated state is not corrected to the normal 5-fraction state even after re-measurement, this will be automatically determined and a message indicating that the fraction will be corrected again will be displayed on the display outside the figure, as well as the corresponding α fraction point at which the fractional point should be entered. An instruction of the fractionation order number corresponding to the part , for example, a message ``PUSH8LOW&[=3'' is displayed, and the sample transport table 11 is placed in the 6th stage state at the measurement start position of sample a. Next, when the fraction correction operation is performed manually using an external key, for example, the "8LOW" key is operated, and the "[=] (equal)" key is operated, the above-mentioned first
As shown in Figure 1 (a), similar data is printed out at an extremely slow measurement speed. Therefore, in the process, the abnormal fraction corresponding to α that did not appear in the remeasurement is manually separated using an external key. If we add the pixels, we get Figure 11(b)
As shown in FIG. 2, a concentration map of the 5 fractions in which the α1 portion has been corrected is obtained, and measurement data after the correction to the normal 5 fractions is obtained, thus completing the remeasurement of the sample a.

Next, the sample transport table 11 is moved in the X-axis direction from the position of the sample a1 by a distance Z2+, that is, a distance of 1 m< from the sample a2.
tt-tt), 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 of sample alI is 6, and therefore, at the same time as this positioning, the fractionation sensitivity corresponding to this number of fractions is 6.
The above CPU command automatically switches and adjusts the level from "high" to "low" within a predetermined range, and then re-measurement of sample a is automatically performed under this fractional sensitivity according to the measurement procedure described above. The measurement results are recorded together with the fractional concentration map, and based on the measurement results, it is automatically determined whether or not the abnormal sample a has been corrected to the normal 5 fractions, as described above. It is determined.

As a result, as shown by the arrow (b) in FIG. 12(a), the abnormal fractionation point f→ that occurred in the β-globulin portion during the first measurement is reduced and compressed, and as shown in FIG. As shown in b), when the fraction is corrected to the 5-fraction state and measurement data for the 5-fraction is obtained, the re-measurement of the sample a1 is completed.

On the other hand, even after re-measurement, the fraction was corrected to the normal 5 fraction of E8.
htly゛&ni&“・l N' 2-,, de'+J”
A process like Q K T ' is performed.

First, an instruction to ``perform fraction correction again,'' such as the message ``P'R, NO, \ (fraction number zero)'' is displayed on the display section. Then,
With the optical system detection unit 12 positioned at the measurement start position of the sample a6, the sample transport table 11 is placed in a standby state, that is, in a "waiting for key human power" state. Next, an operation is performed to erase the abnormal fraction C) occurring in the β globulin portion using the external key manually. In this operation, the fractionation order number corresponding to the abnormal fractionation points e, that is, number (5), which is the fractionation order counted from the left as shown in FIG. ) (This is done by inputting something like ``=E''. With this operation, the fraction calculation is performed again in the CPU for sample a, and the part of the abnormal fraction point (c) is automatically deleted by the calculation. The measurement data corrected into 5 fractions as shown in Figure 12(b) and its concentration i are automatically recorded and printed out.As a result, sample a,
When the measurement data in the 05 fraction state is obtained, the re-measurement is completed.

Thereafter, the trial knee feed table 11 is further controlled to move by a distance 5 in the X-axis direction based on the sample a position, that is, a distance C1m1x from the sample a, and the optical system detection unit 12 performs measurement of the sample av. Then, a fractionation process is performed to correct the sample into 5 fractions using the same procedure as for sample a, and its component concentration is remeasured.

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 by commands from the CPU built in the densitometer based on the memorized values from the first measurement. This is performed by controlling a pulse motor (not shown) that drives the sample transport table 11 in the X-axis and Y-axis directions.

Thus, each abnormal specimen al+al+a? The component concentrations were remeasured for all of these specimens a.

a6. The measurement results of the pattern corrected to the regular 5 fractions for all a7 are recorded and printed out on report paper by the recording device along with the fractional density diagram.

According to the method of this embodiment, a series of remeasurement operations, including switching and adjusting the fractionation sensitivity, are all performed under automatic control, which eliminates most of the manual work required in the past, resulting in labor-saving measurement work. can be achieved. Moreover,
Since the operations of the above-mentioned parts are controlled based on commands from the CPU built into the densitometer, operations can be performed accurately and without errors.

In the above example, the case where abnormal fractionation occurred in 8 samples "!+all+a?" among samples al-ao was explained, but the present invention is not limited to this, and the present invention is not limited to this. Regardless of the method, re-measurement can be performed using the same procedure as above.In addition, in the handwritten example,
After the initial a degree measurement is completed, (Tsutan optical system detection unit 12
Although it was explained that the sample a was positioned at the measurement start position of the first sample a, and then the sample in which the abnormal fraction occurred was remeasured.
It is also possible to configure the optical system detection section 12 to be positioned directly at the position of the first abnormal specimen a2 after the first measurement is completed.

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 that makes up the overall configuration of the device used when implementing the method of the present invention. It is a diagram.

In FIGS. 18 and 14, the start/stop switch provided on the operation panel 101 is operated to drive the device, and the data input keys provided on the keyboard portion (not shown) of the same panel 101 are operated. Operate and convert initial data to CP
When input to U 100, a control signal is sent from this CPU 100 to a mechanical unit 02 consisting of a pulse motor, etc., and the optical system detection unit 12 is operated according to the command from OPU 100. Thereby, the concentration of the fractionated components of each of the samples a to ao electrophoresed on the electrophoresis membrane l is measured. At this time, a detection signal received and detected by the optical system detection section 12 is amplified by the amplifier 103, and then converted into a numerical signal by the A/D converter 104 and taken into the OPU 100. At the same time, from the CPU 100 to the function generator 10
A command signal is sent to 5, and after various arithmetic processing is performed on this converted numerical signal, the results are sent to OP.
The image is printed out from the printer 106 through the U 100 :l. At the same time, 1・'
A fractional concentration map is created. If a fractionation abnormality occurs in any of the samples during the above concentration measurement process,
As mentioned above, the position, fraction number, specimen number, etc. are stored in the OPU 100. Thereafter, once the in-place measurements have been completed for all samples a and -ao, re-measurement of the sample with a fractionation abnormality will be automatically and completely performed in accordance with the above-mentioned procedure based on the instructions from OPU 100. It is carried out in

Then, all of the abnormal samples are remeasured, and when measurement takes are obtained for all of them, all operations are completed and the apparatus is stopped.

In this example, messages accompanying the various operations described above or their operating procedures are displayed on the display of the display unit 107 and printed out from the printer 10B at the same time, so that errors caused by erroneous operations can be detected or debugged. This allows the operation to be performed with certainty. Similarly, in this case, the switching of the fractional sensitivity level, the adjustment, etc. are automatically performed based on the commands of the PU 100, as described above.

'11 As explained above, in the fraction processing power I method according to the present invention, when a fractional abnormality occurs in any of the specimens when measuring the concentration of the specimen sample, the The level of fractionation sensitivity is automatically switched up and down within a predetermined range, and re-measurement for each abnormal sample can be automatically performed sequentially while adjusting the level of fractionation sensitivity up and down. This method has the effect that remeasurement can be performed extremely quickly, reliably, and extremely easily compared to 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, each abnormal sample has a regular 5
Since re-measurement can be reliably performed with the fraction pc fraction corrected, completely reliable measurement data can be obtained for all specimens at all times, which also has the effect of improving the reliability of the measurement data.

In addition, in this invention, the above-mentioned re-measurement operation can be performed almost automatically and completely reliably, making it possible to completely automate the measurement operation in the densitometer, making it possible to significantly save labor. The effect of contributing to the field of clinical testing, which requires rapid processing, is significant.

[Brief explanation of the drawing]

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 8 explains the electrophoresis pattern and sample interval detection operation when the sample intervals are different. Figures 4 to 6 are voltage level diagrams for the case shown in Figure 8, Figure 7(a) and Φ) are explanatory diagrams of the slit used in the method of the present invention, Before switching (b)
indicates after switching. Figure 8 is an explanatory diagram of the main parts of a densitometer to which the method of the present invention is applied, Figures 9 to 12 (a), (b)
Figure 18 is a diagram explaining the re-measurement 1'#4 of an abnormal sample in the method of the present invention, Figure 18 is a basic processing flow diagram when implementing the method of the present invention, and Figure 14 shows the overall configuration of the present densitometer. It is a block diagram centered on the CPU. l... Electrophoresis membrane (support), 11...
...Sample transport table, 12...Optical system detection section, a,
%a,...specimen, am I a@' l a9
・・・・・・Abnormal specimen, (1), etsu ・・・・・・Abnormal fractionation part. Figure 1 - Y × Figure 2 Figure 3 Figure 4 □ □i'' ・1assi Figure 5 Figure 7 (G) (b) Figure 9 Figure 10 Figure 11 (0) (b ) Figure 12 (0) (b) Procedural amendment (method) May 17, 1980 Patent Office Shima 1) Haruki Tono 3, person making the amendment Relationship with the case Patent applicant 4, agent

Claims (1)

[Claims] (1) A plurality of specimens are arranged at regular intervals in the horizontal direction on the same planar support, and a plurality of components of each specimen are migrated in a straight line in the vertical direction. The densitometer measures the fractional concentration of the electrophoresed sample by moving it along the vertical axis and the horizontal axis perpendicular thereto, or by moving the optical detection section along the vertical and horizontal axes relative to the sample. Therefore, a fraction processing method in a densitometer in which the percentage image is obtained by re-measuring the component concentration of an abnormal specimen in which a fraction abnormality has occurred by the methods described in (a) to (d) below. (a) Measure the concentration of all the samples on the support using the fractionation sensitivity selected and set in advance, and after detecting and storing the position of the abnormal sample where the fractionation abnormality has occurred and its sample number, transport the sample. The table or optical system detection unit is moved back to the measurement starting position or positioned at the position of the abnormal specimen. Φ) Next (・), the sample transport table or optical system detection unit is driven and controlled based on the above memorized values, and the fractionation sensitivity is automatically switched up or down depending on the number of fractions of the abnormal sample. The component concentration of the abnormal sample is re-measured using the lever. (C) Based on the re-measurement results, it is automatically determined whether the abnormal sample has been corrected to the regular 5 fractions, and if further abnormal fractions occur. , automatically displays an instruction to that effect and to correct the fraction again. (d) Next, by external input, the fraction of the abnormal sample mentioned above is automatically displayed.
Depending on the number of fraction points, an operation is performed to add a fraction point to the fraction abnormal portion or to delete the fraction point.