JPS5912982B2 - Measuring device for cuvettes - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method and apparatus for automatic reading and recording of the results of a number of virological, bacteriological and hematological reactions.

10 This device can also be used as a spectrophotometer, for example for measuring and recording the results of chemical color reactions.

When investigating the causes of death or determining the concentration of various substances in living organisms, it is often necessary to carry out studies of a large number of reactions in the laboratory15.

These studies may involve hundreds or even thousands of test tubes, for example when serial dilution or chemical colorimetric methods are used. In virus biology, diagnosis can be made by isolating the virus itself or a part thereof, and it is also possible to view the virus with an electron microscope. It is also often possible to make the diagnosis by demonstrating an increased titer of a viral antagonist in the blood, or by demonstrating viral antibodies in the cells25. When discovering the cause of death from a patient's sample using basic virological methods, sample dilution, ie, titer determination, plays an important role in many methods.

Reactions whose results can be read visually from a 30 dilution series are often used in many research methods. This type of reaction is hemagglutination, fixation of complement and inhibition of hemagglutination. If each sample is diluted several times in succession, it is necessary to perform a control titration to improve the reliability of the test results.
In this case, thousands of test tubes are processed and the results are visually read. In current methods, recording of results is primarily done manually. This kind of reading and recording of results is, of course, slow and subject to many errors. In bacteriology, diagnosis is carried out in many ways similar to that in viral science.

Many bacteriological research methods use the serial dilution method, and the results can be read visually. The more common of these methods is the determination of the antistreptococcal lysin titer (AST) of bacteria of the genus Streptococcus, which is based on a hemolytic reaction, and that of bacteria of the genus Staphylococcus (ASTA), which is based on a hemolytic reaction.
) titer measurement. In the Bidal test, a bacterial agglutination-based reaction is used to find out whether salmonella or typhoid types are present in a sample. When determining blood types in hematology, a large number of test tubes must often be handled, and the results must be read visually, especially when several samples are taken.

In testing laboratories, countless measurements are performed every day using spectrophotometers. Only a few commercially available devices of this type have automatic features that allow for manual measurement and recording of results. In some devices, it is possible to automatically measure and record the results of, for example, one sample up to 100 samples. In these available photoelectric photometers, the light beam is passed vertically through the sides of the cuvette and, for example, the light beam cannot reach the sediment at the bottom of the cuvette. In the device according to the invention, the light beam passes through the column of liquid parallel to the longitudinal axis of the cuvette and into the bottom of the cuvette, or the opposite position, which limits the column of liquid.

If the amount of liquid in the cuvette changes, the distance of the light ray passing through the liquid will change at the same time, and the light transmittance or conductivity of the liquid column will therefore change. When the light beam travels parallel to the longitudinal axis of the cuvette, precipitation, turbidity, etc. as a result of serological, bacteriological or hematological reactions can also be measured at the bottom of the cuvette on the basis of certain changes in the intensity of the light beam. It is. Such precipitation or turbidity cannot be measured at the bottom of the cuvette using conventional photometers. For example, in a hemolytic reaction, hemolysis causes the entire reaction mixture to become transparent, which in turn increases the conductivity of light in the liquid. Also in this case, the extent or end point of hemolysis can be determined from certain changes in the intensity of the light beam. The main feature of the method according to the invention is that in the measuring device the light beam is passed from a light source parallel to the longitudinal axis of the cuvette to a detector on one side of the cuvette and on the other side of the cuvette to the measurement of a cuvette or a row of cuvettes or a group of cuvettes. When the light beam passes through the liquid column to be measured, passing through the bottom of the cuvette which is stationary during the measurement and restricts the liquid column, and thus along the longitudinal axis of the cuvette, The results of hematology, bacteriology, serology or chromogenic reactions may be determined by precipitation, turbidity, etc. at the bottom of a fixed cuvette or cuvette that is not fixed in a liquid column, or by precipitation, turbidity, etc. It can be read on the basis of certain changes in the intensity of the light rays caused by changes in the color of the light.

The main feature of the device according to the invention is that the cuvette element can be placed in a stand of the cuvette element, the spacing between the fixedly or removably mounted cuvette being equal to the spacing of the detectors. At the bottom of each cuvette, which is the same and corresponds to the light source in the measuring instrument, and through which the light from the light source passes,
A measurement opening defined by a sleeve-shaped holding part extending from the periphery in a direction opposite to the sample entrance/exit of the cuvette is provided;
A protrusion for stirring is provided and its lower part is tapered such that its cross-sectional area gradually decreases towards the bottom.

Among commercially available single cuvettes, and in which measurements are taken horizontally through the side walls of the cuvette,
The walls of the cuvette are easily scratched and stained when touched with fingers, causing large errors in measurements. Furthermore, in currently available devices where the beam of light travels horizontally, insufficient attention is paid to the position of the cuvette and often this fact is completely overlooked. In the laboratory, preparing a reaction mixture and measuring this mixture in a photoneedle is a daily task.

From a reaction mixture prepared by pipetting a sample and one or several reagents into a cuvette, the concentration of a certain substance, or the kinetics of, for example, an enzyme, is measured photoelectrically. In such measurements, errors in the final results may occur due to defects in the equipment used or due to defects in operation. Ernest McGlynn and his associates (Cll.
nicalChemistryMba8〕832-837
(1973)) reported that in enzyme kinetic measurements, the final results have an error of approximately 40% in the worst case. This error is caused by inaccuracies in the instrumentation, the length of the beam's path through the cuvette, temperature, sample and test collection, time, and the optical and electronic parts of the photometer. . There is a trend today to use as little sample and reagent as possible in preparing reaction mixtures.

A so-called micro-quantity method has been introduced. This method saves on reagent costs and allows several tests to be performed from one sample, which is often not available. When performing the micro-volume method, since the amount to be sampled is small, the error rate in sampling is therefore very large, which in turn causes the final erroneous result.

Similarly, when photoelectrically measuring reaction mixtures prepared using microvolume methods, the percentage of error in the final measurement results increases. The present invention will be described in detail below with reference to the accompanying drawings.

The titration or other reaction is carried out in cuvettes arranged symmetrically at some distance in a stand.

Such stands are made suitable for measuring instruments. In the embodiment shown in FIG.
l, L2, L3) and detector 3 (D,, D2, D3)
are attached to the support stand 1 with an interval corresponding to the interval between the cuvettes C. For example, in this measuring device, the number of the rows of the cubes, the power supply 2 and the detector 3 have the same number.

In that case, the support 1 is set to move automatically from one cuvette to another in the row of cuvettes, or the stand of the cuvettes can be moved and the support 1 is fixed.

The instrument can also be made to have a light source and a corresponding detector in each cuvette, so that the instrument has no mechanically moving parts. For example, when a beam of light travels from the light source 5 through the cuvette 4 to the longitudinal axis of the cuvette, it first encounters the surface of the liquid and then passes through the column of liquid and the bottom of the cuvette to the detector O corresponding to the cuvette O. Proceed to.

Meter 4 records certain changes in the intensity of the light beam. The measuring instrument thus measures each cuvette in the row of cuvettes. Certain changes in the intensity of the light rays occurring in some of the cuvettes of this row, properties due to each reaction and due to precipitation at the bottom of the cuvette, or turbidity, or conductivity of the light rays of the liquid in the cuvette. indicates, for example, the detection point of a change in the dilution series, and it can be read from the recorded results which cubets in this queuet are suspect. In the output section 5 it is possible to use fixed logic elements or a computer which calculates the results in the desired format, receives the results, stores the results in a memory and stores the program. That's what I do.

For each titration, its own specific change in the intensity of the light beam is determined by testing and this is what signifies the resulting point. In the output section it is possible to use the recording card shown in FIG. 3a. This card has the name 6 of the reaction and a code 7 corresponding to the corresponding change in the intensity of the light beam. Another possibility is to attach a regulator to the output, which is mounted at a position corresponding to a standard of the intensity of the light beam and whose characteristics are varied for each reaction. The output section then records the reaction name and result as shown in Figure 3b. The measuring instrument can also be used as a spectrophotometer, since the measuring instrument records the range of light coming from each light source to its corresponding detector, and the detector records the adsorption and/or transmission of liquid in each cuvette. This is the case. FIG. 2 diagrammatically shows another embodiment of the meter using only a single power supply 8. In FIG.

The light beam is guided through the cuvette by an optical fiber 19, from where it passes further into the optical fiber ◎. From here the light rays pass further into the optical fibers surrounding the circle. The fibers coming into the area surrounding this circle are read by the reading head @, and the optical fiber @ sends the light beam to the detector O. The cuvette is the fundamental part of the device according to the invention. As shown in FIGS. 4 and 5, nine cuvettes in this case are arranged in the support plate O of the cuvette element O at certain intervals. These cuvettes are used for measurements according to the principle of vertical measurement, which depends on the absorption capacity of the liquid in the cuvette [phase] or on the precipitation at the bottom of the cuvette [phase]. This is due to changes in the intensity of the light beam or the turbidity of the liquid. When using this principle of vertical measurement, the light beam travels through the bottom of the cuvette [phase] and the intensity of the light beam is recorded across the cuvette by the detector ◎. All the cuvettes [phases] of the cuvette elements [phases] can be closed at the same time by the lid plate 9, while the cuvette elements @ can also be stored or shaken out. The cuvette [phase] of the cuvette element @ is designed in such a way that the principle of vertical measurement can be used with it. The bottom of each cuvette [phase] of the cuvette element @ is provided with a sleeve-like protection part 0 extending from its periphery in the direction opposite to the sample inlet/outlet of the cuvette to prevent the bottom from getting dirty or scratched. The measuring opening of the cuvette [phase] of the cuvette element [phase] is therefore optically first-class.

If the inside of the cuvette is concave, the sediment inside the cuvette will accumulate as a regular "bud" in the center of the bottom of the cuvette. When the inside of the cuvette is concave and the outside is flat, the path of the rays is simply influenced by the plano-concave lens. In this case it is easy to produce cuvettes with similar optical properties. When the device is used as a spectrophotometer, both the inside and outside of the bottom of the cuvette may be flat, so that the path of the light beam is not influenced by the lens. When the bottom of each cuvette [phase] of the cuvette element @ is flat, the bottom of the cuvette can be easily manufactured and no optical errors occur, and the path of the light rays is also controlled by the lens. It won't be affected.

However, the bottom and/or lid of each cuvette of the cuvette element may, of course, be planar, convex or concave, depending on the purpose, on the inner or outer surface to obtain a suitable lens effect. It is towards.
Preferably, the lower part of each cuvette [phase] of the cuvette element O is conical, so that the cuvettes of the element can easily fit into position (see Figure 10), and the liquid in the upper part of the cuvette The particles that settle in the layer settle as appropriate buds at the bottom of the cone. In this way, it is easy to measure deposits or precipitates in small areas. The vertical inner wall of the cuvette [phase] has one or several protrusions O parallel to the longitudinal axis of the cuvette, which improve mixing when the liquid in the cuvette [phase] is shaken. It is something.

When a conical cube is shaken around in an eccentric mixer, the liquid column undergoes a rotational movement which causes the liquid column to rise up along the side of the cube. In such methods, the different liquid layers do not mix well. However, if the above-mentioned protrusions [phases] are attached to the side surface, these protrusions O will cause agitation in the rotating liquid column, and as a result, different liquid layers will be separated even by a slight force. It mixes effectively. The fact that the cube element @ can be placed in only one position in the cube element stand @ is also significant.

The stand of the cuvette element has a projection which fits into a groove or opening in the support plate of the cuvette element. The supporting plate O of the cuvette element [phase] is also marked with the code @, from which the cuvette in question can be identified, either by means of a measuring device or by sight, and at the same time the cuvette element [phase] Each sample in the cuvette of [phase] is identified. This has the advantage that only one code O is used for the identification of nine separate samples, and coding of each sample is avoided. When the element [with] is placed in only one position of the stand @ of the cuvette element and the measurement of the cuvette [phase] of the cuvette element [phase] is carried out in the measuring device, the first cuvette in the measuring device is The bet element [phase] can be used to zero the nine channels of the measuring device, and in subsequent cuvettes the corresponding cuvettes are measured by the same channel.

This is advantageous if small optical defects occur in the measuring aperture 9 of the cuvette when manufacturing the cuvette element O, and even if these defects are repeated in each cuvette, these defects will be eliminated. is removed during the zero point adjustment stage and subsequent stages.
The stand O of the cuvette element is such that one or several cuvettes of the element can be placed therein and pushed into the measuring head of the measuring instrument (see FIG. 4).

The stand for the cuvette element can be used as a stand for the cuvette element when storing or transporting the cuvette, for taking out and adding liquids from the cuvette element, and for culture reactions, measurement reactions, etc. can be used.

The stand of the cuvette element is also thermally adjustable so that the cuvette [phase] of the element [phase] and the liquid therein can be brought to the desired temperature.

When the cuvette element O is in the measurement position of the stand @ of the cuvette element in the measuring end of the measuring instrument, each cuvette of the element is protected from the outside beam and in addition the stand of the cuvette element is It protects Kyuvetsu from the rays of light. When the cuvette element O in the stand @ comes to the measuring point at the measuring end of the measuring instrument, the light flux passes through the opening [phase] in the stand @ of the cuvette element, and this luminous flux has three [phases]. ] between the two points, give an impulse to the fixed position [phase] by means of a detector @r or a light beam diode.

This positioning section [phase] positions each cube of the element [phase] simultaneously to the optical fiber and the detector @ corresponding to the cube [phase].

When this positioning part [phase] rises to the upper position, the lower part of the cone of each cuvette of element O is located in the conical part of the positioning device, and so on. In other words, the cuvette element [phase] is raised at the end of the measurement, so that the upper part of the cuvette element @ stops against the frame of the detection head at the end of the measurement. . Measurements are taken at this location. In this way the cube of elements [phases] will occupy very precise positions. The instrument for positioning the measuring end when some time has passed [
phase] is lowered and lowers the cuvette element @ back into the fixed position of the cuvette element stand @ so that the cuvette element stand can be pushed into the measuring end, but this is not done mechanically. or manually.

When the next cuvette element @ is in the measuring position, the portion (phase) defining the position of the measuring end positions and fixes the next cuvette element in the measuring position. The reagent block (Figure 11) and the cuvette element [phase] (Figures 12 and 13) are various types of equipment, for example containing nine test tubes or cuvettes, from which or 9-channel, multi-step processing of 9 separate samples at the same time
Can be collected using a multiple pipette.
Reagents can be stored as dry substances or solutions in reagent plotters or cuvette elements. Solutions of the reagents are obtained from the dry materials by adding the appropriate liquid to the reagent block or cuvette. It is also possible to store certain ready-made identical or different reagents in reagent blocks or cuvette elements. ．． ―′! ″．′(′T′
? , Rei: Gate? At the time of change, it is simultaneously closed by the reagent block [phase] or the @ lid.

This lid 9 prevents the liquid in the test tube ( ) or cuvette O from evaporating and becoming contaminated. The lid 9 of the cuvette element is also the cuvette element O.
It can also be used to block each cuvette O of the '0S'! ．． It enters below the open surface of the liquid at 2° C. 1+, corresponding to the stopper ←), at which point the light ray passes further from the liquid to the detector 9.

The lid 9 of the cuvette element [phase] is plugged with a single plate (◆
extending downwards from the hole in the plate, and the bottoms of these plugs are closed by a transparent bottom plate O. The cap φ) of the lid of the cuvette element [phase] is shaped such that the lid 9 is placed on top of the cuvette element [phase], and the vented air part is inserted into the plug C of the lid 9. It is left between the vertical or inclined outer wall and the inner wall of the upper part of the corresponding cuvette () of the element [phase]. When the absorption of a liquid in a cuvette of an element [phase] is measured by a method using a photometer according to the principle of vertical measurement according to the invention, the light beam is transmitted through the liquid in the cuvette at the bottom and the lid of the cuvette. The bottom of the stopper 9←) (◆
They travel through the distance between each other.

This distance H. 9 can be adjusted by manufacturing the lid 9 of the cuvette element of different design, so that the length of the stopper O) of the lid 9 can vary or can be adjusted to different heights. This varies by producing a cuvette element [phase] with a Furthermore, the plug φ) of the lid 9 of the cuvette element can also be made opaque, so that the light rays can only pass through the transparent bottom of the lid 9 of the cuvette element. According to the design according to the invention:
Errors in the height of the liquid column are caused by sampling or scattering of the liquid as water droplets [phase] on the inner wall of the cuvette [phase]; errors do not occur in the length of the path of the light beam. It is.

If the surface of the liquid column in the cuvette of the element is curved or sloped, this can cause measurement errors in photometers operating on the principle of vertical measurement.

When the cuvette elements [phases] are closed with the lid 9 of the cuvette element, the bottom [phase] of each cuvette [phase] and the bottom of the stopper () of the lid 9 of the corresponding cuvette element [phase] O are parallel.

Since the light beam then travels perpendicularly through parallel planes and through the liquid column between these planes, the above-mentioned errors are removed from the measurement. Sometimes bubbles O can form on the surface of the liquid released upon shaking or in conjunction with pipetting. Such bubbles become a stumbling block when measuring with a photometer according to the principle of vertical measurement, in which the light beam is directed toward the cuvette O of the element [phase].
through the bottom [phase] of the liquid, then through the liquid column, and finally through the open surface of the liquid and through the air to the detector. When the elements [phases] of the cuvette are closed with the lid 9 of the element, the respective stopper (bottom 0 of the lid) of the lid 9 is exposed to the open surface 9 of the liquid.
It is located under the cuvette O, which corresponds to the plug (O) of the lid 9, which is located somewhat below the cap.

Any bubbles that may form on the liquid surface will then migrate to the open liquid surface between the inner wall of the cuvette ( ) of the element [phase] and the outer wall of the stopper ( ) of the lid 9 of the element [phase]. be. The above defects can also be corrected, but if necessary, instead of the lid of the cuvette element [phase], some below the open surface 9 of the liquid in each of the cuvettes of the element [phase] This is done by dipping the protected point of the detector.

According to Beer's law, liquid adsorption is directly and linearly related to the path of the light beam through the cube.

If the photometer is operated according to this vertical measurement principle, the measurement is carried out inside the cuvette element without using the lid of the element.
are the same as the heights of the liquid columns in ``♂'', and these heights, in turn, have a direct and linear relationship to the amount of liquid collected in the cube.

Therefore, if a small amount of reagent is added, the path of the light beam in the cuvette becomes shorter, but the absorption per unit of length of the light path increases, and the amount of sample is considered to be accurately measured. It is. On the other hand, if
If too much reagent is added, the absorption per unit of beam path length decreases. In this way, in the operation of the photometer according to the principle of vertical measurement, if the measurement is carried out on the lid 9 of the cuvette element [phase], this will keep the path of the light beam in the cuvette () of the element [phase] constant. However, if measurements are taken without using this, as mentioned above, the added reagent may cause errors, but as mentioned above, it will not cause errors in the absorption of the reaction mixture. Errors in the absorption of the reaction mixture are possible only if there are errors in the addition of the sample. In multichannel photometers according to this vertical measurement principle, the detector 9 is located in the immediate vicinity of, or to the right of, the preamplifier @ at the measuring end of the photometer.

In this way, a long conductor from the detector 9 to the pre-amplifier @ is not required. At the measuring end of the photometer, a device for positioning the cuvette element lifts the cuvette element upwards and sends it to a flat part of the measuring end.

At the end of the lifting movement, this flat surface presses the lid 9 firmly onto the cuvette element. This confirms that the lid 9 of the cuvette element [phase] is in its position and that the length of the path of the rays in each cuvette of the element [phase] is now equal. be. The same or different wavelengths can be used in the optical fibers leading to the measuring end.

In this way it is possible to measure cubes of a single element at the same or different wavelengths. The invention is, of course, not limited to the above implementation.

This can be varied in various ways depending on the scope of the claims. Embodiments of the present invention will be described below.

(1) A device according to the claims, characterized in that the cuvettes of the cuvette element can be closed at the same time by a lid, in which the distance between the plugs corresponds to the spacing between the cuvettes of the cuvette element.

(2) in the claims characterized in that the inside and/or outside of the bottom and/or lid of each cuvette of the cuvette element is flat, concave or convex; Device.

(3) An apparatus according to the claims, characterized in that the inside of the bottom of the cuvette or each cuvette of the group of cuvettes used in this apparatus is concave, and the outside of the bottom is flat.

(4) An apparatus characterized in that both the inside and outside of the bottom of each cuvette of the cuvette element are flat.

(5) An apparatus according to the claims, characterized in that the stand of the cuvette element is thermally adjusted.

(6) In the claims, the lid is formed of a single plate with a hollow stopper extending downward, and has transparent bottom plates parallel to the lid plates, each of which is at the same distance from the lid plate. and a device characterized in that the outer diameter of the stopper in the bottom plate is smaller than the inner diameter of the cuvette of the cuvette element corresponding to the lid.

(7) The device according to item 5 above, characterized in that the stopper of the lid is cylindrical.

(8) The device according to item 7 above, characterized in that the cap of the lid has a truncated conical shape that gradually becomes thinner toward the bottom.

(9) The device according to item 5 above, characterized in that the inner wall of the lid stopper is entirely or partially opaque.

[Brief explanation of drawings]

1 and 2 schematically represent a measuring device according to the invention. Figures 3a and 3b show two samples of recording cards used in the output section and on which the results of each patient can be recorded by the recording device. FIG. 4 is a side view of the cuvette element mounted in the cuvette element stand and in the measurement position. FIG. 5 is a top view of the cuvette element mounted on the cuvette element stand. FIG. 6 is a side view of the cuvette element. FIG. 7 is a side view of the lid plate of the cuvette element. FIG. 8 is a top view of the cube element. FIG. 9 is a cross-sectional side view of one cuvette. FIG. 10 shows the cube element and its positioning for measurement. 11th
The figure shows a cross-section of the reagent block in side view. 1st
Figure 2 shows a side view of the cuvette element and the sampling points of the multistage pipette used with the cuvette element. FIG. 13 shows the stage where the cuvette element and the dots on the multi-stage pipette shown in FIG. 12 have reached the point where the liquid container of the multi-stage pipette has reached its rim.
FIG. 14 is a top view of the reagent block, and FIG. 15 is a cross-sectional view of the cuvette element placed in the photometer for measurement and covered with a lid according to the invention. 2, 8... light source, 3, 14...
...detector, 10... cuvette, 12...
...Measurement opening, 16, 18...Cube element, 22...Protection section, 24...
...Protrusion.

Claims (1)

[Claims] 1 A light source arranged on one side of the cuvette and irradiating the interior of the cuvette with a ray of light parallel to its longitudinal axis; In the measuring device for a cuvette, the bottom of each cuvette through which the light from the light source passes is provided with a sleeve-like protective portion extending from the periphery in a direction opposite to the sample entrance/exit of the cuvette. Each cuvette is provided with a defined measuring opening, and the inner peripheral surface of each cuvette is provided with a protrusion for stirring, the lower part of which has a cross-sectional area that gradually decreases towards said bottom. A measuring device for a cube, characterized by being tapered in such a manner.