JPS60173468A - Biological fluid examination system and method - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the measurement of reactive properties of fluid specimens, and more particularly to methods and automated apparatus for the qualitative determination of immunological responses of biological specimens. It is something.

Microscopic slide techniques for manual blood typing tests are not sensitive to subtle reactions and require close attention to observation recording to avoid possible sample misidentification. An alternative to this microscope slide technique is the test tube centrifuge technique.

Additionally, test tube centrifuge techniques have been used to enhance or corroborate the procedures of microscope slide techniques. The test tube centrifuge technique is characterized by being more sensitive to subtle reactions than the microscope slide technique. However, test tube centrifuge techniques are more labor intensive, require judgment by laboratory technicians, and have inherent problems in ensuring accurate sample identification.

The "microwell" test method was developed to obtain maximum utilization of the materials involved in blood typing and to standardize the interpretation and evaluation of biological specimens to provide consistent results throughout. This "microwell" test method uses a plate containing multiple small centrifuge tubes or wells.

Automated reading techniques for blood typing have typically been limited to turbidity measurements. The turbidity results from the destruction of centrifuged red blood cells. The method by which dissociation of red blood cells is initiated is to mix the blood cells with a reagent and vibrate the mixture. Dissociation of the red blood cells is an indication of a negative reaction between the red blood cells and the added reagent. If a positive reaction occurs after shaking, the red blood cells remain tightly associated. The attraction of red blood cells to aggregate when exposed to the reagent is an indicator of a positive reaction. Typically, a positive reaction between the red blood cells and the reagent results in a tightly packed mass of red blood cells and a clear supernatant.

Turbidity can be measured based on the opacity of the specimen.

Transmit a beam of light or energy into the specimen.

A detector measures the decrease in the intensity of the light or energy beam caused by scattering or absorption of the beam by the analyte.

The turbidity of the specimen is measured as a function of the decrease in transmitted light or energy. More specifically, the reduction in the intensity of the transmitted beam occurs due to loss of light or energy due to scattering and absorption of the incident beam by floating transparent blood cells. The effective number of floating red blood cells is directly proportional to the amount of red blood cell dissociation that occurs due to negative reactions.

Despite important advances in the prior art, they reduce subjective judgment of reactions by laboratory personnel, are easy to use, and
There remains a need for automated blood typing systems and methods that require little technical expertise, require fewer personnel, and are relatively inexpensive.

Recognizing the need for improved systems and methods for automatically determining blood type, it is therefore a general attribute of the present invention to provide a novel automated blood typing system and method.

One feature of the invention is to provide a novel automated blood typing method and system that positively identifies specimens from each patient.Another feature of the invention is to provide a novel automated blood typing method and system that unambiguously identifies specimens from each patient. It is an object of the present invention to provide a new device that performs chamber procedures in an automatic and fully reproducible sequence that guarantees consistent results from beginning to end.

Yet another feature of the present invention is to provide a novel automated blood typing method that can be performed in rapid succession with extremely high accuracy.

Yet another feature of the present invention is a novel automated blood type system that accurately determines biological parameters of blood samples based on accurate assessment of the reaction, or lack thereof, between the biological sample and reagents. The objective is to provide a judgment method.

Additional features and advantages of the invention will be set forth in part in the discussion that follows, and in part will be obvious from the description, or may be learned by practice of the invention. The features and advantages of the invention will be realized and obtained by means of the instrumentalities, combinations and steps pointed out individually in the claims.

According to one embodiment of the invention, a method for determining a reactivity characteristic of a biological fluid specimen begins with preparing a sample comprising the specimen and a suitable reagent such that the sample is at least partially opaque. . The sample is centrifuged and a first measurement of the linear dimension of the opaque portion of the sample is taken. Subsequent measurements of the same linear dimension of the sample are then taken. The first measured size is then compared with the second measured size.

According to another embodiment of the invention, an apparatus for measuring reactive characteristics of a biological fluid specimen includes a centrifuge rotor and an energy-transferring flexible belt, the latter moving in unison with the rotor. It can be formed into a cylindrical shape and can be removably mounted. The belt includes elements for receiving multiple specimens. A sensing system illuminates a plurality of analytes contained within the belt and optically measures the reactive properties of the analytes.

To further summarize, blood typing according to the present invention is performed by mixing a biological specimen with a reagent for distinguishing between positive and negative immunological reactions. Accordingly, the present invention generally relates to a method and apparatus for differentiating between positive and negative immunological reactions indicated by the affinity of particles associated with a biological specimen and the combined reagents of interest. It is something. The invention also has applications in other immunoassays using opaque particles other than human red blood cells as carriers for immunological agents (antigens), such as in the determination of hepatitis, rheumatoid arthritis, and infectious mononucleosis. It also has Suitable opaque particles include barium sulfate, latex,
Included are animal red blood cells or particles to which an antigen of interest can be attached and observed.

The drawings, which are incorporated in and constitute a part of this specification, illustrate particularly suitable embodiments of the invention, and are similar to the general description of the invention given above and the details of particularly suitable embodiments given below. The detailed description serves to provide a detailed explanation of the invention.

1 is a perspective view of one embodiment of an automated blood type determination system according to the present invention; FIG. 2 is a cross-sectional view taken generally along section 12-2 in FIG. 1; 3 is a cross-sectional view taken along section line 6-6 in FIG. 2, and FIG. 4 is an enlarged view generally taken from above along section line 4-4 in FIG. , FIG. 5 is a cross-sectional view of the present invention assembly taken generally along section line 5--5 of FIG. 4, and FIG. 6 is an enlarged front view of the segment of the disposable belt shown in FIG. Figure 7 is a vertical view taken along line 7-7 of Figure 6, and Figure 8 is a vertical view taken generally along line 8-8 of Figure 6. 9 is a cross-sectional view taken along line 9 of FIG. 6 at various stages during use of the invention; FIG.
10 is a vertical cross-sectional view taken generally along the line 10-10 of FIG. Figure 11 is a partial enlarged view taken generally along the 9th
12 is a cross-sectional view taken generally along curved arc 11-11 in FIG.
16 is an enlarged cross-sectional view taken generally along section line 13-1'2 in FIG. FIG. 1A is a partial cross-sectional view of a reagent reservoir according to the present invention taken along dividing line 14-14 in FIG. 16 is a schematic diagram illustrating a microprocessor system; FIG. 16 is a schematic diagram of one embodiment of a first input/output board and associated peripherals implemented in one embodiment of the invention; FIG. 18 is a partial schematic diagram (following FIG. 18) of one embodiment of the power driver electrical circuitry and related peripherals implemented in one embodiment of the present invention; FIG. A partial schematic diagram (continued from FIG. 17) of an embodiment of the mechanism of the second motor driver electric circuit and related peripheral equipment, and FIG. 19 is a process flow diagram of an embodiment of the present invention. and FIG. 20 is a flow diagram of one embodiment of the steps for preparing a sample for practicing the present invention.

The general description above and the detailed description below are merely illustrative of the comprehensive invention, and further forms, advantages, and details of the invention depart from the spirit and scope of the invention. It will be readily suggested to those skilled in the art.

Referring to the drawings (similar reference letters are used for similar parts throughout the several drawings), the first
The biological fluid assay device 10 shown in FIG. Dispensing device 200 is included. Biological fluid assay device jif-1
0 also includes a housing 11 having a top structure 12 with a planar working surface 20 on which various control devices are located.
and a bottom structure 14. The biological fluid assay device 10 is electrically actuated by a switch 602 on the front vertical side 22 of the top structure 12. The operator utilizes the keyboard 604 to enter the appropriate test information and obtain the desired test results. The analysis value obtained as a result is printed on the printer 606 or displayed on the display 60.
8 will be shown.

The removable carousel of the giperator-dilutor apparatus 100 receives test tubes 134 in an upright configuration.

134 received in an opening 124 corresponding to the cross section of v 134 along the periphery of carousel 114.
The sample is centrifuged in a conventional centrifuge device and the analytes of interest contained therein are separated by density. An empty dilution cup 134a corresponding to the container tube 134 is received within the inner ring of the opening 122, which corresponds to the cross section of the cup 7° 134a. The dilution cup 134a is used to contain a liquid for diluting the sample taken out from the sample tube 134. Cup 134a may be a conventional clone cup with a v-shaped bottom 122.

As shown in FIG.
Contains 18. Cup 134a is supported on top plate 120 by these edges 7° 124. The test tube 134 is supported on the top of the bottom plate 132 by the center plate 130 and the top plate 132.

Carousel 114 carries biological fluid assay device 121
0 is removably held in a trough 128 below a flat work surface 20. Circular conveyor 114
The central column extends slightly above the flat work surface 20.

At its lower end, the strut 118 fits onto an axle 119 having a radially outwardly pointing barb 121 that engages a hole 123 in the inner surface of the strut 118 .

Test tube 134 is adapted to hold a smaller volume, but is of sufficient height to allow for clarity of the layers within the specimen through the use of removable insert 136. Each insert 136 includes one enlarged diameter funnel portion 138 that frictionally couples with the inner wall of the test tube 134 .

A narrow groove 140 extends downwardly from the conical bottom 139 of this section 138 through the test tube 134.

In this manner, a typical test tube 134 having a capacity of e.g. It is being

A coded bar label 144 fits on the outside facing side of the tube 134. Therefore, optical barcode reader 24 unambiguously identifies any specimens by their labels 114.

The carousel 114 is supported by a support 118 by a mandrel 119.
The reading device 24 reads the continuous tube 114 by rotation of the
The rotary motion is automatically directed to align or orient tube 134 to the desired position.

The orientation of the centrifuge rotor 300, pivot device 110, and reagent dispenser 200 increases the efficiency and availability of the biological fluid assay device 10, as shown in Figure 2. Set it to the maximum. The centrifuge rotor 300 is attached to one of the support plates 350 and is supported by struts 352.

Pivot cover 16 encloses centrifuge rotor 300 inside biological fluid assay device 10 . This cover 1
If you raise 6, you can get the centrifuge rotor 300.

The pipettor-dilution type device 100 is in fluid communication with the syringe 1
02 and conduit needle 106. The needle 106 moves automatically from place to place by the rotation of the shape and the arm 108 that moves back and forth. Arm 108 consists of vertical struts 112 and horizontal struts 110.

To prevent contamination caused by using the conduit needle 106 and syringe 102 with different fluids, a needle cleaning station is used to clean the conduit needle 106 between contact with different fluids.
and wash the syringe 102. The washing place 116 is
As shown in FIG. 1, the circular arc "A" described by needle 106 as it rotates with arm 108 to and from approximately aligned tube 134 and cup 134a.
” located along.

A reagent reservoir 202 is located on top of the flat work surface 20;
Secured by reservoir cover 204.

The reagent reservoir 202 is connected to the reagent pump 2 by an outlet tube 208.
Connected to 06. The reagent bottle 7° 206 is enclosed in an upwardly projecting section of the top structure 12 above the flat work surface 20. Reagent bottle 7° 206 is tube 212
supplies reagent to outlet 210 by.

Electrical components associated with centrifuge rotor 300 are located below support plate 350 as shown in FIGS. 2 and 6. Variable speed motor 304, stepped motor 308
and a solenoid 306 are located below the support plate 350. A variable speed motor 304 is connected to centrifuge rotor 300 by a series of shafts and pulleys. Stepped motor 3
08 is engaged and disengaged by the solenoid 306, as shown in FIG. The solenoid 306 pulls the moving core 322 into an internal coil (not shown) as current flows through the current. When the moving core 322 is drawn into the solenoid 306 by the spring 304, the lever 326 moves to the axial tip point. As the par 326 rotates about the axle point 328, the clutch 330 forces the pulleys 312 and 332 into engagement. The pulley 312 is driven by the variable speed motor 304 using the belt 314.

Pulley 332 is driven by belt 334, the latter operatively cooperating with stepped motor 308.

Stepped motor 308 is connected to pulley 332 by shaft 338 and pulley 336. The main shaft 310 is fixed to the centrifuge rotor 300 by a mounting plate 316.

As shown in FIG. 4, variable speed motor 304 drives pulley 356, belt 314, pulley 312, shaft 310, mounting plate 316, and finally centrifuge rotor 300. Stepped motor 308 drives stepped motor pulley 336, belt 334, pulley 332, shaft 310, mounting plate 316, and finally centrifuge rotor 300.

As shown in FIG. 6, the circulation belt 400 goes around the centrifuge rotor 300. - the circulation belt 400 is held to the stereotaxic lid by belt supports 302 located between the cells 402 of the circulation belt 400;
0 may be an elongated piece folded onto itself so as to assume a cylindrical shape. In general, single-use belt 400 can be made from any material that is translucent, transparent, or transparent to light. Belt 400 is made of a transparent flexible plastic, such as PVC, styrene, cellulose acetate, cellulose butyrate, or a relatively inert, light-transparent material that is compatible with biological reagents used in testing biological specimens. It can be made from any wettable plastic material.

Furthermore, belt 400 is preferably constructed from a suitable translucent material that scatters light and provides more uniform illumination of the sample to enhance optical resolution.

As shown in FIGS. 6, 7 and 8, the belt 4
Each cell 402 of 00 is formed from a flat strip 406 secured to a dimpled strip 408 shaped by vacuum forming or the like. Flat piece 406 and recessed piece 408 form inlet 404 and chamber 418 .

One embodiment of the invention uses a single use belt 400 having, for example, 84 cells 402, but only 7 cells for each series of reactions required to determine the blood type of one patient.
cells 402 are available.

In this way, samples from 12 patients were divided into 40 cells of 84 cells.
Analysis can be performed by using a single-use belt with 2.

Vertically aligned convex chambers 418 extend smoothly radially outward. Each cell 402 is shaped in the shape of an angled inverted test tube section extending upwardly and outwardly from a vertical plane defined by elongate strip 406 . Each cell 402 is generally semi-elliptical when viewed from above, as shown in FIG. Typically, the dimpled strip 408 is heat sealed to form the flat strip 40.
Make it 6. As shown in FIG. 8, a vertical apex 410 is located at the bottom of the chamber 418.

Moderately sloped segment 414 forms radial apex 412
The depth increases towards Segment 414 is connected to piece 406
may be oriented at an angle of about 60° with respect to A spherical radial apex 412 having the shape of the bottom of a normal test tube is connected to the cell 402.
It is the part that extends radially outward. After the radial apex 412, the contour of the cell 402 is segment 41
6 makes a sharp turn back towards piece 406. Accordingly, each cell 402 extends vertically upward along a uniformly shallow, generally vertical segment 420. This uniformly shallow, generally vertical segment 420 is a flat piece 406
Together, they form an inlet 404.

This smooth characteristic of cell 402 significantly enhances the utility of the method of the present invention. The smoothly curved surface associated with the dimpled strip 408 provides little physical resistance to the movement of fluid thereon, and the biological fluid assay system 10
The sensitivity of In this way, surface tension is minimized, flow properties are enhanced, and a better qualitative measurement of the properties of the immunological reaction is possible.

Centrifuge rotor 300 is comprised of a top plate 340, an annular plate 344, and a cylindrical member 342, as shown in FIG. The top plate 340 is fixed at right angles to the central axis of the cylindrical member '342.

Top plate 340 and cylindrical arrangement 342 are secured to annular plate 344 . Annular plate 344 and cylindrical member 342 have a number of aligned apertures 354 . These holes 354 are approximately two widths of cells 402 of belt 400+. Typically, as illustrated in FIG. 4, two of the cells 402 are directly aligned in front of each aperture 354 when the circulation belt 400 is engaged with the centrifuge rotor 300. - The orientation of the circulation belt 400 is maintained by the belt support 402. Belt support 402 carries single use belt 40 on centrifuge rotor 300 adjacent to one or more cells.
0 is positionally certain. In this manner, as shown in FIG. 6, as the centrifuge rotor 300 rotates, each square cell 402 on the single belt 400 passes through the analytical optical system 500. Centrifuge rotor 300 is stably guided by rotor guide 346 and coupling slotted member 348.

Conduit needle 106 and syringe 102 transfer the sample to cell 40.
2 (see Figures 6 to 8) or dilution cup 7''
After sample introduction, conduit needle 106 and syringe 102 are used to place diluent into dilution cup 134a.

The specimen/diluent solution is transferred to conduit needle 106 and syringe 10.
Cell 4 is drawn out from the dilution cup 7°134a by 2.
16' in 02. The conduit needle 106 is connected to the cover 16
By passing through the opening 18 in the single-use belt 4
00 cell 402 can be approached. The diluent is kept in a diluent reservoir 104.

Syringe 102 draws diluent from reservoir 104 and transfers the diluent via conduit needle 106 to dilution cup '134a.

The appropriate angular acceleration of each cell 402 by centrifuge rotor 300 is important. The centrifugal force shapes the red blood cells into small, round, tightly packed clumps 92, as shown in Figures 9B-9E. With sufficient centrifugal force, this mass 92 will form at the radial apex 412 of the cell 402. In the illustrated embodiment, the centrifuge rotor 3
00 is accelerated to a speed sufficient to exert a centrifugal force of approximately 600 G (which corresponds to approximately 2100 revolutions per minute) on the red blood cells in the specimen. centrifuge rotor 3
00 maintains this speed for about 35 seconds.

When the dense mass 92 is sufficiently concentrated and densified by centrifugal force, the motor 304 is automatically de-energized. Without the help of motor 304, centrifuge rotor 300 slows down.

When the centrifuge rotor 300 reaches a certain speed, that is, the rotation of the centrifuge rotor 300 causes gravity to automatically equal the centrifugal force along the inclined segment 414, the stepped motor 308 uses the belt 334. This engages the shaft 310 of the centrifuge rotor 300 . The stepped motor 308 provides a force to drive the centrifuge rotor at a speed that approximately balances the gravitational and centrifugal components of the force along the inclined segment 414 of the cell 402.

Typically, the stepped motor 308 is a centrifuge rotor 30
0 at approximately 60 revolutions per minute or somewhat less than 1 G.

Optical system 500 includes a light source 502 associated with fixed member 506 and extending into member 5060 and optical chamber 504, as shown in FIG. A light beam 509 emitted by light source 502 impinges on lens 508 . 508 in Ren
focuses a beam of light 510 through an aperture 354 in centrifuge rotor 300 and through cell 402 . Furthermore, the lens 508 emits light #i! 510 is focused toward reflector 512. Ray 510 hits reflector 512;
It then assumes its direction again as ray 524.

The light beam 524 passes through the slit 514 and enters the lens box 5.
Proceed to 16. Once inside lens box 516, light ray 524 passes through lens 518. Lens 518 focuses light rays 526 through a transparent surface 520 onto a linear optical detector 522. A linear optical detector 522 is enclosed within a chamber 528 to prevent extraneous readings by external light sources. Since light source 502 is oriented perpendicular to the vertical plane, only the vertical dimensions of the analyte within each cell 402 are imaged.

The reagent dispensing device 200 shown in FIGS. 2, 10, and 11 includes a stopper 224 and a reservoir 202 coupled to a reservoir catch 204.

Air tube 222 and outlet tube 208 protrude from stopper 224 to reagent pump 206. The reagent is in the reservoir 20
2 to reagent l? It is pumped to pump 06 and then goes to outlet 210 via tube 212Z. Outlet 210 is attached to outlet support 214. Exit 210 is Exit 2
The reagents are simultaneously released into the appropriate number of cells 402 directly below 10. The arcuate edges 222 of the outlet support 214 have the same radius of curvature as the centrifuge rotor 300 and extend below the cover 16. Outlet support 21
4 can be rotated at an apex point 218 by arm 216. When rotating at point 218, output D 21
B is located above the flash reservoir 220. Reagent dispensing device 200 is cleaned by flushing solution through device 200 and into flush reservoir 220 .

Microprocessor - or Central Processing Unit (CP)
U) 600, as shown in FIG.
10 controls the R10 board 610, the second R10 board 12, and the third R10 board 646. - as shown in Figure 16
The engineering 10 port is connected to the cPU 600 via the STD bus, and includes the keyboard 604, display 608, and def'-6.
09 and printer 606, and output from these. In the present invention, a printer interface board 654 is used to transfer data between the -I10 board 610 and the printer 606.

The second r10 port 612, as shown in FIG.
Controls power driver board 614 and motor driver board 628. power driver board 6
14 controls a pump assembly 618 that includes an air pump 15, a drive bond 615, an air valve 617, and an air pressure sensor 619. Board 614 also includes gear motor components 620 (including gear motor 621 and gear motor sensor 623), Selenoid 3
06, cover 16 interlock sensor 624, and reagent valve assembly 616 (including valve motor 625,627). Cover movement sensor 624 ensures that cover 16 is closed and - if cover 16 is open, provides appropriate instructions to keyboard 604 (FIG. 16). The air bonder 613 is connected to the air pipe 222 and the drive pump 61 via an air valve 617 and a pressure sensor 619.
5 and controls the dispensing of fluid from the dispensing device 200. Reagent valve assembly 618 is valve motor 625.62
Contains two rotary multivert valves driven by 7.

Valve position sensor 635 allows selective dispensing from reservoir 202. Gear motor 621 serves to direct the carousel from one angular position to the next through the engagement of struts 118 by mandrel 119.

As shown in FIG. 18, the motor driver board 6
28 is a clutch controller 632 and a clutch sensor 633
, power supply 636 for stepped motor 308, light source 5
02, a control device 640 for the linear optical detector 522,
Heat sink 644, control device 6 for stepped motor 30B
42 and servo motor 630.

Servo motor 630 controls the position adjustment of arm 108. The clutch sensor 633 is the current clutch 330
An optical sensor that detects the position of the sensor may be used. The third construction 10 board 646 shown in FIG.
Control 3. Isolation devices 647 are used to optically isolate the various controls and motors from the digital microcontroller components.

The method of the present invention is schematically described in FIG. 19 and is effectively carried out using the apparatus of the present invention. The analysis sequence is initiated by pressing the "Enter" button on the keyboard 604 (step 700). Disposable belt 400 is manually loaded onto centrifuge rotor 300 and secured by belt support 302 (step 702). Cover 16 is lowered to enclose centrifuge rotor 300 and disposable belt 400 (step 7D4).

Approximately 1°Qrnl of blood sample is obtained from each patient and placed manually into the insert 136 of the sample tube 134. However, 6
~7-Samples can be routinely used in test tubes 134 without inserts 136. Each sample tube 134 has a coded bar label 144 attached thereto. The sample tube 134 is centrifuged to separate the blood sample into red blood cells and supernatant. The sample tube 134 containing the separated blood sample (red blood cells and plasma) is manually placed into the opening 124 around the carousel 1140.

Dilution cup 134a is typically placed in internal opening 122 of carousel 114. Carousel 114 is loaded into tank 128 of apparatus 10 (step 706).

The analysis is initiated by pressing the "Start" button on the keyboard 604 (step 708).

The reagent dispensing device 200 simultaneously dispenses the correct reagent into the disposable belt 40007 cells 402 (step 71).
4). The pipettor-dilutor device W10[1 is activated to prepare the sample for analysis (step 710). The operation of this pipettor-dilutor device 100 (step 710) will be described in detail later. During the time that pipettor-dilutor apparatus 100 and reagent dispensing apparatus 200 are operating, extra carousels 114 may be prepared for the next test (step 712).

Reagents are held in reagent reservoir 202. Reagents typically used include antiserum, anti-A1 anti-B1 anti-A-
B, and anti-I), AI reagent erythrocytes, A2 reagent erythrocytes, B reagent erythrocytes, 0 reagent blush spheres, and controls. In addition to this, Rh phenotypic antisera such as anti-CX c, E and e can also be used. Other direct agglutination antisera such as anti-M1 anti-N1 anti-P and anti- can also be used.

After the reagent dispensing device 200 has dispensed the correct amount of reagent into the cell 402 and the pipetter-dilutor device 100 has been prepared and the sample transferred to the cell 402, the carousel 1
14 can be removed from the device 10 (
Step 716). The centrifuge rotor is then accelerated to a speed sufficient to tightly pack the sample into the radial apex 412 at the tip of the cell 402 (step 720). The centrifuge rotor 300 is slowed down to a speed where the gravitational pull is somewhat greater than the centrifugal force exerted on the specimen, thus allowing flow of opaque particles with negative particles (step 722). A baseline reading of the optical properties of the sample in each cell 402 is automatically taken using optical system 500 before any significant flow can occur (step 724). Optical system 500 determines the vertical dimension of the opaque portion of the sample (■ in Figure 9B). Baseline readings are important to determine the correct indication of possible responses. The rotational speed must be low enough to rotate the rotor 300 relative to the optical system 500 while making the necessary measurements in a continuous, grappling fashion. Therefore, the rotation speed is limited by the capabilities of the optical technology used.

After sufficient time (e.g., 25 seconds) has elapsed for flow to complete in a negative test, the optical system 500 automatically reads the reaction that has occurred (step 760), i.e., the opaque portion of the sample (FIG. 9C). ~Determine the vertical dimension in Figure 9E. After the last reading has been taken, the centrifuge rotor 300 stops (step 762), the cover 16 can be raised (step 764), the printer 606 and the display 608 are activated (step 766), and the rotation belt 400 can be removed by hand.

All of the data taken is processed by the microprocessor-600
be stored for analysis.

The baseline reading (step 724) can be automatically compared to the final reading (step 760) by CPU 600 to determine anomalies or malfunctions associated with a particular sample. For example, a baseline reading indicates that the sample has "slipped out" of the radial apex 412 of the cell 402.
I can give instructions on what to do. Such a shift can be interpreted as a flow of the sample caused by agglomeration rather than the fact that the entire opaque part of the sample has slipped into position which may give a false reading. The procedure for determining the pace line characteristics of the sample prevents false readings in this and similar situations.

When the "slip" response 84 shown in FIG.
? ” and asks you to be careful. The weak reactant 88 shown in FIG. 6 is illustrated next to the negative reactant 86. A weak response can be associated with a particular biological parameter by microprocessor-600 and indicated by display 608 or printer 606.

As shown in Figures 9C, 9D and 9E,
The second vertical measurement v2 is compared to the first vertical measurement v1 shown in FIG. 9B by microprocessor-600 by subtracting v2 from v1. If vl and v2
A negative reaction is indicated if the difference exceeds a preset difference determined by microprocessor-600. If this difference does not exceed a preset value determined by microprocessor-600, a positive response is indicated, as shown at 82 in FIG. (no agglomeration, i.e. flow)
indicates a negative test, and agglutination (no flow) indicates a positive test.

It will be appreciated that the present invention provides a baseline drawing for each sample by taking vertical measurements of the closely packed red blood cells of each individual sample. The use of baseline drawing in the present invention provides a standard to which measurements can be compared. Additionally, baseline drawing as shown in Figures 9A through 9E eliminates the effects on response measurements due to variations in analyte volume or opaque particle concentration.

Furthermore, and perhaps even more seriously, such a baseline draw may result in positive and negative reactions due to the extra vertical downward movement of tightly packed red blood cells, as shown at 84 in Figure 6. The influence of the distinction between the two is also removed. This downward movement is known to occur and, without baseline drawing, indicates a false negative response.

Microprocessor 600 evaluates the response by reading the response of the linear optical detector 5220 individual photosensitive elements (picture elements) in an order based on absorbed energy. The image of densely packed red blood cells is made up of individual photosensitive elements (picture elements).
shadow. Linear optical detector 52 of densely packed red blood cells
2. A picture that becomes a shadow when projected onto the surface, the final element has a low energy output. Pixels exposed directly to light beam 509 and not shadowed by dense red blood cells have a high energy output.

The number of low power dark pixels 530 is directly proportional to the length of the closely packed red blood cells in cell 402. If there is a significantly lower power dark pixel due to flowing red blood cells, this should indicate a negative reaction. The strength of the response will be proportional to the increase in the number of low power dark pixels on the linear optical detector 522.

The magnitude of the resulting image or shadow can be used as a comparison to determine the presence and magnitude of a reaction between the red blood cell sample and the reagent. The sum total of such measurements constitutes the determination of blood type.

FIG. 20 illustrates an automated sequence performed by pipettor-dilutor 100.

First, the barcoded labels 144 on the twelve tubes 134 of the carousel 114 are read by a conventional optical code reader 24 (step 800). Arm 10
8 rotates into alignment over the first tube 134 of the carousel 114 (step 802). arm 108
The needle 106 is lowered into contact with the loose supernatant (plasma) remaining on the red blood cells in the tube 134 (step 804).
. Needle 106 draws approximately 90 microliters of supernatant (step 806). Arm 108 is raised away from carousel 114 (step 808). centrifuge rotor 300
Arm 108 is rotated so that it is aligned with aperture 18 in upper cover 16 (step 810). Arm 10
B is lowered so that it is just above the first cell 4020 of the disposable belt 400 (step 812). Pipetter-dilutor apparatus 100 dispenses approximately 30 microliters of supernatant into cell 40.2 (step 814). Centrifuge rotor 300 is automatically directed by microprocessor 600 to position another cell 402 directly below needle 106 (step 816). Pipettor-dilutor 100 repeats the process of dispensing approximately 60 microliters of supernatant into six cells 402.

Six cells 402 (Fig. 19,
After each device (see step 714) receives about 60 microliters of supernatant, the arms 108 are raised away from the cover 16. “Rotate the arm 108 to convey the circular conveyor 1.
The fourteen first tubes 134 and needle 106 are again aligned (step 820).

The arm 108 is brought into contact with the red blood cells at the lower end of the tube 134 by lowering it through the tube 134 (
step 822). The pipettor-dilution device 100 has a needle 10
Approximately 20 microliters of red blood cells are removed from the lower end of tube 134 by 6 (step 824). arm 108
and stay away from carousel 114 (step 826).
. Arm 108 is rotated to align needle 106 with corresponding dilution cup 134a. This dilution cup 134
The red blood cells drawn from the outer tube 134 are diluted using a (step 828). Dilution cup 13 with arm 108
4a (step 860).

A pipettor-dilutor 100 using a syringe 102 and a needle 106 is used to dilute the red blood cells and diluent at 7°13.
4a (step 862).

The dispensing of red blood cells and diluent into the inner cup 70134a is sufficient to provide proper mixing of the red blood cells and diluent. Typically, for human blood, the pipettor-diluent device 100 dispenses about 465 microliters of diluent at about 20 microliters.
Mix with microliters of red blood cells to create a cell suspension with a concentration of approximately 6.5%. This 385% cell suspension is
This provides optimal reading characteristics for presently particularly suitable embodiments of the invention.

Needle 106 withdraws approximately 120 microliters of 6.5% cell suspension from inner cup 134a (step 864).
). Arm 108 is raised and away from carousel 114 (step 866). The arm 108 rotates and the cover 1
6 opening 18 again (step 8)
38). The needle 106 is lowered into this aperture 18 so that it lines up with the fourth cell 402 already containing the appropriate reagent (step 840). A pipettor-dilutor 100 pipets approximately 30 microliters of 3.5% cell suspension into a needle 10.
6 to the cells 402 (step 842). Centrifuge rotor 300 automatically directs to the next appropriate cell (step 844). A total of 4 cells containing the appropriate reagents are 6
．． Continue this dispensing-instruction sequence until you have received 30 microliters of 5% cell suspension. Arm 108 is then raised away from cover 16 (step 846). Centrifuge rotor 300 is directed to the next appropriate cell 402 (step 8
48). The carousel 114 transports the next tube 134 to the needle 10.
Directed to line up with the 6th street (step 852)
The cycle then begins again (step 802) and is repeated for all 12 samples in each tube 134.

The needle 106 is cleaned between patients at a wash station 116 by flushing with the cell suspension contained in the diluent reservoir 104 . The red blood cell suspension solution consists of anhydrous glucose, sodium chloride, sodium citrate, citric acid, deionized water, chloramphenicol, neomycin sulfate, and imocin.

When fluid assay system 10 is not in use overnight, reagent reservoir 202 can be removed and replaced with a bottle containing sterile water. When the reagent dispensing device 200 is not in use, the device flushes the lines with water and the reagent dispensing device 200
Leave water inside. Similarly, the reagent dispensing device 200 can be periodically flushed with white solution and water to clean the internal components of the reagent dispensing device 200.

The number of cells 402 for receiving -h reagents, the number of reagents used, and the number of specimens handled by system 10 can be preprogrammed by the operator. Additionally, the volume of specimen withdrawn from the carousel varies.

The method of the present invention can be used with any analyte that contains an opaque substance that physically reacts with a specific reagent and can be separated by density. Preferably, the reaction renders the opaque portion of the specimen more or less fluid depending on the characteristics of the specimen.Further advantages and modifications of the present invention will be readily apparent to those skilled in the art. It will float. Therefore, the invention in its broader aspects is not limited to the specific details, representative apparatus, and representative examples shown and described herein. Therefore, without departing from the spirit or scope of the general concept of the invention as described herein, it may be understood that
maybe it can be developed.

[Brief explanation of drawings]

FIG. 1 is a perspective view of a specific example of the automated blood tL type determination system according to the present invention, and FIG. 2 is a cross-sectional view generally taken along the dividing line 2-2 in FIG. , Figure 6 is a cross-sectional view taken along the dividing line 6-6 in Figure 2, and Figure 4 is a top view generally taken along the dividing line 4-4 in Figure 6. 5 is a cross-sectional view of the apparatus of the present invention taken generally along section line 5--5 of FIG. 4; and FIG. 6 is an enlarged view of a segment of the disposable belt shown in FIG. 7 is a top view taken along line 7-7 of FIG. 6, and FIG. 8 is a view generally taken along line 8-8 of FIG. 6. 9 is a vertical cross-sectional view taken along line 9 of FIG. 6 at various stages during use of the invention; FIG.
10 is a vertical cross-sectional view taken generally along the line 10-10 of FIG. FIG. 11 is a cross-sectional view taken generally along curved arc 11-11 of FIG. 9; FIG. is a cross-sectional view generally taken along the dividing line 12-12 in FIG. 11, and FIG. 16 is a cross-sectional view taken generally along the dividing line 16-16 in FIG.
14 is an enlarged cross-sectional cutaway view of a reagent reservoir of a reagent dispensing device taken along section line 14-14 in FIG. 16; FIG. FIG. 15 is a schematic diagram of a microprocessor system according to the present invention, and FIG. 16 is a partial cross-sectional view of the present invention. 17 is a schematic diagram of one embodiment of a first power/output board and associated peripherals once implemented in one embodiment; FIG. 18 is a partial schematic diagram of one embodiment of a peripheral device, and FIG. FIG. 19 is a flowchart of the steps of one specific example of the method of the present invention, and FIG. 20 is a flowchart of the steps of one example of sample preparation for carrying out the present invention. Attorney: Manmura Kaoru Procedural Amendment (Voluntary) December 3, 1980 Mr. Commissioner of the Japan Patent Office 1, Indication of the case: 59 years 1:' [Application No. 227612 = 2,
Title of the invention: Biological fluid assay system and method 3; Relationship with the person making the amendment +11'l;'t,'r,'〒Applicant4;
Agent Showa Year Month Date Procedural Amendment (Method) % Formula % 1. Indication of the case Showa t2 year application No. 1-1-7 B/J No. 2, Name of the invention FfS Rigon Ino etc.) 1 H, I ≧ (Mirei)): 1 sinu 2 chi b ah; 1-1-noko ")j' hiro), relationship with the case of the person making the amendment Patent applicant 5, amendment order published in the Nikkan Showa Lo 2019] month Δ4 days 3 Number of inventions to be increased by amendment B Subject of amendment

Claims (9)

[Claims] (1) A method for determining the reaction properties of a biological mixture of analytes and reagents, including the following steps: (a) preparing an at least partially opaque sample containing the analyte and a suitable reagent; and (b) centrifuging the sample. (c) First measure the linear dimension of the opaque part of the sample;
successively measuring the same linear dimension of the sample and (e) evaluating a mathematical coefficient of the measured relationship between the first measured dimension and the second measured dimension. The above method. The method according to claim 1, comprising the step of: (2) performing the first measurement when the centrifugal force on the sample is approximately equal to the gravitational force on the specimen. (3) The 5% scope of claim 2 includes the step of performing subsequent measurements while the sample is rotating at the same speed as the sample is rotating when the first measured value is taken. the method of. (4) The method according to claim 1, wherein the first and subsequent measurements are performed while the sample is rotating. (5) A method according to claim 6, wherein a plurality of samples are centrifuged together and the first field determination and subsequent measurements are performed sequentially on one sample after another. (6) The linear dimension to be measured is vertical. A method according to claim 1. (7) The method according to claim 1, wherein the fluid specimen is blood. 8. The method of claim 1, wherein the step of measuring a linear dimension includes the steps of illuminating a sample and measuring the size of a shadow cast by the illuminated sample. (9) Introducing the sample into a light-transmissive chamber and illuminating the light-transmissive chamber. A method according to claim 1. 49. The method of claim 9, comprising the step of centrifuging the sample to compress the sample into the radial tip portion of the optically transparent chamber. 01) The method according to claim 10, which comprises the step of reducing the centrifugation so as to balance the centrifugal force and gravity acting on the sample before performing the measurement. 12. The method of claim 11, comprising the step of dissociating the opaque portion of the sample by allowing a sufficient amount of time to elapse between the first measurement and subsequent measurements. 0 A device for measuring the reactive properties of a biological mixture of analytes and reagents, comprising: (a) a centrifuge rotor; (b) capable of being formed into a cylinder for movement in unison with said centrifuge rotor; An energy-transparent flexible belt that can be removably worn (
(c) a detection system for irradiating the plurality of specimens contained in the belt and optically measuring the reactive characteristics of the specimens. The above device. α Force A centrifuge rotor has (a) an essentially horizontal circular planar component, (b) an upper end, a lower end, an inner surface, and an outer surface, the upper end having a plurality of recesses, and the outer surface of the lower end being the outer surface of the upper end. a cylindrical component that radiates outward from its outer surface and whose inner surface is the same plane from its upper end to its lower end; a fixed annular component (said annular component being a planar component and disposed at an acute angle with the central axis of the cylindrical component; 1.00 of claim 16, comprising a plurality of recesses and a plurality of recesses aligned in one plane and extending therefrom.
9% W+ which is firmly fixed to the lower end of the carbon-containing structure and further comprises a number of supports extending in the L direction for removably attaching the energy-permeable belt to the centrifuge rotor. The device according to item 14 of the scope of the claim. 0→ The belt is (a) a flat element having an inner surface, an outer surface, an upper end and a lower end; (b) an inner surface, an outer surface, an open upper end and a lower end;
at least one slug formed in the 7c
, a dimpled element with a dimple extending radially outward and vertically oriented (the dimple extends along a moderately inclined slope segment to a radial apex that curves smoothly, extending radially outward toward fJAI+) radial depth gradually increases vertically, then sharply decreases in radial depth along strongly inclined segments, and then uniformly shallower along generally vertical segments, followed by a steeply decreasing radial depth along the generally vertical segments. 14. The device of claim 13, wherein the device comprises: a. Zero force (a) draws fluid from the reservoir and holds the fluid;
and a syringe for dispensing fluid; (b) a conduit needle operatively associated with the syringe for insertion into the fluid to enable said syringe to withdraw or dispense fluid; (c) a carousel; and) a plurality of reservoirs inserted into the carousel from which the conduit needle and the syringe can draw fluid, or into which the conduit needle and the syringe can discharge fluid. 17. The apparatus of claim 16, including dilution devices. θ~ The carousel comprises (a) a column, and (b) a plurality of flat circular plates perpendicular to and rigidly interlocked with said column for receiving and securing a plurality of reservoirs. The plates include a top plate having an aperture corresponding to the relevant cross-section of the reservoir and ensuring its relative position, a bottom plate for supporting the reservoir, and a bottom plate corresponding to the relevant cross-section of the reservoir and ensuring its relative position. 18. The device according to claim 17, comprising an intermediate plate with an opening for securing the position. An element that automatically performs a first reading and taking of the linear dimension of the specimen contained in the belt, an element that automatically performs a second reading of the linear dimension of the specimen contained in the belt after a certain period of time, and 14. The apparatus of claim 13, comprising an element for automatically comparing said first and second readings. (a) the carousel includes a plurality of containers, the containers having coded bar labels on the containers, and the apparatus further including a coded bar label reader configured to read the labels on the containers; 17. The apparatus of claim 16, wherein the apparatus includes an element for directing the carousel past the reader. 01) Contains a number of regularly spaced peripheral containers on the belt, and elements for automatically inserting reagents and specimens in appropriate quantities into said plurality of chambers. An apparatus according to claim 20. (a) an apparatus for measuring the reactive properties of a biological mixture of analytes and reagents, comprising: a centrifuge comprising containers for separate analyte- and reagent-containing samples; a device for automatically measuring the vertical dimension of each of said samples of; a comparison device for comparing a first measurement made by said device with a subsequent measurement made by said device for each sample; It is characterized by The above device. Claim 22, wherein the centrifuge includes a removable flexible belt having a container formed in the belt.
Apparatus described in section. 23. The apparatus of claim 22, wherein the apparatus automatically takes subsequent measurements of each vertical dimension of the sample after a predetermined time period after the first measurement has been fulfilled. The comparison device is adapted to compare the difference between the first measurement and the subsequent measurement to a predetermined difference indicative of a reactive characteristic between the analyte and the reagent. Apparatus according to claim 22.