JP2006171010A - Method of measuring sample - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method of measuring a sample for obtaining precise measurement results by measuring particles, without bubbles, since a cleaning liquid is sprayed to a through hole or close to it and is discharged with the bubbles, before measuring the particles. <P>SOLUTION: The method is used to measure particles in liquid containing the particles obtained, by diluting a sample at least in a diluted solution and comprises a step of removing bubbles by spraying the cleaning liquid to the through hole for detecting particles and collecting the sprayed cleaning liquid, a step of supplying the liquid containing the particles to the through hole, by wrapping the liquid with a sheath liquid made of the diluted solution, after the bubble removal process, and a step of measuring the particles that pass through the through hole. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a sample measurement method for measuring particles in a particle-containing liquid obtained by diluting a sample such as blood with a diluent.

  Prior arts related to this invention include a detection block having a through hole for particle detection, a first cell for supplying a particle-containing liquid to the through-hole in a sheath liquid, and containing particles that have passed through the through-hole. The second cell for receiving and discharging the liquid and the sheath liquid, the electrodes respectively provided in the first and second cells, and one of the first and second cells can be slidably supported to change the gap between them. There is known a particle detector including a sliding member, and a detection block is detachably held in a gap between the first and second cells and is watertightly connected to the first and second cells (for example, Patent Document 1).

Japanese Patent Laid-Open No. 2001-33378

  Conventionally, an electrical detection band method has been used to measure the particle size and number of blood cells in blood or industrial particles such as cement powder, latex, and toner. In the electrical detection zone method, a particle-containing liquid in which a partition wall having one through hole is provided in an electrolyte solution, two electrodes are arranged across the through hole, and particles to be measured are dispersed in the electrolyte solution. Through the through hole.

  As the particles pass through the through-holes, the electrical resistance changes instantaneously and a voltage pulse is generated between the electrodes. Since the pulse height reflects the particle volume, the sphere equivalent diameter of the particle can be measured with almost no influence on the shape, and the volume-based particle size of the sample particle can be obtained based on this result. Further, the number of particles can be obtained from the number of pulses.

By the way, in the particle detector adopting such an electric detection band method, that is, an electric resistance type particle detector, when the particle-containing liquid flows into the through hole, bubbles are generated in the through hole and the vicinity thereof. When the number of bubbles increases, the detection pulse between the electrodes is disturbed, and there is a disadvantage that the particle information is erroneously detected.

  The present invention has been made in consideration of such circumstances, and aims to obtain a highly accurate measurement result by removing bubbles generated in and near the through-hole with a cleaning liquid.

  The present invention relates to a sample measurement method for measuring particles in a particle-containing liquid obtained by diluting a sample with at least a diluent, and spraying the cleaning liquid on the through holes for particle detection and collecting the sprayed cleaning liquid The step of removing bubbles, the step of supplying a particle-containing liquid to the through-hole after being wrapped in a sheath liquid made of a diluent, and the step of measuring particles passing through the through-hole after the bubble removing step A sample measurement method is provided.

  According to this invention, before the measurement of the particles, the cleaning liquid is sprayed to the through-hole and the vicinity thereof and discharged together with the bubbles, so that the particles can be measured without the bubbles, and the measurement is highly accurate. The result can be obtained.

  The sample measurement method of the present invention is a sample measurement method for measuring particles in a particle-containing liquid obtained by diluting a sample with at least a diluent, and is sprayed by spraying a cleaning liquid on a through-hole for particle detection. The step of removing the bubbles by collecting the washing liquid, the step of supplying the particle-containing liquid to the through hole by wrapping the sheath liquid made of a diluent into the through hole after the bubble removing step, and passing through the through hole. And measuring the particles.

  The sample measurement method can be suitably used for a particle analyzer such as a blood analyzer or a urine analyzer.

Hereinafter, the present invention will be described in detail based on embodiments shown in the drawings.
Configuration of Blood Analyzer FIG. 1 is a block diagram showing a configuration of a blood analyzer according to the present invention, and FIG. 2 is an external perspective view thereof. As shown in these drawings, the blood analyzer includes a blood analyzer main body 101 and a personal computer 102.

The main body 101 includes a sampling unit 51, a dilution reaction unit 52, an electric resistance type detection unit 53, a flow cytometer type detection unit 54, and a colorimetric type detection unit 55. The personal computer 102 includes a data processing unit 56 and an output unit 57.
Is provided.

The sampling unit 51 aspirates and quantifies a desired amount of blood sample from the specimen container containing the blood sample to be analyzed, and sends it to the dilution reaction unit 52. The dilution reaction unit 52 dilutes the quantified blood sample at a predetermined magnification and reacts with a necessary reagent, and then sends it to the electrical resistance detection unit 53, the flow cytometer detection unit 54, and the colorimetric detection unit 55. To do.

  The electrical resistance detection unit 53 measures items such as red blood cells and platelets in a blood sample by an electrical detection band method. The flow cytometer type detection unit measures data for counting and classifying white blood cells in a blood sample by flow cytometry. In addition, the colorimetric detection unit 55 measures the hemoglobin concentration in the blood sample by a colorimetric method.

  The data processing unit 56 calculates a desired measurement item from each measurement data obtained by the detection units 53, 54, and 55, and causes the output unit 57 to output it in the form of a numerical list or a scattergram.

The data processing unit 56 is composed of a CPU, ROM, and RAM, and the output unit 57 is composed of a CRT and a printer. The main body 101 includes a metal housing 58, and the metal housing 58 accommodates the sampling unit 51, the dilution reaction unit 52, and the detection units 53, 54 and 55.

However, the particle detector of the electric resistance type detection unit 53 is provided on the outer surface of the front panel 58a of the housing 58 so that the maintenance (maintenance, inspection) can be easily performed, and a metal cover for preventing noise that can be opened and closed. 59
Is housed inside.

(Configuration of particle detector)
3 is a side view of the particle detector 1 accommodated in the cover 59, FIG. 4 is a cross-sectional view taken along the line AA in FIG. 3, FIG. 5 is a cross-sectional view taken along the line BB in FIG. FIG.

  As shown in these drawings, the particle detector 1 includes a plate-like member 3, a first cell 4, a second cell 5, and two through shafts 8 and 9. The plate-like member 3 has a through hole 2 for particle detection. The first cell 4 includes a first lid member 15 and a cylindrical first cell body 16, and the second cell 5 includes a second lid member 18 and a cylindrical second cell body 20.

  As shown in FIG. 4, the shafts 8 and 9 penetrate the first and second cell bodies 16 and 20 sandwiching the plate-like member 3, and tube-like stoppers 10 and 11 are inserted from both ends of the penetrated shafts 8 and 9. , 12, 13 are inserted.

  Then, a nut 31 is screwed onto male threads formed at both ends of the shafts 8 and 9, and the first and second cell bodies 16 and 20 are watertightly coupled to the plate-like member 3 by the tightening force of the nut 31. Yes.

Further, each of the first and second lid members 15 and 18 includes a nozzle 14 and a recovery pipe 17 penetrating the axis. Moreover, the 1st and 2nd cover members 15 and 18 are each provided with the protrusion part 15a, the flange part 15b, the protrusion part 18a, and the flange 18b.

The flange portions 15a and 15b are formed on the annular portions 10a, 11a, 12a, and 13a of the stoppers 10, 11, 12, and 13 with the protruding portions 15a and 18a inserted into the openings of the first and second cell bodies 16 and 20, respectively. It is locked and pressed in the direction of the first and second cell bodies 16 and 20.

  Accordingly, the first and second lid members 15 and 18 are watertightly coupled to the first and second cell bodies 16 and 20 via the packings 21 and 22, respectively. At the same time, the first and second lid members 15 and 18 are positioned by the shafts 8 and 9 so that the nozzle 15a, the through hole 2 and the recovery pipe 17 are coaxial with each other.

As shown in FIG. 3, the insertion hole 32, 33, the locking hole 34, 35, and the insertion hole 32 and the locking hole 34 are communicated with the flange portion 15 b on an arc centered on the axis of the nozzle 14. An arc-shaped communication hole 36, and an arc-shaped communication hole 37 that communicates the insertion hole 33 and the locking hole 35 are formed.

  The insertion holes 32 and 33 have an inner diameter larger than the outer diameter of the annular portions 10a and 12a of the stoppers 10 and 12, and the locking holes 34 and 35 have an inner diameter smaller than the outer diameter of the annular portions 10a and 12a.

Further, strip-shaped rim portions 15c protruding from the surface of the flange portion 15b are formed on the peripheral edges of the insertion hole 32, the communication hole 36, and the locking hole 34 and on the peripheral edges of the insertion hole 33, the communication hole 37, and the locking hole 35. Is provided. And the edge part 15c has the height of the part surrounding the locking hole 35 higher than another part. And the annular parts 10a and 12a are hold | maintained in the state which got on the edge part 15c around the locking holes 34 and 35, as shown in FIG. Thereby, the first lid member 15 is pressed in the direction of the first cell body 16 and is watertightly coupled to the first cell body 16 via the packing 21.

  Further, since the flange portion 18b has the same configuration as the flange portion 15b, the second lid member 18 is also water-tightly coupled to the second cell body 20 via the packing 22 in the same manner.

As shown in FIG. 6, the plate-like member 3 includes a pellet 24 having a through-hole 2 and two ring-like packings 25 and 26 sandwiching the pellet 24 from both sides inside a central opening 23. And the 1st and 2nd cell main-body parts 16 and 20 are watertightly connected with respect to the pellet 24 via packing 25 and 26, respectively.

As shown in FIG. 5, the first cell body 16 includes a stainless steel nipple 6 fitted from above and a nipple 27 integrally formed below. The nipple 6 receives the sheath liquid and supplies it to the through hole 2, wraps the particle-containing liquid ejected from the nozzle 14, and passes through the through hole 2. The tip of the nipple 6 is exposed inside the first cell body 16 and is used as a measurement electrode (negative electrode).

  The nipple 27 has an angle of approximately 45 degrees with respect to the axis of the nozzle 14 so that the cleaning liquid can be received and sprayed to the through-hole 2 obliquely from below.

Further, the second cell body 20 includes nipples 28 and 19 integrally formed above and below, respectively. The nipple 19 has an angle of approximately 45 degrees with respect to the axis of the recovery tube 17 so that the cleaning liquid can be received and sprayed to the through hole 2 from obliquely below.

The nipple 28 is provided for discharging the cleaning liquid in the second cell body 20. The 2nd cell main body 20 is equipped with the electrode 7 for a measurement (FIG. 4) exposed inside. The electrode 7 is a platinum rod-shaped electrode, used as an anode,
The flange 18 is fixed in parallel with the recovery pipe 17.

  The nozzle 14 includes a tube connection portion 29 at the rear end, and the recovery tube 17 also includes a tube connection portion 30 at the rear end.

The nozzle 14 is made of stainless steel and has an inner diameter of 0.2 mm, and the pellet 24 is made of artificial ruby and has a thickness of 1 mm. The diameter of the through-hole 2 varies depending on the particle size of the measurement target particle.
m.

  The first and second cell bodies 16 and 20, the first lid member 15 and the second lid member 18 are all formed by injection-molding a chemical-resistant thermoplastic resin polyetherimide.

The nipple 27 is integrally formed with the first cell body 16, and the nipples 19 and 28 are integrally formed with the second cell body 20. The nozzle 14, the electrode / nipple 6 and the electrode 7 are the first cell body 16 and the second lid member 18 after molding.
Each is press-fitted and fixed with an adhesive.

(Disassembly and assembly of particle detector)
Disassembling and assembling the particle detector A method for disassembling and assembling the particle detector 1 when performing maintenance work for removing the adhering through-hole 2 and the adhering material in the vicinity will be described. First, the operator holds the flange portion 15b shown in FIG. 3 and rotates it by about 45 degrees in the direction of arrow D (counterclockwise). As a result, the annular portions 10a and 12a move relatively to the two insertion holes 32 and 33 of the flange portion 15b. Therefore, the operator can pull out the first lid member 15 from the first cell body 16 as shown in FIG. 7 while grasping the flange portion 15b.

The second lid member 18 can also be pulled out from the second cell body 20 by operating the flange portion 18b in the same manner. Therefore, the operator inserts a fine brush-like brush from each opening of the first and second cell bodies 16 and 20 to remove the through-hole 2 and the adhering material in the vicinity thereof from both sides of the through-hole 2.

Next, when reassembling the particle detector 1, it is performed in the reverse order of the disassembling operation. That is, the flange portion 15b of the first lid member 15 is gripped, and the annular portions 10a and 12a of the stoppers 10 and 12 are inserted into the insertion holes 32 and 33 of the flange portion 15b, respectively. At the same time, the protrusion 15 a is also inserted into the opening of the first cell body 16.

Then, while pressing the flange portion 15b in the direction of the first cell body 16, the flange portion 15b is rotated by about 45 degrees in the opposite direction (counterclockwise direction) to the arrow D in FIG. Accordingly, the annular portions 10a, 12a are connected to the locking holes 3
It moves relatively to 4,35 and rides on the high part of the edge 15c.

As a result, the first lid member 15 is pressed toward the first cell body 16 and is watertightly coupled to the first cell body 16 via the packing 21. The second lid member 18 is also water-tightly coupled to the second cell body 18 by the same procedure. Thereby, the assembling work is completed.

(Structure of particle detector cover)
8 is a perspective view of the cover 59 of the particle detector 1, FIG. 9 is a perspective view showing a state in which the cover 59 is opened, and FIG. 10 is a cross-sectional view of the cover 59. As shown in these drawings, since the cover 59 is fixed by a screw 82, when the screw 82 is loosened and the knob 80 is pulled forward, the cover 59 is opened, and the particle detector 1 installed inside the cover 59 is opened. It is supposed to be exposed.

The cover 59 has a double structure of an outer cover 60 and an inner cover 61. The inner cover 61 includes a corresponding bottom plate 63, and the bottom plate 63 is fixed to the front panel 58 a with an insulating resin screw 65 via an insulating bush 62. The bottom plate 63 includes two lateral auxiliary side plates 63a erected from the periphery and two vertical auxiliary side plates 63b. The auxiliary side plates 63a and 63b are elastic on the inner surface of the inner cover 61 when the cover 59 is closed. Are configured to contact each other.

Also, one auxiliary side plate 64 corresponding to the outer cover 60 is fixed to the front panel 58a with a metal screw 81, and when the cover 59 is closed, the auxiliary side plate 64 elastically contacts the inner surface of the outer cover 60. . The outer cover 60 is fixed to the auxiliary side plate 64 with a metal screw 82.

Further, the outer cover 60 includes four contact plates 66 spot welded to the periphery of the opening. Corresponding to these, four conductive elastic members 67 are fixed to the front panel 58a, and when the cover 59 is closed, the four contact plates 66 elastically contact the corresponding elastic members 67. It has become.

Further, the outer cover 60 is fixed to the inner cover 61 via an insulating member 68 as shown in FIG. As shown in FIG. 9, the inner cover 61 is rotatably supported by two longitudinal auxiliary side plates 63b by two fulcrum pins 69 so that the cover 59 can be opened and closed.

The outer cover 60, the inner cover 61, the auxiliary side plates 63a, 63b, 64, and the contact plate 66 are all made of a metal plate (for example, a stainless steel plate). Therefore, when the cover 59 is closed, the particle detector 1 is covered with a double conductor case insulated from each other. The conductive elastic member 66 can be easily formed by attaching a conductive tape to sponge rubber, for example.

(Configuration and operation of electric circuit of electric resistance type detection unit 53)
FIG. 11 is a circuit diagram showing a main part of an electric circuit of the electric resistance type detection unit 53. As shown in FIG. 11, when a voltage of AC100V is input from the commercial power supply to the switching power supply circuit 71, the switching power supply circuit 71 converts the voltage of AC100V into a voltage of DC5V and DC12V, and uses the ground line G as a common negative electrode. Outputs DC voltage of 5V and 12V.

  The booster circuit 72 boosts DC12V to 56V, and inputs a DC voltage of 56V to the constant current circuit 73 with the ground line G as a negative electrode.

The constant current circuit 73 applies a DC voltage of no-load voltage 56V to the electrodes 6 and 7 of the particle detector 1 so that the electrode 7 is a positive electrode and the electrode 6 is a negative electrode. In the particle detector 1, when the particle-containing liquid wrapped in the sheath liquid passes through the through-hole 2, a constant current is supplied from the constant current circuit 73 between the electrodes 6 and 7, and the electric resistance ( (Or impedance) changes, and the change appears between the electrodes 6 and 7 as a voltage change (pulse).

  The voltage between the electrodes 6 and 7 is amplified by the amplifier 74 and converted into a digital signal by the A / D converter 75. Based on the converted digital signal, the arithmetic circuit 76 calculates data such as the particle diameter and the number of particles and outputs the data to the personal computer 102 (FIG. 1).

  In the circuit shown in FIG. 11, the 5V output voltage of the switching power supply circuit 71 has its positive and negative electrodes connected to the power supply line H and the ground line G, respectively. The amplifier 74, the A / D converter 75, and the arithmetic circuit 76 are connected to the lines H and G. It is driven by receiving a voltage from G.

(Measures against noise in particle detector 1)
8 to 10, the particle detector 1 is covered with a double structure cover 59 including an outer cover 60 and an inner cover 61, and the outer cover 60 is interposed via a front panel 58 a of the housing 58 as shown in FIG. 11. The inner cover 61 is connected to the ground line G of the electric circuit. With such a configuration, the particle detector 1 can be shielded from external noise, and EMC regulations regarding electromagnetic waves can be cleared.

  That is, it has been confirmed that the output of the amplifier 74 is not affected by a radio wave having an electric field strength of 3 V / m obtained by AM modulation of a high frequency of 80 MHz to 1 GHz at 1 KHz.

(Bubble removal operation of particle detector 1)
In the particle detector 1, when particles are measured while flowing the particle-containing liquid together with the sheath liquid from the first cell 4 to the second cell via the through hole 2, bubbles are generated in the through hole 2 and the vicinity thereof. When the number of bubbles increases, the bubbles disturb the detection voltage between the electrodes 6 and 7, and there is a disadvantage that the particle information is erroneously detected.

Therefore, in the present invention, immediately before the measurement of one specimen, the cleaning liquid is sprayed from the nipples 19 and 27 shown in FIG. Thereby, bubbles adhering to or floating in the vicinity of the through-hole 2 and the vicinity thereof are removed and discharged together with the cleaning liquid from the nipples 28 and 6, respectively. Thereby, the particle detector 1 can maintain high detection accuracy.

(Fluid circuit of electric resistance type detection unit 53 and its operation)
FIG. 12 is a main part fluid circuit diagram of the electrical resistance detection unit 53 using the particle detector 1. In the figure, when the valves V1 and V2 are opened, a sample for measuring red blood cells is sucked into the flow path P from the sample chamber SC and stored by the negative pressure in the drainage chamber WC1. The sample chamber SC has a dilution reaction unit 52.
The red blood cell measurement sample diluted in (FIG. 1) is stored in advance.

  When the valves V1 and V2 are closed and the valves V3, V4, V5 and V6 are opened, the diluent is supplied from the diluent chamber DC to the nipples 27 and 19 as a cleaning liquid by positive pressure, and is sprayed to the through-hole 2 and the first and second The second cells 4 and 5 are cleaned and air bubbles are removed, and discharged from the nipples 6 and 28 to the drain chamber WC1.

When the valves V3, V4, V5 and V6 are closed and the valve V7 is opened, the diluent is supplied as a sheath fluid from the diluent chamber DC to the nipple 6 as a sheath fluid, and passes through the through hole 2 and the recovery pipe 17 to drain the fluid chamber. It is discharged to WC2. As a result, a sheath liquid flow passing through the through hole 2 is formed.

When the syringe pump CP is operated in the discharge direction in this state, the sample stored in the flow path P is pushed out and ejected from the nozzle 14 to the through hole 2. The ejected sample is wrapped in the sheath liquid, passes through the through-hole 2, and is discharged to the drainage chamber WC2 through the recovery pipe 17.

  When the sample is wrapped in the sheath liquid and passes through the through-hole 2, the change in impedance between the electrode 6 (FIG. 3) and the electrode 7 (FIG. 2) is measured by the measurement circuit shown in FIG. When the measurement is completed, the syringe pump CP is stopped. Then, the valve V8 is opened, and the residual sample in the sample chamber SC is discharged to the drain chamber WC1.

Next, the valves V1, V2, V8, and V9 are opened, and the diluent is supplied as a cleaning liquid from the diluent chamber DC to the sample suction channel, the sample chamber SC, the syringe pump CP, and the nozzle 14, and after a predetermined time, the valves V1, V2 , V
8, V9 is closed to finish the cleaning.

  In the sample measurement method according to the present invention, since the cleaning liquid is sprayed on the through hole and the vicinity thereof and discharged together with the bubbles before the measurement of the particles, the measurement of the particles can be performed in the absence of the bubbles. As a result, it is possible to obtain a high measurement result and is useful as a sample measurement method.

It is a block diagram which shows the structure of the blood analyzer which concerns on this invention. 1 is a perspective view of a blood analyzer according to the present invention. It is a side view of the particle detector concerning this invention. It is AA arrow sectional drawing of FIG. It is a BB arrow sectional view of Drawing 3. It is the C section enlarged view of FIG. FIG. 5 is an exploded view of the particle detector shown in FIG. 4. It is a perspective view of the cover of the particle detector concerning this invention. It is a perspective view which shows the state which opened the cover shown in FIG. It is a cross-sectional view of the cover shown in FIG. It is a principal part electric circuit diagram of the blood analyzer according to the present invention. It is a principal part fluid circuit diagram of the blood analyzer which concerns on this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Particle detector 2 Through-hole 3 Plate-shaped member 4 1st cell 5 2nd cell 6 Electrode (nipple)
7 Electrode 8 Shaft 9 Shaft 10 Nut 11 Nut 12 Nut 13 Nut 14 Nozzle 15 First Lid Member 16 First Cell Main Body 17 Recovery Pipe 18 Second Lid 19 Nipple 20 Second Cell Main Body 21 Packing 22 Packing 23 Opening 24 Pellet 25 Packing 26 Packing 27 Nipple 28 Nipple 29 Tube connecting part 30 Tube connecting part 59 Cover 60 Outer cover 61 Inner cover 62 Insulating bush 63 Bottom plate 64 Auxiliary side plate 65 Screw 66 Contact plate 67 Elastic member 68 Insulating member 69 Support pin

Claims (6)

A sample measurement method for measuring particles in a particle-containing liquid obtained by diluting a sample with at least a diluent,
A step of removing bubbles by spraying a cleaning liquid on the through holes for particle detection and collecting the sprayed cleaning liquid;
After the bubble removal step, the step of supplying the particle-containing liquid by wrapping it in a sheath liquid made of a diluent into the through hole;
Measuring the particles passing through the through-hole, and a sample measuring method.   The sample measurement method according to claim 1, wherein the cleaning liquid is a diluent.   The sample measurement method according to claim 1, wherein the cleaning liquid is sprayed obliquely with respect to the through hole.   The sample measuring method according to any one of claims 1 to 3, wherein the cleaning liquid is sprayed from the particle-containing liquid supply side and the opposite side thereof.   The sample measurement method according to any one of claims 1 to 4, wherein in the particle measurement step, the particles are measured based on a change in an electric resistance value caused by the particles passing through the through holes. The sample measurement method according to claim 1, wherein the sample is blood or urine.

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