JPH08201267A - Particle analyzer - Google Patents

Abstract

(57) [Summary] [Purpose] A device that analyzes the image and concentration of particles. A sharp image can be obtained even if the flow rate of the sample liquid and sheath liquid is increased and the sampling mechanism is operated during analysis. Be able to analyze the sample well. [Structure] Tubes 40 and 41 for supplying a sample liquid 50 and a sheath liquid 51 are fixed with a fixture 60 so that a resonance frequency thereof is higher than a repetition frequency of imaging. Further, pressure fluctuation absorbing portions 42 and 52 are provided in the paths of both tubes. Then, the delay time of both tubes is made uniform by the pressure fluctuation absorbing section. A flat sheath flow is formed and flown in the flow cell 11, and an image is taken by the image pickup device 30 to analyze the morphology and concentration of particles. [Effect] By increasing the flow rate with the fixture and the pressure fluctuation absorbing section, a clear image can be obtained, and the uniform delay time can shorten the time until the start of analysis.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for analyzing particles by picking up an image of particles suspended in a liquid, for example, a particle analyzer suitable for analyzing particles in blood or urine.

[0002]

2. Description of the Related Art In order to morphologically examine particles, there is a method in which the particles are suspended in a liquid and allowed to flow through a flow cell for optical analysis. For example, in Japanese Unexamined Patent Publication No. 5-296915, a liquid sample is formed into a flat sheath flow and allowed to flow through a flow cell, a passage of particles is detected, a pulse light source is caused to emit light, and a still image of the particles is taken by a CCD camera through a microscope system. A particle analyzer for imaging and analyzing the image is shown.

Further, in order to perform an inspection efficiently with a small amount of liquid sample, a sampling nozzle for sucking the liquid sample and discharging it to the flow cell unit and a mechanism for moving the sampling nozzle to the flow cell unit, that is, a sampling mechanism are provided. There is one described in Kaihei 6-194300.

[0004]

In the particle analyzer, it is necessary to control the flow range of the sample liquid within the thickness of the depth of focus of the microscope system. However, if the flow rate of the sample liquid or the sheath liquid is increased, that is, the volume that flows per unit time is increased, the vibration of the actuator that supplies the sample liquid or the sheath liquid increases. Vibrations not only directly cause pressure fluctuations. The vibration causes the sample tube through which the sample liquid flows and the sheath tube through which the sheath liquid flows to resonate, and indirectly causes a pressure fluctuation in the tube.
Due to these pressure fluctuations, the flow of liquid is disturbed, and the sample flow exceeds the depth of focus, making it impossible to obtain a clear image.

When a sampling mechanism such as that disclosed in Japanese Patent Laid-Open No. 6-194300 is operated during analysis, pressure fluctuations occur in the tube as in the case of the actuator, the flow of liquid is disturbed, and the sample flow is changed. A sharp image cannot be obtained because it exceeds the depth of focus.

The above-mentioned conventional apparatus has not taken this problem and its solution into consideration, and has a drawback that it is impossible to increase the flow rate or operate the sampling mechanism during the analysis.

A first object of the present invention is to provide a particle analyzer capable of obtaining a clear image even if the flow rates of the sample liquid and the sheath liquid are increased.

A second object of the present invention is to provide a particle analyzer capable of obtaining a clear image even if a sampling mechanism is operated during analysis.

A third object of the present invention is to provide a particle analyzer capable of obtaining a clear image by preventing the fluctuation of the flow in the sample tube and the sheath tube.

A fourth object of the present invention is to provide a particle analyzer capable of increasing the flow rates of a sample liquid and a sheath liquid or performing an analysis even if a sampling mechanism is operated during the analysis. It is in.

A fifth object of the present invention is to provide a particle analyzer which can shorten the time until the start of imaging.

[0012]

In order to achieve the above object, a flow cell that forms a sheath flow between a sample liquid containing test particles and a clean sheath liquid, and a sample liquid supply mechanism for supplying the sample liquid to the flow cell. In a particle analyzer including a sheath liquid supply mechanism for supplying a sheath liquid to a flow cell, an imaging mechanism for repeatedly capturing an image of a test particle, and a data analysis mechanism for analyzing the image and outputting the result, Use any means.

For the first and second purposes, a fixing device for fixing the tube forming the path to the path from the sample solution supply mechanism to the flow cell and the path from the sheath solution supply mechanism to the flow cell is provided, and The resonance frequency is set higher than the repetition frequency of the imaging mechanism. ◆ And preferably, the repetition frequency of the imaging mechanism is set to 30 Hz. Further, preferably, as the fixing device, at least one of a fixing device for fixing the tube, or a fixing pipe or a fixing means for fixing the tube so as to give tension to the tube forming a part of the path is used. ◆ Alternatively, a fixture for fixing the tubes forming the path is provided on the path from the sample liquid supply mechanism to the flow cell and the path from the sheath liquid supply mechanism to the flow cell, and the distance between the fixtures is within the range of 5 to 27 cm at the maximum. Set.

For the third and fourth purposes, a pressure fluctuation absorbing portion is provided in the path from the sample liquid supply mechanism to the flow cell. ◆ And, preferably, the pressure fluctuation absorbing portion is a bubble in the path, and the amount of the bubble is set in the range of 5 to 32 microliters. ◆ In addition, a pressure fluctuation absorber is installed in the path from the sheath liquid supply mechanism to the flow cell. And, preferably, the pressure fluctuation absorbing section is provided with at least one of a thin tube thinner than the path, a tube having an air chamber, or a bellows as a damping member. ◆
And preferably, the damping member is a thin tube narrower than the path, and the length of the thin tube is set in the range of 40 to 160 cm.

For the fifth object, a pressure fluctuation absorbing portion is provided between the path from the sample liquid supply mechanism to the flow cell and between the path from the sheath liquid supply mechanism to the flow cell, and the delay time of both paths is roughly the same. Match.

By the above means, each of the above objects can be achieved.

Further, by combining the above means, it is possible to provide a particle analyzer having a clearer image, or a particle analyzer having a clearer image and shortening the time until the start of imaging.

In the present invention, the pressure fluctuation absorbing section means
It is a general term for devices capable of absorbing pressure fluctuations such as air bubbles, thin tubes thinner than the path, tubes with air chambers or damping members such as bellows. Further, in the present invention, the delay time refers to a state in which the flow immediately before the flow cell is steady after the liquid supply is started from the liquid supply mechanism at the start of analysis at a constant flow rate, that is, the flow rate is constant.

[0019]

The sample liquid containing the particles to be tested is collected by the sampling mechanism and then inserted into the sample tube by the insertion mechanism. The sample tube in which the sample liquid is inserted is connected to the sample nozzle of the flow cell. When the driving liquid is supplied at a constant high flow rate from the sample driving liquid supply mechanism connected to the other end of the sample tube, the sample liquid is pushed out by the driving liquid and discharged into the flow cell at a constant high flow rate. Further, the cleaning liquid is supplied to the flow cell through the sheath tube at a constant flow rate higher than that of the sample liquid from the sheath liquid supply mechanism.

At this time, when the flow rate is increased, the vibration of the actuator of the sample liquid and sheath liquid supply mechanism becomes large. Vibration is also generated by operating the sampling mechanism. The vibration is transmitted to the sample tube and the sheath tube, so that pressure fluctuations occur in both tubes. Further, the vibration causes the sample tube and the sheath tube to resonate. If the resonance frequency at this time is low, large pressure fluctuations occur in both tubes. When these pressure fluctuations occur, the flow in the tube also fluctuates.

However, the pressure fluctuation can be made small by preventing vibration by the fixing device for fixing the respective tubes to prevent the resonance of the tubes by making the resonance frequency higher than the imaging repetition frequency. Since the pressure fluctuation becomes small, the flow fluctuation becomes small, and the test particles contained in the sample liquid flow without protruding from the depth of focus of the microscope system, and a clear image can be obtained.

Further, the pressure fluctuation absorbing portion such as air bubbles provided in the middle of the sample tube absorbs the pressure fluctuation directly generated from the vibration, and the flow cell is supplied with the sample liquid having almost no fluctuation in the flow due to the pressure fluctuation. By doing so, there is little or no disturbance in the flow, and a clearer image can be obtained.

Further, a pressure fluctuation absorbing portion such as a damping member provided in the middle of the sheath tube absorbs pressure fluctuations directly generated from the vibrations, and the sheath fluid in which the flow fluctuations hardly occur due to the pressure fluctuations is stored in the flow cell. By supplying it, there will be little or no turbulence in the flow,
A clearer image can be obtained.

Therefore, it becomes possible to increase the flow rates of the sample liquid and the sheath liquid or to operate the sampling mechanism during the analysis.

Further, by making the delay time of both paths of the sample liquid and the sheath liquid approximately coincide with each other by the pressure fluctuation absorbing portion, the time until the start of analysis can be shortened.

Therefore, a stable sheath flow can be formed even if the flow rates of both liquids are increased. At this time, since the flow rate of the sheath liquid is higher than that of the sample liquid, the sample liquid flows between the sheath liquid and the region within the depth of focus of the imaging system. By repeatedly imaging this sample solution, a large number of focused and clear particles can be obtained. This image is analyzed by a data analysis mechanism, converted into information on the type and concentration of particles, and the result is output.

As described above, the flow does not fluctuate even if the sampling mechanism is operated during imaging by the fixed device.
The next sample can be prepared during imaging. Further, similarly, even if the flow rate is increased by the fixing device, there is no fluctuation that adversely affects the measurement, so that the flow rate can be increased. Further, since the pressure fluctuation absorbing portion provided in the sample tube and the pressure fluctuation absorbing portion provided in the sheath tube hardly cause pressure fluctuation in the tube, the image becomes clearer. Furthermore, since the delay times are roughly matched, the time until the start of imaging can be shortened, so that the sample can be measured efficiently.

[0028]

1 is a diagram showing the configuration of a first embodiment of the present invention. ◆ Inside the analyzer 31, a flow cell 11, a sample syringe 14, a sheath syringe 15, and a tank 1
8, a vertical rotation mechanism 20, an imaging device 30, a cleaning device 24, and a sampling mechanism 22 are installed. Vertical rotation mechanism 20
A pipetter 21 is attached to the tip of the above, and the pipette 21 is moved to the positions of the reaction container 25, the washer 24, and the flow cell 11 by the rotation of the vertical rotation mechanism 20. The flexible sample tube 40 passes from the sample syringe 14 through the electromagnetic valve 35 and connects the pipette 21. Similarly, the flexible sheath tube 41 passes from the sheath syringe 15 through the three-way solenoid valve 36 and connects to the flow path 12 of the flow cell 11.

Actuators 16 and 17 are attached to the sample syringe 14 and the sheath syringe 15, respectively, to form a sample liquid supply mechanism and a sheath liquid supply mechanism. A pipette 23 is provided at the tip of the sampling mechanism 22.
Is attached, and the specimen container 26 is placed on the turning radius.
A space for placing and a reaction container 25 are installed.

The tank 18 has a pressurizing function, is connected to the sample syringe 14 by a tube through an electromagnetic valve 34, and the sheath syringe 15 is a three-way electromagnetic valve provided on the sheath tube 41 between the damping member 42 and the sheath syringe 15. A tube is also connected through valve 36.

In this embodiment, tubes having an inner diameter of 1.5 mm are used as the sample tube 40 and the sheath tube 41. The sample tube 40 and the sheath tube 41 are the longest parts and are fixed at intervals of 10 cm.
Is fixed to the housing of the analyzer 31, for example, the chassis. The fixture 60 is not particularly limited in material and shape as long as it can firmly fix both tubes to the chassis or the like. A damping member 42 is inserted in the middle of the sheath tube 41. The damping member 42 of this embodiment uses a thin tube having an inner diameter of 1.0 mm and a length of 100 cm. The interval between the fixtures 60 and how to determine the length of the thin tube will be described later.

Next, the inspection method will be described. The sample 54 to be analyzed is placed in the sample container 26 and placed in the apparatus. The sampling mechanism 22 equipped with an actuator (not shown) rotates to move the pipettor 23 onto the sample container 26, and then descends to move the tip of the pipetter 23 to the sample 54.
Insert the sample into the sample and rotate it to stir the sample 54,
Aspirate 4. The sampling mechanism 22 again rises and rotates to discharge the sample 54 from the pipette 23 onto the reaction container 25 into the reaction container 25. After that, the tip of the pipetter 23 is cleaned inside and outside by a cleaning mechanism (not shown) to prepare for the next sampling. The reaction container 25 is supplied with a dyeing solution from a dyeing solution supply mechanism (not shown), and reacts with the specimen 54 to form the sample solution 50.

Since the sample syringe 14 and the pipettor 21 are connected by the flexible sample tube 40, the pipetter 21 can be moved by the operation of the vertical rotating mechanism 20. The vertical rotation mechanism 20 moves up, rotates, and descends,
First, the pipette 21 is inserted into the washer 24. Washing machine 24
A clean liquid flows inside and cleans the outer surface of the pipettor 21. Then, the solenoid valves 34 and 35 are opened,
The clean liquid pressurized from the tank 18 is supplied to the sample tube 4
The inner surface of the pipetter 21 is cleaned by discharging from the pipetter 21 to the cleaning device 24 through 0.

After that, the pipettor 21 is raised to suck a small amount of air, and then it is moved to the reaction container 25 to suck the sample liquid 50 in which the specimen 54 and the staining liquid have reacted. The amount of air sucked at this time, that is, the amount of bubbles 52 will also be described later. The suction of air or sample liquid 50 by the pipettor 21 is performed by the solenoid valve 3
Actuator 1 with 4 closed and solenoid valve 35 open
This is done by activating 6. With this, pipette 21
Inside the sample liquid 50 and the driving liquid 53 with air bubbles 52 sandwiched between them.
Is satisfied. The driving liquid 53 is played by the cleaning liquid described above.

After the pipette 21 sucks the sample liquid 50,
The reaction container 25 discharges unnecessary liquid and cleans the inside with a cleaning device (not shown) to prepare for the next sample. ◆ Next, the pipettor 21 rotates and descends on the flow cell 11,
The tip is closely attached to the nozzle 13 for connection. During the measurement, the driving liquid 53 is supplied from the sample syringe 14 at a constant flow rate, so that the sample liquid 50 is supplied to the pipette 2 via the bubbles 52.
It is extruded from No. 1 and discharged from the nozzle 13 into the flow path 12. With such a method, the sample liquid 50 only needs to be measured, and the required amount is small. Further, since the portion of the pipettor 21 that contacts the sample liquid 50 is small, the cleaning can be effectively performed in a short time.

After the tank 18 and the sheath syringe 15 are connected to each other by the three-way solenoid valve 36 and the actuator 17 is operated to suck the cleaning liquid into the sheath syringe 15,
By switching the three-way solenoid valve 36 to bring the sheath syringe 15 and the sheath tube 41 into communication with each other and operating the actuator 17 in the opposite direction at a constant speed, the clean liquid flows from the sheath syringe 15 through the sheath tube 41 as the sheath liquid 51 as a flow cell 51. 11 is supplied to the flow path 12.

The flow cell 11 has a shape in which the cross-sectional area is reduced in one direction, and the sample liquid 50 wrapped in the sheath liquid 51 forms a thin flow in the parallel flow channel portion downstream of the reduction portion, that is, in the measurement region. It flows at a constant speed. In the present embodiment, by changing the flow rates of the sample liquid 50 and the sheath liquid 51, the flow velocity can be switched between high speed and low speed in the measurement region in the flow cell 11, and the high speed is 1 m / s and the low speed is 0.2 m / s. Is.

In this embodiment, the flow rate is the volume of the sample solution or the sheath solution flowing in the tube within a unit time, and the flow rate is the flow rate of the sheath flow formed by both solutions in the flow cell 11. That is.

The image pickup device 30 has a particle detecting function,
A pulsed light source is caused to emit light in synchronization with the passage of particles to capture a still image of the particles. The image is captured up to 30 times per second. ◆ The captured image is analyzed by a data analysis mechanism (not shown), converted into information on the type and concentration of particles, and the result is output.

In this embodiment, the sample tube 40
Since the sheath tube 41 is fixed by the fixture 60 at a short interval, the resonance frequency of both tubes is set higher than the imaging frequency so that the imaging is not adversely affected.

Further, the bubble 52 and the damping member 42 absorb the pressure fluctuation directly caused by the vibration to prevent the flow fluctuation. Further, in the analyzer equipped with the fixture 60, the high-frequency pressure fluctuation indirectly generated from the resonance of the tube is absorbed to prevent the fluctuation of the flow.

The fixture 60, the air bubble 52, and the damping member 42 prevent the sample flow flowing through the flow cell 11 from being disturbed.

The distance between the fixtures 60, the amount of bubbles 52, and the length of the damping member 42 are determined as follows.

First, the distance between the fixtures 60 is determined as follows. ◆ Resonance frequency of each tube is 60
The shorter the length between the portion and the fixture 60, the higher the height. FIG. 2 shows the result of investigating the resonance frequency by changing the interval of the fixtures 60 using the tube used in this example. As shown in FIG. 2, if fixed at intervals of about 27 cm,
The resonance frequency of the tube is about 30 Hz, which is the repetition frequency of imaging. The shorter the distance, the higher the resonance frequency, which is convenient. However, it is structurally difficult to fix the distance of 5 cm or less in the vicinity of a moving portion such as the vertical rotation mechanism 20. Therefore, it was obtained that the maximum distance between the fixtures 60 is about 5 to 27 cm, which is appropriate without adversely affecting imaging. Based on this result, in this embodiment, the maximum distance is set to 10 cm as described above. In this case, the resonance frequency is sufficiently higher than the imaging repetition frequency of 30 Hz.

Next, in the pipettor 21, the sample liquid 5
The amount of bubbles 52 existing between 0 and the driving liquid 53 is determined as follows. ◆ FIG. 3 is a diagram showing the fluctuation of the flow when the amount of the bubbles 52 is changed. ◆ As described above, when the bubbles 52 are present in the pipe, there is an effect of absorbing the pressure fluctuation depending on the amount of the bubbles 52 and the resistance coefficient of the flow path. The bubbles 52 prevent fluctuations in the flow by absorbing pressure fluctuations.

Since the same tube is used, the resistance coefficient of the flow path is constant. During measurement, if the fluctuation of the flow is suppressed to 15% or less, the flow becomes more suitable for imaging. As shown in FIG. 3, when the flow velocity in the measurement region in the flow cell 11 is 5 microliters or more at both high speed and low speed, the fluctuation becomes 15% or less, and the condition can be satisfied.

FIG. 4 shows that when the amount of the bubbles 52 is changed,
It is a figure showing a delay time. ◆ This delay time is from the start of liquid transfer from the sample syringe 14 at a constant flow rate at the start of analysis to the steady state of the flow of the sample solution 50 at the nozzle 13, that is, the constant flow rate. Tell the time

As shown in FIG. 4, when the amount of the bubbles 52 increases, a delay time is required this time. This delay time is proportional to only the amount of bubbles 52 regardless of the flow velocity in the measurement area in the flow cell 11. To reduce this time to 1 second or less, it is necessary to reduce the time to about 32 microliters or less.

From the above results, the amount of bubbles 52 should be 5 to 32.
In the case of the microliter range, the pressure fluctuations transmitted from the sample tube 40 were sufficiently absorbed, and the time during which the flow became constant was sufficiently short. Based on this result, in this embodiment, about 20 microliters is sucked regardless of the flow rate.

Finally, the length of the damping member 42 is determined as follows. ◆ FIG. 5 is a diagram showing a variation value when the length of the damping member 42 is changed. The damping member 42 is in the middle of the sheath tube 41 and is an extension of a thin tube having an inner diameter of 1.0 mm. As described above, such a thin tube has a function of absorbing pressure fluctuation.
The damping member 42 absorbs the pressure fluctuation to prevent the flow fluctuation like the bubble 52 described above. As shown in FIG. 5, even if the flow velocity in the measurement area in the flow cell 11 is low, if the damping member 42 is set to about 40 cm or more, the flow fluctuation becomes 15% or less, and the flow becomes more suitable for imaging. For high speed, about 20 cm or more is sufficient.

However, if the damping member 42 becomes long as in the case of the bubble 52 described above, a delay time is required as shown in FIG. ◆ This delay time means that after the liquid is sent from the sheath syringe 14 at a constant flow rate at the start of analysis,
It is the time until the flow of the sheath liquid 51 immediately before being guided to the flow cell 11 reaches a steady state. This delay time is proportional to only the length of the damping member 42, regardless of the flow velocity in the measurement area in the flow cell 11, like the bubble 52.

As in the case of the bubbles 52, it is necessary to set the time to about 160 cm or less in order to reduce this time to 1 second or less.
From the above results, the length of the damping member 42 is 40 to 1
When the pressure is set to about 60 cm, the pressure fluctuations transmitted from the sheath tube 41 are absorbed, and the time during which the flow becomes constant is sufficiently short. Based on this result, in this embodiment, it is set to 100 cm as described above.

The fixing device 60, the air bubble 52, and the damping member 42 can provide the following effects.

Sampling mechanism 22 and actuator 1
The vibrations generated by the operations of 6 and 17 have a large component at the same level as or lower than the repetition frequency of imaging, but due to the effect of the fixture 60, the sample tube 40 and sheath tube 41
Resonance does not occur at a frequency lower than the imaging repetition frequency and does not have a large amplitude. Therefore, the volume change of the pressure fluctuation in the tube is small, the flow fluctuation does not occur in the sample liquid 50 and the sheath liquid 51, a stable and constant sheath flow is formed in the flow cell 11, and a clear image of particles can be captured. .

Further, since a clear image of particles can be obtained even if a device that generates some vibration is used, stepping motors capable of accurately moving a fixed amount of the sample liquid 50 are used for the actuators 16 and 17. be able to. Therefore, the particle concentration can be accurately analyzed.

Furthermore, even if the sampling mechanism 22 is operated during the measurement period, the generated vibration does not interfere, so that the sampling mechanism 22 can be operated during the analysis to carry out the next sample collection and staining reaction. Therefore, it is possible to reduce the time required to analyze a plurality of samples. Further, since it is not affected by vibration, the sheath flow can be sped up and a large amount of sample liquid 50 can be flowed in a short time for analysis.

Moreover, even if all the elements serving as a vibration source such as the sampling mechanism 22 and the pipetter 23 are installed in the same housing, a stable sea flow is formed without being affected by vibration and a clear image is obtained. Therefore, it is possible to supply a small-sized analyzer housed in one housing.

The pressure fluctuation absorbing portion such as the bubble 52 and the damping member 42 can suppress a small fluctuation to obtain a smoother flow, which is more effective in the conventional apparatus and the analyzer having a large flow rate equipped with the fixture. A clear image can be obtained. Further, the analysis can be performed while operating the sampling mechanism 22.

In particular, the time from the start of the operation of the sample syringe 14 and the sheath syringe 15 to the steady state of the flow of the sample liquid 50 and the sheath liquid 51 immediately before being guided to the flow cell 11, that is, the delay time is almost equal to both paths. If they are made equal, in other words, if the damping coefficients of both paths are made approximately equal, the liquid will not flow backward to the slower rising side due to transient imbalance, and the flow will be constant quickly. become. Therefore, the measurement can be started quickly and the time required for analysis can be shortened.

In the present embodiment, as described above,
By setting the amount of the bubbles 52 to 20 microliters and the length of the damping member 42 to 100 cm, both delay times are about 0.5 seconds. As another combination, the amount of the bubbles 52 is 5 microliters, the length of the damping member 42 is 50 cm, and the delay time is 0.25.
It is also possible to arrange for about a second.

In this embodiment, the inner diameter of each tube is 1.5.
mm, the thin tube used for the damping material 42 has an inner diameter of 1.0 mm
Since the above-mentioned ones are used, the intervals of the fixtures 60, the amount of the air bubbles 52, and the length of the damping member 42 are respectively changed. However, when using those having different inner diameters, the ranges different from the above May be the value of.

In this embodiment, both the fixing member 60 and the damping member 42 are used. However, since the fixing member 60 alone can prevent the turbulence of the flow that adversely affects the imaging, the fixing member 60 can be fixed. With the tool 60 alone, an image can be taken while increasing the flow rate or operating the sampling mechanism 22 during analysis. When the damping member 42 is not used, the bubbles 52 may be small enough to partition the sample liquid 50 and the driving liquid 53.

Further, in the particle analyzer having no sampling mechanism 22, the bubbles 52 may be omitted.
In that case, instead of the bubbles 52, a device such as a damping member 42 provided on the sheath tube 41 for absorbing pressure fluctuation may be used.

Similarly, without using the fixture 60, only the bubble 52 and the damping member 42, the bubble 52 or only the damping member 42 are used to increase the flow rate and to perform the analysis while operating the sampling mechanism 22 during the analysis. be able to. It is also possible to make an image clearer in a conventional analyzer.

FIG. 7 shows a second tube using another tube fixing method.
It is a figure which shows the Example of. A sampling mechanism 22, a vertical rotation mechanism 20 and the like which are not shown are the same as those in the first embodiment.

In the case of this embodiment, the sample tube 40
The sheath tube 41 is fixed to the chassis 70 between the sample syringe 14 and the vertical rotation mechanism 20 and between the sheath syringe 15 and the flow cell 11 by using a fixed pipe 71 instead of a flexible tube. Alternatively, each flexible tube is fixed to the chassis 70 using a fixed pipe 71 for fixing. At portions other than the fixed pipe 71, both tubes are fixed by the fixing tool 60 as in the first embodiment. The damping member 42 is
The fixed pipe 71 may be on the sheath syringe 15 side or the flow cell 11 side. Since the tube inside the fixed pipe 71 is fixed, it is almost unaffected by vibration and the same effect as the fixture 60 can be obtained. Therefore, a more stable sheath flow can be obtained in the flow cell 11.

As in the present embodiment, pressure fluctuations are prevented by using fixed pipes except for the portions near the syringes, the vertical rotation mechanism 20 and the damping member 42 where the tubes need flexibility. It is also possible.

FIG. 8 is a diagram showing a third embodiment using another tube fixing method. The sampling mechanism 22 and the sheath tube 41, which are not shown, are the same as those in the first embodiment.

In the case of this embodiment, a guide 76 and pulleys 74 and 75 are attached to a block 72 fixed to the chassis 70. The sample tube 40 comes from the sample syringe 14 to the guide 76 while being fixed to the fixture 60 described in the first embodiment or the fixed pipe 71 described in the second embodiment. Then, after it is hung on the pulleys 74 and 75, it is wound around and fixed to the pillar portion of the vertical rotation mechanism 20. The pulley 74 is pulled by a spring 73 so that tension is applied to the sample tube 40.

In the case of this embodiment, the sample tube 40
Since the tension is applied to the resonance frequency, the resonance frequency becomes high, and it does not resonate at low frequency vibration. Further, since the sample tube 40 is wound around the support column of the vertical rotation mechanism 20 via the pulley 75, the vertical rotation mechanism 20 can rotate vertically, and even if the vertical rotation mechanism 20 moves, it is pulled by the spring 73. It stays in tension because of its presence. The resonance frequency f of the tube is calculated by the following equation.

[0071]

[Equation 1]

Here, S: tension, L: length between fixed parts,
γ: Mass per unit length of the tube, and in the present embodiment, S and L are the tension and distance of the tube between the pulley 75 and the vertical rotation mechanism 20, respectively.

In order to make the resonance frequency higher than 30 Hz which is the imaging repetition frequency, 30 Hz is added to f in the above equation.
Substituted and transformed,

[0074]

[Equation 2]

It suffices to give S, L and γ so as to satisfy the above condition.

Γ is a constant value determined by the tube, but S and L can be freely given. That is, the resonance frequency can be increased by shortening the distance from the pulley 75 to the vertical rotation mechanism 20 or increasing the tension, and thus the operating range of the vertical rotation mechanism 20 can be increased.

FIG. 9 is a diagram showing a fourth embodiment using another damping member. The sampling mechanism 22, sample tube 40, etc., which are not shown, are the same as those in the first embodiment. The damping member 42b provided in the middle of the sheath tube 41 has an air chamber inside,
Absorbs pressure fluctuations due to the compressibility of air.

In the case of this embodiment, the sheath tube 41
Since the pressure fluctuation absorption effect can be obtained without extending the length of the tube, the pressure loss in the tube is small, and a small sheath syringe and a small actuator can be used to supply the sheath liquid 51.

It is convenient to provide an air amount adjusting mechanism in the air chamber of the damping member 42b. In this case, the amount is kept constant even if the volume of the internal air changes due to temperature change or the like. be able to. The amount of this air is set to an appropriate value depending on the flow fluctuation and the delay time, like the amount of the bubbles 52 described in the first embodiment.

FIG. 10 shows a fifth example using another damping member.
It is a figure which shows the Example of. The sampling mechanism 22, sample tube 40, etc., which are not shown, are the same as those in the first embodiment. The damping member 42c provided in the middle of the sheath tube 41 is a bellows and absorbs pressure fluctuation due to volume change.

In the case of the present embodiment, the pressure fluctuation absorbing effect can be obtained without extending the length of the sheath tube 41 as in the damping member 42b described above, so that the pressure loss in the tube is small and the sheath liquid 51 is discharged. A small sheath syringe and small actuator can be used for delivery.

Further, in the case of the present embodiment, since air is not contained inside, it is possible to always obtain a constant pressure fluctuation absorbing effect without being affected by the volume change of the gas due to the temperature change.
The length of the bellows is set to an appropriate value according to the flow fluctuation and the delay time.

FIG. 11 is a diagram showing the structure of another flow cell portion used in the present invention. In this embodiment, the tube fixing method and damping member may be any of those in the above embodiments. ◆ The structure of this flow cell part is
There are two differences from the flow cell part depicted in FIG.

One is the sheath tube 41 and the pipettor 2.
Bubbles entering the flow cell 11 through 1 can be removed at any time by the air bleeder 61 connected to the upper portion of the flow channel 12 of the flow cell 11, so that a sheath flow is formed with excess bubbles remaining in the flow channel. There is nothing to do. Therefore, the pressure fluctuation absorption of the sheath liquid 51 and the sample liquid 50 is not affected by the extra bubbles, and the optimum amount of pressure fluctuation absorption can be performed. Further, it is possible to prevent the movement of the bubbles in the flow cell flow channel from causing the sample flow to fluctuate. Further, it is possible to prevent the position where the sample flow is formed from being changed due to the extra bubbles, and thus the focus of the imaging is deviated.

The other is a drainage tube 62 and a drainage port 6.
A double-structured port 63 is connected between 5 and the waste liquid flowing from the drainage tube 62 is an internal cylinder 64 with a constant liquid level.
And then discharged from the discharge port 65. Therefore, the pressure in the flow path 12 is kept constant, and the sheath flow is kept stable.

[0086]

As described above, according to the present invention, by providing the fixing device in the path, a clear image can be obtained even if the flow rates of the sample liquid and the sheath liquid are increased.

Further, a stable sample flow can be formed even if the sampling mechanism is operating during analysis, and a clear image can be obtained.

Further, by adopting a structure in which a pressure fluctuation device is provided in the sample tube and the sheath tube, it is possible to absorb the pressure fluctuation in both tubes and prevent the fluctuation of the flow to obtain a clear image.

Further, by adopting a structure in which a pressure fluctuation device is provided in the sample tube and the sheath tube, it is possible to increase the flow rates of the sample liquid and the sheath liquid or to perform analysis even when the sampling mechanism is operating during analysis.

In addition, by providing a pressure fluctuation device on the sample tube and the sheath tube and making the delay times of both paths roughly uniform, the time until the start of imaging can be shortened.

[Brief description of drawings]

FIG. 1 is a configuration diagram of an entire particle analyzer showing a first embodiment.

FIG. 2 is a relationship diagram showing a space between fixtures and a resonance frequency.

FIG. 3 is a diagram showing a relationship between a bubble amount and fluctuation.

FIG. 4 is a diagram showing a relationship between a bubble amount and a delay time.

FIG. 5 is a diagram showing a relationship between a length of a damping member and fluctuation.

FIG. 6 is a diagram showing the relationship between the length of the damping member and the delay time.

FIG. 7 is a configuration diagram of a fixing method according to a second embodiment.

FIG. 8 is a configuration diagram of a fixing method according to a third embodiment.

FIG. 9 is a configuration diagram of a damping member according to a fourth embodiment.

FIG. 10 is a sectional view of a damping member according to a fifth embodiment.

FIG. 11 is a configuration diagram of another flow cell unit.

[Explanation of symbols]

11 ... Flow cell, 12 ... Flow path, 13 ... Nozzle, 14 ...
Sample syringe, 15 ... Sheath syringe, 16 ... Actuator, 17 ... Actuator, 18 ... Tank, 2
0 ... Vertical rotation mechanism, 21 ... Pipette, 22 ... Sampling mechanism, 23 ... Pipette, 24 ... Washer, 25 ... Reaction container, 26 ... Specimen container, 30 ... Imager, 31 ... Analysis device,
34 ... Solenoid valve, 35 ... Solenoid valve, 36 ... Three-way solenoid valve, 40
... sample tube, 41 ... sheath tube, 42 ... damping member, 42b ... damping member, 42c ... damping member, 50 ... sample liquid, 51 ... sheath liquid, 52
... Bubbles, 53 ... Driving liquid, 54 ... Specimen, 60 ... Fixing device, 6
1 ... Air bleeder, 62 ... Drainage tube, 63 ... Port,
64 ... Inner cylinder, 65 ... Discharge port, 70 ... Chassis, 71 ...
Fixed pipe, 72 ... Block, 73 ... Spring, 74
... pulley, 75 ... pulley, 76 ... guide.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Hidenori Asai 882 Ichige, Hitachinaka City, Ibaraki Prefecture Stock Company, Hitachi Ltd., Measuring Instruments Division (72) Hideyuki Horiuchi 882, Ichige City, Hitachinaka City, Ibaraki Hitachi, Ltd. Mfg. Co., Ltd.Measuring Instrument Division (72) Masaaki Kurimura Inventor, 882 Ichimo, Hitachinaka City, Ibaraki Prefecture, Ltd. Equipment Division (72) Inventor Masaaki Hanawa 882 Ichige Ichi, Hitachinaka City, Ibaraki Prefecture Hitachi Ltd. Measuring Instruments Division

Claims (13)

[Claims] 1. A flow cell that forms a sheath flow with a sample liquid containing test particles and a clean sheath liquid, a sample liquid supply mechanism that supplies the sample liquid to the flow cell, and a sheath liquid that is supplied to the flow cell. In a particle analyzer including a sheath liquid supply mechanism, an imaging mechanism that repeatedly captures an image of the test particle, and a data analysis mechanism that analyzes the image and outputs a result, from the sample liquid supply mechanism to the flow cell The fixing device for fixing the tube forming the path is provided in the path of the tube and the path from the sheath liquid supply mechanism to the flow cell, and the resonance frequency of the tube is set to be higher than the repetition frequency of the imaging mechanism. Particle analyzer. 2. The particle analyzer according to claim 1, wherein the repetition frequency of the imaging mechanism is set to 30 Hz. 3. The particle analyzer according to claim 1, wherein the fixing device fixes the tube,
Alternatively, the particle analysis device is characterized by using at least one of fixing means for fixing a fixed pipe or a tube forming a part of the path so as to give a tension. 4. A flow cell that forms a sheath flow with a sample liquid containing test particles and a clean sheath liquid, a sample liquid supply mechanism that supplies the sample liquid to the flow cell, and a sheath liquid that is supplied to the flow cell. In a particle analyzer including a sheath liquid supply mechanism, an imaging mechanism that repeatedly captures an image of the test particle, and a data analysis mechanism that analyzes the image and outputs a result, from the sample liquid supply mechanism to the flow cell A fixture for fixing the tube forming the path to the path from the sheath liquid supply mechanism to the flow cell,
A particle analyzer, wherein the maximum distance between the fixtures is set in the range of 5 to 27 cm. 5. A flow cell that forms a sheath flow with a sample liquid containing test particles and a clean sheath liquid, a sample liquid supply mechanism that supplies the sample liquid to the flow cell, and a sheath liquid that is supplied to the flow cell. In a particle analyzer including a sheath liquid supply mechanism, an imaging mechanism that repeatedly captures an image of the test particle, and a data analysis mechanism that analyzes the image and outputs a result, from the sample liquid supply mechanism to the flow cell A particle analysis device characterized in that a pressure fluctuation absorbing section is provided in the middle of the path. 6. The particle analyzer according to claim 1, wherein a pressure fluctuation absorbing section is provided in a path from the sample liquid supply mechanism to the flow cell. 7. The particle analysis device according to claim 5, wherein the pressure fluctuation absorbing portion is a bubble in a path, and the amount of the bubble is set in a range of 5 to 32 microliters. Particle analyzer. 8. A flow cell that forms a sheath flow with a sample liquid containing test particles and a clean sheath liquid, a sample liquid supply mechanism that supplies the sample liquid to the flow cell, and a sheath liquid that is supplied to the flow cell. In a particle analyzer including a sheath liquid supply mechanism, an imaging mechanism that repeatedly captures an image of the test particle, and a data analysis mechanism that analyzes the image and outputs a result, from the sheath liquid supply mechanism to the flow cell A particle analysis device characterized in that a pressure fluctuation absorbing section is provided in the middle of the path. 9. The particle analysis apparatus according to claim 1, 5 or 6, wherein a pressure fluctuation absorbing section is provided in the path from the sheath liquid supply mechanism to the flow cell. 10. The particle analysis device according to claim 8, wherein the pressure fluctuation absorbing portion is provided with at least one of a thin tube narrower than a path, a tube having an air chamber, or a bellows as a damping member. A particle analyzer characterized by being a thing. 11. The particle analysis apparatus according to claim 8, wherein the pressure fluctuation absorbing portion is a damping member made of a thin tube narrower than a path, and the length of the thin tube is 40 to 160c.
A particle analyzer characterized by being set in a range of m. 12. A flow cell that forms a sheath flow with a sample liquid containing test particles and a clean sheath liquid, and a sample liquid supply mechanism that supplies the sample liquid to the flow cell.
In a particle analyzer comprising a sheath liquid supply mechanism that supplies the sheath liquid to the flow cell, an imaging mechanism that repeatedly captures an image of the test particles, and a data analysis mechanism that analyzes the image and outputs a result, Characteristically, pressure fluctuation absorbing portions are respectively provided between the path from the sample liquid supply mechanism to the flow cell and the path from the sheath liquid supply mechanism to the flow cell, and the delay times of the both paths are roughly matched. Particle analyzer. 13. The particle analyzer according to claim 1, wherein a pressure fluctuation absorbing section is provided between a path from the sample liquid supply mechanism to the flow cell and a path from the sheath liquid supply mechanism to the flow cell, respectively. A particle analyzer characterized in that the delay times of the two paths are substantially the same.