JPH06288895A - Particle analyzer - Google Patents

Abstract

(57) [Summary] [Structure] A flat sheath flow is formed in a flow cell 1 having an imaging region and a particle detection region to flow the sample. Sensor 16
The particle passing position is detected to discriminate the particles passing through the imaging area, the pulse light source 21 is caused to emit light to capture a still image of the particles, and the image processor 32 analyzes the type and number of particles. The density corrector 39 corrects the number of imaged particles to obtain the density of the particles. [Effect] Since only particles passing through the imaging region are imaged and the number of particles is corrected to obtain the particle concentration, analysis can be performed with high accuracy.

Description

Detailed Description of the Invention

[0001]

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

[0002]

2. Description of the Prior Art For morphological examination of particles in urine,
In the conventional visual observation, the urine sample was centrifuged, the sediment was dyed to prepare a specimen on a slide glass, and the specimen was observed under a microscope. At that time, the concentration rate of centrifugation is always kept constant, and the amount of sample to be observed is also kept constant, so that it is possible to know what kind of sediment is contained in the original urine sample and at what concentration. It was In addition, there are particles of several μm such as blood cells and bacteria to particles of several hundred μm such as a cylinder in the sediment, and the microscope magnification is switched between high magnification and low magnification for observation. At that time, although the amount of the sample to be observed varies depending on the purpose, it is typically 5 μm at high magnification.
1, the concentrated urine in an amount corresponding to 750 μl of original urine at low magnification was observed, and the number of each sediment component was counted.

As an apparatus for automating these inspections, there is an apparatus which allows particles to be suspended in a liquid and flow in a flow cell for optical analysis. For example, special table Sho 57-500995
In the patent publication, a device is shown in which a fluid sample is passed through a channel having a special shape, in which particles in the sample are confined in a wide image forming region to create a static image. In this device, a magnified image of the flow is formed on the image pickup surface of the CCD camera using a microscope. As the light source, a pulsed light source that periodically emits light in synchronization with the operation of the CCD camera is used. Since the emission time of the pulsed light source is short, a still image can be obtained even when particles are flowing. Image analysis of the obtained static image enables morphological analysis of particles in the liquid. It is also disclosed that the particle concentration can be analyzed by counting how many particles are contained in the imaged sample volume.

Further, as a device for detecting passage of particles without periodically emitting light from a light source and capturing a still image at the timing, there is one disclosed in Japanese Patent Laid-Open No. 63-94156. In the publication, an optical system for particle detection is provided upstream of a particle passage in addition to an optical system for capturing a still image, and it is detected that the particle always crosses a light beam of a light source for a trigger that is turned on. It is described that a stationary image of particles is obtained by causing a pulsed light source to emit light after a certain delay time after detection with a detector.

Further, Japanese Patent Application No. 4-300808 describes a particle concentration analysis method in which the number of particles in an image obtained by detecting particles is counted to calculate the particle concentration. In this method, a particle detection area having the same width as the imaging area is provided upstream of the imaging area, and the width of the fluid sample flowing is equal to or smaller than the width of the imaging area. Can be used to accurately obtain the particle concentration.
Since a sample liquid having a volume significantly larger than the volume of the field of view to be imaged is flowed and particles are imaged while passing through the field of view, analysis can be performed in a short time.

[0006]

When the concentration of particles is analyzed by the method described in Japanese Patent Application No. 4-300808, an image is obtained when the fluid sample supplied to the flow cell passes outside the width of the imaging region. There was a drawback that the efficiency of analysis deteriorated because no particles were captured inside.

Further, there is a drawback that an error occurs in counting particles and the concentration analysis accuracy is lowered.

In order to prevent this, in order to allow the fluid sample to flow strictly within the width of the imaging region, it is necessary to increase the processing accuracy of the flow cell, resulting in a problem of increased manufacturing cost.

Further, in the system in which the particle detector detects that the light beam of the trigger light source which is always turned on is crossed by the particle detector and adjusts the timing, there is unevenness in the light intensity of the light source for detection in the detection area. There was a drawback that the detection sensitivity became uneven and the accuracy of the particle concentration analysis deteriorated.

Further, in the system in which the fluid sample is flown inside the imaging region, the width of the imaging region also changes when the magnification of imaging is changed, so that the width of the flowing fluid sample must be changed, and the apparatus becomes complicated. was there.

Further, there is a case where the position of the flow cell is displaced and the position where the fluid sample flows out of the imaging region, and the accuracy of the concentration analysis is lowered.

Further, when the particles detected at the first portion are imaged at the second portion after a certain delay time, the fluid sample flows at a low speed in the peripheral portion with respect to the center of the flow cell channel.
The particles flowing in the peripheral portion did not reach the second region even after the elapse of a certain delay time, and in some cases, the image was not captured.

As described above, the conventional method has a drawback that the accuracy of the particle concentration analysis is lowered because the position where the fluid sample flows out of the imaging region.

An object of the present invention is to provide a particle analyzer having a high particle concentration analysis accuracy.

[0015]

In order to achieve the above object, a particle analyzer of the present invention detects a passage of a particle in a flow cell in which a sample solution in which a particle is suspended flows and a first portion of the flow cell. And a particle detector that generates a detection signal, an imager that captures an image by capturing an image of the particle at a second portion of the flow cell corresponding to the detection signal, and the number of particles in the image by analyzing the image. In a particle analyzer having an analyzer for counting the number of particles and a particle concentration corrector for estimating the particle concentration in the sample by correcting the number of particles, the particle position corresponding to the position through which the particle detector passes. It is characterized by generating a signal.

[0016] Further, it is characterized in that the image pickup device takes an image when the particle position signal satisfies a predetermined condition.

Further, the particle detector discriminates particles passing inside and outside the width of the first portion corresponding to the width of the second portion, and when the particles pass inside, the image pickup device takes an image. It is characterized by doing.

Further, it is characterized in that the particles are classified into a plurality of particles based on the particle position signal, the particles are counted for each classification, and the particle concentration is corrected using a correction coefficient derived from the count value. is there.

Further, the particle detector irradiates a laser beam on a first portion to detect the action of the particle on the laser beam, and the magnitude of the action and the particle position signal are determined. When the relationship is satisfied, the image pickup device picks up an image.

Further, the particle detector irradiates a laser beam to a first portion to detect the action of the particle on the laser beam, and has detection terminals having a plurality of output terminals on one detection surface. It is characterized by using an element.

The particle detector irradiates a laser beam to a first portion to detect the action of the particle on the laser beam, and uses a detection element having a detection surface divided into a plurality of parts. It is characterized by that.

Further, the particle detector is characterized in that a narrowly focused laser beam is scanned on the first portion to detect the action of the particle on the laser beam.

The particle detector is for detecting the action of the particles on the light irradiated to the first portion, and has a telecentric optical system between the flow cell and the particle detector. It is a feature.

Further, the image pickup device selects one magnification from a plurality of magnifications, and the particle detection device outputs a particle position signal and a particle size signal, and the particle position signal and the particle size signal are output. The image capturing device captures an image when one of a plurality of defined conditions is satisfied, and the condition for capturing an image of a particle is changed in association with selection of a magnification.

Further, it is characterized in that the area imaged by the image pickup device is changed based on the particle position signal.

Further, the present invention is characterized in that the relative positions of the detector, the imager and the flow cell are changed based on the particle position signal.

Further, the image pickup by the image pickup device is performed with a delay time after the detection signal is generated, and the delay time changes depending on the particle position signal.

[0028]

In the analyzer of the present invention, the passage of particles is detected at the first portion of the flow cell. The particle analyzer simultaneously detects the passing position and outputs a particle position signal. The particles that have passed through the first portion flow to the second portion, but some particles may pass outside the second portion. The position where the particle passes the first part and the position where the particle passes the second part are deeply related,
It is possible to determine the condition of the particle position signal so that the particle always passes inside the second portion. Since the image is taken at the second part of the flow cell only when the particle position signal satisfies the condition, the particle is always inside the second part, and the unimaged image of the particle is unnecessarily analyzed. Since it does not occur, the efficiency of analysis can be improved.

Particles that pass outside the second part and are not shown in the image can also be detected and counted at the first part. Therefore, the number of imaged particles can be corrected to correct the particles in the fluid sample. You can get the correct concentration of.

Further, since accurate analysis can be performed even if the fluid sample does not necessarily pass through the inside of the second portion, precise processing of the flow cell is not required and the manufacturing cost can be reduced.

When a laser light source is used, the action on the laser light changes depending on the location even for particles of the same size due to uneven light intensity, but the particle position signal and the action of the particle on the laser light are detected simultaneously. By determining the imaging condition, it is possible to correct the detection sensitivity unevenness and improve the accuracy of the particle concentration analysis.

If the imaging area is reduced by changing the imaging magnification, a part of the fluid sample will protrude from the imaging area, but the inside of the imaging area will be changed by changing the conditions for determining the particles to be imaged from the particle position signal. Since only particles that pass through can be selected, there is no need to change the size of the region where the fluid sample flows, and it is not necessary to use a complicated mechanism for switching the flow.

Further, when the flowing position of the fluid sample is deviated, the particle position signal changes, so that the amount of deviation can be known, and the deviation can be corrected by changing the relative positions of the detector, the imager, and the flow cell. Therefore, high-precision measurement can always be performed in the best condition.

When the particle position signal of the particle detected at the first portion corresponds to the peripheral portion of the flow path of the flow cell, the delay time until the image is taken at the second portion is lengthened to obtain the flow velocity. It is possible to prevent an image from being taken before the particles passing through the slower part of the image reach the second part.

[0035]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A first embodiment of the present invention will be described with reference to FIGS. 1 is a perspective view of a particle analyzer, FIG. 2 is an enlarged perspective view showing a flow cell 1 of a first embodiment, FIG. 3 is a block diagram showing a basic configuration of a detection system of the particle analyzer shown in FIG. FIG. 4 is a block diagram showing a partial configuration of the detection system.

As shown in FIG. 1, the particle analyzer of the present embodiment mainly comprises an optical measuring system 80 arranged on a table 94, a calculator 37, a display 38 and a keyboard 7.
1, a pretreatment device 87 adjacent to the optical measurement system 80, a controller 31 arranged under the table 94, a waste liquid tank 81, a sheath bottle 82, a dyeing liquid bottle 83, and a washing liquid bottle 84. The pretreatment device 87 arranged on the table 94 is covered with a cover 85, and the sample disc 86 can be taken out from the inside by opening the cover 85. The display 38 for displaying the operation state of the device and the result of the analysis is arranged in the vicinity of the optical measurement system 80 so that the operator can perform work while checking the state of the device and the result of the analysis. Has become. A space is provided on the table 94 so that the sample disk 86 can be placed and the sample can be set.

The controller 31 arranged under the table 94 is attached to a rack that can be pulled out for easy maintenance, and has a waste liquid tank 81, a sheath bottle 82, a dyeing liquid bottle 83, and a washing liquid bottle 84.
Is located on the front of the device for easy removal and replacement.

The detection system of this embodiment is constructed as shown in FIGS. 2, 3 and 4.

The flow cell 1 of this embodiment has a structure as shown in FIG. The flow channel portion of the flow cell 1 is made of glass whose upper and lower surfaces are transparent, and the flow cell 1 has a sample liquid inlet 3 located at the center and a seal from the periphery thereof.
Two inlets for the sheath liquid inlet 2 are provided to allow the inlet of the sheath liquid. A detection area 50 is provided in the flow path portion, and imaging areas 10a and 10b are provided downstream thereof. 10a and 10b
Corresponds to the size of the field of view when the imaging magnification is switched.

From the sample liquid inlet 3, the sample liquid 6 containing the test particles 7 is supplied at a constant flow rate. From the sheath liquid inlet 2, a clean sheath liquid containing no particles is supplied from the sheath liquid bottle 82 at a constant flow rate. In the flow cell 1, a sheath flow, which is a steady laminar flow in which the sample liquid is wrapped with the sheath liquid, is formed, and the test particles 7 pass through the flow cell 1 at a constant speed. In this sheath flow,
The sample liquid flows with a flat cross section that is thin and wide. The flow rates of the sample liquid supply unit and the sheath liquid supply unit can be changed, and the cross-sectional shape and speed of the sample liquid 6 in the flow cell 1 can be changed.

The detection system of this embodiment is constructed as shown in FIG.

The flow cell 1 is attached to the stage 78.

A pulse light source 21, is provided on one side of the flow cell 1.
A pulse light irradiation system 75 including a lens 22, a variable diaphragm 42, and a condenser lens 11, a laser 19, a conversion lens 4
3, two optical systems of a mirror 18 and a laser light irradiation system 76 including a condenser lens 11 which is also used as a pulsed light irradiation system are arranged. In the microscope 13 on the opposite side of the flow cell 1, a half mirror 14 is arranged behind the objective lens 12, a sensor 16 is arranged on the transmission side, and a variable lens 41 and a CCD camera 15 are arranged on the reflection side.
A beam trap 48 is arranged near the objective lens 12.

An image processor 32 is connected to the CCD camera 15, and a particle position detector 51 is connected to the sensor 16. The particle position detector 51 is a position shift detector 77,
It is connected to the passing particle counter 36 and the imaging particle discriminator 52. Further, the imaged particle discriminator 52 is connected to the image processor 32 and the pulse generator 34. The image processor 32 is connected to the image pickup particle counter 53, and the image pickup particle counter 53 and the passing particle counter 36 are connected to the concentration corrector 39. Further, the clock 30 is connected to the pulse generator 34 and the CCD camera 15. Further, the pulse generator 34 is the pulse light source 2
1 and the density corrector 39 is connected to the display 38. Further, the displacement detector 77 is connected to the stage 78.

The light emitted from the pulsed light source 21 passes through the pulsed light irradiation system 75 and the imaging area 10a or 10 of the flow cell.
Irradiate the area including b uniformly.

The continuous light 25 emitted from the laser 19 irradiates the area including the detection area 50 of the flow cell 1 through the laser light irradiation system 76. The light transmitted through the flow cell 1 is removed by the beam trap 48 and does not enter the objective lens 12, but scattered light 27 scattered when the particles 7 to be examined pass through the flow cell 1 enters the objective lens 12 and the half mirror 1
After passing through 4, it is detected by the sensor 16.

The particle analyzer of this embodiment can switch between a high magnification mode and a low magnification mode.
Switching between the high-magnification mode and the low-magnification mode is performed by switching the variable lens 41 and the variable diaphragm 42 at the same time. When the variable lens 41 is switched, the size of the visual field imaged by the CCD camera 15 is switched to 10a or 10b.

The CCD camera 15 is controlled by the clock 30 and operates periodically. This CCD camera is of the interlace type and combines two fields to form one screen. The image storage periods of the field S and the field T are shifted by a field period which is half the frame period. Each field has an image storage period and a transfer period during a frame period for obtaining one image. The amount of light incident on the imaging surface during the image storage period is accumulated as electric charge, and the electric charge accumulated during the transfer period is accumulated. Transfer in a relatively short time. After the transfer, the charge amount becomes 0, and the image storage period starts again.

The structures of the sensor 16 and the particle position discriminator 51 are shown in FIG. The sensor 16 has an elongated detection surface, and electrodes are provided at both ends thereof. The longitudinal direction of the detection surface is oriented in a direction orthogonal to the flow direction of the flow cell 1. When the scattered light 27 is incident on the portion having the detection surface, a photoelectromotive force is generated, and photocurrents 29a and 29b are formed and flow to the electrodes at both ends to be extracted. Photocurrent 29a and 29b
The ratio of the magnitude of is associated with the position where the scattered light 27 is incident on the detection surface of the sensor 16. The extracted current is iv
Each of the converters 54a and 54b converts the voltage. The sum of the respective voltages is obtained by the adder 55, and the iv converter 5
The value obtained by dividing the output of 4a by the output of the adder 55 is the divider 56.
Can be obtained at. The calculation result of the divider 56 is output as a particle position signal, and the calculation result of the adder 55 is output as a particle size signal.

The image pickup particle discriminator 52 detects the moment when the output of the adder 55 peaks, and when the value of the output of the adder 55 and the value of the output of the divider 56 at that time satisfy the predetermined conditions, The signal is sent to the image processor 32 and the pulse generator 3
Send to 4. The conditions are set as follows.
That is, first, the output value of the divider 56 is a fixed upper limit,
Except when the value is out of the lower limit range. Also, adder 5
The case where the output value of 5 is out of the specified upper and lower limits is also excluded. However, the upper limit and the lower limit of the adder 55 are set high when the output value of the divider 56 is close to the center of the range, and are set low when the output value is far from the center of the range. Further, the upper limit and the lower limit of the output value of the divider 56 are set separately for the high magnification mode and the low magnification mode.

When the pulse generator 34 receives the timing signal, it generates a pulse signal after a certain delay time to make the pulse light source 21 emit light. A signal is input from the clock 30 to the pulse generator 34, and the CCD camera 1
Even if the timing signal is inputted twice or more during two consecutive field periods of 5, the pulse signal is limited to be emitted only once at the first time.

The CCD camera 15 accumulates the image of the particles 7 to be inspected passing through the image pickup area 10 as electric charge at the moment when the pulsed light source emits light, and transfers it to the image processor 32. When the image processor 32 receives the timing signal from the imaged particle discriminator 52, it analyzes the image data, morphologically classifies individual particles in the image, and counts each class.

The passing particle counter 36 is a particle position discriminator 51.
The particle size signal and the particle position signal output by are distinguished and classified into a plurality of numbers, and counting is performed for each classification.

After repeating the operation for a certain period of time, the imaging particle counter 53 and the passing particle counter 36 send the respective counting results to the concentration corrector 39. The concentration corrector 39 calculates the counting results, outputs the concentration as the classification and type of the test particles in the fluid sample, and displays it on the display 38.

The position shift detector 77 is a particle position discriminator 51.
The particle position signals from are averaged for a certain period, and when the average result is out of the predetermined range, an instruction is issued to the stage 78 to move the flow cell 1. The direction and distance of movement is determined according to the average result.

In the case of the present embodiment, when the test particles 7 contained in the sample liquid 6 pass through the detection region 50 in the flow cell 1, the continuous light 25 is scattered and the scattered light 27 is formed on the detection surface of the sensor 16. Image. The position of image formation is determined by the position in the detection area 50 through which the particles 7 to be detected pass. The intensity of the scattered light 27 corresponds to the size of the test particle 7. Therefore, the output value of the adder 55 in FIG. 4 has a peak when the test particles 7 pass through the detection region 50, and the output value of the adder 55 at that time represents the size of the test particles 7, and the divider 56 The output value represents the position of the test particle 7 in the detection region 50.

Since the image pickup particle discriminator 52 determines the condition for outputting the timing signal by the output values of both the divider 56 and the adder 55, some of the particles contained in the sample liquid 6 are in the detection region 50. The timing signal is transmitted only for particles having a certain range of sizes that pass through the range.

In the present embodiment, in the high magnification mode, the sample liquid 6 has a width wider than that of the imaging region 10a.
Some particles do not pass through the imaging region 10a because they flow through the inside. Since the detection region 50 is wider than the width of the sample liquid 6, all the particles in the sample liquid 6 pass through the detection region 50.

Since the flow is in a regular laminar flow state in the flow cell 1, the detection region 50 to the imaging region 10a,
Up to 10b, the particles flow almost parallel to the inner wall of the flow cell 1. Therefore, particles flowing in the same width as the imaging region 10a or 10b at the position of the detection region 50 are
Or flow in 10b.

By making the condition that the image pickup particle discriminator 52 outputs a timing signal correspond to the case where particles pass through the same width as the image pickup region 10a in the detection region 50, the image pickup region
An image can be taken only when particles pass through 10a.

Therefore, even if the width in which the sample liquid 6 flows is set to be wider than the width of the imaging region 10a, the imaging region 10
Particles passing outside a will not be detected and imaged, and since the particles are not reflected in the image, an error will occur in counting the number of particles in the image and an error in the concentration analysis.

Further, in the case of the low magnification mode in this embodiment, the width of the image pickup area 10b is wider than the width of the image pickup area 10a, but since the discrimination condition of the image pickup particle discriminator 52 is changed depending on the mode, the width of the image pickup area 10b is changed. The timing signal can be set to be output when the particles pass, and the particles that pass through any part of the imaging region 10b can be detected and imaged, so that a large number of particles can be imaged and the accuracy of concentration analysis can be improved. Get higher

In the case of the present embodiment, particles that do not pass through the imaging area 10 are not imaged, but all particles are detected in the detection area 50 and counted by the passing particle counter 36.
Since the passing particle counter 36 classifies the particles according to the position of the particles and the size of the particles and counts for each classification, the total number of particles in the sample liquid 6 and the number of particles that have passed through the imaging region 10 are known. The imaged particle counter 53 counts the particles in the image for each classification analyzed by the image processor 32. Therefore, the imaging particle counter 53 obtains the morphological classification of particles in the image, and the passing particle counter 36 obtains the total number of particles and the number of particles in the imaging region. Therefore, the density corrector 39 combines the information. By performing the calculation, the total number of particles for each morphological classification can be estimated, and the particle concentration can be obtained with high accuracy.

When the magnification of the microscope is switched, the depth of focus and the size of the field of view change at the same time. In this case, the particle concentration can be analyzed with high accuracy without adjusting the width of the sample liquid 6 to the imaging region 10. As a result, a special mechanism for switching the width of the sample liquid 6 is not required, and the device can be simplified. Further, to control only the thickness of the sample liquid 6 without controlling the width is to control the speed of supplying the sample liquid 6 from the sample liquid inlet 3 and the speed of supplying the sheath liquid from the sheath liquid inlet 2. It is possible, and by adjusting the thickness of the sample liquid 6 to match the depth of focus of the microscope, the out-of-focus of the imaged particles is eliminated, the accuracy of image analysis is improved, and the accuracy of concentration analysis is increased. Since the thickness of the sample liquid 6 can be adjusted to the thickness of the depth of focus without reducing the thickness, a large amount of sample can be flowed and analyzed within a fixed time, and the analysis time can be shortened. .

Further, in the case of the present embodiment, in the case of a sample having a low particle concentration, the number of particles passing through the image pickup region 10 is small, but when the particles do not pass, no image is picked up and the particle detection region is detected. Since the imaging is performed after passing through 50, the number of opportunities for imaging the particles is increased, and a large number of particles can be imaged. Further, when the flow velocity of the sample solution 6 is increased or a plurality of particles pass continuously when the particle concentration is high, the pulse generator 34 is in synchronization with the operation of the CCD camera 15 and has a 2-field cycle. Even if the timing signal is received a plurality of times in the meantime, the pulse light source 21 emits light only once at the first time. The CCD camera 15 accumulates the image of the particles illuminated by the pulsed light 26 and the image processor 3
Since the image is transferred to No. 2, an image can be obtained without multiple exposure. The number of particles captured in the image obtained in this way,
There is a non-linear relationship between the particle concentrations of the sample liquid 6,
Using the relationship, the concentration of the sample liquid 6 can be derived from the number of particles imaged. Further, the particle concentration can be more accurately estimated by correcting the number of particles passing through the inside of the field of view and the number of particles passing through the outside from the output of the passing particle counter 36. Therefore, since the particle concentration can be correctly obtained even when the sample liquid 6 is flown at a high speed, the time required for analysis can be shortened. Furthermore, since a large number of particles in a sample can be analyzed and counted at a fixed time, it is possible to analyze the particle concentration with high accuracy.

Further, in the case of this embodiment, since the irradiation intensity distribution of the continuous light 25 in the detection region 50 is as shown in FIG. 5, the magnitude of scattered light is large when the particles pass through the center, It becomes smaller when passing through both ends. Therefore, even for particles of the same size, the output value of the adder 55 is high when the particles pass through the center and low when the particles pass through both ends. However, the discrimination condition for the imaging particle discriminator 52 to output the timing signal is that the output range of the adder is high when the particles pass through the center, and the output range of the adder is low when the particles pass through both ends. Since the particle size is set, particles in a certain size range are imaged regardless of the particle passage position. Therefore, the number of particles that are imaged does not differ depending on the position where the particles pass, so that the particle concentration can be analyzed with high accuracy.

Furthermore, there is no need to use a special optical system to make the intensity distribution of the continuous light 25 uniform in the detection region 50, and the apparatus can be made inexpensive.

Further, in the case of this embodiment, the position shift detector 7
7 averages the particle position signals from the particle position discriminator 51 for a certain period of time, and when the averaged result is out of the predetermined range, a command is issued to the stage 78 to move the flow cell 1, so that the sample solution 6 When the flowing position of is shifted with respect to the microscope 13, it can be automatically corrected. Therefore, the number of particles that cannot enter the imaging region 10 and cannot be analyzed can be minimized, and analysis can always be performed with high accuracy.

Even if the stage 78 moves the microscope 13 instead of the flow cell 1, the same effect can be obtained.

FIG. 6 shows a second embodiment of the present invention.
The difference from the first embodiment is that the sensor 16b for detecting the scattered light 27 emitted by the particles is a combination of a plurality of detection elements. Although the number of elements is three in the figure, the number of elements can be increased if necessary. An electrode is emitted from each detection element, and the passage position of the particle can be known depending on which electrode outputs a signal.

In the case of this embodiment, when the scattered light is detected by the central element, the particles pass through the inside of the imaging area 10, and when the scattered light is detected by the outside element, the particles are outside the imaging area 10. Since it passes, the particles can be easily discriminated by imaging only when the particles are detected by the central element, and the circuit for the discrimination can be simply configured.

Further, in the case of this embodiment, even when a plurality of particles pass through the detection area 50 at the same time, the imaging area 10
As long as there are no particles passing through the inside, scattered light does not irradiate the central element of the sensor 16b, so that it is possible to analyze with high accuracy without erroneously capturing an image.

FIG. 7 shows a third embodiment of the present invention.
The difference between the first embodiment and the second embodiment is that the sensor 16c that detects the scattered light 27 emitted by the particles is a combination of a large number of detection elements, and the light intensity distribution on the detection surface is determined by scanning periodically. Is to detect.

In the case of this embodiment, even if a large number of particles are simultaneously passing through the detection region 50, they can be detected separately, so that the passing particle counter 36 can count correctly. The accuracy of particle concentration analysis can be improved.

Further, in the case of this embodiment, not only the average position where the particles are passing but also the width of the particles can be known, and the particles to be analyzed can be discriminated and imaged more accurately. , The efficiency of analysis can be increased.

As means for detecting the passage position of the particles, in addition to the method using the sensor described above, a small spot of the laser beam is used, and scanning is performed at high speed in the detection area 50 to change the intensity of scattered light. A method of detecting can also be used.
In this case, since the laser beam is narrowed down, the intensity of scattered light is increased, and there is an advantage that even small particles can be detected. Further, since the laser beam is scanned, the detection area 50
It also has the advantage that the sensitivity of particle detection does not change at any position inside.

FIG. 8 is a diagram showing a detection optical system according to the fourth embodiment of the present invention. In this case, a telecentric optical system in which a mask 23 is provided at the back focal position of the objective lens 12 is adopted.

In the case of this embodiment, even if the position where the particles pass in the flow cell 1 is shifted in the optical axis direction, the image formation of the scattered light causes blurring and the image formation position does not change. Therefore, the position where the particles pass can be detected without being affected by the positional deviation in the optical axis direction, and the particle concentration can be accurately analyzed.

FIG. 9 shows the flow in the flow cell of the fifth embodiment of the present invention. In this embodiment, there is a distance between the detection area 50 and the imaging area 10. The flow in the flow cell has a flow velocity distribution 88, which is fast in the center and slow in the periphery. The pulse generator 34 receives the timing signal from the imaging particle discriminator 52, and after the delay time, the pulse light source 21
In this embodiment, the delay time is controlled by the passage position of the particles, and the delay time is shortened when the particles flow in the central part and is increased when the particles flow in the peripheral part. .

In the case of this embodiment, even if there is a flow velocity distribution in the flow cell, it is possible to take an image at the moment when the particles pass through the imaging region 10, so the number of particles to be taken becomes large,
Highly accurate particle concentration analysis is possible. In addition, since there is no particle imaging error due to the flow velocity distribution, the distance between the detection region 50 and the imaging region 10 can be lengthened, and the continuous light 25 can be prevented from affecting the imaging. It is possible to analyze with high accuracy.

In addition, the sample liquid 6 is applied to the peripheral portion where the flow velocity is slow.
Since the analysis can be performed by flowing the liquid, the width of the flow path of the flow cell can be narrowed, and the required amount of the sheath liquid can be reduced. By reducing the amount of sheath liquid used, the sheath bottle 82 and the waste liquid tank 81 can be reduced in size,
The device can be downsized.

Another embodiment of the present invention uses a CCD camera 15 having a wide field of view. The image processor 32 inputs a particle passage position and performs image processing only on that area, and performs particle morphology analysis and particle counting.

In the case of this embodiment, since the image processing is performed only on the part where the particles are present, the time required for the image processing can be short and the time required for the analysis can be shortened.

Further, in the case of this embodiment, since the image processing only needs to be performed on a narrow area, the capacity of the image processing apparatus can be small and the image processing speed can be slow, so that the apparatus can be constructed at low cost. be able to.

[0085]

According to the present invention, a particle detector detects a position where a particle passes, and an image is analyzed in accordance with the timing of the particle passing through the image pickup area to analyze the image, and the number of the imaged particles is corrected. Since the particle concentration is analyzed in this manner, it is possible to provide an analyzer that can analyze the particle concentration with high accuracy.

[Brief description of drawings]

FIG. 1 is a perspective view showing the overall structure of a first embodiment of the present invention.

FIG. 2 is a perspective view showing the configuration of a flow cell.

FIG. 3 is a block diagram showing a basic configuration of a detection system according to the first embodiment of the present invention.

FIG. 4 is a block diagram showing a basic configuration of a detector of the first embodiment.

FIG. 5 is a characteristic diagram showing a laser intensity distribution.

FIG. 6 is a sectional view showing the structure of a detector according to a second embodiment.

FIG. 7 is an explanatory view showing the structure of the detector of the third embodiment.

FIG. 8 is an explanatory diagram showing a configuration of a detection system of a fourth embodiment.

FIG. 9 is an explanatory view showing the flow in the flow cell of the fifth embodiment.

[Explanation of Codes] 1 ... Flow cell, 6 ... Sample solution, 7 ... Test particle, 10
... Imaging area, 12 ... Objective lens, 13 ... Microscope, 15 ...
CCD camera, 16 ... Sensor, 19 ... Laser, 21 ... Pulse light source, 32 ... Image processor, 34 ... Pulse generator, 3
7 ... Calculator, 38 ... Display, 39 ... Concentration corrector, 50 ...
Detection area, 78 ... Stage.

Claims (1)

[Claims] 1. A flow cell in which a sample solution in which particles are suspended flows, a particle detector that detects passage of the particles at a first portion of the flow cell and generates a detection signal, and the particle detector corresponding to the detection signal. An imager that captures an image of the particles at a second portion of the flow cell to obtain an image, an analyzer that analyzes the image to count the number of particles in the image, and a particle in the sample that corrects the number of particles. A particle analyzer having a particle concentration corrector for estimating the concentration, wherein the particle detector generates a particle position signal corresponding to a position where the particle passes.