JPH075093A - Flow type particle image analyzer - Google Patents

Abstract

PURPOSE:To realize switching of a magnification of imaging in each measuring mode by a simple mechanism in a particle image analyzer for imaging a still image of a particle suspended in flowing liquid and analyzing the particle. CONSTITUTION:A sample flows to a flow cell 202, passage of particles is detected via a laser light 211, a pulse lamp 205 emits a light based on the detection, a still image of the particle is picked up by a TV camera 207, and the image is processed. In this case, a magnification change-over lens 208 is inserted in response to a measuring mode.

Description

Detailed Description of the Invention

[0001]

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

[0002]

2. Description of the Related Art Conventionally, cells and particles existing in blood and urine have been classified by preparing a sample on a slide glass and observing with a microscope. In the case of urine, since the particle concentration in urine is low, the sample is observed after being centrifugally concentrated by a centrifugal separator.

As an apparatus for automating these observation and inspection operations, in the case of a sample sample such as blood, it is set on a microscope after being spread on a slide glass, and the microscope stage is automatically scanned so that particles are present. There is a method in which the microscope stage is stopped at a position, a still image of a particle is photographed, and a feature extraction by an image processing technique and a pattern recognition method are used to classify particles in a sample.

However, in the above method, it takes time to prepare a sample, and it is necessary to find particles while mechanically moving the microscope stage and move the particles to an appropriate image capturing area. Therefore, there are problems that analysis takes time and the mechanical mechanism becomes complicated.

A flow cytometer method is known in which a smear sample is not prepared, but a sample sample is allowed to flow in a flow cell while being suspended in a liquid, and is optically analyzed. The method using a flow cytometer is for observing the fluorescence intensity and scattered light intensity from each particle in a sample.
It has a processing capacity of 00. However, it is difficult to observe the feature amount that reflects the morphological features of the particles, and it is impossible to classify the particles by the morphological features that have been conventionally performed under a microscope.

Further, when urine is used as a sample, the urinary sediment examination method for classifying urinary sediment components of cells and particles present in urine requires many procedures as follows.

In the case of urine, since the concentration of formed components in urine is low, it is necessary to centrifuge and concentrate the sample in advance with a centrifuge before testing. First, urine collected from a patient is collected from a urine cup into a Spitz tube, the urine sample is centrifuged, and the precipitate is unstained or stained to prepare a sample on a slide glass. After that, the specimen is observed under a microscope to classify the sediment components that can be observed within a certain visual field. At that time, it is possible to know what kind of sediment component is contained in the original urine sample and at what concentration by keeping the concentration rate of centrifugation constant and the amount of the sample to be observed constant.

Since the sediment components include particles of several μm such as blood cells and bacteria to particles of several hundred μm such as a cylinder, it is necessary to observe all particles with a constant microscope magnification. I can't. Therefore, it is necessary to switch the magnification of the microscope between high magnification and low magnification and observe the particles at the respective magnifications.

Since many such operations are required, the formed material is destroyed and the concentration is varied. Also, because the number of particles to be visually observed is small and there are many working processes,
It will put a great burden on the laboratory technologist.

Therefore, recently, as disclosed in Japanese Patent Application Laid-Open No. 3-105235, as a device for automating these observation and inspection operations, a smear sample is not prepared and a sample sample is suspended in a liquid. , A sample sample is passed through a flow cell having a specially shaped channel, in which the particles in the sample are flowed through a wide imaging area, a static image is taken by a flash lamp, and the particle analysis is performed using the image. ing. When a magnified image of sample particles is projected on a CCD camera using a microscope, a flash lamp, which is a pulse light source, periodically emits light in synchronization with the operation of the CCD camera. The light emission time of the pulse light source is short, a still image can be obtained even when particles are continuously flowing, and the CCD camera can take 30 still images per second.

In Japanese Patent Laid-Open No. 3-105235, two types of measurement modes, a high magnification measurement mode for measuring small particles and a low magnification mode for measuring large particles, are provided in the same way as the conventional method, and during measurement It is switching. However, since the means for switching the magnification realizes the image of the objective lens by using a projection lens corresponding to two kinds of different magnifications, one for high magnification and the other for low magnification, two kinds of lenses are required. Further, in the high magnification mode, it is necessary to lengthen the optical path to adjust the magnification, which increases the number of parts such as a reflection mirror and complicates the switching mechanism.

[0012]

When the particles in the urinary sediment component are imaged, since the particle sizes are various,
In order to efficiently perform the number and classification of plural types of particles in a sample, it is necessary to change the measurement mode and change the magnification of the particle image to be acquired.

However, as described in the prior art, the method of switching between high magnification and low magnification requires two types of projection lenses with different magnifications, which has a drawback that the switching mechanism becomes complicated.

When the imaging magnification is changed,
The amount of light on the image pickup surface of the TV camera changes, and the image signal level changes greatly. Therefore, it is necessary to control the amount of light incident on the TV camera.

An object of the present invention is to provide a method of switching the imaging magnification between high-magnification and low-magnification measurement modes without exchanging the objective lens. And the switching mechanism is simply configured.

[0016]

To achieve the above object, in a particle image analyzer having a plurality of measurement modes having different particle detection systems and different sample liquid flow conditions, this occurs when switching the measurement modes. It is intended to provide means for solving the problem of changing the optical system, the mechanical system, and the image processing system.

To this end, the magnification switching lens of the optical system has only a low magnification, and the switching mechanism can be easily realized.

Further, in order to achieve the above object, a mechanism for controlling the light emission amount of the light source and for adjusting the light amount is provided in the optical system.

[0019]

According to the present invention, as a method of switching the imaging magnification between the high magnification and the low magnification measurement modes, the magnification switching lens is only a low magnification, so that the number of types of lenses to be inserted is reduced and the switching mechanism is simplified. It is composed of.

[0020]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment of a flow type particle image analysis apparatus according to the present invention will be described below with reference to the drawings. FIG. 1 is a basic configuration diagram of a flow type particle image analysis apparatus including an embodiment of the present invention.

The flow type image analysis apparatus of the present invention comprises a sample supply unit 1, an analysis unit for detecting a formed component and acquiring an image, an image processing unit for processing the acquired image 3, and data for analyzing image data. It is composed of a processing unit 4.

Next, as an embodiment of the present invention, an analysis unit of a urine sediment inspection apparatus for analyzing particle components in urine will be described in detail with reference to FIG.

The sample collected from the sample container 11 set in the sample supply unit 1 by the sample nozzle 12 is discharged to the dyeing tank 13, and then dyed by the dyeing liquid 14 discharged in a fixed amount. The dyed sample flows stably in the center of the flow cell so as to be surrounded by the sheath liquid 15 so as to form a flat sample flow in the central portion of the flow cell 202.

A flash lamp 205 which is a light source of a microscope
The emission timing of is controlled by a detection signal emitted by the particles passing through the particle detection area. The semiconductor laser 211 is always lit and is always focused on the particle detection region in the sample. The scattered light when the particles to be measured pass through the detection area is reflected by the beam splitter 213 and converted into an electric signal according to the scattered light intensity in the particle detector 212. If the particle detection electric signal has a predetermined signal level or higher, it is determined that the particles to be image-processed have passed, and when the particles reach a predetermined position in the image pickup area of the TV camera, the flash lamp 2
It is controlled so that 05 lights up. This is determined by the distance between the particle detection area and the imaging area and the sample flow velocity.

On the other hand, the pulsed light emitted from the flash lamp 205 travels on the optical axis of the microscope, passes through the condenser 210, and is focused on the sample in the flow cell 202. The pulsed light applied to the sample flow in the flow cell 202 is condensed by the microscope objective lens 206, projected on the CCD image forming position 209 in the TV camera 207, and converted into an image data signal. At this time, the magnification of the particle image is controlled by the magnification switching lens determined by the measurement mode. Further, since the flash lamp 205 emits light for a short time, the image of the particles passing through the imaging area is captured by the TV camera 207 as a still image. Still images are
The image processing unit 3 classifies particles. The data and the number of the classified data are combined into a final result by the data processing unit 4 after the measurement is completed.

Next, an embodiment of the magnification switching system of the present invention will be described in detail with reference to FIGS.

Normally, when changing the magnification of the microscope, the magnification of the objective lens is changed, but the present invention is characterized in that the imaging magnification is changed without changing the objective lens.

In the high magnification mode, which is a measurement mode for picking up images of small particles, the magnification is determined by the magnification of the objective lens 206, and the image formation of the particles by the objective lens is performed by the TV camera 20.
No. 7 CCD image pickup surface 209 is directly projected. Therefore, the magnification switching lens 2 for adjusting the magnification
It is not necessary to insert 08.

In the low-magnification mode, which is a measurement mode for imaging large particles, a magnification switching lens 208 is inserted between the objective lens 206 and its image plane to reduce the magnification from the high magnification, and the TV camera 207 has a smaller magnification. CCD image forming position 20
9 is projected. In this case, the objective lens and TV
Since the position of the camera is configured in the same way as the high magnification, it is not necessary to change the optical path length, and it is not necessary to readjust the focus.

Next, a light quantity control method on the TV camera image plane 209 will be described. This can be realized by controlling the output voltage of the flash lamp control unit 204 so as to control the amount of light suitable for that mode when the measurement mode changes.

As another method, as shown in FIG. 4, an optical filter 214 is added to the front surface or the rear surface (the front surface in the figure) of the switching lens 208 inserted in the low magnification mode, and the light amount is adjusted at the same time as the lens switching. It is possible to adjust the light amount at low magnification.

The flow-type particle image analyzer of the present invention is used for classification and analysis of biological samples and cells suspended in a liquid, blood cell components such as red blood cells and white blood cells in blood, or urinary sediment components present in urine. It is valid. In particular, when counting the number of particles of urinary sediment components in urine and classifying particles, the number of particles existing in each sample may differ by several digits or more, so image processing should be performed on the particles detected by the particle detection means. Is effective in knowing the information on the number of particles in the sample liquid.

[0033]

Since the flow-type particle image analysis apparatus of the present invention has the configuration described above, it has the following effects.

In the high magnification mode, the magnification switching lens is not necessary because the image forming position of the objective lens coincides with the image pickup surface of the TV camera. In the low magnification mode, since the magnification switching lens is inserted between the objective lens and its image forming surface, one type of magnification switching lens can be configured, so that a simple optical system can be realized. Further, since the light amount adjustment is controlled by the same mechanism as the magnification switching lens, it can be realized by a simple mechanism.

[Brief description of drawings]

FIG. 1 is an overall configuration diagram of a flow type particle image analysis device according to the present invention.

FIG. 2 is a configuration diagram of a urine sediment analysis unit in FIG.

FIG. 3 is a diagram showing a magnification switching means (high magnification).

FIG. 4 is a diagram showing a magnification switching means (low magnification).

[Explanation of symbols]

1 ... Sample supply unit, 2 ... Analysis unit, 3 ... Image processing unit, 4
... data processing unit, 11 ... sample container, 12 ... sampling nozzle, 13 ... dyeing tank, 14 ... dyeing liquid, 15 ... sheath liquid, 202 ... flow cell, 204 ... flash lamp control unit, 205 ... flash lamp, 207 ... TV camera , 208 ... Magnification switching lens, 209 ... CCD, 21
0 ... Condenser lens, 211 ... Semiconductor laser, 212
... Particle detector, 213 ... Half mirror, 214 ... Optical filter.

Claims (6)

[Claims] 1. A particle sample suspended in a liquid is flown in a flow cell as a flat flow, particles in the particle sample passing through a particle detection region in the flow cell are detected, and detected in an imaging region in the flow cell. A still image of particles is captured, and in a flow-type particle image analyzer that performs morphological classification of particles by image-analyzing the captured particle image, depending on the flow condition of the measurement sample, it has a plurality of measurement modes, A flow-type particle image analyzer having a mechanism capable of realizing a plurality of imaging magnifications according to respective measurement modes. 2. The flow type particle image analyzer according to claim 1, wherein there are two types of imaging magnifications, high magnification and low magnification, depending on the measurement mode. 3. The flow type particle image analysis device according to claim 1, wherein the amount of image pickup light can be changed depending on the measurement mode. 4. The flow type particle image analysis device according to claim 1, wherein the measurement target is a biological cell. 5. The flow-type particle image analyzer according to claim 1, wherein the measurement target is a blood cell component existing in blood. 6. The flow-type particle image analyzer according to claim 1, wherein the object to be measured is urine, particularly a urinary sediment component present in urine.