JPH04110639A - Particle fractionating apparatus - Google Patents

Abstract

PURPOSE:To ensure the fractionation of particles with a simple structure by discharging particle floating liquid through an opening, measuring the property of the particles, and discharging the fluid to a passing path so that the discharged liquid droplets are displaced in response to the result of measurement. CONSTITUTION:Discharged liquid droplets containing cells undergo inelastic collision by the discharge of the fractionating liquid droplets from discharge nozzles 18. The falling direction is changed. Thus, the liquid droplets are dropped not into a chamber E but into a chamber C in a separate pickup container 19 and sampled. At this time, when there is dispersion in masses of the cell liquid droplets, the dropped position is deviated to the right and left directions. The droplets are fractionated and sampled in five kinds of A - E in response to the properties of the cells. The time period from the time point when the liquid droplets which are discharged from a discharging nozzle 1 pass a part to be measured to the time point when the droplets reach the fractionating point is considered to be the constant specified time when the discharging speed is stable. Therefore, the specified time obtained by subtracting the time until the liquid reaches the fractionating point from the nozzle 18 from this time is stored. When the stored time is elapsed from the detection of the light, the nozzles 18 are driven. Thus, the discharged liquid can be made hit on the liquid droplets containing the cells accurately.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a particle sorting device that separates and measures individual analyte particles in a sample liquid, and sorts the analyte particles based on the results.

[Prior Art] As an example of a conventional particle sorting device, a device as shown in FIG. 4 has been put into practical use under the name of cell sorter.

A sample liquid and a sheath liquid, which are cell suspension liquids, are stored in a sample container 1 and a sheath container 2, respectively, and are pressurized by a compressor or a nitrogen gas cylinder and a regulator, etc., and guided to a nozzle 5, from which they are ejected into the atmosphere as a trickle 6. Due to the vibration of the vibrator 7 attached to the nozzle 5, the trickle 6 later turns into droplets 8 and falls. The trickle 6 is irradiated with laser light from a laser light source 9, and the intensity of scattered light and fluorescence emitted from the cells in the trickle are measured by photodetectors 14 and 17. From this result, the properties of the cells are analyzed in real time, and depending on the results, a charging means (not shown) applies a positive, negative, or zero charging voltage to the fluid, so that the droplet 8 is positively or negatively charged. Charged negatively or to zero. High-voltage electrostatic deflection plates 26a and 2 are placed on the falling trajectory of the droplet.
6b are arranged facing each other, and the falling cells fim are deflected in a direction according to their electric charge, and are placed in different containers 2728.
．． 29 and was collected. In this way, cells can be sorted and collected according to their properties.

However, in the conventional particle sorting device described above, the target cells must be charged just before they become droplets, requiring delicate adjustments and making it difficult to maintain stable operation at all times.

In addition, high voltage must be handled, which poses the problem of danger.

Also, basically positively charged things, negatively charged things,
It can only be separated into three types, although they are not charged in either direction. It is possible to change the charging voltage into two types, strong and weak, and divide it into five types using the magnitude of the deflection depending on the strength of the charge, but even if charging can be performed accurately, the size of the droplet, that is, the mass, will always be the same. There is a problem that the amount of deflection is not necessarily constant, and as a result, the amount of deflection changes, and accurate classification cannot always be performed.

Therefore, in order to solve these problems, the applicant of the present invention proposed a new particle sorting device that can reliably separate many types of particles with a simple configuration in Japanese Patent Application No. 1-264553.

[Object of the Invention] The object of the present invention is to further improve the device proposed above, and to provide a device that can obtain the same effect with a simpler structure without using a large-scale mechanism such as a pressurizing mechanism. .

[Means for achieving the object] The particle sorting device of the present invention that achieves this object has a particle thickness i
! i! In a particle sorting device that separates individual particles in a liquid according to their properties, means for discharging a small amount of particle suspension containing individual particles from an opening by the action of discharge energy, and measuring the properties of the individual particles. and a means for ejecting fluid 8 toward a path through which the ejected droplet passes so as to displace the ejected droplet, depending on the result of the measurement.

[Examples] Examples of the present invention will be described in detail below with reference to the drawings. FIG. 1 is a diagram showing a configuration diagram of an embodiment of the present invention. Note that the same reference numerals as in FIG. 4 of the prior art example represent the same or similar members.

In this example, the particle suspension to be separated is a cell suspension such as blood, but the target object is not limited to this.

In FIG. 1, the particle suspension liquid is guided to a droplet discharge nozzle 1, and by the droplet discharge nozzle 1, droplets 8 containing individual particles are discharged downward and fall. FIG. 2 shows the details of the discharge mechanism inside the discharge nozzle 1, and (a) to (d) of the same figure show that heat energy is applied by the heater 2 to generate air bubbles 3, and droplets 4 containing cells S are formed. The process up to discharge is shown. A more detailed explanation is provided in Japanese Patent Application No. 1-26 filed by the applicant.
No. 0707.

A measuring optical system is arranged on the falling path of the droplet 8, and around the path below it, ejection nozzles 18a to 18d, which have the same principle as the above-mentioned ejection nozzle 1, are provided point-symmetrically in four directions. ing. It is installed facing each discharge port or sorting point so that it intersects at one point (hereinafter referred to as the "sorting point") the axis of the fall path of the droplet containing the sorting liquid or cells discharged from each discharge nozzle. . Note that each discharge nozzle is directed slightly downward toward the separation point to prevent the discharged liquid from splashing onto the opposing nozzle and to improve sampling stability.

A circular sorting container]9 is arranged below the discharge nozzle 18. The preparative container 19 is divided into five chambers, with a small chamber E in the center and four chambers A, B, and C in a circular ring around it.
, D.

These four chambers correspond to the directions of the four discharge nozzles 18a to 18d.

The laser light emitted from the laser light source 9 is focused into an elliptical shape by cylindrical lenses 10 and 11, and is irradiated onto droplets containing individual cells passing through the test area. At this time, scattered light and fluorescence are generated from the cells due to the passage of the droplet. This scattered light and fluorescence are collected by light receiving lenses 13 and 15,
The intensity or pressure is detected by photodetectors 14 and 17, respectively. A beam stopper 12 is provided in front of the light receiving lens 13 to block direct light from the laser light source, and only scattered light is detected by the photodetector 14.

Further, an optical filter 16 that transmits only fluorescence wavelengths is placed behind the light receiving lens 15, and a photodetector 17 that transmits only fluorescence wavelengths is placed.
Detected in

Next, the operation of the apparatus of this embodiment will be explained.

In an arithmetic circuit (not shown), cell properties such as cell size, type, and properties are analyzed in real time from detected values obtained each time each cell passes. Examples of analysis methods include:
Common methods include determining the size of cells according to the intensity of scattered light, or staining cells in advance with a fluorescent reagent and observing the generation of fluorescence to determine the nature and type of cells.

A control circuit (not shown) selects a predetermined discharge rate among the four types according to the desired property conditions such as the size, type, or properties of the cells determined based on the results of analysis performed in real time by the arithmetic circuit. Control the nozzle to drive or not to drive at all.

If the ejected droplet containing cells falls as it is, it will fall into the sorting container 19.
When a droplet falls into the chamber E or is discharged from the discharge nozzle 18, as shown in FIG. 2, it collides completely inelastically with the droplet containing the target cells, Change the falling direction of the cell fluid layer. As a result, the droplets fall into the chamber C of the separation container 19 instead of the chamber E and are collected. At this time, if there is some variation in the mass of the cell droplets, the dropping position may shift slightly in the horizontal direction of the page, or the cell droplet may be placed in the room A around the sorting container 19.
Since B, C, and D have a large area, they can be reliably separated. In this way, cells can be collected into five types, A to E, according to their properties.

From the time when a droplet containing individual cells discharged from the discharge nozzle 1 passes through the test area, that is, when scattered light or fluorescence is generated and detected, until the droplet reaches the sorting point. The time is considered to be a constant predetermined time T1 as long as the ejection speed is stable. Therefore, from this T1, a certain predetermined time T3 (
=TI 72) is stored, and the discharge nozzle 18 is driven when a time T3 has elapsed since light was detected. This allows the ejected liquid to accurately hit the droplet containing the targeted narrow chest.

In addition, in the above embodiment, four discharge nozzles 18 for sorting were provided to perform five types of sorting, but the number of discharge nozzles is not limited to this, but n (n is an integer of 1 or more). ) can perform classification of n+1 types.

Further, as the droplet discharge nozzle 1 or 18, an on-demand type droplet discharge nozzle using an electrostrictive vibrator may be used instead of one that discharges the droplet using thermal energy as described above.

[Effects of the Invention] According to the present invention, particle separation can be reliably performed with a simple configuration without using a large-scale device such as a pressure mechanism.

[Brief explanation of drawings]

FIG. 1 is a configuration diagram of an embodiment of the present invention, Figure 2 is an explanatory diagram of the principle of separation. Figure 3 is an explanatory diagram of the principle of droplet ejection, Figure 4 is a configuration diagram of the conventional device. The main symbols in the diagram are: 1... Particle suspension liquid discharge nozzle, 9... Laser light source, 18...discharge nozzle for separation, 19...Preparative container (C) (b) (d)

Claims (1)

[Claims] (1) In a particle sorting device that separates individual particles in a particle suspension according to their properties, means for discharging from an opening a minute amount of a particle suspension containing individual particles by the action of discharge energy; A particle sorting device comprising: means for measuring the properties of the liquid; and means for ejecting a fluid toward a path through which the ejected droplets move so as to shift the ejected droplets according to the measurement results. .