JPS63151855A - Flowcell/nozzle - Google Patents

Abstract

PURPOSE:To secure a sorting with a stable photometry, by building up a jet nozzle of a light transmitting material to irradiate the coaxial flow of a sample liquid and a sheath liquid flowing therethrough with a laser beam. CONSTITUTION:A flowcell/nozzle 22 is mounted at the lower end of a flow chamber 21, which is provided with a sample tube 23 and a sheath tube 24. The tip of the sample tube 23 is extended to the center of the chamber 21, a sample suspension 25 is wrapped with the sheath liquid 26 and jetted from the nozzle 22 in a coaxial flow 27. The nozzle 2 is built up of a light transmitting material and irradiated with a laser beam 32 to measure optical characteristic of fine particle of the sample suspension. An ultrasonic vibrator 20 is provided at the upper end of the chamber 21 to turn the coaxial flow 27 from the nozzle 22 into drops 28 and sorting of the sample particle and the liquid is performed with a deflection plate to which a voltage is applied. This achieves the stability of photometry as well as the sureness of sorting by employing the flowcell/nozzle 22 made of a light transmitting material.

Description

Detailed Description of the Invention The present invention relates to a flow cell nozzle used in a cell sorter that analyzes the optical properties of microparticles such as cells and sorts the cells according to the properties, and a cell sorter equipped with the nozzle. It relates to a flow cytometer.

Before or after a coaxial flow containing a sample suspension wrapped in a sheath liquid is ejected from a jet nozzle, it is irradiated with laser light, and the sample passes through the laser light to emit scattered light, fluorescent light, etc. In addition to measuring and analyzing each cell in the sample, the entire jet stream is vibrated to form droplets, and based on the above measurement and analysis, the droplets are charged and charged at the droplet formation point. The so-called cell sorter, which deflects and separates each charged droplet by passing it between deflection plates to which a high voltage is applied, can separate and collect cells at high speed based on the optical characteristics found as a result of analysis, so it is useful for general cell analysis and DNA analysis. Various research on cells such as measurement, chromosome measurement, cell fusion, etc. 1. It is widely used in the fields of cytology (blood cell tests), medical diagnosis such as cancer screening, and gene biology.

In the so-called in-air measurement method shown in Figure 1, two methods are used to irradiate the sample with laser light using a conventional cell sorter. A coaxial flow 3 containing a liquid 1 surrounded by a sheath liquid 2 is ejected from a jet nozzle 4 of a vibrated flow chamber, and the ejected flow is irradiated with a laser beam 5 to perform optical measurement. The charged droplets are divided into droplets 6 and charged according to the optical measurement, and are distributed by the deflection plate 7. In the figure, 8 is a sample cell, 9 is a container for collecting separated sample cells, and 10 is a waste liquid receiver. However,
In this way, with the method of shining light on the part of the liquid column ejected from the jet nozzle, minute distortions and vibrations on the surface of the liquid column cause
The optical signal is easily affected, and in order to obtain good data,
Cleaning of the nozzle and optical adjustment to avoid optical noise are required.

On the other hand, in an example of the so-called flow cell method shown in FIG.
A laser beam 5 is irradiated onto the coaxial flow in the middle of the first phase, and optical measurement is performed. The coaxial flow exiting the flow cell 11 is ejected into droplets and fractionated in the same manner as described above. In the figure, 12 is a piezoelectric element for applying vibration. However, with this method of shining light in a flow cell and then narrowing the flow and ejecting it as a jet, there are fewer problems with measurement, and it is easy to obtain good data with good reproducibility. Since the flow is constricted later, the moving speed of the sample particles tends to vary, and the distance between the measurement point and the droplet point is also larger than in the above-mentioned in-air measurement method, making it difficult to sort (
The reliability of preparative separation becomes considerably worse.

Therefore, an object of the present invention is to provide a flow cell nozzle that eliminates the drawbacks of the two jet methods described above and takes advantage of their advantages, thereby achieving both stability of optical measurement and reliability of sorting.

In order to solve the problems of the conventional technology and achieve the above objectives,
The flow cell nozzle according to the present invention is used in a cell sorter by forming the jet nozzle with a light-transmitting material to have the function of a flow cell, and by irradiating the coaxial flow of sample liquid and sheath liquid passing through the jet nozzle with a laser beam, it is possible to perform optical measurement. It is characterized by the following.

The following is a third embodiment of the flow cell nozzle according to the present invention.
This will be explained based on FIGS.

FIG. 3 shows a flow chamber 21 portion of a cell sorter to which the flow cell nozzle of the present invention is applied, and the flow cell nozzle 22 of the present invention is attached to the lower tip of the flow chamber 21. In the figure, 23 and 24 are a sample tube and a sheath liquid tube, respectively. The tip of the sample tube 23 extends to the center of the flow chamber 21, and a coaxial flow 27 in which a sample suspension 25 is surrounded by a sheath liquid 26 flows through the nozzle 2.
A droplet 28 is ejected from the droplet 2 and falls a certain distance.
become. Note that 20 is an ultrasonic vibrator.

An enlarged vertical cross-sectional view of an example of the flow cell nozzle 22 is shown in FIG.
As shown in the figure. That is, in the present invention, the jet nozzle portion for forming the jet flow is formed separately from the main body of the flow chamber 21 using a light-transmitting material, and the flow cell is provided with a central flow path 29, an entrance surface 30, and an exit surface 31. Dual-purpose nozzle 2
2. This nozzle 22 is fixed to the lower end of the flow chamber 21 by, for example, welding, and generates a jet flow as a jet nozzle, and also functions as a flow cell by irradiating a sample passing through the nozzle with a laser beam 32.

Next, an example of the shape of the flow cell nozzle 22 will be described.

The nozzle 22 shown in the partial vertical cross-sectional view of the nozzle in FIG. 5 has a recess 33 on the lower end surface, so that the length of the flow path 29 is shortened.

The nozzle 22 shown in the cross-sectional view of FIG. 6 has a cylindrical shape, and has a concentric flow passage 29 formed in the center. If the refractive index of the material forming the nozzle satisfies the relationship: nozzle refractive index>sample/sheath liquid>air, and if the inner and outer diameters are appropriately selected, the laser beams 32 that are incident in parallel can be emitted in parallel. . The advantage is that it is easy to measure forward small-angle scattered light.

The nozzle 22 shown in the cross-sectional view of FIG.
is incident. This example has the advantage that even if the sample flows away from the center of the nozzle, measurements can be performed under the same conditions by moving the nozzle in parallel.

The nozzle 22 shown in the cross-sectional view of FIG. 8 is also approximately rectangular, but
The flow path 29 has a circular cross section. When the convergent laser beam 32 is incident on the nozzle of this example, there is an advantage that the reflection loss of the incident light on the nozzle surface can be reduced.

As described above, according to the present invention, stable measurement can be performed in a measurement area surrounded by a hard light-transmitting wall surface such as a flow cell, and the sample and sheath liquid passing through the measurement area can be coaxially Since the flow remains as it is (without narrowing the sample or changing the flow velocity) and becomes a jet flow and becomes droplets, it is possible to obtain the same sorting reliability as in the air measurement method.

In other words, by obtaining a nozzle that has the functions of both a jet nozzle and a flow cell, the drawbacks of the conventional air measurement method in terms of light measurement and the drawbacks in sorting of the flow cell method can be solved simultaneously, resulting in stable light measurement. It is possible to achieve both accuracy and reliability of sorting.

[Brief explanation of the drawing]

Fig. 1 shows a conventional in-air measurement method, Fig. 2 shows a conventional flow cell method, Fig. 3 shows a flow chamber part equipped with a flow cell nozzle of the present invention, and Fig. 4 5 to 8 are enlarged longitudinal cross-sectional views of the nozzle of the present invention, which are cross-sectional views showing examples of different shapes of the nozzle of the present invention. 21... Flow chamber, 22... Flow cell nozzle, 25... Sample suspension, 26... Sheath liquid, 27... Coaxial flow, 28... Droplet, 29... Distribution Path, -32...Laser light.

Claims (1)

[Claims] 1. A coaxial flow containing a sample suspension surrounded by a sheath liquid is irradiated with laser light before or after it is ejected from a jet nozzle, and scattered light is emitted when the sample passes through the laser light; In addition to analyzing each cell in the sample by measuring fluorescence, etc., the entire jet stream is vibrated to cause droplets to fall, and based on the above measurement and analysis, an electric charge is applied to the droplets at the droplet formation point. In the cell sorter, the jet nozzle is made of a light-transmitting material to have the function of a flow cell, and the sample passes through the jet nozzle. A flow cell nozzle that is characterized by being able to perform optical measurements by irradiating a coaxial flow of liquid and sheath liquid with laser light. 2. Irradiate the sample suspension with a laser beam before or after it is ejected from the jet nozzle, and measure the scattered light, fluorescence, etc. emitted when the sample passes through the laser beam. At the same time, each cell in the sample is analyzed one by one, and the droplet is dropped by vibrating the entire jet flow, and based on the above measurement analysis, the droplet is charged at the point where the droplet is formed, and a high voltage is applied. In a cell sorter that deflects and separates each charged droplet by passing it between deflection plates to which a A cell sorter that uses a nozzle that also serves as a flow cell, and is characterized by being able to measure light by irradiating a laser beam onto the flow. 3. Irradiate the sample suspension with a laser beam before or after it is ejected from the jet nozzle, and measure the scattered light, fluorescence, etc. emitted when the sample passes through the laser beam. At the same time, each cell in the sample is analyzed one by one, and the droplet is dropped by vibrating the entire jet flow, and based on the above measurement analysis, the droplet is charged at the point where the droplet is formed, and a high voltage is applied. In a cell sorter that deflects and separates each charged droplet by passing it between deflection plates to which a A flow cytometer is a specialized analysis device that uses a nozzle that also functions as a flow cell, and is characterized by the ability to measure light by irradiating a laser beam onto a flow.