JPH04152245A - Particle analyzing device - Google Patents

Abstract

PURPOSE:To facilitate analysis of particle to be inspected using a simple configuration by utilizing the photo-trapping phenomenon, and photographing and analyzing the image of particle seized in a certain place on the part to be inspected. CONSTITUTION:In the condition that a beam waist of a certain intensity is formed on a transparent plate 11, a good diluted particle floating liquid L for measurement is fed from a supply hole 24b followed by necessary treatment, and particle D to be inspected is discharged together with water contained in the liquid L to outside a nozzle 24a. By means of photo-trapping phenomenon a high pressure acts on the particle S in the direction of greater photo intensity, so that the particle S in the dripping liquid LO having hopped out of a particle separator device 24 will be trapped in the center of the beam waste on the transparent plate 11. In this condition the particle image is photographed. The image is taken into a frame memory 20 and a VTR 21 and stored as image information in the form of digital values and analog values, and analysis of the particle S is performed by an image processing device 22 on the basis of the image in the frame memory 20.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a particle analysis device that analyzes individual test particles in a suspended liquid in which minute particles are suspended based on images thereof.

[Prior art] As shown in FIG.
into the suspended liquid, and the image of the particles to be examined is exposed to CO
The image is formed on a two-dimensional image sensor 4 such as D, and the image of the particle to be inspected is used to analyze the particle to be inspected. In this case,
First, stage 1 is moved on the X and Y axes perpendicular to the optical axis of the apparatus to grasp one sample particle, and then stage 1 is moved along the Z axis of the optical axis.
Move in the axial direction to focus.

[Problems to be Solved by the Invention] However, in the conventional example described above, the operation to focus on individual particles to be examined is troublesome, and the drive mechanism for moving the stage 1 three-dimensionally is complicated and large. do. Also,
In order to automatically and sequentially obtain images of test particles, a particle search means and an automatic focusing means are also required.

An object of the present invention is to provide a particle analysis device that has a simple configuration and can easily perform analysis using images of particles to be examined.

[Means for Solving the Problems] In order to achieve the above-mentioned object, the particle analysis device according to the present invention includes a storage part for storing a particle suspension containing test particles, and a small amount of water from the storage part. The present invention is characterized by having a discharging means for discharging a particle suspension liquid onto the subject part, a condensing means for condensing a laser beam onto the subject part, and an imaging means for taking an image of the subject part. That is.

[Function] The particle analysis device having the above-mentioned configuration discharges a small amount of particle suspension from the storage section onto the object to be examined on which the laser beam is focused, and uses the optical trap phenomenon to emit light to a certain place on the object to be examined. An image of the trapped test particle is captured and analyzed.

[Embodiment J] The present invention will be explained in detail based on the embodiment shown in FIGS. 1 and 2.

FIG. 1 shows an embodiment of the present invention. For example, the optical axis 0 of a semiconductor laser light source 5 that emits a laser beam of 830 nm is shown in FIG.
A concentration filter 6, a beam expander 7, and a beam splitter 8. A dichroic mirror 9 that transmits only visible light beams of 800 nm or less is arranged on the optical axis 02 of the dichroic mirror 9 in the reflection direction. Number NA=1.

An immersion type objective lens 10 having a diameter of 25, a transparent plate 11, a lens 12, and an elucidation light source 13 that emits visible light flux are arranged in this order, and an immersion oil 14 is placed between the objective lens 10 and the transparent plate 11. Injected. On the other hand, a condensing lens 15 and a photodetector 16 are arranged in the reflection direction of the beam splitter 8, and a concentration filter 17, a condensing lens 18, and a two-dimensional imaging device such as a CCD are arranged on the optical axis 02 behind the dichroic mirror 9. Element 19 is arranged. The output of the image element 19 is connected to a frame memory 20 and a VTR 21, respectively.
The output of the frame memory 20 is connected to the image processing device 22, and the outputs of the VTR 21 and the image processing device 22 are connected to the CRT 23.
It is connected to the. Further, a particle separator 24 is arranged to face the transparent plate 11, and this particle separator 24
As shown in Figure 2 (a), one side with one open end is 5
It has a structure in which a supply port 24b for supplying a particle suspension liquid is attached near the opening of a nozzle 24a with a square cross section of about 0 μm, and a small heater 24c is provided inside the nozzle 24a to absorb the heat. In order to prevent the temperature near the opening from increasing due to conduction, a heat dissipating member 24d is attached to the outside near the opening of the nozzle 24a.

In the above configuration, the laser beam emitted from the semiconductor laser light source 5 also travels on the optical axis 01, and the density filter 6
, a part of the beam passes through the beam expander 7, passes through the beam splitter 8, is reflected by the dichroic mirror 9, passes through the objective lens 10 and the immersion oil 14, and reaches the transparent plate 11, where a beam having an intensity gradient is formed. It forms the waist. A part of the laser beam is reflected by the beam splitter 8 and sent to the photodetector 1 by the condensing lens 15.
Based on the received light amount, a control circuit (not shown) controls the amount of light emitted from the semiconductor laser light source 5 so that the actual output of the laser beam emitted from the semiconductor laser light source 5 becomes a set value.

In this way, with a beam waist of a desired intensity formed on the transparent plate 11, a sufficiently diluted measurement particle suspension liquid is supplied from the supply port 24b as shown in FIG. 2 (at). When the heater 27c is turned on to generate heat after the inside of the supply port 24b is filled, the water in the heated particle suspension liquid instantly vaporizes, and bubbles B are generated as shown in FIG. 2 tb+.

Since the volume of the particle suspension liquid expands due to the air bubbles B, one test particle S near the opening is discharged to the outside of the nozzle 24a together with the water in the particle suspension liquid, as shown in FIG. 2 (cl). At this point, when the heater 27c stops generating heat, the bubbles B are cooled and contracted, so a pulling force acts on the particle suspension liquid L° discharged from the opening, and the particle suspension liquid containing the test particles S is L becomes a droplet LO, which flies out into the air and reaches the transparent plate 11, as shown in Fig. 2 fdl.Generally, microparticles suspended in a liquid have a high light intensity due to the optical trapping phenomenon. Since high pressure is applied in the direction, droplets LO fly out from the particle separator 24.
The particles S to be detected inside are always optically trapped at the center position of the beam waist of the transparent plate 11.

In this way, when the illumination light source 13 is turned on with the test particles S suspended on the transparent plate 11, the visible light beam travels on the optical axis 02 and passes through the lens 12 to the test particles S on the transparent plate 11. is illuminated, and the image of the particles to be detected is formed on the image sensor 19 via the immersion oil 14, the objective lens 1o, the dichroic mirror 9, the density filter 17, and the condensing lens 18. This image is taken into the frame memory 20 and VTR 21 and stored as image information of digital values and analog values, and the image processing device 22 analyzes the test particles S from the image in the frame memory 20, and the results are displayed on the CRT 23. Alternatively, it is output to a printer (not shown) or the like. If the focal point of the imaging optical system is adjusted in advance to match the center position of the beam waist of the transparent plate 11, the particle image to be examined will always be in focus, so there is no need to provide a focusing means. It is possible to grasp the droplets LO that successively fly out from the particle separator 24 at the focal position and perform analysis from a clear image of the test particles S.In addition, the image of the VTR 21 can also be output to the CRT 23. -The images stored in the VTR 21,
The data may be played back later, subjected to A/D conversion, and then stored in the frame memory 20.

Note that the droplet ejection method is not limited to the above-mentioned method of ejecting bubbles by generating bubbles using thermal energy. may be discharged.

The light intensity of the laser beam needs to be controlled so as to move the test particle S to the center position of the beam waist, but not to destroy the test particle S. Further, the wavelength of the laser beam from the semiconductor laser light source 5 needs to be set in consideration of the particles to be detected, for example, in a range that does not harm living cells as in this embodiment.

[Effects of the Invention] As explained above, the particle analysis device according to the present invention discharges a small amount of particle suspension liquid from the storage section onto the specimen on which the laser beam is focused, and uses the optical trap phenomenon to Since an image of the target particle grasped at a fixed location above is captured and analyzed, a focusing operation is omitted, making analysis easier. Furthermore, the structure is simple, with no need for a drive means for the test section, a test particle search means, or a focusing means.

[Brief explanation of drawings]

Drawings 1 and 2 show an embodiment of the particle analysis device according to the present invention, FIG. 1 is a configuration diagram of the embodiment of the present invention, FIG. 2 is an explanatory diagram of the operating principle, and FIG. is a configuration diagram of a conventional example. 5 is a semiconductor laser light source, 6.17 is a concentration filter, 7 is a beam expander, 8 is a beam splitter, 9
13 is a dichroic mirror, 13 is an illumination light source, 14 is immersion oil, 19 is an image sensor, 20 is a frame memory, 2
2 is an image processing device, 21 is a VTR, 23 is a CRT, 24
2 is a particle separator, 24a is a nozzle, 24b is a supply port, 2
4c is a heater, and 24d is a heat radiation member. Patent applicant Canon Co., Ltd. Figure 1 Figure 3

Claims (1)

[Claims] 1. A storage part for storing a particle suspension containing test particles, a discharge means for discharging a small amount of the particle suspension from the storage part onto a test part, and a laser beam focused on the test part. A particle analysis device comprising: a condensing means for emitting light; and an imaging means for taking an image of the test area.