JPS63141587A - Laser processing method for raw samples - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to processing of a raw sample such as a living cell or microorganism by irradiating the raw sample with laser light.

[Conventional technology]

2. Description of the Related Art Conventionally, it has been known to process living cells or microorganisms (raw samples) by irradiating them with laser light. This is 1
For example, it is disclosed in Japanese Patent Application Laid-Open No. 60-83583. Japanese Patent Application Laid-Open No. 60-83583 discloses that living cells are irradiated with laser light to perforate them, and DNAf is extracted from the perforated parts.
have been disclosed that translocate into live f1 cells.

The configuration of the apparatus for processing a raw sample using the laser described above is shown in FIG.

In FIG. 2, 1 is a drilling laser, and 2 is a reference laser provided to view the optical path of 1, considering the case where 1 is not visible light. Further, 3 is a microscope, and 4 is an optical path deflection device built into the microscope. The above-mentioned drilling laser 1 passes through the shutter 5, and this reference laser enters the optical interface 6 as it is, where both lasers are converted into desired characteristics, and then both are The light beams are made coaxial and incident on the optical path deflection device No. 4. The coaxial lasers are minutely positioned here, focused by an objective lens 7, and incident on a sample 10 in a sample bottle 9 placed on a stage 8. As shown in FIG. 3, the optical path deflecting device has a structure in which two mirrors 11.11' and a scanner 12.12' are incorporated, and the laser beam incident thereon can be deflected in the X and Y directions of the sample 10. One method of using the device configured as described above is to fix the mirror 11.11"k and scan the stage (8 in Fig. 2) in the X and Y directions. In this case, the perforating laser When is a pulsed laser, the laser is irradiated onto the sample as indicated by the white circle in Fig. 4.The probability of hitting the target at this time depends on the distribution state of the cells 13 and the laser irradiation density, and the irradiation density is It depends on the number of pulses of laser light and the moving speed of the stage.

[Problem that the invention seeks to solve]

In the above conventional technology, when the distribution density of target cells is low as shown in FIG. 5, and when it is high as shown in FIG.

However, as a matter of course, the probability of hitting a cell is higher when the distribution density of cells is higher.

However, in the conventional technology, no consideration was given to controlling the distribution density of cells, and when the distribution density of cells was low, the probability of a hit was low. Furthermore, the position of the raw sample is not constant in the height direction of the sample container. Therefore, even if the laser hits a live sample at a position that is far away from the focus of the objective lens, the energy density will be low and it will not be possible to round-perforate the sample.

Thus, in the case of the prior art, the probability that a raw sample would be perforated was very low.

An object of the present invention is to provide a method for laser processing a raw sample, which increases the probability that the raw sample will be perforated, and can process the raw sample with high efficiency.

c) Means for Solving Problems] The above purpose is to make a living sample suspended in a container, and to transfer this floating living sample to a predetermined area using electrophoresis (electrophoresis or magnetophoresis). This can be achieved by irradiating the raw sample within this predetermined area with a laser beam.

[Effect]

Since the raw sample is in a floating state within the container, the raw sample migrates by applying an electric field or a magnetic field to the XY plane in the direction perpendicular to the laser beam irradiation direction of the container. This electrophoresis causes the raw sample to move to a predetermined area,
The raw sample distribution density within this region increases. Therefore, when a laser beam is irradiated onto the raw sample within this high-density area.

Irradiation efficiency increases.

〔Example〕

The sixth factor is the overall configuration diagram in one embodiment of the present invention, the plan view of the sample container 9 in FIG. 6 is shown in FIG.
-A and BB sectional views are shown in FIG.

In FIG. 1, 9 is a sample container, 8 is a culture solution containing genes, etc., 13 is a raw sample, and 19 is an L-shaped electrode fixed to the diagonal of the sample container, so that a DC power source 16 is loaded. It has become. 20 is a porous isolation membrane that prevents changes in pH and temperature of the electrolytic solution near the electrodes from reaching the entire sample bottle, and facilitates electrophoresis of the raw sample.

Illusion 1 Further, 201d is a support supporting the isolation membrane.

The lid of the sample container is perpendicular to the optical axis of the objective lens 70, as shown at 9-1 in FIG. on the other hand.

The bottom surface 9-2 of the sample container is inclined with respect to the lid. In a device configured as described above.

When the electrode 19 is energized, the raw sample moves as indicated by the arrow in FIG. 1 due to electrophoresis. As a result, the distribution density of the raw sample within the plane shown in FIG. 1 increases.

On the other hand, since the cross-sectional area of the sample container decreases in the direction of movement as shown in FIG. 7, the distribution density of the 0 part raw sample becomes very high. In this high-density state, the movement of the cells is restricted, so that they are substantially fixed.

On the other hand, the average diameter of such a raw sample is measured using the TV left camera 4 and the raw sample diameter detector shown in FIG. The irradiation density of the pulsed laser depends on the moving speed of the stage 8 when the number of pulse repetitions of the drilling laser is constant, and the optimal stage speed according to the average diameter of the raw sample is as follows. The speed is determined by the stage controller 17, and the stage is controlled to that speed. Note that it is also possible to keep the moving speed of the stage constant and control it by increasing or decreasing the number of pulse repetitions of the drilling laser 1.

The above uses electrophoresis to move raw samples.

By the way, if we replace the pair of electrodes 19 with a pair of electromagnets,
If a 0.0 isolation membrane is removed, it becomes a magnetophoresis device, and the raw sample can be moved by magnetophoresis to provide the same effect as the electrophoresis described above.

〔Effect of the invention〕

According to the present invention, the floating raw sample is moved to a predetermined area, the distribution density lft of the raw sample within the area is increased, and laser irradiation is performed, so that efficient processing can be realized.

[Brief explanation of the drawing]

FIG. 1 shows an embodiment of the present invention, and FIG. 2 shows a conventional example. FIG. 3 is a conceptual diagram of the optical path polarization device of FIG. 2, and FIGS. 4 and 5 are diagrams showing the laser irradiation state. 6th
The figure is an overall configuration diagram of an embodiment of the present invention, and FIG. 7 is a sectional view of FIG. 1. 9... Sample container, 15... Raw sample diameter detector, 16.
...Power supply, 17...Stage controller, 19...
·electrode. 20... Isolation membrane.

Claims (1)

[Claims] 1. In a method for laser processing a raw sample in which the raw sample is processed by irradiating the raw sample with laser light, the raw sample is brought into a floating state in a container, and the raw sample in the suspended state is subjected to electrical or 1. A method for laser processing a raw sample, comprising: collecting the raw sample in a predetermined area by magnetophoresis, and irradiating the raw sample within the predetermined area with the laser beam.