JPH085579A - Micro area X-ray irradiation device - Google Patents

Abstract

PURPOSE:To obtain an X-ray irradiation system for microarea by which an X-ray irradiation test can be conducted conveniently and quickly for a microarea on a biological sample. CONSTITUTION:The microarea X-ray irradiation system comprises an X-ray source 20, a visible light source 13, an optical system 25 for condensing the light from the X-ray source or the visible light source toward a sample in a sample case 26, an optical system 27 for imaging a pattern condensed through the condenser optical system 25, an image pickup means 29 disposed at the imaging position of the imaging optical system 27, a vessel 18 for evacuating the optical path between the X-ray source 20 and the image pickup means 29, and means 30 for evacuating the vacuum vessel 18. In such X-ray irradiation system, an X-ray and visible light transmissive window is made through one of two faces intersecting the optical path of the sample case 26 and a visible light transmissive window is made through the other face. The area of the visible light transmissive window is set larger than that of the X-ray and visible light transmissive window.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a sample (such as a biological sample).
The present invention relates to a small area X-ray irradiating device that irradiates X-rays to the inside.

[0002]

2. Description of the Related Art Recent developments in biotechnological technology largely depend on observation means such as optical microscopes and electron microscopes.
However, the optical microscope is characterized in that it can handle a living biological sample in a liquid, but its spatial resolution is limited to the wavelength of visible light. Further, although the electron microscope has a high spatial resolution, the sample must be placed in a vacuum, and since there is no window material through which the electron beam passes, the biological sample cannot be observed as it is alive.

Therefore, an X-ray microscope, which has the possibility of observing a living biological sample with high resolution, has attracted attention and has been developed. With the development of fine precision engineering, the performance of X-ray optical elements for X-ray microscopes has been improved, and a tester for X-ray microscopes has been manufactured. The window material of the sample container is suitable for observing biological samples 2.4 ~ 4.4nm
A silicon nitride film or the like that transmits X-rays is used.

Biological samples are easily damaged by X-ray irradiation, and there is a concern that a living sample cannot be observed. Therefore, the practical application of an X-ray microscope has been questioned. Therefore, an experiment for collecting data on the limit amount of X-rays that a biological sample can withstand is performed using an X-ray microscope tester. The sample container for X-ray microscope as shown in FIG. 4 is used as it is.

Further, the damage to the living body due to radiation exposure has become a social problem in various situations such as nuclear reactors and nuclear experiments, and it is urgent to clarify the action of radiation on the living body and collect data for radiation therapy. It is peeling. The experiment on the damage of the biological sample by the X-ray irradiation using the microscopic area X-ray irradiation apparatus also plays a role in this.

[0006]

A sample container 26 'which has been conventionally used as shown in FIG.
Back etching the silicon substrate 32 on which
Two members having a line-transparent window are used. The thickness of the sample solution 40 sandwiched between the two members is adjusted by the spacer 34 formed on one or both of the members. In a vacuum environment, the X-ray transmission window receives an internal pressure (a pressure difference from the vacuum), and therefore has a limited size. That is, if it is too large, the window material (silicon nitride film) 31 is bent outward by the internal pressure,
The sample solution becomes thick and X-rays cannot be transmitted.

In the X-ray microscope, the thickness of the sample solution is 10 μm.
It should be less than m, but a silicon nitride film with a thickness of about 0.1 μm and a 200 μm square that transmits about 60% of 2.4 to 4.4 nm X-rays will bend about 3 μm. The thickness of the solution is 10 μm or less. The assembly of the sample container is performed under an optical microscope. However, since the size of the X-ray transmission window provided on each of the two members forming the sample container is only 200 μm square, the positions of the two X-ray transmission windows are different. Performing optical adjustment (optical path adjustment) by accurately matching the above is a complicated task involving trial and error.

According to the present invention, it is not necessary to exactly match the positions of the two light transmission windows provided facing the sample container,
Therefore, it is an object of the present invention to provide a micro-area X-ray irradiating device which is extremely easy to assemble a sample container and which can easily and quickly perform an X-ray irradiation test on a microscopic area of a biological sample.

[0009]

Therefore, the first aspect of the present invention is to provide "at least an X-ray source that generates X-rays, a visible light source that emits visible light, and a sample that emits light emitted from the X-ray source or the visible light source. Condenser optical system for condensing on a sample in a container or at a place where the sample is installed, image forming optical system for forming an image of a condensing pattern by the condenser optical system, and image pickup means arranged at an image forming position by the image forming optical system. A micro-region X-ray irradiator comprising: a vacuum container for evacuating an optical path from the X-ray source to the imaging means; and an evacuation means for evacuating the vacuum container to a vacuum. Of the two surfaces intersecting the optical path, one surface is provided with a transparent window for X-rays and visible light, and the other surface is provided with a visible light transparent window, and
There is provided a minute area X-ray irradiator (claim 1), characterized in that the area of the visible light transmitting window is made larger than the area of the X-ray and visible light transmitting window.

A second aspect of the present invention is that "the condenser optical system and the imaging optical system are each optical system using a reflection type optical element, and the minute area X-ray irradiation is performed. Apparatus (claim 2) ". Further, the present invention thirdly provides a "small area X-ray irradiating apparatus (claim 3)" in which the condenser optical system is an optical system using a reflection type optical element. .

A fourth aspect of the present invention is that the reflective optical element is a spheroidal multilayer mirror, a rotating parabolic multilayer mirror, a Schwarzschild mirror, or a Walter mirror. Alternatively, the minute area X-ray irradiator according to claim 3 is provided.

[0012]

Unlike the case of the X-ray microscope, the sample container of the micro-area X-ray irradiator according to the present invention needs only one window for transmitting X-rays on the light source side. You don't have to use one. That is, in the microscopic area X-ray irradiator of the type capable of focusing with visible light, of the two surfaces of the sample container intersecting the optical path extending from the light source to the imaging means, the surface on the light source side transmits X-rays and visible light. A window and a visible light transmitting window may be provided on the surface on the image pickup means side.

Since the visible light transmitting window only needs to pass visible light, a thick glass plate can be used for the window. Since the thick glass plate does not bend due to internal pressure, the size is not limited. If the visible light transmitting window is made sufficiently larger than the X-ray transmitting window, the window alignment is not necessary.
It is preferable that the size and the thickness of the glass plate are the same as those of the silicon substrate with the X-ray transmission window because the base and lid of the conventional sample container can be used as they are.

The material of the visible light transmitting window is not limited to glass, and any material that allows visible light to pass, such as transparent plastic resin, may be used. Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples.

[0015]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 is a schematic block diagram of a micro-area X-ray irradiator using a Walter mirror for imaging with X-rays and visible light as a first embodiment. The infrared pulsed laser light 12 from the YAG laser device 11 is reflected by the infrared light reflecting mirror 15,
The laser light condensing lens 16 condenses the laser light through the laser light introduction window 17 onto the surface of the target plate 19 installed in the vacuum container 18 to generate plasma (an X-ray source, an example of a visible light source) 20.

The plasma 20 emits plasma light 21 such as X-rays, ultraviolet rays, and visible light. Only the X-rays pass through the X-ray transmission filter 22, and are condensed at the position of the sample container 26 by a Wolter mirror (an example of condenser optical system) 25 for condensing.
Irradiate the sample in the container. An image of the condensing pattern at the position of the sample container 26 is formed on an imager (an example of an image pickup unit) 29 by an image-forming Walter mirror (an example of an image-forming optical system) 27. A vacuum exhaust device (an example of exhaust means) 30 is connected to the vacuum container 18.

Alignment of the optical arrangement of the irradiation device is performed using the X-ray with the sample container 26 removed.
That is, alignment is performed so that the image pickup device 29 can obtain a good X-ray focusing pattern. Next, after the alignment, the visible laser light 14 from the HeNe laser device (an example of a visible light source) 13 is irradiated with an infrared light reflecting mirror in a state in which a good X-ray focusing pattern is formed on the image pickup device 29. 15 is transmitted, and the laser beam focusing lens 16 focuses the laser beam on the surface of the target plate 19 through the laser beam introduction window 17.

The filter operating unit 24 is used to transmit the X-ray transmission filter 2
When 2 is retracted from the optical path, the scattered light from the surface of the target plate 19 is condensed by the condensing Walter mirror 25, and an image of the condensing pattern is formed on the imager 29 by the imaging Walter mirror 27. It The position of the HeNe laser device 13 is adjusted so that the position of this light focusing coincides with the position of the X-ray focusing. By doing so, the condensing position of the X-ray can be known using visible light.

When irradiating a sample with X-rays, first, X
While the line transmission filter 22 is retracted from the optical path, the sample container 2 is placed at the visible light condensing position while observing with the image pickup device.
The positions of 6 are aligned and the irradiated portion of the sample is confirmed. Next, the X-ray transmission filter 22 is inserted in the optical path, and infrared pulsed laser light is emitted from the YAG laser device, so that X-rays are focused and emitted at a desired position on the sample.

FIG. 2 shows, as a second embodiment, a schematic configuration diagram of a minute area X-ray irradiating apparatus using an objective lens for image formation with visible light. A target plate 19 in which the infrared pulsed laser light 12 from the YAG laser device 11 is set in a vacuum container 18 through a laser light introduction window 17 by a laser light focusing lens 16
The light is focused on the surface of and the plasma 20 is generated. Plasma 2
0 emits plasma light 21 such as X-rays, ultraviolet rays, and visible light, but the light that passes through the filter differs depending on the filter selected using the filter operating unit 24. A vacuum exhaust device 30 is connected to the vacuum container 18.

The alignment of the optical arrangement of the irradiation device is performed by using visible light contained in the plasma light 21 emitted by the plasma 20 with the sample container 26 removed. The visible light extinction filter 23 is inserted into the optical path by the filter operating device 24 so that the visible light that has been dimmed to the extent that the observation sample and the image pickup device 29 are not damaged is transmitted. Similar to the case of using X-rays in the first embodiment, alignment is performed using dimmed visible light, and when aligned, the sample container 26
While observing with an image pickup device, the position of the sample container 26 is overlapped with the position where the visible light is condensed, and the irradiated portion of the sample is confirmed.

Next, the X-ray transmission filter 22 is inserted into the optical path by the filter operating unit 24, and the infrared pulsed laser light is irradiated from the YAG laser device, so that the X-ray is focused and irradiated at a desired position on the sample. It FIG. 3 shows an example of the sample container. The base portion 35 of the sample container is provided with one ring-shaped concave portion to expose a rubber O-ring 37, and the lid 36 is provided with two ring-shaped concave portions to expose a part of the rubber O-rings 37 ′ and 39. It is embedded in.

A 0.4 mm thick silicon window substrate 32 having a 3 μm thick spacer 34 made of a photoresist material and a 0.1 μm thick silicon nitride film which is an X-ray transparent window material is back-etched to form an X-ray transparent window ( It is also a visible light transmission window) 3
1 has been produced. To prepare the sample container, the sample container base 35 is placed horizontally, the window substrate 32 is installed, the sample solution is dropped on the window, and a glass plate of the same size and the same thickness as the window substrate 32 (visible light transmission) is used. 33, which corresponds to the window, is placed. When the excess sample solution is wiped with filter paper, covered with the lid 36 and fastened with the screw 38, the sample solution 40 is sealed by the O-ring and the sample solution 40 does not leach even in vacuum.

According to the sample container of this embodiment, the positions of the X-ray and visible light transmission window 31 and the visible light transmission window 33 having a larger area than that of the transmission window 31 are provided so as to face each other. Therefore, it was extremely easy to assemble the sample container. In this example, yeast was used as a sample for X-ray irradiation. Sample container 2 with dry yeast reconstituted with water and trypan blue stain added
It was put in No. 6, and it was confirmed by an optical microscope that yeast was not stained.

The sample container 26 was attached to a microscopic area X-ray irradiator and focused with visible light before irradiating with X-rays.
The sample container was taken out and the sample was observed with an optical microscope. It was observed that only yeasts exposed to strong X-rays were stained and surrounding yeasts not exposed to X-rays were not stained, and it was found that yeasts lose their dye exclusion ability when exposed to strong X-rays.

In the first and second embodiments described above, the Wolter mirror is used as the X-ray focusing mirror and the X-ray imaging mirror, but these are not limited to the Walter mirror and the focal points of X-ray and visible light coincide with each other. You can also use Schwarzschild mirror, spheroid multilayer mirror, etc. Further, a micro-area X-ray irradiator using synchrotron radiation as a light source can be manufactured, and synchrotron radiation can be regarded as parallel light. A mirror is used.

[0027]

According to the micro-area X-ray irradiator of the present invention, it is not necessary to accurately align the positions of the two light transmission windows provided on the sample container so as to face each other, and therefore the sample container can be assembled extremely easily. It is easy, and moreover, an X-ray irradiation test on a minute area of a biological sample can be performed simply and quickly.

[Brief description of drawings]

FIG. 1 is a first embodiment (schematic configuration diagram) of a micro-area X-ray irradiator of the present invention, which uses a Walter mirror for imaging with X-rays and visible light.

FIG. 2 is a second embodiment (schematic configuration diagram) of a microscopic area X-ray irradiator of the present invention, which uses an objective lens for imaging with visible light.

FIG. 3 is an example of a sample container for a micro-area X-ray irradiator of the present invention.

FIG. 4 is an example of a sample container for a conventional micro area X-ray irradiation device.

[Explanation of symbols]

11 YAG Laser Device 12 Infrared Pulse Laser Light 13 HeNe Laser Device (Example of Visible Light Source) 14 Visible Laser Light 15 Infrared Light Reflecting Mirror 16 Laser Light Condensing Lens 17 Laser Light Introducing Window 18 Vacuum Container 19 Target Plate 20 Plasma (Examples of X-ray source and visible light source) 21 Plasma light 22 X-ray transmission filter 23 Visible light dimming filter 24 Filter exchanger 25 Condensing wolter mirror (an example of condenser optical system) 26 Sample container 27 Imaging wolter mirror (Example of Imaging Optical System) 28 Imaging Lens for Visible Light (Example of Objective Lens, Imaging Optical System) 29 Imager (Example of Imaging Unit) 30 Vacuum Evacuation Device (Example of Exhaust Unit) 31 X-ray and Visible light transmission window 32 Window substrate 33 Glass plate (an example of visible light transmission window) 34 Spacer 35 Sample container base 36 Sample container lid 37 O phosphorus 37'O ring 38 threaded 39 O-ring 40 sample solution than on

Claims (4)

[Claims] 1. At least an X-ray source for generating X-rays, a visible light source for generating visible light, and light emitted from the X-ray source or the visible light source on a sample in a sample container or at a sample installation location. A condenser optical system, an image forming optical system for forming an image of a condensing pattern by the condenser optical system, an image pickup means arranged at an image forming position by the image forming optical system, and an optical path from the X-ray source to the image pickup means. A micro-region X-ray irradiator having a vacuum container for creating a vacuum, and an exhaust unit for exhausting the vacuum container to a vacuum, wherein one surface of two surfaces intersecting the optical path of the sample container Is provided with an X-ray and visible light transmission window and the other surface is provided with a visible light transmission window, and the area of the visible light transmission window is X.
A micro-area X-ray irradiator characterized by being made larger than the area of a transmission window for rays and visible light. 2. The micro area X-ray irradiator according to claim 1, wherein the condenser optical system and the imaging optical system are each optical system using a reflection type optical element. 3. The micro area X-ray irradiator according to claim 1, wherein the condenser optical system is an optical system using a reflection type optical element. 4. The micro area X according to claim 2, wherein the reflective optical element is a spheroidal multilayer mirror, a paraboloid multilayer mirror, a Schwarzschild mirror, or a Walter mirror. X-ray irradiation device.