JPS6068312A - Photographing method of optical microscope image - Google Patents

Abstract

PURPOSE:To obtain a clear optical microscope image of a sample even when its surface is not flat by varying the relative position relation between the lens barrel of an optical microscope and the sample. CONSTITUTION:The surface of the sample 4 on a stage 5 at the side of the lens barrel 2 is not flat. The stage 5 is moved in the direction Z of the optical axis of the optical microscope 1 to bring the highest projection part of the sample 4 into focus within a photographic visual field. The distance between the sample 4 and lens barrel 2 at this time is denoted as Z1. Then, the stage 5 is moved to bring the lowest recessed part of the sample 4 into focus. The distance at this time is denoted as Z3. The distances Z1 and Z3 are stored in a control part 10. Photography is started at the distance Z1 and the stage 5 is scanned in the optical axis direction at an equal speed until the sample 4 is at the distance Z3. A shutter 9 is open during this period associatively with a synchronous motor 8, which stops at the distance Z3 to close the shutter 9. The sample 4 is scanned in the optical axis direction to vary the quantities of light corresponding to respective positions of the sample 4 with time. Therefore, images are integrated on the film in a camera 3 to expose corresponding positions of the focusing position one after another.

Description

DETAILED DESCRIPTION OF THE INVENTION (a) Field of Industrial Application The present invention relates to a method of taking a microscopic image of a sample surface using an optical microscope.

(b) Prior art In general, optical microscopes are widely adopted because they are easy to prepare the sample to be observed, are easy to operate, can distinguish colors, and are inexpensive. has been done. Although optical microscopes have many advantages as described above, they have the disadvantage that clear optical microscope images cannot be obtained by simply photographing samples with curved or uneven surfaces due to their shallow depth of focus. For this reason, conventionally, only samples with relatively flat surfaces have been observed. When observing samples with curved or uneven surfaces, stereoscopic microscopes and scanning electron microscopes are mainly used. However, in the former case, the magnification is at most several ten times, which is not sufficient. In the latter case, a high magnification of tens of thousands of times can be obtained, but there are drawbacks such as complicated preparation of the sample and operation of the device, inability to distinguish colors, and high price.

(C) Purpose The present invention has been made in view of the above problems, and
To solve the conventional problems by making it possible to take a clear optical microscope image even for a sample whose surface is curved or uneven.

As shown in Figure 1, the light intensity of an optical microscope image is at its maximum when it is in focus on the sample surface, and when it deviates from the focus position to a certain extent, the background intensity remains constant regardless of the sample position. The present invention focuses on this point, and in order to achieve the above object, the distance between the sample and the lens barrel when focusing on the most convex part of the sample within the field of view of the sample. , between the distance between the sample and the lens barrel when focusing on the most concave part of the sample, and move at least one of the sample and the lens barrel of the optical microscope from one distance to the other within the photographic exposure time. Scanning is performed in the optical axis direction f. For example, if the amount of light is sequentially accumulated on the film, only the in-focus areas will be exposed accordingly, resulting in a photographed image that is in focus as a whole.

(E) Examples Hereinafter, the present invention will be described in detail with reference to the drawings.

FIG. 2 is a schematic diagram of an optical microscope to which the present invention is applied. In Fig. 2, 1 is an optical microscope, 2 is a lens barrel with a built-in magnifying lens (not shown), 3 is a camera attached to this lens barrel 2, 4 is a sample to be observed, 5 is a sample This is a stage for placing 4. A rack portion 5a is formed on this stage 5, and a pinion 6 is meshed with the rack portion 5a. 8 is a synchro motor for rotationally driving the pinion 6, 9 is a shutter of the camera 3, and 10 is a control unit that controls the operations of the synchro motor 8, the shutter 9, and the like.

Next, a method of taking an optical microscope image of the sample 2 by applying the optical microscope 1 having the above configuration will be explained.

First, the sample 4 is placed on the stage 5. It is assumed that the surface of the sample 4 on the lens barrel 2 side is not flat but has irregularities as shown in FIG. 3(a). Next, the stage 5 is moved in the optical axis direction of the optical microscope 1 (Z direction in FIG. 2). Then, while looking through the lens barrel 2, the user focuses on the most convex portion of the sample 4 (point A1 in the figure) within the field of view of the sample 4. The distance between the sample 4 and the lens barrel 2 at this time is defined as Zl. Subsequently, the stage 5 is moved to focus on the most concave portion of the sample 4 (point A2 in the figure) within the same field of view. The distance between the sample 4 and the lens barrel 2 at this time is 73 degrees.

The distances Z+ and Za between the sample 4 and the lens barrel 2 are determined by the control unit 1.
Store it at 0. Subsequently, the sample 5 is positioned at the initially set distance Z1, and from this state the control unit 10 is operated to open the shutter 9 and photographing is started. At the same time as the shutter 9 is opened, the synchro motor 8 is also driven,
Rotate pinion 6. Then, stage 5 is set to sample 4.
is scanned in the optical axis direction at a constant speed until it reaches a distance Z8 set later (in this example, it is moved downward). During this time, the shutter 9 remains open in conjunction with the synchro motor 8, and when a predetermined distance Z8 is reached, the synchro motor 8 stops and the shutter 9 closes. In this way, the sample 4 is scanned in the optical axis direction from one distance Zl to the other distance Z8 within the photographic exposure time, so as time passes, the amount of light corresponding to each part of the sample 4 changes as shown in Figure 3 ( b)～(
It changes sequentially as shown in d). Therefore, the images are accumulated on the film of the camera 3, and the areas corresponding to the in-focus areas are exposed one after another. As a result, a photographed image that is in focus within the field of view is obtained as a whole.

Background fog occurs during scanning, but this can be removed by adjusting the sensitivity of the film, the aperture of the camera 3, the brightness of the illuminator (not shown), the photographic paper, the printing conditions, etc. be able to. In addition, in the above embodiment, the sample 4 was scanned from the position Z1 where the focus was on the most convex part of the sample 4 to the position Z8 where the focus was on the most concave part of the sample 4, but the reverse case may also be used. Furthermore, instead of moving the sample 4 side, the lens barrel 2 may be moved. Furthermore, the sample 4 and the lens barrel 2 can be moved closer to or farther apart from each other.

(f) Effects As described above, according to the present invention, a clear optical microscope image can be obtained even if the sample surface is not flat by changing the relative positional relationship between the lens barrel of the optical microscope and the sample. Furthermore, it can be realized at a much lower cost than a scanning electron microscope. Furthermore, the excellent effect of being able to obtain photographed images even at a magnification of several hundred times, which was not possible with a stereomicroscope, is exhibited.

[Brief explanation of drawings]

Fig. 1 is a characteristic diagram explaining the relationship with the amount of light when focused on the sample surface, Figs. 2 and 3 show embodiments of the present invention, and Fig. FIG. 3, which is a block diagram of the optical microscope, is a characteristic diagram showing the relationship between the shape of the sample surface and the amount of light when focused on the sample. 1... Optical microscope, 2... Lens barrel, 3... Camera,
4...Sample. Applicant: Shimadzu Corporation Representative Patent Attorney: Oka 1) Hide Kazu/ (7) Figure 1 Figure 2

Claims (1)

[Claims] ([) A method of taking an optical microscope image of a sample whose surface is not flat, which measures the distance between the sample and the lens barrel when focusing on the most convex part of the sample within the field of view of the sample, and the sample. During the exposure time, move at least one of the sample and the lens barrel from one distance to the other along the optical axis of the optical microscope. A method for taking an optical microscope image, characterized by scanning in a direction.