JP5838492B2 - Simple microscope, dark field observation method and video recording method using it - Google Patents

Description

  The present invention relates to a simple microscope with an illumination device capable of dark field observation by oblique illumination, a dark field observation method using the microscope, and a photographing recording method using a digital camera using the microscope.

The following Patent Document 1 is attributed to the present inventor, and the present invention is an improved invention of claim 4 of the present invention. In the fourth aspect of the invention, the lens holding member and the sample holding member are overlapped with each other by a magnetic force, but dark field observation is difficult because no illumination device is attached.
The following Patent Document 4 is also related to the present inventors. This is a thing that can adjust the focus by changing the distance between the lens holding member and the sample holding member, but it is difficult to observe the dark field because the illumination device is not attached, and the present invention is to improve this point. In addition to the lens holding member and the sample holding member, an illumination device is provided.

It is an unmistakable fact that the 17th-century Dutch Levenhook (Lehwenhook) observed bacteria with a single-lens microscope with a single lens. However, it is believed that the observation was made by the dark field method, although the device itself and the method used for the observation were not known because it was kept secret (Non-Patent Document 1).
The present inventor has tried to confirm this, but at last, the inventors succeeded in observing bacteria using a dark field and recently considered that there is a patentability. One objective lens is noticeable, but the illumination light of different angles is obviously different because the illumination light angle changes continuously while the illumination light of different angles is switched. is there.

Japanese Patent No. 3806828 JP 9-197289 A Japanese Patent Laying-Open No. 2005-003909 (Claim 4) JP 2003-098440 A (FIGS. 3 to 8) JP 2006-119557 A (paragraph numbers 0009 to 0015)

Kazuaki Tengo “Levenhook's Letter” Kyushu University Press 2004, page 327

  The problem we want to solve is a simple portable microscope that can be used for bright-field and dark-field observations where bacteria and microalgae of about 1 micron can be seen, and even elementary school students can manufacture easily. It is to provide.

In the present invention, a lens holding plate holding a spherical lens and a sample holding plate in which a hole for light transmission is opened to hold a sample on the outer peripheral portion thereof, and an illumination device is provided on the back surface of the sample holding plate. The lens holding plate and the sample holding plate can be adjusted by adjusting the distance between the spherical lens of the lens holding plate and the sample of the sample holding plate by adsorbing and fixing in a state where it can be slid by the magnetic force of a permanent magnet. The two holding plates are bonded together by being overlapped so as to be rotatable around the fulcrum of the end, or are bonded together via a permanent magnet spacer.
In the text, attaching and detaching means that the connected part is rotated on a two-dimensional surface, and both objects are separated or combined, or both objects are separated or combined in the Z direction in three dimensions. It is.

By doing so, the illumination device can be slid by hand to change the position of the light source, and the incident angle of the illumination light to the sample can be continuously changed.
However, with this structure, there is no problem to use it fixedly, but when it is portable, the lighting device gets in the way and puts it in a pocket or when it is placed in the pocket. Therefore, in addition to the sample holding plate, another plate-like body called a slide plate is provided under the sample holding plate so that the lighting device can be adsorbed and fixed by magnetic force on the slide plate so that it can be moved. .

In order to make the lighting device as compact as possible, a button battery was used as a power source. However, because of the property that the electrode of the button battery attaches to the magnet, the electrode plate is attached by the magnetic force of a permanent magnet as a switch mechanism. The lamp was turned on by energizing the battery electrode in close contact with the battery electrode, thereby reducing the size and manufacturing cost.

Furthermore, the spacers between each holding plate are magnets so that they can be assembled without tools when manufacturing at event venues.

To use the microscope of the present invention, first move the illuminator close to the lens, adjust the focus by bright field observation, move the illuminator with your finger while looking through the lens with your eyes, and move away from the lens. Since it gradually darkens from the outer periphery of the field of view, it is possible to easily perform dark field observation by directing the viewer's eyes toward the outer periphery. At this time, by adjusting the focus while tilting the microscope main body as shown in FIG. 2, dark field observation can be performed by oblique illumination with the darkness and resolution desired by the observer within the possible range of this microscope where distortion is small. Become.

With the naked eye of the simple microscope of the present invention, it is possible to achieve a resolution equivalent to that of a conventional abdominal microscope by only one spherical lens, at least one light source, movement of the illumination device, and adjustment of the observer's line of sight. Bright field observation and dark field observation are possible. Furthermore, because it is extremely lightweight, about 100 grams, it can be used for portable observation of algae such as mushroom spores and green insects in the field.

Furthermore, an unexpected and remarkable effect is that the microscope of the present invention can record and record a three-dimensional feeling with color without using a high-sensitivity digital camera to stain minute objects of the order of 1 micron. It is a thing.
Regarding the visual recognition with this colored image, the present applicant has previously applied for the above-mentioned Patent Document 5, and this phenomenon is regarded as a light interference phenomenon between the lens and the observation sample, and the diameter of the spherical lens is 0. It was explained in paragraph No. “0015” of the specification of Patent Document 5 that the phenomenon occurs when the thickness is 2 mm or less (about 2300 times). However, since the spherical shape is small, the production is not possible due to high cost and mass production. However, this time, it was found that even if the diameter of the spherical lens is 0.2 mm or more, it can be easily colored by using the microscope of the present invention and a digital camera equipped with a high-sensitivity image sensor that is commercially available.

Examining variously, for color imaging of minute objects, the product of the magnification of the spherical lens and the zoom magnification of the high sensitivity digital camera is preferably 3000 times or more and 30000 times or less, and the magnification of the spherical lens is 100 It is understood that a value that is more than double and less than 6000 times is a value suitable for colorization, and in particular, from the viewpoint of manufacturing, it has been found that 100 times or more and 1000 times or less are particularly good. In this case, when the refractive index is about 0.3 mm or more, the diameter is naturally more than that.
Once this figure is understood, it becomes possible to manufacture a fixed-focus digital camera for exclusive use of a fixed magnification fixed bacteria test as an application of this simple microscope.
In addition, in order to confirm the micron size of the observation sample, and to see the degree of resolution by oblique illumination by the movement of the illumination light of the present invention and the degree of change in coloration, the sample mounting plate (cover It was also found that the plate of transparent diffraction grating is very good as the role of glass.

As a result of this series of research, we have uploaded videos taken with this microscope and digital camera on the “Youtube (registered trademark)” color-coded videos, so “Tadao Sato Bacteria”, “Tadao Sato Blood” Search with keywords such as "Colorize unstained bacteria", "Bacteria 2".

In addition, the video on “Youtube” shows the lens magnification and the magnification of Cybershot DSC-WX200 (trade name) using Sony's high-sensitivity back-illuminated image sensor close to the 600 × spherical lens of the simple microscope of the present invention. The product with the zoom magnification of the camera was changed between 3000 to 30000 times and recorded, and the illumination light was “a white LED (OSPW5111-YZ) manufactured by OptoSupply Co., Ltd. was emitted with a 3V battery. This bullet-type LED was shot and recorded by irradiating from 2 mm below the sample.

Samples are food kimchi juice and seashell weathered powder dissolved in tap water, applicant's blood, natto, etc. These samples are about 0.5 μm wide and 0.1 μm deep groove 1 μm intervals on a transparent plastic plate as an objective micrometer, the diffraction grating is a kind of slide glass, a sample is applied thereto, and a 0.04 mm thick low-density polyethylene film is covered as a cover glass. It is the moving image which zoomed up and image | photographed by making the spherical lens of the microscope of this invention approach, attaching the opening part of the said camera to a lens holding plate. Small round platelets appear to move in the blood, gonococcus moves like fish, and cocci, yeast, and octopus are falling under their own weight while performing Brownian motion in the solution. )

As can be seen from this video, the coloration of unstained microscopic objects can be regarded as a kind of spectroscopic image with an optical lens. Therefore, if the distance between the spherical lens (objective lens) and the sample is fixed, the wavelength and intensity of the illumination light are changed, and the corresponding image is analyzed, the physical properties of the unstained minute object can be understood. Gram-positive or negative bacteria may be identified without complicated treatment with a staining solution.
In addition, since it is a magnification of 3000 times or more, details beyond the resolution cannot be seen, but since it is colored, the contrast is improved and the movement of minute objects is well understood, and the space is expanded by increasing the magnification, Since the time is not expanded, the speed of moving bacteria etc. can be observed on the monitor very fast, and there is a possibility that the type of bacteria can be specified by the speed and appearance. Even with non-motile bacteria, the growth rate can be observed quickly on the monitor, and the quantitative speed of colony formation can be understood on the monitor, so I think that it can also play an auxiliary role in the official method of bacterial testing.

The coloration of the unstained minute object is mainly due to the chromatic aberration (axial chromatic aberration and lateral chromatic aberration) of the lens. The minute object is colored (the chromatic aberration also occurs when the minute object itself is transparent and spherical like a lens).
Enlarging beyond the resolution from 3000 times to 30000 times will cause blurring of the outline, but you can still recognize what it feels like because it improves the contrast by making it color. . Therefore, when a moving bacterium moves, the color tone (tone) and hue change depending on the distance from the lens (distance in the focal direction), and the movement of the minute object in three dimensions and the softness of the object. Can be realized.

The simple microscope of the present invention has the effects as described above, and even elementary school students can make it if they have all the parts, so the value is really desirable for education and contributes to the health of developing countries. There is a high possibility that we can expect it.
In addition, the visual field is narrower than that of an expensive practical training microscope of a famous manufacturer, but the field of view is not inferior in terms of resolution. I can observe. And since it's lightweight, sleeping and observing in the dark field makes you feel like you are looking at outer space and you can get a restful sleep, so you want to give it to children hospitalized for illness The conventional microscope is too heavy to sleep and cannot be observed very much. Therefore, the present invention is an excellent microscope from this point.

1 is a schematic side view of an embodiment of the present invention. Illustration of dark field observation method using oblique illumination Side schematic diagram with the lighting device turned off Side view showing an example of lighting device Side view showing an example of lighting device 1 is a schematic side sectional view of an embodiment of the present invention.

The details of the present invention will be described with reference to the drawings.

FIG. 1 is a schematic side view showing an example of the present invention. A lens mounting hole is formed at a substantially central portion of the plane of the lens holding plate 2, the spherical lens 1 having a diameter of about 1 mm is mounted, and a coupling screw hole is formed at one end. An opening 14 for light transmission is formed at the center of the plane of the sample holding plate 4, and a screw screw for coupling is opened at one end. A screw hole for coupling is also formed at one end of the slide plate 7. Between the lens holding plate 2 and the sample holding plate 4 and between the sample holding plate 4 and the slide plate 7, spacers are provided in the screw holes, and they are coupled with screws and nuts.

The spacer provided between the lens holding plate and the sample holding plate is preferably a soft material such as elastic rubber for focusing. The spacer provided between the sample holding plate and the slide plate is preferably a hard material such as wood, metal, or resin in order to maintain a constant spacing. By turning the nut 11, the spacer 8 expands and contracts, and the focus can be adjusted by changing the distance between the lens and the sample. The spacer provided between the sample holding plate and the slide plate must be thick enough to move the illuminator, but it can be made thin by bending the slide plate by metal pressing, or it can be eliminated in some cases. Is possible.

The lighting device is composed of a light bulb, a battery, an electrode plate, and a plate-shaped magnet. The light-emitting device 5 is attached to the upper surface of the plate-shaped magnet 6 which is arranged at the bottom and is attached to the end, and the electrode plate 23 is attached near the center. Then, one wire of the bulb and the electrode plate 23 are joined. The electrode plate 21 is attached to the bottom of the plate magnet 20 to be attached from above, and is joined to the other wire of the bulb. By sandwiching the battery between the plate magnet 20 and the plate magnet 6, the electrode plate is brought into close contact with the battery by the magnetic force, and the light bulb is turned on. When not in use, the light bulb is extinguished by sticking the plate magnet 20 to the bottom surface of the sample holding plate as shown in FIG. The lighting device can be configured with only a light bulb and a plate magnet without the battery, but in this case, a power source such as a battery must be prepared outside, which is inconvenient for observation in the field.

The size of the lens holding plate 2, the sample holding plate 4 and the slide plate 7 is about 25 mm wide × about 100 mm long and about 1 mm thick. The material is preferably a metal, but is not limited thereto. The sample holding plate 4 and the slide plate 7 are preferably made of a ferromagnetic material such as iron or stainless steel such as SUS430 so that the magnet of the illumination device is attached.

The plate-like magnet 20 of the lighting device has a circular or rectangular shape with a diameter of about 30 mm, and the plate-like magnet 6 has a width of about 30 mm and a length of about 50 mm, and is larger than the width of the slide plate 7. The magnet can be easily grasped by hand, and adjustment by movement becomes easy.
A light bulb, an organic EL, an LED light bulb, or the like can be used as the light bulb of the lighting device, a button battery can be used as the battery, and a conductive foil such as copper can be used as the electrode plate.

FIG. 2 is a schematic side view for explaining a dark field observation method using oblique illumination. When the illumination device 30 is slid on the slide plate 7 away from the lens from directly below the spherical lens 1, oblique illumination is obtained. As the resolution of the moving direction increases and further moves, the illumination light is blocked by the edge of the opening 14 of the sample holding plate 4 and the light does not directly enter the spherical lens 1 so that it gradually becomes darker. When looking toward the light bulb of the illuminator, there is a little distortion of the image of the sample, and there is a dark field observation of the shape of the sample (an intermediate image between bright and dark fields depending on the moving position of the illuminator). Can do. The tilting direction may vary depending on the sample. This is because, for example, when a sample is placed in water for observation, it is affected by the refraction of light by water. When observing, if the microscope main body is not moved, it is necessary to move the eyes, which is accompanied by eye fatigue. Since this microscope is extremely light and small, it is easy to observe if the microscope main body is held by hand and is used with a slight tilt as shown in FIG. Incidentally, sliding the illumination device on the slide plate is also a key point of the present invention, but this is intended to move the light-emitting bulb of the illumination device between the sample holding plate 4 and the slide plate 7. Therefore, even if the plate magnet 6 of the lighting device is slid on the back surface of the sample holding plate 4, the gist of the present invention is the same, and there is a light bulb such as a flat and thin LED. The present invention can be configured even if a lens holding plate, a sample holding plate, and a plate-like magnet provided with only a light bulb are attached to the back surface of the sample holding plate as an illumination device.

As an embodiment of the illumination device, as shown in FIG. 4, the positive line of the light bulb 5 is not connected to the electrode plate 21, but only the electrode plate 21 is attached to the plate magnet 20, and the electrode plate 42 is attached to the insulating sheet 41. The electrode plate 43 can be attached to the positive side of the battery 22 so as to be detachably attached with an adhesive or the like, and the electrode plate 43 can be connected to the positive side of the bulb.

In addition, as shown in FIG. 5, the lighting device is obtained by attaching an electrode plate 52 attached to an insulating sheet 51 such as a thin PP sheet having a little elasticity away from the electrode surface of the battery 22 to the plate magnet 6. Alternatively, the plate-like magnet 20 can be attracted to or separated from the electrode surface side of the battery 22, and the illumination light can be turned on and off by the electrode plate 52 contacting or leaving the battery electrode.

FIG. 6 shows an object in which the distance between the lens holding plate 102 and the sample holding plate 104 is variable by a focus adjustment screw 111, and further an illumination device D is provided between the slide plate 107 and the sample holding plate 104. Movement and fixation are facilitated by the plate magnet 6 of the substrate, and oblique illumination and dark field illumination are possible. In FIG. 6, a space is provided between the sample holding plates by the magnet spacers 109 and 119 so that the illumination device D can move.

As a method of forming this space portion, if the slide plate 107 is subjected to hat bending with a metal press or the like, the thickness of the magnet spacer can be reduced. In some cases, it is possible to secure a sufficient space even if it is not used. It is.

The lens holding plate 102, the sample holding plate 104, and the slide plate 107 are substantially the same size as the lens holding plate 2, the sample holding plate 4 and the slide plate 7 in FIG. The M3 screw tap is cut, and the fixing / rotating shaft screw 110 and the focus adjusting screw 111 are screwed in. Further, only a screw penetration hole is formed in the left end portion of the lens holding plate 102, and a portion where the tip end portion of the focus adjusting screw 111 hits is slightly recessed in the right end portion. Further, a through hole for the focus adjusting screw 111 is formed at the right end portion of the slide plate 107.

When the focus adjustment screw 111 is turned to the right, the lens holding plate 102 is separated from the magnet 108 and the distance between the lens 1 and the preparation 3 is increased. When the focus adjustment screw 111 is turned to the left, the thickness is about to be reversed. The attractive force by the magnetic force of a 4 mm square with a side of 20 mm or a circular magnet with a diameter of about 20 mm and an inner hole with a diameter of about 4 mm and a magnet spacer 118 of the same shape with a thickness of 2 mm is always working. The focus adjustment screw 111 of the lens holding plate (the material of the screw is preferably a magnetic material) is slightly recessed and is not attracted greatly by the magnetic force of the screw. When the lens holding plate 102 is greatly separated from the magnet spacer 108 due to the rotation of the magnetic field, the magnetic force becomes weak, and this is not a problem when observed horizontally. When observing in the vertical may be lens-holding plate 102 deviates.

Therefore, as shown in FIG. 6, a spacer 117 made of a soft material such as elastic rubber or urethane foam and a magnet spacer 118 are used in an overlapping manner. In this way, when the screw 110 is tightened, the force is applied to the tip of the focus adjustment screw 111 so that the lens holding plate 102 does not come off even when viewed vertically, and the fixed and rotating shaft screw 110 is focused. It also works as a fine adjustment. When the magnetic force of the magnet 118 is made strong, the spacer 117 is not always necessary, but the fixed and rotating shaft screw 110 does not play a role of fine adjustment of the focus.

Further, the fixed and rotating shaft screw 110 of FIG. 6 has a length that passes through the screw hole tapped in the sample holding plate 104 but does not reach the slide plate 107, so that the slide plate has the focus adjusting screw 111. The illumination device D and the button battery in the illumination device D can be easily removed by rotating it as an axis, and the lens holding plate 102 is rotated around a fixed and rotating shaft screw 110 as an axis, so that the sub-sample holding plate 103 can be easily replaced. When returning, the magnetic force of the magnet spacer 108 and the recessed portion 102 are firmly attracted and fixed.

Further, in the embodiment of FIG. 6, the sample 3 is placed on the sample holding plate 104 via the sub sample holding plate 103 having a hole of about 10 mm in a circular plate magnet having a thickness of 1 mm and a diameter of about 30 mm. Thus, the observation range of the sample 3 can be expanded by moving the sub-sample holding plate 103. This method can also be applied to the first embodiment.
In this embodiment, the lens holding plate 102, the sample holding plate 104, and the slide plate 107 are preferably made of ferromagnetic iron or stainless steel such as SUS430 at least for the portion where the magnet is attracted.

Since the simple microscope of the present invention can be mass-produced industrially and can be provided at a low price, it is optimal for use as a school teaching material.
And most of all, the maximum possibility is just the outer shape, and details are not known due to the diffraction limit. It is possible to directly observe the moment of the eye, and the identification of bacteria and the like becomes possible by the color, and an auxiliary role of shortening the culture method by the conventional official method of long-time bacteria can be expected.

DESCRIPTION OF SYMBOLS 1 Spherical lens 2 Lens holding plate 3 Sample (The sample mounting plate was illustrated)
4 Sample holding plate 5 Light bulb 6 Plate magnet 7 Slide plate 8 Spacer 9 Spacer 10 Screw 11 Nut 14 Opening portion 20 Plate magnet 21 Electrode plate 22 Battery 23 Electrode plate 30 Illuminating device 41 Insulating sheet 42 Electrode plate 43 Electrode plate A , B Human eyes C Virtual line D of the eye D Illuminating device 51 Insulating sheet 52 Electrode plate 102 Lens holding plate 103 Sub sample holding plate 104 Sample holding plate 107 Slide plate 108 Magnet spacer 109 Magnet spacer 110 Fixing and rotating shaft screw 111 Focus adjustment screw
117 Spacer (soft material)
118 Magnet spacer 119 Magnet spacer

Claims (11)

It has a lens holding plate that holds a spherical lens and a sample holding plate that has a light transmission opening and holds the sample. The illumination device can be slid by the magnetic force on the back of the sample holding plate. The lens holding plate and the sample holding plate are detachably coupled via a spacer in a state where the distance between the spherical lens of the lens holding plate and the sample of the sample holding plate can be adjusted. A simple microscope characterized by things. A lens holding plate holding a spherical lens; a sample holding plate formed with an opening for light transmission and holding a sample; and a slide plate. The spherical lens of the lens holding plate and the sample The lens holding plate and the sample holding plate are coupled via a spacer in a detachable state with the distance between the holding plate and the sample being adjustable, and a space in which the illumination device can move is provided between the sample holding plate and the slide plate. A simple microscope characterized in that the illumination device is attached in a detachable state through a spacer, and the illumination device is adsorbed and fixed on a slide plate by a magnetic force in a slidable state. The simple microscope according to any one of claims 1 to 2, wherein lighting and extinguishing of the light bulb of the illuminating device are performed by closely contacting or separating the electrode plate from the battery electrode by a magnetic force of a permanent magnet. Simple microscope according to 3 any one of claims 1, characterized in that the material of the spacer to the magnet. The simple microscope according to any one of claims 1 to 4, wherein a sub-sample holding plate is provided in an opening of the sample holding plate. 6. The simple microscope according to claim 5 , wherein a transparent plate-like diffraction grating is provided as a sample mounting plate in an opening of the sub-sample holding plate. The sample can be observed in the darkness desired by the observer within a possible range by sliding the illumination device away from the lens and changing the line of sight. Dark field observation method using the simple microscope described. In the case where the simple microscope according to any one of claims 1 to 6 is used to record an image of a digital camera equipped with a high-sensitivity image sensor close to a spherical lens portion of a lens holding plate, the spherical shape is recorded. Using a simple microscope characterized in that the magnification image of the lens and the zoom magnification of the camera are set to 3000 times or more and 30000 times or less and a magnified image of a minute object of the observation sample is visually recognized by a color image by using chromatic aberration. Shooting and recording method. 9. The photographing and recording method using a simple microscope according to claim 8, wherein the spherical lens has a magnification of 100 to 1000. 9. The method of photographing and recording using a simple microscope according to claim 8, wherein the high-sensitivity image sensor is a back-illuminated CMOS. A spherical lens for recording and recording with a digital camera in a single microscope comprising a single spherical lens with transmitted illumination light by applying a sample to a diffraction grating using the simple microscope according to any one of claims 1 to 6. The magnification of the zoom lens of the spherical lens and the zoom magnification of the camera is set to 3000 times or more and 30000 times or less, and the enlarged image of the minute observation object is used as a color image by using chromatic aberration. A video recording method using a simple microscope characterized by visual recognition.

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