SEMITRANSPARENT SLIDE, COVER GLASS AND METHOD TO MAKE PREPARATION

Abstract: A slide is arranged to be semitransparent to scatter the light emitted from a microscope to optically eliminate the outlines of pores of a filter. Material such as cell, nuclei or extracellular materials is captured on the top surface of the filter by filtration. A specimen preparation can be made by placing the filter with the material thereon on the semitransparent slide. A metal evaporation layer, so-called multicoating, can be positioned on the surface of a cover glass, so that the material can be more clearly recognized, especially at high magnification.

Description: SPECIFICATION SEMITRANSPARENT SLIDE, COVER GLASS AND METHOD TO MAKE PREPARATION Technical Field This invention relates to a slide which is arranged to be semitransparent, a cover glass whose top surface is multicoated, and a method to prepare a specimen preparation. By placing a filter, which, in advance, has collected material such as cells by filtration, on a semitransparent slide, a specimen preparation can be prepared. Background Art In the past, a preparation is usually made by sme.arling material such as cells on a slide glass. According to this method, the material is captured by excoriation or centesis and they are smeared on a slide glass. Then the slide glass with the material thereon is preserved in such as dyeing solution, thereafter a mounting medium is applied thereon to seal the material, and a cover glass is placed thereon. In this prior art, the work to s eare the material on the slide glass is completed by hands, so that the ma erial could easily be destroyed and deformed. Further, when the material is preserved in the solution, much of it drop from the slide glass into the solution. Also, a lot of expreience and good skill are required to accomplish this smear method. To make the preparation easier, Japanese Patent Application No. 61-250188, for example, introduces a new method wherein a suspension medium including material is. drawn through a polycarbonate membrane filter, which is transparent and a thickness thereof is about 5 to 10 μ m and innumerable microscopic pores are created through the filter. After the material is captured on the filter as the suspension medium is drawn through the pores, number of needed solutions such as dyeing solutions are applied to the material. According to this new method, since it is unnecessary to smear the material on the slide glass, the problem that the material is destroyed and deformed can be eased and the outlines of the material can better be recognized as they naturally are. After the dyeing process is completed, the material is supposed to be placed on a slide glass to prepare a specimen preparation. To make the preparation with this new method, however, either the material has to be transferred to the slide glass (see U.S. Patent No. 4,395,493), or the filter with the material thereon should be placed on the slide glass and the filter has to be melted by a melting liquid, eliminating the pores and leaving the material on the slide glass. When the material collected on the filter is transferred on the slide glass, which is done by pressing the filter to the slide glass as to transcribe the material to the slide glass, much of it is destroyed and deformed by the pressure. Further, by this so-called transcribing process, it is possible that only a small percentage of the whole material can be transcribed on the slide glass. As far as the other method is concerned, the material would be shrinked and faded by an affect of the melting liquid. Conerning these problems, it is desirable that a specimen preparation can be prepared only by placing a filter with material thereon onto a slide glass, unnecessiating the transcribing and melting processes. However, suppose a specimen preparation is prepared by this method, the outlines of the pores of the filter are recognized, and they double with the outlines of the material, so that an examination of the material will become inaccurate. Therefore, the major object of the present invention is to optically eliminate the pores of the filter, to unnecessiate the transcribing and melting processes, to provide a slide (not necessarily a glass slide) and a cover glass that enable you to accurately examine the material, and to provide a method to prepare a specimen preparation using the semitransparent slide. Brief Description of Drawings Fig. 1 is a perspective view of a preferred embodiment of a semitransparent slide according to the present invention. Fig. 2 is a perspective view of another preferred embodiment of a semitransparent slide according to the present invention. Fig. 3 is a sectional view of another preferred embodiment of a se itransparent slide according to the present invention. Fig. 4 is a sectional view of a preparation made by a method to prepare a specimen preparation according to the ^ present invention. Fig. 5 is a sectional view of a preferred embodiment of a cover glass according to the present invention. Disclosure of Invention To accomplish the object of the present invention, a slide 1 is arranged to be semitransparent to scatter the light emitted from a microscope to optically eliminate the outlines of the pores 4 of a filter 3. That is, optically speaking, the pores 3 are lighted so that the degree of lightness of the outlines of the pores 4 becomes similar to that of the surrouding area, so that the pores 4 cannot be recognized. According, to the present invention, semitransparency is obtained by actively refracting and scattering the light (that is to say that the light beams are prevented from forwarding in the straight direction and they are bent in many directions), which is emitted from a microscipe while the transmittancy of the light is kept still high. It is not declining a transmi ttency of the light. There may by many ways to do so. For example, it is accomplished by including in a slide a large number of parcticles which have such specific characteristics that the light tramsmi ttancy thereof is lower than that of a slide or the light refraction thereof is much larger than that of a slide. Of course, the particles could possesses both characteristics at the same time. It is possible to adjust a degree of semitransparency by changing a roughness of the particles. Another way to effectively refract and scatter the light of a microscope is arranged such that a semitransparent layer 2, the light refraction thereof is larger than that of a slide 1, is positioned on the surface of a si ide 1. The semitransparent layer 2 can be made by including in a transparent macromo 1 ecu 1 e resin a innumerable number of articles 6, an inorganic materials made of such as ceramics. To further effectively refract the light of a microscope, the border surface of the slide 1 and the semitransparent layer 2 can be arranged rougher. Of courese, the light refraction of the slide 1 and the semitransparent layer 2 in this case must be arranged dissimilar. There may be many ways to make the border surface rougher. One of them, for example, is such that the surface of a roller, that presses and produces a slide 1 or a semitransparent layer 2, can be arranged rough. A specimen preparation according to the present invention can be prepared by at least placi g a filter 3 with material thereon on a semitransparent slide. To make a more complete preparation, an organic solven t should be appl ied on the semitransparent slide, then the filter 3 with the material 5 thereon, a mounting medium, and a cover glass 8 should respectively be placed thereon. This method prevents a formation of air bubbles in the specimen preparation. According to the present invention, when material on a filter 3 which is placed on a semitransparent slide is examined with a microscope, the slide is lighted -from underneath by the light of a microscope. In such circumstances, since the slide 1 is arranged to be semitransparent, the light emitted thereto is refracted and scattered in various directions. The refracted light emits pores 4 of the filter 3 from various directions, so that the degree of the lightness of the pores 4 becomes similar to that of the surrounding area, resulting the outlines of the pores 4 optically eliminating, though they phisically stil 1 exist. In the prior art, a slide is transparent and the light trans i ttancy thereof is high, therefore the light of a microscope advances straight ahead so that the pores 4 of a filter 3 are clealy observed, resluting the acuurate examination of material 5 i possiblet. According to the present invention, such problem is eliminated since the light from a microscope refracts and scatters in many directions. As stated previously, the effective light refraction can be obtained by arranging the border surface of a slide 1 and a semitransparen layer 2 a roug surface 7, because in this way, the light firstly refractes at the rough surface 7 and secondly at the semitransparent layer 2. According to this method, since the light can be refracted effectively, the semitransparent layer 2 can be made thinner and the number of particles included in the semitransparent 2 can be minimized, which is. advantageous when productivities are concerned. A semitransparent slide according to the present invention can be made mechanically stronger and therefore it is used for a longer time. This can be accomplished by making the semitransparent layer 2 with a macromol ecu 1 e resin and the particles 6 included in a semitransparent layer 2 with an inorganic materials such as ceramics. By -doing so, the chemical, heat and weather resistance of the resin of the semitransparent layer 2 improves. As it is known, the resin can easily be faded by chemicals and a long time use. Also, the mechanical strength of the resin is weaker than glass. Therefore, to include inorganic particles 6 in the resin to eliminate the weakness of the resin is extremely effective to make the semitransparent slide as strong as a slide made of glass. Best Mode for Carrying Out of the Invention The details of the present invention is described hereinafter. Fig. 1 shows a preferred, embodiment of the present invention. In the preferred embodiment, an unnumerable number of particles, made of ceramics having the light refraction dissimilar with the slide 1 and a characteristic of a remarkable light refraction, are included in the material of slide 1 to make the transparent slide 1 a muddy white color. The light emitted from a microscope is refracted inside the semitransparent slide, and the refracted light emits the pores 4 of a filter 3 from various directions, so that the pores 4 cannot be recognized when material on the filter 3 are examined. The slide 1 can be colored to other than white such as light blue. Fig. 2 shows another preferred embodiment of the present invention. A semitransparent layer 2, in this case, is positioned on a transparent slide 1 made of macromo 1 ecu 1 e resin. The method to arrange the macromolecule resin semitransparent is not limited to one, but, for example, it is done by crystallizing the macromolecule to white or by blending other resins in the resin. According to this preferred embodiment, the light is refracted by the semitransparent layer 2 and the similar effect pursued by the first preferred emobodiment can be achieved. This semitransparent layer 2 is crystallized in white and an unnumerab.le number of particles 6 of ceramics are contained, so that the refraction and the scattering of the light can effectively be accomplished. By adding an emitting material or a layer to the surface of each particle, the light can more effectively be refracted and scattered. Fig. 3 shows an another preferred embodiment of the present invention, wherein the border surface of the slide 1 and the semitransparent layer 2 is arranged to be a rough surface 7. By arranging so, the light is refracted by the rough surface 7, and it is further refracted by the semitransparent layer 2, so that the light can actively be refracted and scattered. Also, the slide 1 and the semitransparent layer 2 can more firmly stick each other. The rough surface 7 can be made by blasting sands to the surface of the slide 1 or by soaking the slide 1 in chemicals such as a fluoride acid. The semitransparent layer 2 can be positioned on the surface of the slide 1 by such methods that a macromolecule resin is pressed and hardened on the slide 1, it is applied or sprayed to the surface of the slide 1, it is transcribed onto the slide 1, or the slide 1 is soaked in the macromolecule resin. In this case, for example, the thickness of the slide 1 may be arranged about 1 mm and that of the semi tranparent layer 2 less than 0.2 mm to arrange the total thickness of the semitransparent slide less than 1.2 mm to meet the requirements of J I S (Japanese Industrial Standard). To pursue the object of the present invention, a high transmittancy and an active refraction and scattering are required to a transparent slide. The semitransparent slide described above possesses more than 60 ~80 percent of trans i ttancy and a highly active light refraction caused by the semitransparent layer 2, so that the pores 4 of the filter 3 can completely be erased optically. It is a highly remarkable slide. The shape of the semitransparent slide according to the present invention Js not limited to those described as the preferred embodiments. A semitransparent slide can be arranged such as a sandwich- 1 ike shape by making another transparent layer on the semitransparent layer 2. In this case, the thickness of the semi transparenet layer 2 may be prepared to be about 0.2 mm and that of transparent layer may be about 0.01 mm, arranging the total thickness of the semitransparent slide including the transparent slide 1 about 1.2 mm. By making the semitransparent slide a sandwich- 1 ike shape and adjusting a thickness of the transparent layer, a resolution becomes better and the outlines of material can clearly be recognized, so that an examination of the material becomes easier and accurate. Further, a semitransparent layer 2 can be positioned not only on the top surface of the slide 1 but also under surface or both top and under surfaces of the slide 1. The characteristics of the present invention is that a slide 1 is arranged to\be semitransparent to efficiently refract and scatter the light emitted from a microscope to optically erase the pores 4 of a filter 3. A method to make the slide semitransparent is not limited at all. Semitransparent described here inludes one that looks opaque by naked eyes but transmits the light therethrough. A slide 1 here includes such that can place things such as material and filter 3 thereon, and it is not limited to be a glass slide. Therefore, it includes, for example, products of macromolecule resin and ceramics. Further, a slide here includes one that can be recognized as opaque or transparent depending on the visual angle. Fig. 4 shows a specimen preparation made with a semitransparent slide according to the present invention and a filter 3. It is prepared by the following procedures. First, xylol is applied on the semitransparent slide and a filter 3 with material thereon is placed on it. The xylol flows into the pores 4 and push the air out of the pores 4. Therefore, the whole material 5 can be positioned at the same level on the filter 3 and air bubbles can not be formed, so that examination of the material become easier and accurate. Then, a mounting medium 9 is applied on the filter 3 and a cover glass 8 is placed thereon, completing the procedure. At this point, the xylol applied previously evaporates and it is replaced by the mounting medium 9, so that the filter 3 and the slide 1 are firmly sticked each other, and the material 5 can completely be sealed. A method to make a preparation is not limited to the one explained above. For example, any other appropriate mediums such as immersion oil can be accordingly applied thereon. A characteristic of a method to make a preparation according to the present invention is that a filter 3 with material collected thereon is pla-ced on the semitransparent slide. The effect is that the light emitted from a microscope is actively refracted and scattered, so that the pores 4 of the filter 3 are opticall-y erased, making the accurate and easier examination of the material 5 possible. The material of a filter 3 is not limited. A polyester membranf i 1 ter, a polycarbonate membrane filter, a thickness thereof is about 10/_rm with an unnumerable number of pores 4 therethrough, the diameter of the pores 4 is a few micron, and so-called a honeycomb- 1 ike filter which is made of a thin alumina having unnumerable munber of honeycomb- 1 ike pores therethrough are some of the filters preferably used as a membrane filter 3. Fig. 5 shows a cover glass 8 according to the present invention. A metal evaporation layer 10, a thickness thereof is in microns to angstroms and is formed of metal molecules such as magnesium fluoride, aluminum and zirconium oxide, is placed on the transparent cover glass 8 by a vacuum evaporation, so- called multicoating. According to a test, this cover glass is especially effective when material is to be examined at high magnification, since the multi coating layer, the metal evaporation layer 10, affects to prevent the light reflection when the light comes out from the cover glass 8 to the air. This improves the light trans i ttancy of the cover glass 8. As stated previously, according to the present invention, a slide is arranged to be semitransparent, so that. the outlines of the pores of a filter cannot be observed. Therefore, neither a so-called transcription or a mel ting procedure become unnecessary, and material on the filter can be recognized clearly. By arranging a border surface between a slide and a semi tranparent layer, the light emitted from a microscope can be more effectively refracted and scattered. By making a semitransparent layer, with a macromolecule resin and including inorganic particles therein, the degree of the light refraction becomes higher, the chemical, heat and whether resistance improves, and mechanical strength advances, so that the durability of the semitransparent slide becomes as tough as the one made of glass. According to a method to make a specimen preparation, a filter with material thereon is placed on a semitransparent slide, which unnecess ia te ,a smearing process, so that the material cannot be destroyed or deformed and they do not even drop in a dyeing from the filter. This enables you to examine the material without an erroneous. Another effect of a method to make a preparation according to the present invention is that it does not require neither a lot of expreience nor good skill and anyone could make the preparation without much difficulties. This improves an efficiency of preparing specimen preparations. Further, by making a metal evaporation particle layer on a cover glass, material can be examined more clearly especially when they are recognized at high magnification.

Claims: Claims: (1) A semitransparent slide, a part or a whole thereof is arranged to be semitransparent to refract and scatter the light. (2) A semitransparent slide claimed in claim 1, comprising a transparent slide and a lot of particles included in said transparent slide, the light transmi ttancy of said particles being lower than that of said transparent slide. (3) A semitransparent slide claimed in claim 1, wherein a transparent slide is crystallized in white. (4) A semitransparent slide claimed in claim 1, comprising a transparent slide, and a semitransparent layer positioned on the surface of said transparent slide. (5) A semitransparent slide claimed in claim 4, wherein the border surface between said transparent slide and said semitransparent layer is a rough surface. (6) A semitransparent slide claimed in claim 4, wherein said semitransparent layer is made of macromolecule resin, said macromolecule being crystallized in color. (7) A semitransparent slide claimed in claim 4, wherein said semitransparent layer is made of macromolecule resin, said macromolecule layer includes a lot of inorganic particles such as globular shape powder, pigments, powdered bones, ceramics and so on. (8) A cover glass comprising a metal evaporation layer thereon. (9) A method to make a preparation which comprises: at least placing a filter on a slide, said filter having material thereon. (10) A method to make a preparation which comprises: placing a filter with material thereon on a semitransparent slide, dropping a mounting medium on said filter, and thereafter placing a cover glass on said filter.

Inventors: SHIBASAKI MICHIRO, SHIBASAKI, MICHIRO
Applicants: SHIBASAKI MICHIRO, SHIBASAKI, MICHIRO
Application year: 1989
Main classification: G02B21
IPC codes: G01N1, G02B21