JP2006023644A - Container for microscopic observation - Google Patents

Container for microscopic observation Download PDF

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JP2006023644A
JP2006023644A JP2004203358A JP2004203358A JP2006023644A JP 2006023644 A JP2006023644 A JP 2006023644A JP 2004203358 A JP2004203358 A JP 2004203358A JP 2004203358 A JP2004203358 A JP 2004203358A JP 2006023644 A JP2006023644 A JP 2006023644A
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aqueous solution
well
transparent
microscope observation
container
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Motohiro Suzuki
基弘 鈴木
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Nikon Corp
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Abstract

<P>PROBLEM TO BE SOLVED: To achieve appropriate microscopic observation regardless of height of the level of an aqueous solution in a container is high or low. <P>SOLUTION: A well 2a contains a biological sample S and an aqueous solution W. As shown in figure (a), when the level of the aqueous solution W is high, a transparent disk 3 floats on the aqueous solution W and the upper surface P1 and the under surface P2 of a disk body 4 keep parallelism with the bottom face of the well 2a. As shown in figure (b), when the level of the aqueous solution W is low, protrusions 5 of the transparent disk 3 get contact with the bottom face of the well 2a, therefore the upper surface P1 and the under surface P2 of the disk body 4 keep parallelism with the bottom face of the well 2a, and the disk body 4 is prevented from touching and pressing the biological sample S. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

本発明は、生体試料および水溶液を収納して顕微鏡ステージに設置される顕微鏡観察用容器に関する。   The present invention relates to a microscope observation container that contains a biological sample and an aqueous solution and is placed on a microscope stage.

生物細胞を培養しながら顕微鏡観察したり、生物細胞に試薬を添加して反応状態を顕微鏡観察するのに用いられる容器として、例えばマイクロプレート（ウエルプレート）がある。マイクロプレートに培養液や染色液などの水溶液を注入すると、表面張力の作用でマイクロプレート壁面近くで液面が彎曲し、そのレンズ作用により顕微鏡像が歪んで見える。特に、口径が１０ｍｍ程度以下のマイクロプレートでは、この悪影響が顕著に現れる。そこで、この液面彎曲を防止するために、透明平板を水溶液に浮かせて観察する方法が知られている（例えば、特許文献１参照）。   For example, a microplate (well plate) is used as a container for observing a biological cell while culturing it, or adding a reagent to the biological cell and observing the reaction state under a microscope. When an aqueous solution such as a culture solution or staining solution is injected into the microplate, the liquid surface is bent near the wall surface of the microplate due to the surface tension, and the microscopic image appears to be distorted due to the lens action. In particular, this adverse effect is prominent in microplates having a diameter of about 10 mm or less. Therefore, in order to prevent this liquid surface bending, a method of observing a transparent flat plate in an aqueous solution is known (for example, see Patent Document 1).

特開平５−１８１０６８号公報（第２頁、図３，５）Japanese Patent Laid-Open No. 5-181068 (2nd page, FIGS. 3 and 5)

マイクロプレート中の水溶液の液面の高さは、常に一定とは限らない。水溶液の量が減ってしまうと、上記の特許文献１の技術では、透明平板の下面が生物細胞（試料）に接触したり、生物細胞を押圧することになり、正常な状態での顕微鏡観察を妨げるという問題がある。   The level of the aqueous solution in the microplate is not always constant. When the amount of the aqueous solution decreases, in the technique of Patent Document 1 described above, the lower surface of the transparent flat plate comes into contact with the biological cell (sample) or presses the biological cell. There is a problem of obstructing.

（１）上記課題を解決するために、請求項１の発明に係る顕微鏡観察用容器は、生体試料および水溶液を収納する容器本体と、容器本体中の水溶液に浮いた状態で上面が水平面と平行になる透明平板とを備え、透明平板は、上面と平行な下面の外周部に突起を有することを特徴とする。
請求項２の発明に係る顕微鏡観察用容器は、請求項１の顕微鏡観察用容器において、突起が透明平板よりも密度が大きいことが好ましい。
請求項３の発明に係る顕微鏡観察用容器は、請求項１または２の顕微鏡観察用容器において、突起が３個以上設けられていることが好ましい。
（２）請求項４の発明に係る顕微鏡観察用容器は、請求項１〜３の顕微鏡観察用容器において、透明平板の上面に、疎水性処理が施されていてもよい。
請求項５の発明に係る顕微鏡観察用容器は、請求項１〜４のいずれかの顕微鏡観察用容器において、透明平板の下面に、親水性処理が施されていてもよい。 (1) In order to solve the above-mentioned problem, a microscope observation container according to the invention of claim 1 includes a container main body for storing a biological sample and an aqueous solution, and a top surface parallel to a horizontal plane in a state of floating in the aqueous solution in the container main body. The transparent flat plate has a protrusion on the outer peripheral portion of the lower surface parallel to the upper surface.
In the microscope observation container according to the second aspect of the present invention, in the microscope observation container according to the first aspect, the protrusions preferably have a density higher than that of the transparent flat plate.
The microscope observation container according to the invention of claim 3 is preferably provided with three or more projections in the microscope observation container of claim 1 or 2.
(2) The microscope observation container according to the invention of claim 4 is the microscope observation container of claims 1 to 3, wherein the upper surface of the transparent flat plate may be subjected to hydrophobic treatment.
The microscope observation container according to the invention of claim 5 is the microscope observation container according to any one of claims 1 to 4, wherein the lower surface of the transparent flat plate may be subjected to a hydrophilic treatment.

本発明によれば、透明平板の下面の外周部に突起を設けたので、水溶液の液面が下がっても透明平板が試料に触れることなく適正な観察ができる。   According to the present invention, since the protrusion is provided on the outer peripheral portion of the lower surface of the transparent flat plate, proper observation can be performed without the transparent flat plate touching the sample even when the liquid level of the aqueous solution is lowered.

以下、本発明の実施の形態による顕微鏡観察用容器について、図１〜５を参照しながら説明する。
図１は、本発明の実施の形態による顕微鏡観察用容器が顕微鏡観察のためにセットされる倒立型顕微鏡の構成を模式的に示す全体構成図である。 Hereinafter, a microscope observation container according to an embodiment of the present invention will be described with reference to FIGS.
FIG. 1 is an overall configuration diagram schematically showing the configuration of an inverted microscope in which a microscope observation container according to an embodiment of the present invention is set for microscope observation.

図１に示される倒立型顕微鏡１０は、基台１１、ステージ１２、照明系支柱１３、鏡筒１４および対物レンズ１５を備える。ステージ１２は、顕微鏡観察用容器１を載置し、図中、Ｘ−Ｙ方向に移動できるとともに、焦準ハンドル１６によりＺ方向（光軸方向）に移動できるようになっている。照明系支柱１３の上部には、光源１７ａを収納するランプハウス１７が設けられ、照明系支柱１３の内部には、照明光学系が収納されている。鏡筒１４の先端には、接眼レンズ１８が設けられ、鏡筒１４内部には、透過観察用の光学系が収納されている。   An inverted microscope 10 shown in FIG. 1 includes a base 11, a stage 12, an illumination system support 13, a lens barrel 14, and an objective lens 15. The stage 12 mounts the microscope observation container 1 and can be moved in the XY direction in the drawing, and can be moved in the Z direction (optical axis direction) by the focusing handle 16. A lamp house 17 that houses a light source 17 a is provided above the illumination system support 13, and an illumination optical system is stored inside the illumination system support 13. An eyepiece 18 is provided at the tip of the lens barrel 14, and an optical system for transmission observation is accommodated inside the lens barrel 14.

光源１７ａから発する照明光Ｌ１は、顕微鏡観察用容器１（以下、ウエルプレート１）の上側からウエルプレート１へ入射する。倒立型顕微鏡１０においては、照明光Ｌ１が不図示のコンデンサレンズによって視野全体を均一に照射する、いわゆるケーラー照明が採用されている。ウエルプレート１中の観察対象物を透過した透過光Ｌ２は、ウエルプレート１の下側に位置する対物レンズ１５へ入射し、対物レンズ１５により顕微鏡像が形成される。この顕微鏡像は、接眼レンズ１８により観察される。なお、倍率や開口数などが異なる複数種類の対物レンズ１５をレボルバ１５ａに装着し、観察時にはレボルバ１５ａの回転により任意の対物レンズ１５を光路に挿入してもよい。   The illumination light L1 emitted from the light source 17a enters the well plate 1 from the upper side of the microscope observation container 1 (hereinafter, well plate 1). The inverted microscope 10 employs so-called Koehler illumination in which the illumination light L1 uniformly irradiates the entire field of view with a condenser lens (not shown). The transmitted light L2 that has passed through the observation object in the well plate 1 is incident on the objective lens 15 located on the lower side of the well plate 1, and a microscopic image is formed by the objective lens 15. This microscopic image is observed by the eyepiece 18. A plurality of types of objective lenses 15 having different magnifications and numerical apertures may be attached to the revolver 15a, and the arbitrary objective lenses 15 may be inserted into the optical path by rotating the revolver 15a during observation.

図２は、本発明の実施の形態によるウエルプレート１の外観を模式的に示す斜視図である。ウエルプレート１は、ウエルと称する多数の円柱状の孔がマトリックス状に形成された透明プレート２と、各々のウエル２ａにそれぞれ対応する複数個の透明円板３とから成る。透明円板３は、通常、その外径がウエル２ａの内径よりもやや小さく、その厚さがウエル２ａの深さよりも薄く作製される。透明円板３の厚さは、ウエルの寸法にもよるが、０．１〜１ｍｍの範囲が好ましい。   FIG. 2 is a perspective view schematically showing the appearance of the well plate 1 according to the embodiment of the present invention. The well plate 1 includes a transparent plate 2 in which a large number of cylindrical holes called wells are formed in a matrix, and a plurality of transparent disks 3 corresponding to the respective wells 2a. The transparent disk 3 is usually manufactured with an outer diameter slightly smaller than the inner diameter of the well 2a and a thickness smaller than the depth of the well 2a. The thickness of the transparent disc 3 is preferably in the range of 0.1 to 1 mm, although it depends on the dimensions of the well.

ウエルプレート１の大きさを一定とすると、各々のウエル２ａの内径を大きくすれば、ウエル２ａの個数は少なくなり、各々のウエル２ａの内径を小さくすれば、ウエル２ａの個数は多くなるので、目的に応じて種々のウエルプレート１が使い分けられている。各々のウエル２ａには、観察対象物として様々な種類の生物試料Ｓと水溶液Ｗ（培養液、染色液など）が収容される。生物試料Ｓと水溶液Ｗがウエル２ａの中に注入された後に、透明円板３がウエル２ａの中に挿入され、顕微鏡観察が行われる。   If the size of the well plate 1 is constant, if the inner diameter of each well 2a is increased, the number of wells 2a decreases. If the inner diameter of each well 2a is decreased, the number of wells 2a increases. Various well plates 1 are properly used according to the purpose. Each well 2a accommodates various types of biological samples S and aqueous solutions W (culture solution, staining solution, etc.) as observation objects. After the biological sample S and the aqueous solution W are injected into the well 2a, the transparent disk 3 is inserted into the well 2a, and microscopic observation is performed.

図３を参照して透明円板３について詳細に説明する。図３は、透明円板３の外観を模式的に示す斜視図である。透明円板３は、円板本体４と突起部５から成る。円板本体４の上面Ｐ１と下面Ｐ２は、いずれも平面であり、互いに平行である。下面Ｐ２の外周部分Ｒ（２点鎖線の外側領域）には、寸法形状および重量が等しい３個の円柱状の突起部５が設けられ、これらの３個の突起部５は、等間隔に配列している。突起部５の長さは、生物試料Ｓの大きさ（厚さ）にもよるが、２０μｍ以上が望ましい。円板本体４と突起部５とを同一のプラスチック材料で作製する場合は、例えば一体成型で加工する。突起部を円板本体４よりも高い密度の材料で作製する場合は、例えば接着により円板本体４に固定する。なお、透明円板３の形状は、ウエル２ａの形状と同じにせずに、例えば多角形や星型としてもよい。   The transparent disk 3 will be described in detail with reference to FIG. FIG. 3 is a perspective view schematically showing the appearance of the transparent disk 3. The transparent disk 3 includes a disk body 4 and a protrusion 5. The upper surface P1 and the lower surface P2 of the disc body 4 are both flat and parallel to each other. Three columnar projections 5 having the same size and weight are provided on the outer peripheral portion R (outer region of the two-dot chain line) of the lower surface P2, and these three projections 5 are arranged at equal intervals. is doing. Although the length of the protrusion 5 depends on the size (thickness) of the biological sample S, it is preferably 20 μm or more. When manufacturing the disc main body 4 and the projection part 5 with the same plastic material, it processes by integral molding, for example. When the protrusion is made of a material having a higher density than that of the disk main body 4, it is fixed to the disk main body 4 by adhesion, for example. The shape of the transparent disk 3 may be, for example, a polygonal shape or a star shape, without being the same as the shape of the well 2a.

透明円板３は、水溶液Ｗに浮かせて用いられるので、水溶液Ｗの密度よりも小さいものであればよい。水溶液Ｗの密度は、溶質の種類や含有率によって異なるが、一般に１ｇ／ｃｍ３以上である。簡単のために、透明円板３の体積の大半を占める円板本体４の密度と水溶液Ｗの密度とを比較することにより、透明円板３が水溶液Ｗに浮くか否かを説明する。円板本体４の材質としては、例えばポリエチレン、ポリプロピレン、ポリスチレンが挙げられる。また、突起部５を円板本体４よりも高い密度の材料で作製する場合は、突起部５の材質として、ＡＳ樹脂（スチレン−アクリロニトリル共重合体：密度１．０８ｇ／ｃｍ３）、ＡＢＳ樹脂（密度１．０６ｇ／ｃｍ３）が挙げられる。 Since the transparent disk 3 is used while being floated on the aqueous solution W, any material that is smaller than the density of the aqueous solution W may be used. The density of the aqueous solution W varies depending on the type and content of the solute, but is generally 1 g / cm 3 or more. For simplicity, whether or not the transparent disk 3 floats in the aqueous solution W will be described by comparing the density of the disk body 4 that occupies most of the volume of the transparent disk 3 and the density of the aqueous solution W. Examples of the material of the disc main body 4 include polyethylene, polypropylene, and polystyrene. Further, when the protrusion 5 is made of a material having a higher density than that of the disc body 4, the material of the protrusion 5 is AS resin (styrene-acrylonitrile copolymer: density 1.08 g / cm 3 ), ABS resin. (Density 1.06 g / cm 3 ).

ポリエチレンとポリプロピレンは、密度が１ｇ／ｃｍ３以下であるので水溶液Ｗに浮くが、ポリスチレン（密度１．０５ｇ／ｃｍ３）は、密度が１ｇ／ｃｍ３よりわずかに大きいので、水溶液Ｗの密度によって浮く場合と浮かない場合がある。従って、水溶液Ｗの密度がポリスチレンの密度を越えていれば、ポリスチレン製の円板本体４でも使用できる。なお、透明円板３は、水溶液Ｗに浮くという条件を満たせばよいので、例えば円板本体４を中空構造として、見かけの比重を水溶液Ｗの密度より小さくすれば使用可能となる。 Since polyethylene and polypropylene have a density of 1 g / cm 3 or less, they float in the aqueous solution W, but since polystyrene (density 1.05 g / cm 3 ) has a density slightly higher than 1 g / cm 3, it depends on the density of the aqueous solution W. May float or not float. Therefore, if the density of the aqueous solution W exceeds the density of polystyrene, the disk body 4 made of polystyrene can be used. In addition, since the transparent disk 3 should just satisfy | fill the conditions that it floats in the aqueous solution W, for example, if the disk main body 4 is made into a hollow structure and an apparent specific gravity is made smaller than the density of the aqueous solution W, it can be used.

円板本体４の上面Ｐ１には疎水性処理が施されている。疎水化の方法には、ふっ素系樹脂の塗布、ふっ素系化合物のプラズマ重合処理、カップリング剤による処理がある。透明円板３の上面Ｐ１は、疎水性を呈しているので、水溶液Ｗとの濡れ性が悪く、水溶液Ｗの液滴が上面Ｐ１の上に留まることがない。従って、上面Ｐ１は、常に露出した状態に保たれるので、照明光Ｌ１は常に平面に入射することになる。   The upper surface P1 of the disc body 4 is subjected to a hydrophobic treatment. Hydrophobization methods include application of a fluorine-based resin, plasma polymerization of a fluorine-based compound, and treatment with a coupling agent. Since the upper surface P1 of the transparent disk 3 is hydrophobic, the wettability with the aqueous solution W is poor, and droplets of the aqueous solution W do not stay on the upper surface P1. Therefore, since the upper surface P1 is always kept exposed, the illumination light L1 always enters the plane.

また、円板本体４の下面Ｐ２には親水性処理が施されている。親水性付与の方法には、界面活性剤の塗布やグラフト重合、プラズマ重合、紫外線照射などの処理がある。透明円板３の下面Ｐ２は、親水性を呈しているので、水溶液Ｗとの濡れ性が良く、透明円板３の姿勢を安定させる効果がある。   Further, the lower surface P2 of the disc body 4 is subjected to hydrophilic treatment. Examples of methods for imparting hydrophilicity include treatments such as application of a surfactant, graft polymerization, plasma polymerization, and ultraviolet irradiation. Since the lower surface P2 of the transparent disc 3 exhibits hydrophilicity, it has good wettability with the aqueous solution W and has an effect of stabilizing the posture of the transparent disc 3.

以下、図４を参照しながら、ウエル２ａ内の生物試料Ｓ、水溶液Ｗおよび透明円板３の状態を説明する。図４（ａ）は、水溶液Ｗの液面Ａの水位が高い場合のウエル２ａ内の状態を模式的に示す部分断面図である。図４（ｂ）は、水溶液Ｗの液面Ａの水位が低い場合のウエル２ａ内の状態を模式的に示す部分断面図である。図４（ａ）と図４（ｂ）とが異なる点は、水溶液Ｗの液面Ａの水位だけである。   Hereinafter, the state of the biological sample S, the aqueous solution W, and the transparent disc 3 in the well 2a will be described with reference to FIG. FIG. 4A is a partial cross-sectional view schematically showing a state in the well 2a when the water level of the liquid surface A of the aqueous solution W is high. FIG. 4B is a partial cross-sectional view schematically showing the state in the well 2a when the water level of the liquid surface A of the aqueous solution W is low. The difference between FIG. 4A and FIG. 4B is only the water level of the liquid surface A of the aqueous solution W.

図４（ａ）を参照すると、生物試料Ｓは、水溶液Ｗの中にあり、ウエル２ａの底面に接触している。透明円板３は、水溶液Ｗに浮いた状態で、円板本体４の上面Ｐ１、下面Ｐ２がウエル２ａの底面と平行を保っている。突起部５を付設することにより、水溶液Ｗに浮いた状態で重心位置が下方（図１のＺ方向）へ移動するが、水平面（図１のＸ−Ｙ方向）での重心移動はないので、透明円板３が傾斜することはない。また、突起部５を円板本体４よりも密度が高い材料で作製すれば、水溶液Ｗに浮いた状態で透明円板３の安定性がより向上し、透明円板３が上下反転するような事態を防ぐ効果が大きくなる。   Referring to FIG. 4A, the biological sample S is in the aqueous solution W and is in contact with the bottom surface of the well 2a. In the state where the transparent disk 3 floats in the aqueous solution W, the upper surface P1 and the lower surface P2 of the disk body 4 are kept parallel to the bottom surface of the well 2a. By attaching the protrusion 5, the center of gravity moves downward (Z direction in FIG. 1) while floating in the aqueous solution W, but there is no center of gravity movement in the horizontal plane (XY direction in FIG. 1). The transparent disk 3 does not tilt. Further, if the protrusion 5 is made of a material having a higher density than the disk body 4, the stability of the transparent disk 3 is further improved in the state of being floated on the aqueous solution W, and the transparent disk 3 is turned upside down. The effect of preventing the situation is increased.

上述したように、３個の突起部５は、同一重量であり、下面Ｐ２の外周部分に等間隔に配列しているので、透明円板３全体としてのバランスは崩れない。突起部５の個数は、４個以上であっても、下面Ｐ２の外周部分に等間隔に配列すればバランスは維持される。   As described above, since the three protrusions 5 have the same weight and are arranged at equal intervals on the outer peripheral portion of the lower surface P2, the balance of the entire transparent disk 3 is not lost. Even if the number of the protrusions 5 is four or more, the balance is maintained if they are arranged at equal intervals on the outer peripheral portion of the lower surface P2.

図中、上方からの照明光Ｌ１は、水平面である上面Ｐ１へ垂直に入射し、円板本体４、水溶液Ｗ、生物試料Ｓを透過し、透過光Ｌ２として透明プレート２の底部から下方へ射出する。照明光Ｌ１の光束を上面Ｐ１の外周部分Ｒの内側に入射させることにより、照明光Ｌ１が突起部５に遮蔽されることなく、生物試料Ｓはケーラー照明の条件で照明される。   In the figure, illumination light L1 from above enters perpendicularly to the upper surface P1, which is a horizontal plane, passes through the disc body 4, the aqueous solution W, and the biological sample S, and is emitted downward from the bottom of the transparent plate 2 as transmitted light L2. To do. By causing the luminous flux of the illumination light L1 to enter the inside of the outer peripheral portion R of the upper surface P1, the biological sample S is illuminated under the Koehler illumination conditions without the illumination light L1 being shielded by the protrusions 5.

図４（ｂ）では、図４（ａ）に比べて水溶液Ｗの水位が下がったため、透明円板３の突起部５の先端がウエル２ａの底面に接している。透明円板３は、水位の変化に応じて上下するからである。透明円板３には突起部５があるので、円板本体４の下面Ｐ２が生物試料Ｓに接触したり押圧することは回避される。透明円板３は、水溶液Ｗに浮いた状態ではなくなっているが、円板本体４の上面Ｐ１、下面Ｐ２がウエル２ａの底面と平行を保っていることには変わりはない。従って、図４（ｂ）でも図４（ａ）と同様に、生物試料Ｓは照明光Ｌ１によりケーラー照明の条件で照明される。このように、透明円板３を使用することにより、水溶液Ｗの水位によらず、生物試料Ｓを適切な照明条件で観察することができる。   In FIG. 4B, since the water level of the aqueous solution W is lower than that in FIG. 4A, the tip of the protrusion 5 of the transparent disk 3 is in contact with the bottom surface of the well 2a. This is because the transparent disk 3 moves up and down according to the change in the water level. Since the transparent disk 3 has the protrusions 5, it is avoided that the lower surface P <b> 2 of the disk body 4 contacts or presses the biological sample S. The transparent disk 3 is no longer in the state of floating in the aqueous solution W, but the upper surface P1 and the lower surface P2 of the disk main body 4 remain unchanged from the bottom surface of the well 2a. Accordingly, in FIG. 4B, as in FIG. 4A, the biological sample S is illuminated by the illumination light L1 under the Koehler illumination condition. Thus, by using the transparent disc 3, the biological sample S can be observed under appropriate illumination conditions regardless of the water level of the aqueous solution W.

ここで、図５を参照して、透明円板３を使用しない場合のウエル２ａ中の生物試料Ｓと水溶液Ｗの状態を説明する。図５は、ウエル２ａの内部の状態を模式的に示す部分断面図であり、図４の場合と比較するための図である。図５においても図４と同様に、生物試料Ｓは、水溶液Ｗの中にあり、ウエル２ａの底面に接触している。しかし、透明円板３を使用しないために、水溶液Ｗの液面Ａは、表面張力の作用で図示のように彎曲している。このように液面Ａが彎曲していると、そのレンズ作用によってケーラー照明の条件を満たさなくなり照明ムラが生じる。また、液面Ａが彎曲していると、位相差顕微鏡観察においては、輪帯照明光を形成する遮光板と対物レンズの瞳の位置にある位相リングとが共役であるという条件を満たさなくなり、適正な観察に支障をきたすことになる。このような顕微鏡観察に及ぼす悪影響は、ウエル２ａの内径が小さくなるほど顕著になるが、図４で説明したように、透明円板３を使用することにより、ウエル２ａの内径の大小を問わず適正な顕微鏡観察が可能となる。   Here, the state of the biological sample S and the aqueous solution W in the well 2a when the transparent disk 3 is not used will be described with reference to FIG. FIG. 5 is a partial cross-sectional view schematically showing the internal state of the well 2a, for comparison with the case of FIG. In FIG. 5, as in FIG. 4, the biological sample S is in the aqueous solution W and is in contact with the bottom surface of the well 2a. However, since the transparent disk 3 is not used, the liquid level A of the aqueous solution W is bent as shown in the figure due to the action of surface tension. When the liquid surface A is curved in this way, the lens action does not satisfy the conditions of Kohler illumination, resulting in illumination unevenness. Further, when the liquid surface A is curved, in the phase-contrast microscope observation, the condition that the light shielding plate for forming the annular illumination light and the phase ring at the position of the pupil of the objective lens are conjugate is not satisfied, It will interfere with proper observation. Such an adverse effect on microscopic observation becomes more pronounced as the inner diameter of the well 2a becomes smaller. However, as described with reference to FIG. 4, the use of the transparent disk 3 makes it appropriate regardless of the inner diameter of the well 2a. Microscope observation is possible.

以上説明したように、本実施の形態の透明円板３は、次のような効果がある。（１）透明円板３には突起部５があるので、水溶液Ｗの水位が下がっても円板本体４の下面Ｐ２が生物試料Ｓに接触したり押圧することを回避できる。（２）透明円板３を水溶液Ｗに浮かせることにより、液面Ａの彎曲によるレンズ作用を除去できる。（３）透明円板３が水溶液Ｗの液面Ａのほとんどを覆っているので、水溶液Ｗの蒸発や水溶液Ｗ中への塵埃、異物の混入を防止できる。（４）透明円板３を取り外さずに、透明円板３の上から水溶液Ｗを補充したり他の試薬を添加することができるので、作業性を低下させることがない。（５）透明円板３の上面Ｐ１は、疎水性を呈しているので、補充した水溶液Ｗなどの液滴が上面Ｐ１の上に留まることがない。（６）透明円板３の下面Ｐ２は、親水性を呈しているので、水溶液Ｗとの濡れ性が良く、透明円板３の姿勢が安定する。   As described above, the transparent disk 3 of the present embodiment has the following effects. (1) Since the transparent disk 3 has the protrusions 5, it can be avoided that the lower surface P <b> 2 of the disk body 4 contacts or presses the biological sample S even if the water level of the aqueous solution W is lowered. (2) By floating the transparent disk 3 in the aqueous solution W, the lens action due to the curvature of the liquid surface A can be removed. (3) Since the transparent disk 3 covers most of the liquid surface A of the aqueous solution W, evaporation of the aqueous solution W and entry of dust and foreign matter into the aqueous solution W can be prevented. (4) Since the aqueous solution W can be replenished or another reagent can be added from above the transparent disk 3 without removing the transparent disk 3, workability is not lowered. (5) Since the upper surface P1 of the transparent disk 3 exhibits hydrophobicity, the replenished liquid droplets such as the aqueous solution W do not stay on the upper surface P1. (6) Since the lower surface P2 of the transparent disk 3 is hydrophilic, the wettability with the aqueous solution W is good, and the posture of the transparent disk 3 is stabilized.

本発明は、顕微鏡観察用容器（ウエルプレート）の透明円板の下面の外周部に突起を設けたことに特徴がある。本発明は、その特徴を損なわない限り、以上説明した実施の形態に何ら限定されない。例えば、この顕微鏡観察用容器を正立型顕微鏡による観察に用いることもできる。   The present invention is characterized in that protrusions are provided on the outer peripheral portion of the lower surface of the transparent disk of the microscope observation container (well plate). The present invention is not limited to the embodiments described above as long as the characteristics are not impaired. For example, the microscope observation container can be used for observation with an upright microscope.

本発明の実施の形態に係るウエルプレートが顕微鏡観察のためにセットされる倒立型顕微鏡の構成を模式的に示す全体構成図である。It is a whole block diagram which shows typically the structure of the inverted microscope in which the well plate which concerns on embodiment of this invention is set for microscope observation. 本発明の実施の形態に係るウエルプレートの外観を模式的に示す斜視図である。It is a perspective view which shows typically the external appearance of the well plate which concerns on embodiment of this invention. 本発明の実施の形態に係るウエルプレートの透明円板の外観を模式的に示す斜視図である。It is a perspective view which shows typically the external appearance of the transparent disc of the well plate which concerns on embodiment of this invention. 本発明の実施の形態に係るウエルプレートのウエル内の状態を模式的に示す部分断面図である。図４（ａ）は、水溶液Ｗの液面Ａの水位が高い場合、図４（ｂ）は、水溶液Ｗの液面Ａの水位が低い場合を示す。It is a fragmentary sectional view which shows typically the state in the well of the well plate which concerns on embodiment of this invention. 4A shows a case where the level of the liquid surface A of the aqueous solution W is high, and FIG. 4B shows a case where the level of the liquid level A of the aqueous solution W is low. 本発明の実施の形態の比較例であり、ウエルプレートのウエル内の状態を模式的に示す部分断面図である。It is a comparative example of embodiment of this invention, and is a fragmentary sectional view which shows typically the state in the well of a well plate.

符号の説明Explanation of symbols

１：ウエルプレート
２：透明プレート
２ａ：ウエル
３：透明円板
４：円板本体
５：突起部
１０：倒立型顕微鏡
１１：基台
１２：ステージ
１３：照明系支柱
１４：鏡筒
１５：対物レンズ
Ａ：液面
Ｌ１：照明光
Ｌ２：透過光
Ｐ１：上面
Ｐ２：下面
Ｓ：生物試料
Ｗ：水溶液 1: Well plate 2: Transparent plate 2a: Well 3: Transparent disc 4: Disc main body 5: Projection 10: Inverted microscope 11: Base 12: Stage 13: Illumination system support 14: Lens barrel 15: Objective lens A: Liquid surface L1: Illumination light L2: Transmitted light P1: Upper surface P2: Lower surface S: Biological sample W: Aqueous solution

Claims (5)

生体試料および水溶液を収納する容器本体と、
前記容器本体中の水溶液に浮いた状態で上面が水平面と平行になる透明平板とを備え、
前記透明平板は、前記上面と平行な下面の外周部に突起を有することを特徴とする顕微鏡観察用容器。 A container body for storing a biological sample and an aqueous solution;
A transparent flat plate whose upper surface is parallel to the horizontal plane in a state of floating in the aqueous solution in the container body,
The said transparent flat plate has a processus | protrusion in the outer peripheral part of the lower surface parallel to the said upper surface, The container for microscope observation characterized by the above-mentioned. 請求項１に記載の顕微鏡観察用容器において、
前記突起は、前記透明平板よりも密度が大きいことを特徴とする顕微鏡観察用容器。 In the microscope observation container according to claim 1,
The projection has a density higher than that of the transparent flat plate. 請求項１または２に記載の顕微鏡観察用容器において、
前記突起は、３個以上設けられていることを特徴とする顕微鏡観察用容器。 In the microscope observation container according to claim 1 or 2,
A microscope observation container, wherein three or more protrusions are provided. 請求項１〜３のいずれか一項に記載の顕微鏡観察用容器において、
前記透明平板の上面には、疎水性処理が施されていることを特徴とする顕微鏡観察用容器。 In the microscope observation container according to any one of claims 1 to 3,
A microscope observation container, wherein an upper surface of the transparent flat plate is subjected to hydrophobic treatment. 請求項１〜４のいずれか一項に記載の顕微鏡観察用容器において、
前記透明平板の下面には、親水性処理が施されていることを特徴とする顕微鏡観察用容器。 In the container for microscope observation as described in any one of Claims 1-4,
A microscope observation container, wherein the lower surface of the transparent flat plate is subjected to hydrophilic treatment.
JP2004203358A 2004-07-09 2004-07-09 Container for microscopic observation Pending JP2006023644A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010286510A (en) * 2009-06-09 2010-12-24 Nikon Corp Observing apparatus
WO2012117647A1 (en) * 2011-02-28 2012-09-07 三洋電機株式会社 Observation program and observation device
JP2015004801A (en) * 2013-06-20 2015-01-08 Ｊｆｅスチール株式会社 Fixing jig of sample for microscopic observation
WO2015198866A1 (en) * 2014-06-24 2015-12-30 日本写真印刷株式会社 Culture container

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010286510A (en) * 2009-06-09 2010-12-24 Nikon Corp Observing apparatus
WO2012117647A1 (en) * 2011-02-28 2012-09-07 三洋電機株式会社 Observation program and observation device
US9189677B2 (en) 2011-02-28 2015-11-17 Panasonic Healthcare Holdings Co., Ltd. Recording medium having observation program recorded therein and observation apparatus
JP2015004801A (en) * 2013-06-20 2015-01-08 Ｊｆｅスチール株式会社 Fixing jig of sample for microscopic observation
WO2015198866A1 (en) * 2014-06-24 2015-12-30 日本写真印刷株式会社 Culture container
US10138454B2 (en) 2014-06-24 2018-11-27 Nissha Printing Co., Ltd. Culture container

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