JP2002277365A - Sample holder for high-pressure freezer - Google Patents
Sample holder for high-pressure freezerInfo
- Publication number
- JP2002277365A JP2002277365A JP2001391608A JP2001391608A JP2002277365A JP 2002277365 A JP2002277365 A JP 2002277365A JP 2001391608 A JP2001391608 A JP 2001391608A JP 2001391608 A JP2001391608 A JP 2001391608A JP 2002277365 A JP2002277365 A JP 2002277365A
- Authority
- JP
- Japan
- Prior art keywords
- sample
- sample holder
- frame member
- frame members
- pressure
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000010432 diamond Substances 0.000 claims abstract description 28
- 229910003460 diamond Inorganic materials 0.000 claims abstract description 23
- 238000007710 freezing Methods 0.000 claims abstract description 12
- 230000008014 freezing Effects 0.000 claims abstract description 12
- 125000006850 spacer group Chemical group 0.000 claims description 15
- 229910052751 metal Inorganic materials 0.000 claims description 14
- 239000002184 metal Substances 0.000 claims description 14
- 238000012856 packing Methods 0.000 claims description 7
- 238000001816 cooling Methods 0.000 abstract description 19
- 239000007788 liquid Substances 0.000 abstract description 7
- 239000000523 sample Substances 0.000 description 67
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000012472 biological sample Substances 0.000 description 4
- 230000005540 biological transmission Effects 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 3
- 230000002093 peripheral effect Effects 0.000 description 3
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000002826 coolant Substances 0.000 description 2
- 229910052802 copper Inorganic materials 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 229910052737 gold Inorganic materials 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 230000001143 conditioned effect Effects 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000004017 vitrification Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、高圧凍結装置用試
料ホルダに関する。更に、本発明は、互いに連結・分離
可能な少なくとも２つのフレーム部材を有し、かつ連結
されたフレーム部材が試料の収容部を形成する、高圧凍
結用試料ホルダに関する。[0001] The present invention relates to a sample holder for a high-pressure freezing apparatus. Furthermore, the present invention relates to a sample holder for high-pressure freezing, which has at least two frame members that can be connected to and separated from each other, and the connected frame members form a sample storage portion.
【０００２】本発明における高圧凍結装置の意義は、高
圧下において生物試料その他の試料（その処理物ないし
中間体を含む）、とりわけ含液試料（主として水（H
2O）を含む試料であるが、アルコール等の液体により水
の全部ないし一部を置換した試料も含まれる）を高速に
凍結する（冷凍保存ないしガラス化：Vitrifizieren）
ための凍結装置を言う。The significance of the high-pressure freezing device in the present invention is that a biological sample or other sample (including its processed product or intermediate), especially a liquid-containing sample (mainly water (H
A sample containing 2 O), but the sample obtained by replacing all or part of the water by a liquid such as alcohol included) and frozen quickly (frozen or vitrified: Vitrifizieren)
For freezing equipment.
【０００３】[0003]
【関連する技術】上述の種類の装置は、DE-PS-180674
1、EP0853238A1及びEP0637741A1に記載されている。[Related Art] An apparatus of the type described above is disclosed in DE-PS-180674.
1, EP0853238A1 and EP0637741A1.
【０００４】そのような高圧凍結装置は、本出願人によ
り“Leica EM HPF”という名称で販売されており、また
刊行物“LEICA EM HPF, High Pressure Freezer, 1.K.-
LEICA EM HPF-E-6/94,Juni 1994”に記載されている。[0004] Such a high-pressure freezing device is sold by the applicant under the name "Leica EM HPF" and the publication "LEICA EM HPF, High Pressure Freezer, 1.K.-
LEICA EM HPF-E-6 / 94, Juni 1994 ".
【０００５】高圧凍結装置“Leica EM HPF”は、およそ
2000bar（2×108Pa）の圧力下、103〜105Ｋ（ケルビ
ン）／secの冷却速度で従来の試料をガラス化すること
ができる。この既知の装置では、室温から−100℃への
重要な冷却過程（フェーズ）は、試料の表面においてお
よそ10msec続く（冷却速度104Ｋ／sec）。[0005] The high-pressure freezing device "Leica EM HPF"
A conventional sample can be vitrified under a pressure of 2000 bar (2 × 10 8 Pa) at a cooling rate of 10 3 to 10 5 K (Kelvin) / sec. In this known device, the important cooling phase from room temperature to −100 ° C. lasts approximately 10 msec at the surface of the sample (cooling rate 10 4 K / sec).
【０００６】試料の内部における冷却速度は、専ら試料
の物理的性質に依存する。厚い生物ないし含液試料をガ
ラス化するために、試料は高圧下に置かれる。[0006] The cooling rate inside the sample depends exclusively on the physical properties of the sample. To vitrify thick biological or liquid samples, the samples are placed under high pressure.
【０００７】印加圧力を上昇することにより、ガラス化
のための冷却速度はより小さいもので十分となる。生物
試料をガラス化するための冷却速度は、常圧では、およ
そ10 5〜106Ｋ／secであるが、2000bar（2×108Pa）未満
の圧力では、冷却速度は、10 3〜104Ｋ／secに過ぎな
い。即ち、生物試料は、高圧下では100倍小さい冷却速
度でもガラス化することができる。By increasing the applied pressure, vitrification occurs.
A lower cooling rate for suffices. Creature
At normal pressure, the cooling rate for vitrifying the sample
So 10 Five~Ten6K / sec, but 2000 bar (2 × 108Pa)
At a pressure of 10, the cooling rate is 10 Three~TenFourIt's just K / sec
No. In other words, the cooling rate of a biological sample is 100 times smaller at high pressure.
It can be vitrified as much as possible.
【０００８】試料を冷却するために、互いに連結・分離
可能な少なくとも２つのフレーム部材を有する金属製ホ
ルダが使用される。互いに連結されたフレーム部材は、
試料収容室を形成する。上述の刊行物“LEICA EM HPF,
High Pressure Freezer, 1.K.-LEICA EM HPF-E-6/94,Ju
ni 1994”及び刊行物“Balzers, Hochdruck-Gefriermas
chine HPM 010, Balzers Union Aktiengesellschaft”
（刊行日不詳）にこの種の試料ホルダが記載されてい
る。In order to cool the sample, a metal holder having at least two frame members which can be connected and separated from each other is used. The frame members connected to each other
A sample storage chamber is formed. The above publication "LEICA EM HPF,
High Pressure Freezer, 1.K.-LEICA EM HPF-E-6 / 94, Ju
ni 1994 ”and the publication“ Balzers, Hochdruck-Gefriermas
chine HPM 010, Balzers Union Aktiengesellschaft ”
(Published date unknown) describes a sample holder of this kind.
【０００９】試料を凍結するために、試料ホルダは、一
方では試料の喪失を防ぎ、他方では冷却剤（例えば液体
窒素）を吹き付けるために、（試料の）表面の十分な
（広い）部分が障害なく開放された状態に維持されるよ
うに、高圧凍結装置に拡げ張った状態に組み込まれる
（eingespaunt）この種の装置は、本願ドイツ優先権出
願時には未公開のドイツ出願DE 100 15 733（その後ド
イツ公開DE 100 15 733A）に記載されている。In order to freeze the sample, the sample holder must on the one hand prevent a loss of the sample and on the other hand a sufficient (wide) part of the surface (of the sample) to obstruct the coolant (eg liquid nitrogen). This type of device is installed in an extended state in a high-pressure freezing device in such a way that it remains open and unopened, as is the case with the German application DE 100 15 733, which was unpublished at the time of the German priority application. It is described in published DE 100 15 733A).
【００１０】[0010]
【発明が解決しようとする課題】冷却及びその物理的過
程に関する既知の技術からは、生物試料の中心部におけ
る冷却速度は、専ら試料表面での冷却速度に制限されて
（ないし条件付けられて）決まることも既知である。こ
の場合、本質的特徴は、表面での冷却速度が大きい場
合、一種の「飽和効果」が生じ、他方、表面での冷却速
度が小さいと必然的に冷却の結果は悪くなる、というこ
とである。それゆえ、重要なことは、装置によって達成
可能な冷却速度が、試料保持装置によってできるだけ低
減されず、生物試料の表面に伝達するということであ
る。From the known techniques of cooling and its physical processes, the rate of cooling at the center of a biological sample is limited (or conditioned) solely to the rate of cooling at the surface of the sample. It is also known. In this case, the essential feature is that if the cooling rate on the surface is high, a kind of "saturation effect" occurs, whereas if the cooling rate on the surface is low, the cooling result necessarily worsens. . What is important, therefore, is that the cooling rate achievable by the device is not reduced as much as possible by the sample holding device and is transferred to the surface of the biological sample.
【００１１】それゆえ、本発明の課題は、達成可能な冷
却速度をできるだけ低減せずに試料の表面に伝達する高
圧凍結装置用試料ホルダを創出することである。[0011] It is therefore an object of the present invention to create a sample holder for a high-pressure freezer that transmits to the surface of the sample with as low a reduction in the achievable cooling rate as possible.
【００１２】[0012]
【課題を解決するための手段】この課題は、互いに連結
・分離可能な少なくとも２つのフレーム部材を有し、か
つ連結されたフレーム部材が試料の収容部を形成する、
高圧凍結用含液試料ホルダにより解決される。この試料
ホルダは、フレーム部材の少なくとも１つが、ダイヤモ
ンド部材を具えることを特徴とする。The object of the present invention is to provide at least two frame members that can be connected and separated from each other, and the connected frame members form a sample receiving portion.
It is solved by a liquid-containing sample holder for high-pressure freezing. The sample holder is characterized in that at least one of the frame members includes a diamond member.
【００１３】[0013]
【発明の実施の形態】以下に本発明の好ましい実施の形
態を示すが、これらは従属請求項の対象でもある。試料
ホルダは、更に、複数のフレーム部材の間に、金属製の
リング状スペーサが設けられていることが好ましい。試
料ホルダは、更に、２つのフレーム部材が、ディスク状
のダイヤモンドとして構成されていることが好ましい。
なお、ディスク状とは完全な円盤状に限らず、多角形そ
の他近似の形状を含むものとする。試料ホルダは、更
に、少なくとも１つのフレーム部材が、ディスク状に構
成されており、それ以外のフレーム部材は、金属製であ
り、かつ周縁部に（環状に）延在して形成されたウェブ
を有することが好ましい。試料ホルダは、更に、２つの
フレーム部材が、内側へ配向するそれらの面部分が平面
状に構成されており、これら面部分の間には、該面部分
の周線部に（環状に）延在するスペーサがパッキングと
して、かつ試料収容部を形成するために設けられている
ことが好ましい。DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Preferred embodiments of the present invention are described below, which are also the subject of the dependent claims. It is preferable that the sample holder is further provided with a metal ring-shaped spacer between the plurality of frame members. In the sample holder, the two frame members are preferably configured as disk-shaped diamonds.
The disk shape is not limited to a perfect disk shape but includes a polygon and other approximate shapes. In the sample holder, at least one frame member is formed in a disk shape, and the other frame members are formed of metal and extend (annularly) around the peripheral edge of the web. It is preferred to have. In the sample holder, the two frame members are further configured so that their inwardly oriented surface portions are planar, and between these surface portions, a (circular) extension is formed on the peripheral portion of the surface portion. It is preferable that the existing spacer is provided as a packing and for forming the sample accommodating portion.
【００１４】本発明の特徴は、フレーム部材がダイヤモ
ンドから形成されているか、或いは少なくともダイヤモ
ンド部材を有していることである。ダイヤモンドは、そ
の硬度が非常に大きいということの他に、比熱が低くか
つ熱伝導度が非常に良好であるという長所を持つ。ダイ
ヤモンドは天然、合成いずれも含み、単結晶、多結晶の
いずれでもよく、工業上ないし商業上合成ダイヤモンド
と称するものを含む。A feature of the invention is that the frame member is made of diamond or at least has a diamond member. In addition to its very high hardness, diamond has the advantages of low specific heat and very good thermal conductivity. Diamonds include natural and synthetic ones, and may be single crystal or polycrystalline, including those industrially or commercially referred to as synthetic diamonds.
【００１５】本発明の更なる実施形態では、フレーム部
材（好ましくはダイヤモンド製のフレーム部材）の少な
くとも１つが、ディスク状に形成されている。２つのフ
レーム部材の間には、試料を収容するために必須の深さ
（厚み）を規定する金属製リング状スペーサが設けられ
ている。このリング状スペーサは、この場合、パッキン
グとしても機能することが可能である。In a further embodiment of the invention, at least one of the frame members (preferably a diamond frame member) is formed in the shape of a disk. Between the two frame members, there is provided a metal ring-shaped spacer that defines a depth (thickness) essential for accommodating the sample. This ring-shaped spacer can also function as a packing in this case.
【００１６】フレーム部材の１つのみをダイヤモンドか
ら形成すること、或いはダイヤモンドが嵌め込まれたフ
レーム部材を形成することも勿論可能である。この場
合、試料収容部の深さ（厚み）の形成は、ダイヤモンド
の嵌合により、又は他方のフレーム部材の相応の形状に
より行われる。好ましい実施形態によれば、試料ホルダ
は、１つのダイヤモンド製ディスク状フレーム部材と１
つの金属製ディスク状フレーム部材とを有する。金属製
フレーム部材は、その縁部に、環状に延在して（密着な
いし一体）形成したウェブを有する。It is of course possible to form only one of the frame members from diamond, or to form a frame member in which diamond is fitted. In this case, the formation of the depth (thickness) of the sample accommodating portion is performed by fitting a diamond or by a corresponding shape of the other frame member. According to a preferred embodiment, the sample holder comprises one diamond disc-shaped frame member and one
And two metal disk-shaped frame members. The metal frame member has an annularly extending (closely or integrally formed) web at an edge thereof.
【００１７】試料ホルダの更なる実施形態では、ダイヤ
モンド、ないしフレーム部材の一方に高圧を伝達・導入
（印加）するための孔を形成することも可能である。In a further embodiment of the sample holder, it is possible to form a hole for transmitting and introducing (applying) a high pressure in one of the diamond or the frame member.
【００１８】この種の試料ホルダでは、金属として、
金、アルミニウム又は銅等を使用すること、及びダイヤ
モンドとして工業的に入手可能ないわゆる（ＣＶＤ法等
で製造された）多結晶合成ダイヤモンドを使用すること
が有利であることがわかっている。In this type of sample holder,
It has been found to be advantageous to use gold, aluminum or copper or the like, and to use so-called polycrystalline synthetic diamond (manufactured by CVD or the like) which is commercially available as diamond.
【００１９】本発明の更なる実施形態では、２つのフレ
ーム部材が、内側へ配向するそれらの面部分が平面状に
構成されている。これら面部分の間には、該面部分の周
線部に（環状に）延在するスペーサがパッキングとし
て、かつ試料収容部を形成するために設けられている。
スペーサの寸法を種々採ることにより、試料を収容する
空間の寸法（深さないし厚み等）も簡単に種々異なるよ
うに形成することができる。この場合、フレーム部材の
少なくとも一方は、その表面を不規則な形状にすること
ができる。In a further embodiment of the invention, the two frame members are configured such that their inwardly oriented surface portions are planar. Between these surface portions, a spacer (annularly) extending around the peripheral portion of the surface portion is provided as a packing and for forming a sample container.
By adopting various dimensions of the spacer, the dimensions (depth or thickness, etc.) of the space for accommodating the sample can be easily and variously varied. In this case, at least one of the frame members can have an irregular surface.
【００２０】[0020]
【実施例】本発明の実施例を図面を参照して詳細に説明
する。なお、請求の範囲に付した図面参照符合は、発明
の理解の容易化のために過ぎず、本発明を図示の態様に
限定することを意図しない。また、以下の実施例も発明
の理解の容易化のためであって、本発明の技術的思想を
逸脱しない範囲において当業者により実施可能な修正・
変更も本発明に含まれることも当然である。DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be described in detail with reference to the drawings. It should be noted that the reference numerals in the appended claims are merely for facilitating the understanding of the present invention, and are not intended to limit the present invention to the illustrated embodiments. The following embodiments are also for the purpose of facilitating the understanding of the present invention, and can be modified and implemented by those skilled in the art without departing from the technical idea of the present invention.
Naturally, modifications are included in the present invention.
【００２１】図１には、フレーム部材２と、それに対向
配置するフレーム部材３とを有する試料ホルダ１が示さ
れている。この２つのフレーム部材２及び３は、ダイヤ
モンドで製造されており、かつ円盤状小板として構成さ
れている。２つのフレーム部材２及び３の間には、リン
グ状スペーサ９が設けられているが、このスペーサ９
は、好ましくは金属製である。この金属としては、好ま
しくは、金、銅又はアルミニウムが使用される。リング
状スペーサ９の大きさ（寸法）は、試料収容部６の大き
さ（寸法）を規定する。そのため、リング状スペーサ９
の寸法を種々変化させることにより試料収容部６の大き
さを容易に種々変化させることができる。FIG. 1 shows a sample holder 1 having a frame member 2 and a frame member 3 arranged opposite to the frame member. The two frame members 2 and 3 are made of diamond and are configured as disk-shaped platelets. A ring-shaped spacer 9 is provided between the two frame members 2 and 3.
Is preferably made of metal. Preferably, gold, copper or aluminum is used as this metal. The size (dimension) of the ring-shaped spacer 9 defines the size (dimension) of the sample accommodating section 6. Therefore, the ring-shaped spacer 9
The size of the sample storage section 6 can be easily changed by changing the dimensions of the sample storage section 6 in various ways.
【００２２】試料収容部６は、高圧凍結装置の固定装置
（これは図１には描かれていない）に嵌め込まれる（な
いし挟み込まれる）とき、リング状スペーサ９によって
（外部から）隔離される。When the sample container 6 is fitted (or sandwiched) in a fixing device (not shown in FIG. 1) of the high-pressure freezing device, it is isolated (from the outside) by a ring-shaped spacer 9.
【００２３】図２には、ダイヤモンド製フレーム部材２
と該フレーム部材２に対向配置する金属製フレーム部材
３とを有する試料ホルダの他の実施例が示されている。
フレーム部材３は、その縁部に（密着ないし一体）形成
されかつ該縁部に環状に延在するウェブ８を有するが、
このウェブ８は、図１の実施例のリング状スペーサ９と
同等の機能を持つ。ウェブ８の寸法を種々異なるものと
することにより、試料収容部６の大きさないし寸法も種
々異なるものとすることができる。（密着ないし一体形
成）ウェブ８により、固定的な試料収容部６を形成する
ことができ、これにより試料の挿入及び除去が容易にな
る。FIG. 2 shows a frame member 2 made of diamond.
Another embodiment of the sample holder having a metal frame member 3 disposed opposite to the frame member 2 is shown.
The frame member 3 has a web 8 formed (closely or integrally) at an edge thereof and extending annularly at the edge.
This web 8 has the same function as the ring-shaped spacer 9 of the embodiment of FIG. By making the size of the web 8 different, the size or the size of the sample container 6 can also be made different. With the web 8 (close contact or integral formation), a fixed sample storage portion 6 can be formed, thereby facilitating insertion and removal of the sample.
【００２４】図３には、図１の実施例に対応する試料ホ
ルダ１が示されているが、ここでは、フレーム部材３に
は、後置の高圧（生成）装置（これは、図３には描かれ
ていない）のため（が圧力を伝達即ち、圧力媒体を導入
するため）の内孔１０が設けられている。フレーム部材
３には、更に、高圧伝達（印加）孔５を有するフレーム
部材４が配される。高圧伝達（印加）孔５は、試料収容
部６で終端し導通する内孔１０と連接する。これによ
り、試料収容部６に必要な圧力を直接形成（印加）する
ことができる。FIG. 3 shows a sample holder 1 corresponding to the embodiment of FIG. 1. Here, a frame member 3 is provided with a high-pressure (generation) device provided downstream (this is shown in FIG. 3). (Not shown), an inner hole 10 is provided (for transmitting pressure, ie for introducing a pressure medium). The frame member 3 is further provided with a frame member 4 having a high-pressure transmission (application) hole 5. The high-pressure transmitting (applying) hole 5 is connected to an inner hole 10 that terminates in the sample container 6 and conducts. Thereby, the required pressure can be directly formed (applied) to the sample storage unit 6.
【００２５】図４には、図１の実施例に対応する試料ホ
ルダの他の実施例が示されているが、ここでは、フレー
ム部材４がパッキング９を介してフレーム部材２と直接
（即ち、図３の実施例のフレーム部材３を介さずに）接
続している。この実施例では、フレーム部材３は、完全
に（フレーム部材４で）代替されている。FIG. 4 shows another embodiment of the sample holder corresponding to the embodiment of FIG. 1, in which the frame member 4 is directly connected to the frame member 2 via the packing 9 (ie, in other words, the sample holder is not shown). (Not via the frame member 3 in the embodiment of FIG. 3). In this embodiment, the frame member 3 is completely replaced (by the frame member 4).
【００２６】図５には、図４の実施例に対応する試料ホ
ルダ１の他の実施例が示されているが、ここでは、環状
に延在するパッキング用リング状スペーサ９が、底部７
と該底部７に環状に延在する密着（ないし一体）形成ウ
ェブ８とを有する金属製フレーム部材３によって置き換
えられている。このフレーム部材は、図２の実施例のフ
レーム部材３に対応するが、ここでは、底部７は可能な
限り薄く形成されている。フレーム部材３は、この場合
も分離可能に試料ホルダ１に配されているので、フレー
ム部材３を別個に（それだけを）分離、取り外すことが
できる。FIG. 5 shows another embodiment of the sample holder 1 corresponding to the embodiment of FIG. 4, in which a ring-shaped packing spacer 9 extending annularly is provided with a bottom portion 7.
And a metal frame member 3 having an intimate (or integral) forming web 8 extending annularly on the bottom 7. This frame member corresponds to the frame member 3 of the embodiment of FIG. 2, but here the bottom 7 is formed as thin as possible. In this case as well, the frame member 3 is separably disposed on the sample holder 1, so that the frame member 3 can be separated and removed separately (only).
【００２７】このため、試料をより容易に試料ホルダ１
ないし試料収容部６内に挿入することができ、更に再び
より容易に取り出すことができる。For this reason, the sample holder 1 can more easily hold the sample.
Alternatively, it can be inserted into the sample container 6, and can be taken out again more easily.
【００２８】上述の及びダイヤモンド製（好ましくはＣ
ＶＤ多結晶合成ダイヤモンド製）のフレーム部材には、
１つ又は複数のダイヤモンド（部材）が嵌め込まれてい
ることも勿論可能である。The above and diamond (preferably C
(VD polycrystalline synthetic diamond)
It is of course possible that one or more diamonds (members) are fitted.
【００２９】液体窒素等の冷却剤が吹き付けられる面
（ないし面積）をより大きくするために、フレーム部材
の外表面を不規則な（凹凸等をつけた）形状に構成する
ことも本発明の技術的思想の範囲内に含まれることも自
明である。In order to further increase the surface (or area) on which a coolant such as liquid nitrogen is sprayed, the outer surface of the frame member may be formed in an irregular shape (with irregularities or the like). It is self-evident that they fall within the scope of the spiritual idea.
【００３０】[0030]
【発明の効果】本発明の独立請求項１により、対応する
所定の課題として掲げた効果が得られる。即ち、本発明
の高圧凍結装置用試料ホルダにより、達成可能な冷却速
度をできるだけ低減せずに試料の表面に伝達することが
できる。各従属請求項により、更に付加的な効果が、そ
れぞれ達成される。According to the independent claim 1 of the present invention, the effects listed as the corresponding predetermined problems can be obtained. That is, the sample holder for the high-pressure freezing apparatus of the present invention can transmit the cooling rate that can be achieved to the surface of the sample without reducing the achievable cooling rate as much as possible. Further dependent advantages are achieved by the respective dependent claims.
【図１】２つのダイヤモンド製フレーム部材と、環状金
属製リングとを有する試料ホルダの縦断面図。FIG. 1 is a longitudinal sectional view of a sample holder having two diamond frame members and an annular metal ring.
【図２】ダイヤモンド製フレーム部材と、金属製フレー
ム部材とを有する試料ホルダの縦断面図。FIG. 2 is a longitudinal sectional view of a sample holder having a diamond frame member and a metal frame member.
【図３】２つのダイヤモンド製フレーム部材と、高圧伝
達（印加）孔を有する付加的フレーム部材とを有する試
料ホルダの縦断面図。FIG. 3 is a longitudinal sectional view of a sample holder having two diamond frame members and an additional frame member having a high-pressure transmission (application) hole.
【図４】ダイヤモンド製フレーム部材と、高圧伝達（印
加）孔を有する他のフレーム部材とを有する試料ホルダ
の縦断面図。FIG. 4 is a longitudinal sectional view of a sample holder having a diamond frame member and another frame member having a high-pressure transmission (application) hole.
【図５】ダイヤモンド製フレーム部材と、試料収容部及
びパッキングを形成するフレーム部材とを有する試料ホ
ルダの縦断面図。FIG. 5 is a longitudinal sectional view of a sample holder having a diamond frame member, and a frame member forming a sample container and a packing.
１ 試料ホルダ ２ フレーム部材 ３ フレーム部材 ４ フレーム部材 ５ 高圧伝達孔 ６ 試料収容部 ７ 底部 ８ ウェブ ９ リング状スペーサ １０ 内孔 DESCRIPTION OF SYMBOLS 1 Sample holder 2 Frame member 3 Frame member 4 Frame member 5 High-pressure transmission hole 6 Sample storage part 7 Bottom part 8 Web 9 Ring-shaped spacer 10 Inner hole
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ テーマコート゛(参考） Ｇ０１Ｎ 1/28 Ｊ ──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. 7 Identification symbol FI Theme coat ゛ (Reference) G01N 1/28 J
Claims (5)
フレーム部材を有し、かつ連結されたフレーム部材が試
料の収容部を形成する、高圧凍結用試料ホルダにおい
て、 フレーム部材（２；３）の少なくとも１つが、ダイヤモ
ンド部材を具えることを特徴とする試料ホルダ。1. A high-pressure freezing sample holder having at least two frame members which can be connected to and separated from each other, and wherein the connected frame members form a sample accommodating portion. A sample holder, wherein at least one comprises a diamond member.
に、金属製のリング状スペーサ（９）が設けられている
ことを特徴とする請求項１に記載の試料ホルダ。2. The sample holder according to claim 1, wherein a metal ring spacer is provided between the plurality of frame members.
ク状のダイヤモンドとして構成されていることを特徴と
する請求項１又は２に記載の試料ホルダ。3. The sample holder according to claim 1, wherein the two frame members are configured as disk-shaped diamonds.
が、ディスク状に構成されており、 他のフレーム部材（３）は、金属製であり、かつ周縁部
に延在して形成されたウェブ（８）を有することを特徴
とする請求項１又は２に記載の試料ホルダ。4. At least one frame member (2; 3).
Is configured in the form of a disk, and the other frame member (3) is made of metal and has a web (8) formed to extend around the periphery. 3. The sample holder according to 2.
配向するそれらの面部分が平面状に構成されており、こ
れら面部分の間には、該面部分の周線部に延在するスペ
ーサ（９）がパッキングとして、かつ試料収容部（６）
を形成するために設けられていることを特徴とする請求
項１〜４のいずれかに記載の試料ホルダ。5. The two frame members (2; 3) are configured such that their inwardly oriented surface portions are planar, and extend between these surface portions in a circumferential portion of the surface portion. The existing spacer (9) serves as a packing, and the sample container (6)
The sample holder according to any one of claims 1 to 4, wherein the sample holder is provided in order to form the sample holder.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE10065143.7 | 2000-12-23 | ||
| DE10065143A DE10065143B4 (en) | 2000-12-23 | 2000-12-23 | Sample holder for a high pressure freezing device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2002277365A true JP2002277365A (en) | 2002-09-25 |
| JP4146638B2 JP4146638B2 (en) | 2008-09-10 |
Family
ID=7669084
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2001391608A Expired - Fee Related JP4146638B2 (en) | 2000-12-23 | 2001-12-25 | Sample holder for high pressure freezer |
Country Status (3)
| Country | Link |
|---|---|
| US (1) | US20020079318A1 (en) |
| JP (1) | JP4146638B2 (en) |
| DE (1) | DE10065143B4 (en) |
Families Citing this family (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9518899B2 (en) * | 2003-08-11 | 2016-12-13 | Sakura Finetek U.S.A., Inc. | Automated reagent dispensing system and method of operation |
| US7767152B2 (en) * | 2003-08-11 | 2010-08-03 | Sakura Finetek U.S.A., Inc. | Reagent container and slide reaction retaining tray, and method of operation |
| US7744817B2 (en) * | 2003-08-11 | 2010-06-29 | Sakura Finetek U.S.A., Inc. | Manifold assembly |
| US7501283B2 (en) * | 2003-08-11 | 2009-03-10 | Sakura Finetek U.S.A., Inc. | Fluid dispensing apparatus |
| DE102004041965B4 (en) | 2004-08-31 | 2009-08-13 | Leica Mikrosysteme Gmbh | Device and method for freeze substitution and embedding of biological samples |
| US7293426B2 (en) * | 2004-10-05 | 2007-11-13 | Washington University | Apparatus for freezing a biological sample |
| DE102005021962B4 (en) * | 2005-05-12 | 2009-08-13 | Leica Mikrosysteme Gmbh | Sample holder for high pressure freezing samples and high pressure freezer with sample holder |
| US8459509B2 (en) | 2006-05-25 | 2013-06-11 | Sakura Finetek U.S.A., Inc. | Fluid dispensing apparatus |
| WO2009155430A2 (en) * | 2008-06-18 | 2009-12-23 | The Cleveland Clinic Foundation | Systems and methods for vitrifying tissue |
| US8752732B2 (en) | 2011-02-01 | 2014-06-17 | Sakura Finetek U.S.A., Inc. | Fluid dispensing system |
| US8580568B2 (en) | 2011-09-21 | 2013-11-12 | Sakura Finetek U.S.A., Inc. | Traceability for automated staining system |
| US8932543B2 (en) | 2011-09-21 | 2015-01-13 | Sakura Finetek U.S.A., Inc. | Automated staining system and reaction chamber |
| EP3287775B1 (en) | 2016-08-26 | 2019-04-03 | Leica Mikrosysteme GmbH | Modular specimen holders for high pressure freezing and x-ray crystallography of a specimen |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NL135905C (en) * | 1967-11-14 | |||
| US5044165A (en) * | 1986-12-03 | 1991-09-03 | Board Of Regents, The University Of Texas | Cryo-slammer |
| US5255783A (en) * | 1991-12-20 | 1993-10-26 | Fluoroware, Inc. | Evacuated wafer container |
| CH686534A5 (en) * | 1993-08-03 | 1996-04-15 | Martin Wohlwend | Method and apparatus for the vitrification of samples, in particular biological samples. |
| ATE224536T1 (en) * | 1997-01-13 | 2002-10-15 | Daniel Dr Studer | SAMPLE HOLDER FOR WATER CONTAINING SAMPLES AND METHOD FOR USE THEREOF |
-
2000
- 2000-12-23 DE DE10065143A patent/DE10065143B4/en not_active Expired - Fee Related
-
2001
- 2001-12-20 US US10/022,528 patent/US20020079318A1/en not_active Abandoned
- 2001-12-25 JP JP2001391608A patent/JP4146638B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| DE10065143A1 (en) | 2002-06-27 |
| US20020079318A1 (en) | 2002-06-27 |
| JP4146638B2 (en) | 2008-09-10 |
| DE10065143B4 (en) | 2007-12-27 |
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