JPS6079246A - Low-temperature fixing device - Google Patents
Low-temperature fixing deviceInfo
- Publication number
- JPS6079246A JPS6079246A JP59186666A JP18666684A JPS6079246A JP S6079246 A JPS6079246 A JP S6079246A JP 59186666 A JP59186666 A JP 59186666A JP 18666684 A JP18666684 A JP 18666684A JP S6079246 A JPS6079246 A JP S6079246A
- Authority
- JP
- Japan
- Prior art keywords
- cooling bath
- specimen
- holder
- light source
- cooling
- 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.)
- Pending
Links
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
-
- F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
- F25—REFRIGERATION OR COOLING; COMBINED HEATING AND REFRIGERATION SYSTEMS; HEAT PUMP SYSTEMS; MANUFACTURE OR STORAGE OF ICE; LIQUEFACTION SOLIDIFICATION OF GASES
- F25D—REFRIGERATORS; COLD ROOMS; ICE-BOXES; COOLING OR FREEZING APPARATUS NOT OTHERWISE PROVIDED FOR
- F25D3/00—Devices using other cold materials; Devices using cold-storage bodies
- F25D3/10—Devices using other cold materials; Devices using cold-storage bodies using liquefied gases, e.g. liquid air
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
Landscapes
- Physics & Mathematics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Mechanical Engineering (AREA)
- Thermal Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Optics & Photonics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Combustion & Propulsion (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Microscoopes, Condenser (AREA)
- Sampling And Sample Adjustment (AREA)
- Investigating Or Analyzing Materials Using Thermal Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【発明の詳細な説明】 ［産業上の利用分野］ 本発明１±、低温固定装置に関する。[Detailed description of the invention] [Industrial application field] The present invention 1± relates to a cryogenic fixation device.
［従来の技術］
小さな標本を低温固定する前にこの小標木（これはサス
ペンションの小滴又は培養組織の薄い層から成る）は標
本ホルダに貼着される。ホルダ上の標本を取扱ったり、
又ある場合にホルダに微小電極を接続したり、又はホル
ダを他の方法で変えたりすることは、低温固定操作を成
功裏に行う際重要な工程である。このため低温固定用標
本を調製する際に一般にステレオ式ＷＪ微競が使用され
る。低温固定操作の次に冷却浴より標本を注意深く取出
すが、この操作も注意を要する。BACKGROUND OF THE INVENTION Before cryofixing a small specimen, a small marker (which consists of a drop of suspension or a thin layer of cultured tissue) is affixed to a specimen holder. Handle the specimen on the holder,
Also, in some cases connecting the microelectrode to the holder or otherwise modifying the holder is an important step in performing a successful cryofixation operation. For this reason, a stereo WJ micrograph is generally used when preparing specimens for cryogenic fixation. Following the low-temperature fixation operation, the specimen is carefully removed from the cooling bath, but this operation also requires care.
［発明が解決しようとする問題点］
調製および低温固定後の操作は、標本ホルダの注意深い
操作を要するので、これら操作をするにほ、標本ホルダ
と冷却浴を良好に照明しなければならない。この冷却浴
は、一般に冷却液用の円筒チャンバを構成する金属容器
から成り、チャンバの寸法は・Ａ″）に径か約２０ ｍ
ｍで高さがｌ ＯＯｍｍであり、浴内の冷却液の温度
は、一定温度に制ｆｌ’ｌｌ ３れる。標本調製には、
両手を要し、注入装、＋、−／ｉ、ステレオ式顕微鏡お
よび低温媒体供給ラインにも空間が必要なため、必要な
照明をすることができる装置を１投置することは困難で
あった。特に標本ホルタの照明と冷却浴の全照明を組合
わせることは困難であった。[Problems to be Solved by the Invention] Preparation and post-cold-fixation operations require careful manipulation of the specimen holder, and the specimen holder and cooling bath must be well illuminated for these operations. This cooling bath generally consists of a metal container constituting a cylindrical chamber for the cooling liquid, the dimensions of which are approximately 20 m in diameter.
m and the height is lOOmm, and the temperature of the cooling liquid in the bath is controlled to a constant temperature. For specimen preparation,
It was difficult to set up a single device that could provide the necessary illumination because both hands were required and space was also required for the injection device, +, -/i, stereo microscope, and cryogenic medium supply line. . In particular, it was difficult to combine the illumination of the specimen holder with the total illumination of the cooling bath.
Ａ、、発明の［！的は、簡単でかつ一般に低温固定装置
にも適用できる態様で標本ホルダを照明すると共に冷却
浴の内部全体を照明する装置を提供するにある。A. Invention [! The aim is to provide a device for illuminating a specimen holder and for illuminating the entire interior of a cooling bath in a manner that is simple and generally applicable also to cryofixation devices.
［問題点を解決するための手段及び作用効果］未発明に
よれば、冷却液用冷却浴と、該冷却浴の下面に設けろえ
たウィンドと、冷却浴の内部を照明するための光ビーム
を一ヒ方に向はウィンドを通過させる光源とから成る低
温固定装置が提供される。[Means and effects for solving the problem] According to the invention, a cooling bath for cooling liquid, a window provided on the lower surface of the cooling bath, and a light beam for illuminating the inside of the cooling bath are integrated. A cryofixation device is provided comprising a light source passing through a window in the opposite direction.
本装置を使用するとき、光ビームはウィンドを通って下
方から冷却浴へ進入するが、このウィンドはカラス又は
他の耐冷かつ極耐極低温材料からなり、冷却浴の下面に
シールされる。標本ホルダは通常冷却浴の上方に位置し
、光ビームは、浴内部を照明するよう向けられるという
よりも標本ホルダおよびホルダ上の標本に向けられる。When using the device, the light beam enters the cooling bath from below through a window, which is made of glass or other cold- and cryogenic-resistant material and is sealed to the underside of the cooling bath. The specimen holder is typically located above the cooling bath and the light beam is directed at the specimen holder and the specimen on the holder rather than being directed to illuminate the interior of the bath.
従って、光源は一つだけ必要であり、標本をホルダに取
付けるとき顕微鏡によって標本ホルダを見ることができ
、本装置は、標本ホルダおよび標本を両手で操作するこ
とに制限を課するものではない。Thus, only one light source is required, the specimen holder can be viewed by the microscope when the specimen is attached to the holder, and the device does not impose restrictions on two-handed manipulation of the specimen holder and specimen.
本発明の一実施例では、冷却浴の直下に光源が配置され
る。冷却浴にビームが進入する前に光ビームの赤外線成
分を除去するため熱遮断フィルタを設けてもよい。この
熱遮断フィルタはウィンドで構成してもよい。In one embodiment of the invention, a light source is placed directly below the cooling bath. A heat blocking filter may be provided to remove the infrared component of the light beam before it enters the cooling bath. This heat cutoff filter may be constructed of a window.
本発明の別の実施例では、冷却浴の一側面に配置され、
光ビームを上方に向け、ウィンドを通過させる反射手段
を更に含む。この配列では、電球の交換な容易にする。In another embodiment of the invention, located on one side of the cooling bath,
It further includes reflective means for directing the light beam upwardly and through the window. This arrangement makes replacing light bulbs easy.
この反射手段は、例えば、プリズム・鏡又は光ガイドか
ら構成でき、光源は好ましくは弱い赤外線成分を有する
光を放出する反射式ランプから構成するか、又はビーム
から赤外線成分の大部分を除去するタイプの光ファイバ
光力ネトを設けてもよい。The reflecting means can, for example, consist of a prism-mirror or a light guide, and the light source preferably consists of a reflector lamp emitting light with a weak infrared component, or of a type that removes a large part of the infrared component from the beam. A fiber optic power network may be provided.
冷却浴は、冷却浴を横断してきた光ビームが透過するポ
ー）・を有するふたを含むことができる。The cooling bath may include a lid having a port through which a light beam has traversed the cooling bath.
このポー）・は１冷却浴の検査を可能としかつ冷却浴か
カバーｙれた状態で標本ホルダ上での標本調製を、Ｔ（
能とする。以下添附図面を参明ルで本発明の実施例を従
来例と対比しつつ詳述する。This port allows inspection of the cooling bath and allows specimen preparation on the specimen holder with the cooling bath covered.
Noh. DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be described in detail below with reference to the accompanying drawings in comparison with conventional examples.
［実施例］
第３図は、公知のタイプの低温固定装置を示す。窒素４
のような低温液体を含むポット状容器３は、ハウジング
１内に収容された断熱材２に囲まれており、液体窒素４
は図示するようなスタンバイモード時に金属製円筒体５
状の浴を直接又は間接的に冷却する。この円筒体５は、
冷却媒体６を含み、この媒体に標本７を浸漬して低温固
定するようになっている。この冷却媒体の温度はサーモ
スタット装置により維持される。EXAMPLE FIG. 3 shows a cryofixation device of a known type. nitrogen 4
A pot-shaped container 3 containing a cryogenic liquid such as
The metal cylindrical body 5 is in standby mode as shown in the figure.
cooling the bath directly or indirectly. This cylindrical body 5 is
It contains a cooling medium 6, and the specimen 7 is immersed in this medium to be fixed at a low temperature. The temperature of this cooling medium is maintained by a thermostatic device.
標本７は、標本ホルダ８に取付けられ、ホルタは支拍体
１３−ヒの挿入ロッド９に取付けられている。この挿入
ロッドはレバー１０を操作すれば降下して冷却液６内に
進入できる。標本ホルダ８に標本７を取付けるのを観察
するためにステレオ式顕微鏡１１が使用されるが、この
取付操作は極端に困難であることが多い。The specimen 7 is attached to a specimen holder 8, and the holder is attached to the insertion rod 9 of the support body 13-hi. This insertion rod can be lowered and entered into the coolant 6 by operating the lever 10. A stereo microscope 11 is used to observe the attachment of the specimen 7 to the specimen holder 8, but this attachment operation is often extremely difficult.
本操作をスタンバイ操作モードにした状態で標本数句操
作をしている間、冷却浴５を含む容器３の断熱ふた１２
は、閉１Ｆ位置に維持され、窒素の消費を制限すると共
にホルダ８上にある標本７の予冷却を悪影響を与えない
ようにしている。During the sample count operation with this operation in standby operation mode, the heat insulating lid 12 of the container 3 containing the cooling bath 5 is closed.
is maintained in the closed 1F position to limit nitrogen consumption and to avoid adversely affecting the precooling of the specimen 7 on the holder 8.
このような公知の装置の配列では、照明が不充分のまま
標本の取付けが行なわれる。ステレオ式顕微鏡１１は、
ユーザに面する装置の一側面を占める一方１反対の側面
すなわち後方の側面は支持体１３に占められている。標
本の調製をするには、−船に両手を使用しなければなら
ないので１、それ以外の２つの側面も光源を設置する余
裕はない。冷却浴５に標本７を浸漬した後、冷却液６を
含む浴により構成される円筒空間内にある標本はかろう
じて観察・取扱いできるにすぎない。In such known device arrangements, mounting of specimens takes place with insufficient illumination. The stereo microscope 11 is
One side of the device facing the user is occupied, while the opposite or rear side is occupied by a support 13. To prepare the specimen, one must use both hands on the vessel, and there is no room for light sources on the other two sides. After immersing the specimen 7 in the cooling bath 5, the specimen within the cylindrical space defined by the bath containing the cooling liquid 6 can only be barely observed and handled.
［実施例］
第１図に示す装置では、ガラスウィンド１４は冷却液６
を含む冷却浴５の底部を形成するが、それ以外の点で冷
却浴および挿入装置は第３図に示した配列と同じである
。冷却液６の直下には、高出力石英−よう素ランプ１５
から成る光源が設けられている。このランプはホルダ１
６内に取付けられているか、ホルダは装置を傾ければ取
外すことができ、こうして電球を容易に交換できるよう
になっている。集光レンズ１７は、ランプからの光線を
合焦する。ランプからの光のうちの赤外線成分をフィル
タで除去すなわち吸収するために熱遮断フィルタが設け
られているが、このフィルタはウィンド１４又はランプ
の反射器により構成される。ウィンドは冷却液６により
冷却される。[Example] In the device shown in FIG.
forming the bottom of the cooling bath 5 containing the cooling bath 5, but otherwise the cooling bath and insertion device are the same as the arrangement shown in FIG. Directly below the coolant 6 is a high-power quartz-iodine lamp 15.
A light source is provided. This lamp is in holder 1
6, the holder can be removed by tilting the device, thus making it easy to replace the bulb. A condenser lens 17 focuses the light beam from the lamp. A heat blocking filter is provided to filter out or absorb the infrared component of the light from the lamp, this filter being constituted by the window 14 or the reflector of the lamp. The window is cooled by a cooling liquid 6.
第２図に示す装置の実施例でｌオ、ふた１２゛のうちの
透明インサート１８状の部分は、ふたを閉じている時で
も標本ホルダ７を開明できるように光ビームを冷却浴の
外へ通過できるようにしている。In the embodiment of the apparatus shown in FIG. I am allowing it to pass.
後部壁に取付けられたホルダ２０内に反射ランプ１９が
装置の外に位置するように取付けられている。ホルダ２
０は、図示した例とは別に２つの側壁のいずれかに数句
けることもできる。反射ランプから孜出されたビーム２
１は、プリズム２２から成る反射装置により、冷却浴に
より構成されたチャンバ内へ進入するようヒ方へ向けら
れる。A reflector lamp 19 is mounted outside the device in a holder 20 mounted on the rear wall. Holder 2
0 can also be written several times on either of the two side walls apart from the example shown. Beam 2 emitted from the reflective lamp
1 is directed towards it by means of a reflector consisting of a prism 22 into a chamber constituted by a cooling bath.
第１図および第２図に示す装置は、種々の変更をするこ
とがでさる。例えば、光源と標本ホルダの間の光路内に
種々の光素子を導入できる。特に反射装置はプリズムの
かわりに鏡又は光ファイ／へのようなガイドかも構成で
きるし、種々の光源も使用できるう冷却浴から光を放出
できるポートは、固体透明体、２つの透明カバープレー
トを有する管、光線を更に合焦するレンズ、又は吸熱ガ
ラスから構成できる。The apparatus shown in FIGS. 1 and 2 is capable of various modifications. For example, various optical elements can be introduced in the optical path between the light source and the specimen holder. In particular, the reflector can be a mirror or a guide to an optical fiber instead of a prism, and a variety of light sources can be used.The port for emitting light from the cooling bath can be made of a solid transparent body, with two transparent cover plates. It can consist of a tube with a tube, a lens to further focus the light beam, or an endothermic glass.
第１図は、冷却浴の直下に光源を設けた本発明に係る装
置の一実施例を示す略式横断面図、第２１４は本発明に
係る装置の別の実施例を示す略式横１・・・ハウジング
、２・・・断熱材。
３・・・容器、 ４・・・・・・液体窒素。
５・・・・・・冷却浴、 ７・・・・・・標本。
８・・・・・・標本ホルダ、１４・・・・・・ウィンド
。
１５・・・・・・ランプ、 １９・旧・・反射式ランプ
。
２０・・・・・・ホルダ、２２・・・・・・反射手段。
出縁ｎ人 ラニー・ライヘルド、オブティフンエ・ヴエ
ルケφアーゲー
代理入 弁理士 加藤 朝道Fig. 1 is a schematic cross-sectional view showing one embodiment of the apparatus according to the present invention in which a light source is provided directly below the cooling bath, and Fig. 214 is a schematic cross-sectional view showing another embodiment of the apparatus according to the present invention.・Housing, 2...insulation material. 3... Container, 4... Liquid nitrogen. 5...Cooling bath, 7...Specimen. 8... Specimen holder, 14... Window. 15... Lamp, 19. Old... Reflective lamp. 20...Holder, 22...Reflection means. N person Lanny Reicheld, Obtifune Werke φ AG Patent attorney Asamichi Kato
Claims (6)
たウィンドと 冷却浴の内部を照明するため光ビームを
上方に向はウィンドを通過させる光源とから成る低温固
定装置。(1) A cryogenic fixation device consisting of a cooling bath for cooling liquid, a window provided on the lower surface of the cooling bath, and a light source that passes a light beam upward through the window to illuminate the inside of the cooling bath.
の範囲第１項記載の装置。(2) The apparatus according to claim 1, wherein a light source is disposed directly below the cooling bath.
ムを−１一方へ向はウィンドを通過させる反則手段を更
に含む特許請求の範囲第１項記載の装置。(3) The apparatus according to claim 1, further comprising a light source disposed on one side of the cold 1'B bath, and a counter means for causing the light beam to pass through a window in one direction.
求の範囲第２項記載の装置。(4) The apparatus of claim 2, wherein the reflecting means comprises a fiber optic light guide.
射器はランプにより放出される光の赤外線成分の小なく
とも一部を吸収する特許請求の範囲第１項記載の装置。5. The apparatus of claim 1, wherein the light source comprises a reflector lamp, the reflector of which absorbs at least a portion of the infrared component of the light emitted by the lamp.
ムを透過するポートが設けられている特許請求の範囲第
１項記載の装置。(6) The apparatus according to claim 1, wherein the cooling bath lid is provided with a port that transmits the light beam that has passed through the cooling bath.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE3332741.6 | 1983-09-10 | ||
| DE19833332741 DE3332741C2 (en) | 1983-09-10 | 1983-09-10 | Device for illuminating the sample and sample holder as well as the cooling bath on a device for immersion cryofixation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS6079246A true JPS6079246A (en) | 1985-05-07 |
Family
ID=6208770
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59186666A Pending JPS6079246A (en) | 1983-09-10 | 1984-09-07 | Low-temperature fixing device |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JPS6079246A (en) |
| DE (1) | DE3332741C2 (en) |
| FR (1) | FR2551870A1 (en) |
| GB (1) | GB2146262B (en) |
| SE (1) | SE8404487L (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013181987A (en) * | 2012-03-02 | 2013-09-12 | Leica Mikrosysteme Gmbh | Light stimulus and cryopreservation device of biologica sample |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3532606C1 (en) * | 1985-09-12 | 1986-11-13 | C. Reichert Optische Werke Ag, Wien | Device for metal-mirror cryofixation of, in particular, biological objects |
| AT508017B1 (en) * | 2009-07-22 | 2010-10-15 | Leica Mikrosysteme Gmbh | TRANSFER OF A SAMPLE CARRIER IN CORRELATIVE ELECTRONIC MICROSCOPY |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AT343941B (en) * | 1976-02-06 | 1978-06-26 | Reichert Optische Werke Ag | METHOD AND DEVICE FOR PRODUCING PREPARATIONS FOR MICROSCOPY, IN PARTICULAR ELECTRON MICROSCOPY FROM NATIVE HISTOLOGICAL OBJECTS AND PHYSICOCHEMICAL SIMILAR PRODUCTS BY CRYOFIXATION |
-
1983
- 1983-09-10 DE DE19833332741 patent/DE3332741C2/en not_active Expired
-
1984
- 1984-08-30 FR FR8413433A patent/FR2551870A1/en not_active Withdrawn
- 1984-09-04 GB GB08422330A patent/GB2146262B/en not_active Expired
- 1984-09-06 SE SE8404487A patent/SE8404487L/en not_active Application Discontinuation
- 1984-09-07 JP JP59186666A patent/JPS6079246A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013181987A (en) * | 2012-03-02 | 2013-09-12 | Leica Mikrosysteme Gmbh | Light stimulus and cryopreservation device of biologica sample |
Also Published As
| Publication number | Publication date |
|---|---|
| SE8404487L (en) | 1985-03-11 |
| FR2551870A1 (en) | 1985-03-15 |
| DE3332741C2 (en) | 1986-09-04 |
| GB8422330D0 (en) | 1984-10-10 |
| GB2146262B (en) | 1986-10-29 |
| GB2146262A (en) | 1985-04-17 |
| DE3332741A1 (en) | 1985-04-04 |
| SE8404487D0 (en) | 1984-09-06 |
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