EP0377721B1 - Procédé pour la réalisation des réactions avec des quantités minimes d'echantillons fluides - Google Patents

Procédé pour la réalisation des réactions avec des quantités minimes d'echantillons fluides Download PDF

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Publication number
EP0377721B1
EP0377721B1 EP89907732A EP89907732A EP0377721B1 EP 0377721 B1 EP0377721 B1 EP 0377721B1 EP 89907732 A EP89907732 A EP 89907732A EP 89907732 A EP89907732 A EP 89907732A EP 0377721 B1 EP0377721 B1 EP 0377721B1
Authority
EP
European Patent Office
Prior art keywords
sample
receiving tube
end part
lid
insert
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.)
Expired - Lifetime
Application number
EP89907732A
Other languages
German (de)
English (en)
Other versions
EP0377721A1 (fr
Inventor
Dieter Korf
Dagmar Dr. Flach
Stephan Dr. Diekmann
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Walter Sarstedt Geraete und Verbrauchsmaterial fuer Medizin und Wissenschaft
Original Assignee
Walter Sarstedt Geraete und Verbrauchsmaterial fuer Medizin und Wissenschaft
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Walter Sarstedt Geraete und Verbrauchsmaterial fuer Medizin und Wissenschaft filed Critical Walter Sarstedt Geraete und Verbrauchsmaterial fuer Medizin und Wissenschaft
Publication of EP0377721A1 publication Critical patent/EP0377721A1/fr
Application granted granted Critical
Publication of EP0377721B1 publication Critical patent/EP0377721B1/fr
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5082—Test tubes per se
    • B01L3/50825—Closing or opening means, corks, bungs
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices

Definitions

  • the invention relates to a method for carrying out reactions at temperatures above 50 ° C. of components to be heated, which emit components that evaporate when heated, in particular fluid samples.
  • reaction In particular in molecular biological work, reactions often have to be carried out at temperatures above 50 (up to 100 ° C.) (heat inactivation of proteins, denaturation of nucleic acids, restriction digestion with Taq I and the like). It is common to carry out these reactions in standard reaction vessels that are used in preheated water baths or in bores of heated metal blocks.
  • the volumes of the reaction solutions are usually between 10 and 50 ⁇ l, the volumes of the reaction vessels between about 1000 and 2500 ⁇ l.
  • the reaction vessels therefore contain a large excess of volume, in which water evaporates from the reaction solutions and is deposited on the inside of the lid. This increases the concentration in the reaction solution, which can sometimes lead to the sample completely drying out. This is extremely cumbersome for the examinations and treatments and can only be prevented by special measures, such as repeated centrifugation of the reaction vessels or overlaying the reaction solution with oil.
  • the object of the invention is to provide a method of the type mentioned, in which on the one hand the handling of the reaction vessel is ensured by a sufficiently voluminous design, but on the other hand one substantial concentration or even drying out of the sample is avoided during the reaction.
  • the procedure according to the invention ensures that the gas volume with the part of the reaction vessel filled with the reaction solution or the like is reduced so much that it is no longer possible to evaporate the reaction solution into larger spaces above it, so that the sample dries out heat treatment is effectively avoided.
  • the volume within the reaction vessel which is not required for receiving the sample, is reduced to such an extent that the evaporation space is as small as possible above the sample, which limits the evaporation of liquid from the sample.
  • the approach ensures as close as possible above the sample arranged in the sample receiving tube that all-round sealing is ensured in such a way that the remaining volume above the sealing area is separated from the actual reaction space in which the sample is located in a gas-tight manner. In this way, the resulting steam is limited to the relatively small reaction space.
  • a major problem is to make the seal between the neck and the sample tube so that when the neck is inserted into the sample tube, as little as possible and if possible no pressure is built up in the reaction space. This is important on the one hand, because otherwise the insertion of the approach into the sample tube is only possible with increased effort. It could also build up pressure in the reaction space be harmful to the sample or the tests to be performed.
  • a pressure build-up when inserting the approach into the sample tube can be avoided particularly advantageously by the features of claim 2.
  • a container made of thermoplastic material with a lid having an extension and a second closure is known from CH-A-330 179; a spoon for receiving a stool sample or the like is provided at the bottom of the second closure, which is introduced into the container after the stool sample has been taken and is closed off from the outside twice by the lid and the second closure.
  • the method carried out with this container thus consists in enclosing a stool sample taken with a spoon separated from the environment by a double lock and then transporting it to the site of the examination.
  • a reaction is to be carried out in a sample collection tube.
  • a sample receiving tube 11 provided with a circular cross section has a region 11 '''' which widens conically upwards above a rounded-off tip which is formed as a rounded tip, followed by a relatively short circular-cylindrical region 11 ''', followed by a short extension section 11 '' and finally an upper circular cylindrical section 11 ', which is relatively long and has the largest diameter relative to the other parts. Its upper end represents the upper open end of the sample tube 11.
  • An external thread 26 is provided on the outer wall of the sample receiving tube 11, onto which a screw cap 12 provided with an internal thread 27 is screwed.
  • the cover 12 On its inside, the cover 12 has a circular cylindrical recess 28 which is open at the bottom and which is concentric with the central axis 23 and into which a hollow cylindrical extension 24 is inserted from below in a sliding or preferably clamped fit.
  • the attachment 14 could also be fixed in the recess 28 by suitable fastening means, for example adhesive.
  • the extension 14 can be pulled out again when the cover 12 is unscrewed, so that replacement is possible.
  • the circumferential flange 29 forming the recess 28 is radially outside a slight distance 30 from the inner wall 15 of the sample tube 11, so that in a state between the upper end wall 31 and a lower ring wall 32 of the lid 12 there remains a clearance 33 when fully screwed on adjoins a lateral vent opening 25 in the threaded peripheral edge 34 of the cover 12. In this way, a pressure equalization can take place between the interior 13 of the sample receiving tube 11 and the atmosphere.
  • the hollow-cylindrical extension 14 has a slightly smaller outer diameter than the circular-cylindrical region 11 ′′ ′′ and merges at the lower end into an end part 20 that tapers slightly downwards and has a bottom wall 17 at its lower end.
  • the conical end part 20 is in a sealing connection 35 with the inner wall 15 of the sample receiving tube 11, which also tapers downward in this area, within the area 11 ''''.
  • the end part 20 with the bottom wall 17 at the lower end of the sample tube 11 separates a reaction space 19 into which a sample 18 is filled, which is to be subjected to heat treatment by inserting the sample tube 11 into a heating apparatus.
  • the attachment 14 could be replaced by an attachment 14 which is somewhat longer and has a narrower end part 20 at the bottom, which is indicated by dashed lines in FIG. 1. In this way, a substantially smaller reaction space 19 'could be separated from the total internal volume of the sample receiving tube 11.
  • attachment 14 Due to the interchangeability of the attachment 14, one and the same cover 12 could be provided with attachments 14 of different lengths and different sizes of reaction spaces.
  • a corrugation 21 which facilitates unscrewing and screwing is provided on the outer periphery of the screw cap 12.
  • a further corrugation 36 is located on the outer circumference of the upper region 11 'of the sample receiving tube 11 in order to also provide increased resistance when unscrewing the other hand.
  • the method according to the invention can be carried out as follows: With the cover 12 unscrewed and the extension 14 removed, the sample 18 is first filled into the sample tube 11 in the desired amount. Then, depending on the amount filled in, a suitable length 14 is selected and inserted into the screw cap 12 from below. Then the neck 14 is inserted into the interior of the sample tube 11 until finally the lid 12 can be screwed onto the external thread 26.
  • the lid 12 must have sufficient freedom of movement in the axial direction so that the conical end part 20 of the attachment 14 comes into contact with the inner wall of the conical part 11 '''' of the sample tube 11 and by turning the lid 12 further in the closing direction the lower wall 37 of the cover 12 and the upper end edge 38 of the extension 14 a sufficient axial force in the direction of the arrow can be generated for the production of the sealing connection 35.
  • a labeling insert 40 is inserted into an upper recess 39 of the cover 12.
  • the entire lower conical end part 20 of the projection 14 does not abut the conical inner wall 15 of the region 11 '' '', but only a circumferential bead 20 'provided in the region of the bottom wall 17.
  • both the end part 20 and the circumferential bead 20 have a clear distance from the inner wall 15 when the extension 14 is pushed in from above until it comes into engagement with it, this can be immediate venting of the reaction chamber 19 takes place before the sealing connection 35 is produced, as a result of which a substantial pressure build-up in the reaction chamber 19 is avoided when the sealing connection 35 is being produced.
  • an inwardly projecting ring step 22 is provided at the lower end of the circular cylindrical region 11 ′′ ′′, with which the conical end part 20 of the projection 14 cooperates like a plug. This also makes it possible to separate the reaction space 19 in a gas-tight manner from the residual volume above it.
  • FIG. 4 differs from that of FIG. 1 in that instead of the hollow cylindrical extension 14, a relatively thin rod-shaped extension 14 'made of solid material is provided, which carries at its lower end a sealing plate 17' forming the end part, which at Screwing on the cover 12 comes into sealing engagement 35 with the inner wall of the conical region 11 ''''.
  • the rod-shaped extension 14 ' can also be separated from the cover 12 by pulling it out and replaced, for example, by a longer or shorter extension 14'.
  • ventilation is carried out in the exemplary embodiment according to FIG. 1 through a transverse bore 25 in the cover 12, in the exemplary embodiment according to FIG. 4 a groove 25 'is provided on the internal thread 26, which via the clearance 33 and the circumferential gap 30 is used to vent the interior 13 guaranteed.
  • a ring step 22 can also be provided at the lower end of the circular cylindrical region 11 ′′ ′′ when using a rod-shaped extension 14 ′, which cooperates with a stopper 20 ′′ forming the end part and attached to the lower end of the rod-shaped extension 14 ′.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Devices For Use In Laboratory Experiments (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)

Abstract

Un réacteur pour recevoir des quantités minimes d'échantillons fluides à chauffer, qui produisent lorsqu'ils sont chauffés des substances évaporables, comporte une éprouvette d'échantillonnage (11) dont le volume est nettement supérieur à celui desdites quantités minimes, qui est fermée en bas et ouverte en haut et dont le haut peut être fermé par un couvercle (12). Une garniture (14) est fixée audit couvercle (12) et se prolonge dans l'espace intérieur (13) de l'éprouvette d'échantillonnage (11), dont elle réduit considérablement le volume, la partie inférieure de la garniture (14) étant placée de manière hermétique contre une partie conique (11'''') de l'éprouvette d'échantillonnage (11) de manière à délimiter une petite chambre de réaction (39) inférieure séparée.

Claims (7)

  1. Procédé pour réaliser des réactions à des températures supérieures à 50°C d'échantillons (18), en particulier d'échantillons fluides, que l'on doit chauffer et qui libérent lors du chauffage des composants vaporisables, dans lequel
    - on prépare un tube (11) de prélèvement pour échantillons, susceptible d'être manipulé, le tube (11) présentant une paroi intérieure (15), une extrémité inférieure fermée (24) et une extrémité supérieure ouverte,
    - on remplit dans le tube de prélèvement (11) uniquement une quantité d'échantillon (18) correspondant à une petite fraction du volume du tube de prélèvement et on place cette quantité d'échantillon sur l'extrémité fermée (24),
    - on introduit à travers l'extrémité ouverte du tube de prélèvement et dans l'espace intérieur (13) du tube de prélèvement un insert (14, 14') qui présente une partie d'extrémité inférieure (17', 20, 20''), l'air déplacé de ce fait pouvant s'échapper vers le haut dans l'atmosphère environnante,
    - on rapproche la partie d'extrémité inférieure (17', 20, 20'') de l'insert (14) aussi près de l'extrémité fermée (24) du tube de prélèvement (11) qu'il ne reste au-dessus de l'échantillon (18) qu'un espace de vaporisation aussi petit que possible en excluant un séchage total de l'échantillon (18) au cours du traitement thermique,
    - on étanche simultanément la périphérie de la partie d'extrémité (17', 20, 20'') de l'insert (14, 14') vis-à-vis de la paroi intérieure (15) du tube de prélèvement (11), et
    - on chauffe ensuite l'échantillon (18).
  2. Procédé selon la revendication 1, caractérisé en ce que l'on prévoit un passage d'air entre l'insert (14, 14') et sa partie d'extrémité (17', 20, 20'') d'une part et la paroi intérieure (15) du tube de prélèvement (11) avant d'atteindre la position finale inférieure de la partie d'extrémité (17', 20, 20'').
  3. Procédé selon l'une ou l'autre des revendications 1 et 2, caractérisé en ce que l'on referme au moyen d'un couvercle (12) l'extrémité ouverte du tube de prélèvement (11) au même moment où l'on assure l'étanchéité entre la périphérie de la partie d'extrémité (17', 20, 20'') de l'insert (14, 14') et de la paroi intérieure (15) du tube de prélèvement (11).
  4. Procédé selon la revendication 3, caractérisé en ce que l'on prévoit un échappement d'air (25, 25'; 30) dans le couvercle (12) et/ou entre le couvercle (12) et la paroi intérieure (15) du tube de prélèvement (11) au moins pendant la mise en place du couvercle (12).
  5. Procédé selon la revendication 4, caractérisé en ce que l'échappement d'air (25, 25'; 30) est également maintenu après mise en place complète du couvercle (12).
  6. Procédé selon l'une quelconque des revendications 3 à 5, caractérisé en ce que, après réalisation de l'étanchéité entre la périphérie de la partie d'extrémité (17', 20, 20'') de l'insert (14), on applique au moyen du couvercle (12) une pression axiale sur la partie d'extrémité (17', 20, 20'') dans un sens tel que l'on renforce l'effet d'étanchéité entre la partie d'extrémité (17', 20, 20'') et la paroi intérieure du tube de prélèvement (11).
  7. Procédé selon l'une quelconque des revendications précédentes, caractérisé en ce que l'on utilise un tube de prélèvement ayant un volume de 1000 à 2000 µl et un échantillon ayant un volume de 10 à 50 µl.
EP89907732A 1988-07-07 1989-07-07 Procédé pour la réalisation des réactions avec des quantités minimes d'echantillons fluides Expired - Lifetime EP0377721B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE8808738U 1988-07-07
DE8808738U DE8808738U1 (de) 1988-07-07 1988-07-07 Gefäß zur Druchführung von Reaktionen bei erhöhter Temperatur

Publications (2)

Publication Number Publication Date
EP0377721A1 EP0377721A1 (fr) 1990-07-18
EP0377721B1 true EP0377721B1 (fr) 1993-09-15

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Family Applications (1)

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EP89907732A Expired - Lifetime EP0377721B1 (fr) 1988-07-07 1989-07-07 Procédé pour la réalisation des réactions avec des quantités minimes d'echantillons fluides

Country Status (4)

Country Link
US (1) US5167929A (fr)
EP (1) EP0377721B1 (fr)
DE (2) DE8808738U1 (fr)
WO (1) WO1990000442A1 (fr)

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Also Published As

Publication number Publication date
DE58905630D1 (de) 1993-10-21
EP0377721A1 (fr) 1990-07-18
US5167929A (en) 1992-12-01
DE8808738U1 (de) 1988-09-01
WO1990000442A1 (fr) 1990-01-25

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