WO1994019097A1 - Procede et dispositif permettant de melanger des liquides dans des plaques de microtitrage - Google Patents

Procede et dispositif permettant de melanger des liquides dans des plaques de microtitrage Download PDF

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Publication number
WO1994019097A1
WO1994019097A1 PCT/DE1994/000139 DE9400139W WO9419097A1 WO 1994019097 A1 WO1994019097 A1 WO 1994019097A1 DE 9400139 W DE9400139 W DE 9400139W WO 9419097 A1 WO9419097 A1 WO 9419097A1
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WO
WIPO (PCT)
Prior art keywords
gas
liquid
cavities
mixing
cavity
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.)
Ceased
Application number
PCT/DE1994/000139
Other languages
German (de)
English (en)
Inventor
Gunter Siegfried Fischer
Gerhard Peter Wilhelm KÜLLERTZ
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.)
Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Original Assignee
Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
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 Max Planck Gesellschaft zur Foerderung der Wissenschaften eV filed Critical Max Planck Gesellschaft zur Foerderung der Wissenschaften eV
Publication of WO1994019097A1 publication Critical patent/WO1994019097A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/40—Mixers using gas or liquid agitation, e.g. with air supply tubes
    • B01F33/407—Mixers using gas or liquid agitation, e.g. with air supply tubes by blowing gas on the material from above
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/30—Micromixers

Definitions

  • the invention relates to a method for the contact-free mixing of solutions in the cavities of microtiter plates.
  • microtiter plates Homogeneous solutions in microtiter plates are necessary if chemical reactions with measurements of the optical density in the individual cavities of a microtiter plate have to take place in a defined manner.
  • Processes for the contact-free mixing of liquids are known to a large extent.
  • the methods used are based on the one hand on the transmission of vibrations to the liquid-containing vessels or use flowing liquids or gases in order to achieve mixing by swirling.
  • Processes for producing homogeneous solutions in microtiter plates are based on the mechanical transmission of accelerated movements to the microtiter plate.
  • the microtiter plate is made to vibrate by a vibrator.
  • Vibration generator is usually one
  • the vibration generator can also be a carriage which can be influenced in a targeted manner and moved linearly and on which the microtiter plate is placed, as is the case, for example, in the microtiter plate evaluation system (MR 7000) from Dynatech (14340, Syllyfield Circle, Chantilly, VA 22021, USA). is realized.
  • MR 7000 microtiter plate evaluation system
  • the microtiter plate is accelerated in a circular manner so that the liquid in the individual cavities is set into a circular motion due to its inertia. Mixing is to be achieved by this circular movement.
  • the motion sensor is usually a shaker, as is also used in chemical laboratories for mixing larger quantities of liquid.
  • the aim of the invention is the rapid mixing of liquid quantities in microtiter plates using a non-contact method, which is particularly suitable for the subsequent, also time-dependent, optical determination of the concentration in the liquids.
  • the object of the invention is to provide a novel method and a device for the rapid, contact-free mixing of liquids in microtiter plates.
  • a gas stream is directed onto the liquid surface in such a way that only a part of the liquid surface is hit by the gas stream. This displaces the liquid that is exposed to the gas flow. If the gas pressure drops, the liquid can flow back again. By repeating this process several times, the liquid in the microtiter plate is mixed in the desired manner.
  • gases or gas mixtures which do not show any undesired interactions with the liquid present in the microtiter plate are suitable as the gas type. Air, but also nitrogen and argon, have been found to be particularly suitable for aqueous solutions.
  • the frequency of the intermittent or pulsating gas flow must be so low that at least part of the displaced liquid can flow back. This depends on the viscosity of the liquid.
  • the frequency must be like this be large that the mixing of the liquids in the microtiter plates takes place in a time which is sufficiently fast for the user and is preferably chosen to be about 6 bursts per second. In general, however, one can say that a frequency of 1 to 10 is suitable.
  • the intensity of the pulsating gas flow must be chosen so that it is strong enough to displace part of the liquid which is exposed to the gas flow. But it must not be so strong that liquid from the individual
  • Microtiter plate cavity is pressed out.
  • the strength must also be chosen so that gas bubbles do not form due to the turbulence that forms on the liquid surface exposed to the gas jet.
  • the direction of the gas jet is preferably perpendicular to the liquid surface. This procedure leads to a pulsating mixing behavior of the liquid. However, it can also be advantageous to point the gas jet obliquely at the liquid surface. With a suitable inclined arrangement of the pulsating gas jet on the edge zone of the liquid surface, circular movements of the liquid can occur and mixing can be achieved.
  • the angle at which the gas jet strikes the liquid surface in a non-vertical arrangement must be chosen such that no liquid is torn out of the individual microtiter plate cavity by the gas flow.
  • the gas flow should be such that about 30-60% of the
  • Liquid surface are exposed to the gas flow. However, it can also be advantageous to expose more than 60% or less than 30% of the liquid surface to the gas stream. If the cross-section is chosen too small, in addition to a reduction in the mixing speed, it may be undesirable
  • the cross section of the gas jet should preferably be circular. Large deviations from this circularity can also lead to the inclusion of gas bubbles in guide the liquid medium.
  • Fig. 1 Scheme of the mixing device for microtiter plates
  • the cavities in rows D1 to D12 were empty.
  • the cavities of lines E1 to E12, FI to F12, Gl to G12 and Hl to Hl2 were not mixed. Mixing was carried out immediately after the last one had been added
  • FIG. 1 In order to be able to direct the gas onto the liquid surface according to the invention, the arrangement shown in FIG. 1 was used selected. A gas flow at a pressure of 5 bar was passed from a commercially available pressure bottle with nitrogen via a 1 m long hose line (d - 100 mm) to a roller hose pump. The pump hose 5 has an inner diameter of 80 mm. The pump hose then went straight to that
  • Gas distributor 1 of the mixing device shown in Fig. 1 performed.
  • the pressure of the pump hose against the pump roll bracket and the conveying speed were set so that the gas flow could impinge on the liquid surface from the 8 nozzles 2 at a frequency of about 6 gas bursts per second.
  • Buffer solution 0.035 m HEPES buffer Ph 7.6 stock solution A: Stock solution A is a solution of human hemoglobin
  • 10 ⁇ l of stock solution A were pipetted into 8 cavities 4 of a microtiter plate 3.
  • an 8-channel pipette was used to pipette 200 ⁇ l of the buffer solution in one step to the 10 ⁇ l provided.
  • the extinctions of these 8 cavities were then measured over a period of 600 seconds at intervals of approximately 10 seconds at 390 nm using a commercially available microtiter plate reader at 6 ° C.
  • FIG. 2 show two columns of a microtiter plate with the cavities AI to Hl (column 1) and A2 to H2 (column 2).
  • the absorbance timings in column 2 were obtained using a two-second mixing step.
  • the absorbance timings in column 1 were obtained without mixing.
  • the final absorbances of the unmixed samples do not reach the exact final value even after more than 10 minutes. Visually, inhomogeneities in the individual cavities can still be observed even after 20 minutes. All mixed samples have the same end absorbances after the mixing time.
  • Compressed air was used as the mixed gas, which was produced using a commercially available laboratory radial fan.
  • the delivery rate was 9 m 3 / h at a pressure head of 450 mm water column.
  • the compressed air is fed to a gas distributor 1 via a hose feed 5 of 100 cm in length with an inner diameter of 90 mm.
  • the cycle frequency of the motor-driven 12-channel valve located in the gas distributor can be controlled via the engine speed.
  • the compressed air coming from the valve is directed via nozzles 2 onto the liquid surface of the individual cavities 4.
  • the nozzles 2 were 30 mm long and had a circular cross section.
  • the nozzles 2 had a diameter of 2 mm at the tapered end and 5 mm at the air inlet opening.
  • the distance between the nozzles 2 of the chamber shown in FIG. 1 and the microtiter plate 3 was 5 mm.
  • reaction solution A 120 ⁇ l are automatically pipetted into each cavity. No reaction solution was pipetted into the cavities D1, D2, D3, p4, DS, D6, D7, D8, D9, D10, DU and D12.
  • the 8-fold pipetting automatically pipettes 20 ⁇ l substrate solution per cavity within 7 seconds. No substrate solution was pipetted into the cavities D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, DU and Dl2.
  • the microtiter plate is moved into the "waiting position" for 7 seconds.
  • microtiter plate is mixed in the manner described above.
  • Microtiter plate measured 70 times in approximately 9 seconds.
  • the extinction changes obtained, the curve of which corresponds to a first-order reaction, can be represented graphically. 3 shows the result of such a measurement.
  • the "do not mix" was achieved by interrupting the corresponding nozzles.
  • the measurement curves can be evaluated as described in EP 0 360 029 AI.
  • Rate constants the mean values and the standard deviations of the rate constants of the same enzyme concentrations, with and without a mixture, are summarized in Table 1.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

L'invention concerne un procédé et un dispositif permettant de mélanger des liquides. Selon l'invention, si on utilise un type de gaz, une fréquence, une intensité, une direction et une section appropriés, un courant de gaz produisant périodiquement des pulsations et agissant sur la cavité d'une plaque de microtitrage permet d'opérer un mélange rapide dudit liquide dans la cavité de la plaque microtitrage, sans qu'il y ait formation de bulles dans le liquide, ce qui risquerait de gêner l'étape suivante qui consiste en une mesure optique du degré de concentration.
PCT/DE1994/000139 1993-02-26 1994-02-10 Procede et dispositif permettant de melanger des liquides dans des plaques de microtitrage Ceased WO1994019097A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DEP4306085.4 1993-02-26
DE4306085A DE4306085A1 (de) 1993-02-26 1993-02-26 Verfahren und Vorrichtung zum Durchmischen von Flüssigkeiten in Mikrotiterplatten

Publications (1)

Publication Number Publication Date
WO1994019097A1 true WO1994019097A1 (fr) 1994-09-01

Family

ID=6481476

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1994/000139 Ceased WO1994019097A1 (fr) 1993-02-26 1994-02-10 Procede et dispositif permettant de melanger des liquides dans des plaques de microtitrage

Country Status (2)

Country Link
DE (1) DE4306085A1 (fr)
WO (1) WO1994019097A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6890485B1 (en) 1997-05-16 2005-05-10 Aurora Discovery, Inc. High throughput chemical handling system

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3087840A (en) * 1958-06-16 1963-04-30 Macrosonic Process Corp Methods and means for producing physical, chemical and physicochemical effects by large-amplitude sound waves
DE1299507B (de) * 1964-10-21 1969-07-17 Kawasaki Jakogyo Kabushiki Kai Verfahren zum pneumatischen Mischen von Mischgut mittels periodisch verlaufender Blasvorgaenge
DE1782092A1 (de) * 1968-07-18 1971-07-08 Inst Chemii Ogolnej Verfahren zum Mischen von pulverfoermigen und feinkoernigen Feststoffen mit Fluessigkeiten und Vorrichtung zur Durchfuehrung des Verfahrens
EP0176014A2 (fr) * 1984-09-22 1986-04-02 Eppendorf-Netheler-Hinz Gmbh Procédé et dispositif pour mélanger un échantillon d'un liquide à analyser
EP0281958A2 (fr) * 1987-03-07 1988-09-14 Hoechst Aktiengesellschaft Dispositif de régulation de température et de mélange du contenu d'une plaque de micro titration

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3087840A (en) * 1958-06-16 1963-04-30 Macrosonic Process Corp Methods and means for producing physical, chemical and physicochemical effects by large-amplitude sound waves
DE1299507B (de) * 1964-10-21 1969-07-17 Kawasaki Jakogyo Kabushiki Kai Verfahren zum pneumatischen Mischen von Mischgut mittels periodisch verlaufender Blasvorgaenge
DE1782092A1 (de) * 1968-07-18 1971-07-08 Inst Chemii Ogolnej Verfahren zum Mischen von pulverfoermigen und feinkoernigen Feststoffen mit Fluessigkeiten und Vorrichtung zur Durchfuehrung des Verfahrens
EP0176014A2 (fr) * 1984-09-22 1986-04-02 Eppendorf-Netheler-Hinz Gmbh Procédé et dispositif pour mélanger un échantillon d'un liquide à analyser
EP0281958A2 (fr) * 1987-03-07 1988-09-14 Hoechst Aktiengesellschaft Dispositif de régulation de température et de mélange du contenu d'une plaque de micro titration

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6890485B1 (en) 1997-05-16 2005-05-10 Aurora Discovery, Inc. High throughput chemical handling system
US7105132B2 (en) 1997-05-16 2006-09-12 Aurora Discovery, Inc. Liquid chemical distribution method and apparatus

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Publication number Publication date
DE4306085A1 (de) 1994-09-01

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