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EP0562358B1 - Procédé de correction d'erreur de volume a la conception d'un système de pipetage - Google Patents

Procédé de correction d'erreur de volume a la conception d'un système de pipetage Download PDF

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Publication number
EP0562358B1
EP0562358B1 EP93103815A EP93103815A EP0562358B1 EP 0562358 B1 EP0562358 B1 EP 0562358B1 EP 93103815 A EP93103815 A EP 93103815A EP 93103815 A EP93103815 A EP 93103815A EP 0562358 B1 EP0562358 B1 EP 0562358B1
Authority
EP
European Patent Office
Prior art keywords
pipette
liquid
volume
tip
dead volume
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
EP93103815A
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German (de)
English (en)
Other versions
EP0562358A2 (fr
EP0562358B8 (fr
EP0562358A3 (en
EP0562358B2 (fr
Inventor
Dieter Dr. Husar
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.)
Eppendorf SE
Original Assignee
Eppendorf Netheler Hinz GmbH
Eppendorf Geraetebau Netheler and Hinz GmbH
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Application filed by Eppendorf Netheler Hinz GmbH, Eppendorf Geraetebau Netheler and Hinz GmbH filed Critical Eppendorf Netheler Hinz GmbH
Publication of EP0562358A2 publication Critical patent/EP0562358A2/fr
Publication of EP0562358A3 publication Critical patent/EP0562358A3/de
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    • 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
    • B01L3/02—Burettes; Pipettes
    • B01L3/021—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
    • B01L3/0217—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids of the plunger pump type

Definitions

  • the invention relates to a method for correcting the volume error in the design of a pipetting system with a piston stroke pitt and a pipette tip which can be plugged thereon.
  • the piston stroke pipette has a cone for attaching the pipette tip.
  • the pipette tip has a strongly conical section and then, up to the push-on opening, a weaker conical section with respect to the inner contour.
  • Such a pipette tip has a deep and in particular slim when immersed Vessel has the disadvantage that a pipette carrying it can come into contact with the vessel and become contaminated. This is fundamentally not the case with a pipette tip which, according to EP-A-0 182 943, has a long cylindrical nose, starting from a tip opening, followed by a strongly conical and then a plug-on opening.
  • the setting range of absorbable amounts of liquid extends in each case from the tip opening over all different sections of the pipette tip. If a small piston stroke is set, liquid is only drawn into the strongly conical (or cylindrical) initial area. With a larger piston stroke, the weaker conical (or the conical and then the cylindrical) area is reached.
  • the pipetting systems have in common that, in cooperation with an adjustable piston stroke pipette, they absorb liquid volumes that can unacceptably deviate from the displayed liquid volume, the deviation being possible differently across the setting range.
  • the pitch of a spindle of the piston stroke pipette for the adjustment of the piston stroke is tentatively determined so that the correctness deviation between the liquid volume taken up and the one indicated is tolerable across the entire setting range.
  • the resulting deviations in accuracy are accepted and can only be determined on the basis of subsequent measurements.
  • US Pat. No. 5,024,109 already discloses a method and an apparatus for correcting the error of a piston stroke pipette.
  • the height of the liquid in the pipette tip for the desired liquid volume, the changes in the dead volume due to the liquid column in the tip and the required piston displacement for the desired liquid volume and the change in the dead volume are determined.
  • the height of the liquid column for a desired volume is based on the tip geometry determined, especially for a conical tip.
  • the change in dead volume is determined on the basis of hydrostatic considerations that are linked to the height of the liquid column.
  • the piston displacement is determined from the sum of the desired volume and the change in the dead volume.
  • This technology uses an electromechanical pipette that is connected to a control computer. Before use, data from the pipette tip, the pipetting system and the liquid must be entered into the computer. This pipetting technique is complex.
  • the object of the invention is to improve the pipetting system of the type mentioned at the outset, so that the pipetting liquid to be absorbed can be worked easily and precisely over the entire setting range. Furthermore, the pipetting system is intended to enable the piston-stroke pipettes or pipette tips to be exchanged, with the aim of continuing to work precisely and simply.
  • the invention is based on the knowledge that the pipette correction factor, cross-sectional profile along the tip, dead volume and, if appropriate, zero point shift of the display are decisive for the amount of the accuracy deviations over the setting range.
  • By coordinating these influencing factors with one another it is consequently possible to implement a predefined correctness deviation from the displayed liquid volume in the setting range, which can also be zero, for each liquid volume taken up.
  • deviations in accuracy are no longer minimized by selecting the spindle pitch according to the specification of the other influencing variables, but rather all the influencing variables mentioned above are coordinated with one another in such a way that predetermined deviations in accuracy are achieved.
  • pipette tips can be adapted to conventional piston-stroke pipettes in such a way that predetermined deviations in accuracy are achieved.
  • a pipetting system can be equipped with an extended pipette tip, which avoids wall contact and accompanying contamination.
  • the deviation in accuracy can be defined in different ways.
  • the specification of a constant absolute or relative correctness deviation is particularly favorable because the difference between the liquid volume absorbed and the displayed liquid volume is then easiest to determine.
  • a vote can be carried out for each volume of liquid absorbed requires a predefined deviation from the volume of liquid absorbed by the conventional pipetting system instead of the volume of liquid displayed. Then the differences of the pipetted liquid volume from the conventionally pipetted volume are known with the same display. This knowledge can be interesting for comparative investigations with a modified pipette tip.
  • the liquid level in the setting range of receivable liquid volumes always rises up to a working range of the pipette tip.
  • the adjustment can include a zero point shift of the displayed liquid volume relative to the piston stroke.
  • values of pipette correction factor, dead volume and possibly zero point shift are sometimes specified, in particular if the pipetting system is to be implemented by adapting a pipette tip to an existing piston stroke pipette. To achieve the specified deviation from the correctness, it is then sufficient to adjust the cross-sectional profile along the tip to the specified influencing variables.
  • the working area of the pipette tip is essentially cylindrical. Then, in particular, a constant absolute deviation in accuracy can be achieved over the entire setting range of absorbable liquids. For a small tip opening and a sufficiently large volume area for receiving adjustable amounts of liquid, this can have a gradually widening transition area.
  • the transition area preferably has a transition radius or transition cone, the cone angle being kept as small as possible to avoid undesired effects (fountain effect).
  • a work area is preferably arranged downstream of the transition area.
  • the pipetting system has a pipette tip with a thin suction tube adjacent to the tip opening. Then the top preferably has an expanding transition and a subordinate work area.
  • Its pipette correction factor can be adjustable in order to adapt the pipetting system to different liquid densities or air pressures. In the simplest case, this can be done by replacing a spindle for adjusting the piston stroke.
  • a display device with an expandable scale can also be used.
  • the display can also be shifted to zero by means of similar measures.
  • An adjustable dead volume can also be provided. This is possible in particular by exchanging pipette tips, a pipette tip for lighter liquids having the same diameter having to be made longer. In order to change the dead volume, intermediate pieces between the piston-stroke pipette and the pipette tip can also be used.
  • the influencing variables can be matched experimentally to achieve the specified correctness deviations.
  • an accuracy deviation from the displayed liquid volume can first be specified for each liquid volume taken up.
  • These deviations can now be realized by coordinating the pipette correction factor, cross-sectional profile along the tip and dead volume. Since the pipette correction factor and the dead volume are usually no longer changed after the selection has been made, the values for these influencing parameters are initially at least provisionally determined, it being possible to orientate on values of known pipetting systems. Then the last one remains Influencing variable of the cross-sectional shape along the pipette tip, with which the specified accuracy deviations can be ensured. Trials, for example, result in clear values for this if the other influencing factors have been selected beforehand. If the cross-sectional profile found cannot be realized, the predefined influencing variables would have to be corrected or even a different correctness deviation had to be specified.
  • the piston stroke volume V piston of a piston stroke pipette is larger than the absorbed liquid volume V liquid by the volume error ⁇ V. .
  • V piston V Liquid + ⁇ V
  • V piston V Liquid + ⁇ gh p O V t
  • V No. (V Liquid + ⁇ gh p O ⁇ V t - b) 1 a
  • the pipette has a housing 1. At the upper end of the housing there is an actuating button 2 which can be moved against the spring 14.
  • the button 2 is connected to a piston rod 7 which extends through a spindle 6.
  • the lower end of the rod 7 carries a connecting piece 8 which has a carrier 9 for a piston 10.
  • a spring 18 presses a lower sleeve 19 for guiding the piston 9, 10 against an inner conical surface 21 of the housing 1.
  • the sleeve 19 is supported on the housing via a flexible sealing ring 20.
  • the piston rod 7 is provided with a flange 22 on which the spindle 6 rests in the initial position of the rod 7 and the piston 9, 10.
  • This flange 22 defines the upper stop for the piston 9, 10.
  • the upper stop is adjustable as follows:
  • the button 2 is connected to a downwardly extending sleeve 5 which is rotatably held in the housing.
  • the spindle 6, which surrounds the rod 7, is rotatably connected to the rotatable sleeve 5.
  • the outer thread of the spindle 6 works together with an internal thread of a part 36 which is fastened in the housing 1. When the knob 2 is turned, the spindle 6 is screwed into or out of the threaded part 36.
  • the spindle 7 is axially adjusted, whereby the setting of the stop for the flange 22 of the rod 7 is set, which takes effect when the piston 9, 10 returns to an initial position, when the button 2 is released and reset by the spring 14 .
  • the stroke of the piston is limited by the upper stop of the piston rod in its initial position.
  • a wheel 50 is adjusted via a worm gear.
  • the sleeve 5 has teeth 52 on the outside.
  • the rotation of the wheel 50 is indicated by a scale which is visible through a window 53.
  • the scale is calibrated to adjust the upper stop so that a desired amount of liquid is drawn into the pipette tip, which is attached to the lower end of the tube 12.
  • FIGS. 4 and 5 show a part of the dead volume 11 and 12 as well as the setting means and display means 51 for setting and displaying the desired amount of liquid.
  • curves A 1 and A 2 represent the behavior of cylindrical pipette tips P 1 according to FIG. 2.
  • the negative volume error - ⁇ V increases linearly with increasing piston stroke.
  • the curve A 1 corresponds to a pipette tip with a smaller one and the curve A 2 of a pipette tip with a larger cross section.
  • the volume error - ⁇ V of the pipette tip is compensated for by tuning the piston stroke pipette.
  • the pipette correction factor By setting the pipette correction factor, the difference between the displayed liquid volume V Qty. and the piston stroke V piston in dependence on the piston stroke just chosen so that the indicated liquid volume V Anz. the absorbed volume of liquid V Liquid. corresponds.
  • the piston-stroke pipette with the curve K 1 and the larger pipette correction factor a is assigned to the pipette tip with the curve A 1 and the piston-stroke pipette with the curve K 2 to the pipette tip with the curve A 2 .
  • the graph illustrates that for each piston stroke V piston the difference between the indicated liquid volume V Qty . and the absorbed volume of liquid V Liquid. is zero.
  • Curve B in FIG. 1 is assigned to pipette tip P 2 from FIG. 2. This has a thin suction tube adjacent to the tip opening, which initially causes a relatively high rise in the liquid column with a small piston stroke. As a result, the negative volume error - ⁇ V is also relatively high in this initial range. Fills the liquid column with the suction tube a smaller increase in the negative volume error is observed when rising into the adjacent cylindrical working volume.
  • the curve C in FIG. 1 corresponds to the pipette tip P 3 of FIG. 2.
  • This has a conical transition area to a cylindrical working volume adjacent to a thin suction tube. 1, a region of attenuated slope of the volume error for the transition area adjoins the steep initial section, which finally leads to a region of weak slope for the cylindrical working section.
  • the curve C corresponds to a piston pipette in which the difference between the indicated liquid volume V Qty. is represented by the curve K 3 for the piston stroke V piston over the piston stroke.
  • This has a zero point shift NV on the ordinate, ie with the piston in the starting position, a liquid volume V Qty. displayed.
  • the zero point shift is selected so that with a piston stroke V piston , which causes an increase in the liquids in the working area of the pipette tip, a compensation of the volume error is just achieved.
  • the system thus has an error curve F that of maximum initial value falls to a value zero, which is reached when the liquid column rises into the working range.
  • the pipette tip in FIG. 3 has a tip opening 1 'at the bottom for liquid passage, which is followed by a thin suction tube 2'.
  • the suction tube 2 ' opens into a transition cone 3', which communicates at the other end with an expanded working area 4 '.
  • the working area 4 ' opens at the top into a push-on cone 5' with a push-on opening 6 'for a piston stroke pipette with an adjustable piston stroke.
  • the liquid In the setting range of absorbable amounts of liquid, the liquid always rises through the tip opening 1 and the suction tube 2 and the transition cone 3 up to the working area 4.
  • the pipette tip in the working area 4 is shaped approximately circularly on the inside.
  • the transition cone has a clear cone angle, which is approx. 7 ° to avoid a fountain effect.
  • a typical pipette tip has an overall length of approximately 100 mm, of which approximately 25 mm are allotted to the suction tube 2. The diameter increases from approx. 0.5 mm in the tip opening 1 to approx. 3 mm at the beginning of the working area 4.

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  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Devices For Use In Laboratory Experiments (AREA)
  • Sampling And Sample Adjustment (AREA)

Claims (14)

  1. Procédé de correction de l'erreur ΔV sur le volume lors du dimensionnement d'un système de pipettage, comportant une pipette fonctionnant par la course d'un piston, et une pointe de pipette pouvant être enfoncée sur cette pipette, la pipette à course de piston comportant des dispositifs de réglage pour modifier la course du piston, ainsi que des dispositifs d'affichage du volume de liquide pris chaque fois par la pipette, le rapport entre la course du piston et le volume affiché du liquide, étant déterminé par un facteur de correction a, la pointe de pipette présentant une ouverture de pointe, une zone de volume reliée à elle, pour recevoir des volumes réglables de liquide, et une ouverture d'enfichage, reliée à elle, pour la pipette à course de piston, tandis qu'entre l'ouverture de la pointe et le piston, est réalisé un volume mort Vt, l'erreur de volume ΔV à corriger résultant de ce que le volume mort Vt, entre le niveau du liquide et le piston de la pipette augmente sous l'effet du poids de la colonne de liquide dans la pointe de pipette, la correction de l'erreur de volume ΔV se faisant par détermination, par construction, du volume mort Vt, du facteur de correction de pipette a, et de l'évolution de la section transversale Q de la pointe de pipette qui présente une zone de travail cylindrique, de telle façon que le volume de liquide VAnz indiqué par le dispositif d'affichage corresponde au volume de liquide reçu ou à une valeur dérivée prédéfinie.
  2. Procédé suivant la revendication 1, caractérisé en ce que le facteur de correction de pipette, l'évolution de la section transversale intérieure et le volume mort sont adaptés à un décalage du point zéro du volume affiché du liquide, par rapport à la course du piston.
  3. Procédé suivant la revendication 1 ou la revendication 2, caractérisé en ce que l'évolution de la section transversale intérieure le long de la pointe est adaptée à des valeurs prédéfinies du facteur de correction de pipette, du volume mort et, le cas échéant, du décalage du point zéro.
  4. Procédé suivant l'une des revendication 1 à 3, caractérisé en ce qu'à la zone de réglage des volumes de liquide à recevoir, est associée une zone de travail de la pointe de pipette, dans laquelle le niveau du liquide effectue un mouvement ascendant dans toute la zone de réglage.
  5. Procédé suivant l'une des revendication 1 à 4, caractérisé en ce que le facteur de correction de pipette, l'évolution de la section transversale intérieure, le volume mort et, le cas échéant, le décalage du point zéro sont adaptés les uns aux autres en tenant compte des relations: R abs. = V Flüss. - (V Flüss. + ρ gh Po . V t - b) 1 a
    Figure imgb0026
     ou R rel = R abs. /V Flüss. = écart relatif de précision,
    Figure imgb0027
     avec: R abs = V Flüss. - V Anz. = écart absolu de précision V Flüss. = o h Q(y) dy = volume de liquide reçu
    Figure imgb0028
    Q(y)   = Section transversale à la distance y de l'ouverture de la pointe
    VAnz.   = volume de liquide affiché
    ρ   = Densité du liquide à aspirer par la pipette
    g   = accélération due à la pesanteur
    h   = hauteur de la colonne de liquide dans la pointe de pipette
    Vt   = volume mort
    Po   = pression ambiante
    a   = facteur de correction de pipette
    b   = décalage du point zéro
  6. Procédé suivant l'une des revendication 1 à 5, caractérisé en ce qu'on utilise une pointe de pipette comportant une zone de travail essentiellement cylindrique.
  7. Procédé suivant l'une des revendications 1 à 6, caractérisé en ce qu'on utilise une pointe de pipette dans laquelle une zone de transition s'élargissant progressivement est située en amont de la zone de travail de la pointe de pipette.
  8. Procédé suivant la revendication 7, caractérisé en ce que la zone de transition est déterminée par un angle le plus faible possible.
  9. Procédé suivant l'une des revendication 6 à 8, caractérisé en ce qu'on utilise une pointe de pipette disposant d'un fin tube d'aspiration adjacent à l'ouverture de pointe.
  10. Procédé suivant l'une des revendication 6 à 9, caractérisé en ce qu'on règle le facteur de correction de pipette.
  11. Procédé suivant l'une des revendication 6 à 10, caractérisé en ce qu'on règle le décalage du point zéro de l'affichage.
  12. Procédé suivant l'une des revendication 6 à 11, caractérisé en ce qu'on règle le volume mort.
  13. Procédé suivant la revendication 12, caractérisé en ce qu'on règle le volume mort en échangeant des pointes de pipette et/ou des pièces intermédiaires entre la pipette à course de piston et la pointe de pipette.
  14. Procédé suivant l'une des revendication 1 à 13, caractérisé en ce qu'on règle le volume mort en faisant varier le réglage de la butée inférieure de la course du piston.
EP93103815A 1992-03-25 1993-03-10 Procédé de correction d'erreur de volume a la conception d'un système de pipetage Expired - Lifetime EP0562358B8 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE4209620A DE4209620C1 (de) 1992-03-25 1992-03-25 Verfahren zur Korrektur des Volumenfehlers ïV bei einem Pipettiersystem
DE4209620 1992-03-25

Publications (5)

Publication Number Publication Date
EP0562358A2 EP0562358A2 (fr) 1993-09-29
EP0562358A3 EP0562358A3 (en) 1994-05-18
EP0562358B1 true EP0562358B1 (fr) 1997-07-16
EP0562358B2 EP0562358B2 (fr) 2005-08-24
EP0562358B8 EP0562358B8 (fr) 2005-10-26

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EP93103815A Expired - Lifetime EP0562358B8 (fr) 1992-03-25 1993-03-10 Procédé de correction d'erreur de volume a la conception d'un système de pipetage

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US (1) US5481900A (fr)
EP (1) EP0562358B8 (fr)
DE (2) DE4209620C1 (fr)

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DE10118875C1 (de) * 2001-04-18 2002-09-12 Eppendorf Ag Verfahren zum kontrollierten Dosieren von Flüssigkeiten unter Verdrängung eines Gaspolsters
EP2286921A1 (fr) 2006-07-14 2011-02-23 Eppendorf AG Dispositif de dosage électronique destiné au dosage de liquides

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10118875C1 (de) * 2001-04-18 2002-09-12 Eppendorf Ag Verfahren zum kontrollierten Dosieren von Flüssigkeiten unter Verdrängung eines Gaspolsters
EP2286921A1 (fr) 2006-07-14 2011-02-23 Eppendorf AG Dispositif de dosage électronique destiné au dosage de liquides

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Publication number Publication date
EP0562358A2 (fr) 1993-09-29
DE59306907D1 (de) 1997-08-21
EP0562358B8 (fr) 2005-10-26
DE4209620C1 (de) 1993-12-16
EP0562358A3 (en) 1994-05-18
US5481900A (en) 1996-01-09
EP0562358B2 (fr) 2005-08-24

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