EP0562358B2 - 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
EP0562358B2
EP0562358B2 EP93103815A EP93103815A EP0562358B2 EP 0562358 B2 EP0562358 B2 EP 0562358B2 EP 93103815 A EP93103815 A EP 93103815A EP 93103815 A EP93103815 A EP 93103815A EP 0562358 B2 EP0562358 B2 EP 0562358B2
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EP
European Patent Office
Prior art keywords
pipette
volume
liquid
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
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EP93103815A
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German (de)
English (en)
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EP0562358B8 (fr
EP0562358A2 (fr
EP0562358A3 (en
EP0562358B1 (fr
Inventor
Dieter Dr. Husar
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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
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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 and a pipette tip that can be plugged in.
  • the Kolbenhubpipette has a cone for attaching the pipette tip.
  • the pipette tip has one starting from the tip opening strongly conical and then up to the slip on a weaker conical section based on the inner contour on.
  • Such a pipette tip when immersed in a deep and especially slim vessel has the disadvantage that it carrying a pipette wall contact with the vessel get and be contaminated.
  • This is basically a pipette tip not the case, according to EP-A-0 182 943 from a tip opening a long cylindrical nose, then a strong conical and then a Has slip-on.
  • the adjustment range extends more absorbably Amounts of liquid each from the tip opening over all different sections away from the pipette tip. If a small piston stroke is set, fluid is only in the strongly conical (or cylindrical) initial area sucked in. For larger Piston stroke becomes the weaker conical (or the conical and then the cylindrical) Area reached.
  • the pipetting systems have in common that they interact with an adjustable piston stroke pipette to accommodate fluid volumes that are unacceptable may differ from the indicated liquid volume, the deviation over the Setting range is possible in different ways.
  • the slope of a Spindle of the piston stroke pipette for the adjustment of the piston stroke on a trial basis determines that the correctness deviation between recorded and displayed liquid volume can deviate, with the deviation is possible differently over the adjustment range. Therefore, the pitch of a spindle of Kolbenhubpipette for the adjustment of the piston stroke tentatively determined so that the corrective deviation between recorded and displayed volume of liquid over the entire adjustment range is bearable. The resulting accuracy deviations are accepted and are possibly due to subsequent measurements ascertainable.
  • U.S. Patent No. 5,024,109 already discloses a method and apparatus for correcting the failure of a piston-stroke pipette.
  • the height of the liquid column for a desired volume is determined by the tip geometry, especially for a conical tip.
  • the change The dead volume is determined on the basis of hydrostatic considerations related to the height of the liquid column tie.
  • the piston displacement becomes the sum of the desired volume and the change in the dead volume determined.
  • This technique uses an electromechanical pipette connected to a control computer is. Before use, data of the pipette tip, the pipetting system and the liquid must be entered into the computer be entered. This pipetting technique is expensive.
  • the invention has the object, the pipetting system of the type mentioned above improve so that over the entire adjustment range of pipetting liquid to be taken over easily and accurately can be worked. Furthermore, the pipetting system is intended to replace the piston stroke pipettes or pipette tips while continuing to work accurately and simply.
  • the invention is based on the finding that pipette correction factor, cross-sectional profile along the tip, Dead volume and, if necessary, zero offset of the display for the amount of the corrective deviations the setting range are relevant. By matching these influencing factors to each other, it is therefore possible in the Adjustment range for each liquid volume taken a given accuracy deviation from the displayed To realize liquid volume, which can also be zero.
  • the spindle pitch according to the invention according to specification minimized the remaining predictors, but all o.g. Influencing variables coordinated so that predetermined Correctness deviations are achieved. In such a pipetting system is then for each indicated liquid volume the exact value of the liquid volume taken known.
  • the correctness deviation can be defined in various ways. For practical handling the specification of a constant absolute or relative accuracy deviation is particularly favorable, because then the Difference of the recorded liquid volume is the easiest to determine.
  • the liquid level always rises in the adjustment range of absorbable liquid volumes up into a cylindrical working area of the pipette tip.
  • the vote can be a zero offset of the displayed Include fluid volume with respect to the piston stroke.
  • the working area of the pipette tip is substantially cylindrical. Then, in particular, a constant Absolute accuracy deviation over the entire range of absorbable liquids realized away become.
  • the transition area preferably has a transition radius or transition cone, wherein the cone angle to avoid unwanted Effects (fountain effect) is kept as small as possible.
  • the transition region is preferably one Subordinate workspace. Especially for use with relatively long and slim vessels has the pipetting system a pipette tip with a thin aspiration tube adjacent to the tip opening. Then the Tip also prefers an expanding transitional as well as a downstream workspace.
  • pipette correction factor may be be adjustable. This is in the simplest case by replacing a spindle for adjusting the Piston strokes feasible. Also comes for a display device with a spreadable scale into consideration.
  • an adjustable dead volume can be provided. This is especially the replacement of pipette tips possible, with a pipette tip designed for lighter liquids with the same diameter longer must become. To change the dead volume can also intermediate pieces between Kolbenhubpipette and pipette tip be usable.
  • a vote of the influencing variables to achieve the predetermined corrective deviations done experimentally. It can first for each recorded fluid volume a corrective deviation be specified by the indicated liquid volume. These deviations are now through Adjustment of pipette correction factor, cross-sectional profile along the tip and dead volume to realize each other. Since the pipette correction factor and the dead volume are usually no longer changed after making the selection, First, the values for these influence parameters are at least provisionally determined, with an orientation to Values of known pipetting systems is possible. Then remains as the last predictor of cross-sectional shape along the pipette tip, with the specified accuracy deviations can be ensured. For that result For example, by experiments unique values, if the other influencing factors have been previously chosen. If the cross-sectional profile found could not be realized, the predetermined influencing variables would have to be corrected or even specify another correctness deviation.
  • V piston V Liq. + ⁇ gh p O V t
  • V Num. (V Liq. + ⁇ gh p O ⁇ V t - b) ⁇ 1 a
  • R Section. V Liq. - (v Liq. + ⁇ gh p O ⁇ V t - b) ⁇ 1 a
  • R Section. V Liq. - V Num.
  • the pipette according to FIGS. 4 and 5 is disclosed in the applicant's US-A-5511433. This disclosure is referred to.
  • the pipette has a housing 1. At the top of the housing is a Actuating knob 2 attached, which is movable against the spring 14.
  • the knob 2 is connected to a piston rod 7, which extends through a spindle 6.
  • Connecting piece 8 As shown in Fig. 5, the lower end of the rod 7 enters Connecting piece 8, which has a support 9 for a piston 10.
  • a spring 18 presses a lower sleeve 19th 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 resilient sealing ring 20 from.
  • the piston 9, 10 When the button 2 is pressed down vertically, the piston 9, 10 is pressed into the cylinder chamber 11, wherein he presses air from the cylinder chamber 11 through the tube 12.
  • a attachable Attached pipette tip At the lower end of the tube 12 is a attachable Attached pipette tip, which is shown in Fig. 3 on an enlarged scale.
  • the pipette tip is on a conical Seat attached to the lower end of the tube 12 in a manner not shown.
  • the cylinder chamber 11, the tube 12 and the pipette tip define the dead volume of the pipette.
  • the piston rod 7 is provided with a flange 22 on which the spindle 6 in the initial position of the rod. 7 and the piston 9, 10 rests.
  • This flange 22 defines the upper stop for the piston 9, 10.
  • the upper stop is adjustable as follows:
  • the knob 2 is connected to a downwardly extending sleeve 5, which is rotatable in the housing is held.
  • the spindle 6, which surrounds the rod 7, is rotatably connected to the rotatable sleeve 5.
  • the outer Thread of the spindle 6 cooperates with an internal thread of a part 36 which is fixed in the housing 1. When the knob 2 is rotated, the spindle 6 is screwed into or out of the threaded portion 36.
  • the spindle 7 is thereby adjusted axially, whereby the setting of the stop for the flange 22 of the rod 7 is set which becomes effective upon return of the piston 9, 10 to an initial position when the button 2 is released and is reset by the spring 14.
  • the stroke of the piston is determined by the upper stop of the piston rod in limited to its starting position.
  • a wheel 50 is adjusted via a worm gear.
  • the sleeve 5 has outside a toothing 52.
  • the rotation of the wheel 50 is indicated by a scale, the through a window 53 is visible.
  • the scale is calibrated to adjust the top stop so that one desired amount of liquid is sucked into the pipette tip, attached to the lower end of the tube 12 is.
  • FIGS. 4 and 5 show part of the dead volume 11 and 12 as well as the adjusting means and Display means 51 for setting and displaying the desired amount of liquid.
  • the 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 and the curve A 2 of a pipette tip with a larger cross-section.
  • the volume error - ⁇ V of the pipette tip is compensated by tuning the piston stroke pipette. Namely, by adjusting the pipette correction factor, the difference between the indicated liquid volume V Anz. and the piston stroke V piston in response to the piston stroke just chosen so that the indicated liquid volume V no. the recorded liquid volume V liquid. equivalent.
  • the piston stroke pipette with the curve K 1 and the larger pipette correction factor a of the pipette tip with the curve A 1 and the piston stroke pipette with the curves K 2 of the pipette tip with the curve A 2 are assigned.
  • the diagram illustrates that for each piston stroke V piston the difference between the indicated liquid volume V no . and the received liquid volume V liquid. is equal to zero.
  • the curve B in FIG. 1 is assigned to the pipette tip P 2 of FIG. 2. This has adjacent to the tip opening a thin suction tube, which initially causes a small piston stroke a relatively high increase in the liquid column. As a result, in this initial region, the negative volume error - ⁇ V is relatively high. If the liquid column fills the suction tube, a slight 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 adjacent to a thin suction tube a conical transition region to a cylindrical working volume. Thus, in FIG. 1, adjacent to the steep initial section, there is an area of attenuated slope of the volume error for the transition region, which eventually terminates in a region of weak slope for the cylindrical working section.
  • the curve C corresponds to a Kolbenhubpipette, in which the difference of the indicated liquid volume V no. for the piston stroke V piston is represented over the piston stroke through the curve K 3 .
  • This has on the ordinate a zero offset NV, ie the piston in the starting position is already a liquid volume V Anz. displayed.
  • the zero offset is chosen so that in a piston stroke V piston , which causes an increase of the liquids in the working range of the pipette tip, just a compensation of the volume error is achieved.
  • the system thus has an error curve F, which falls from the maximum initial value to a value zero, which is reached when the liquid column rises to the working range.
  • the pipette tip in Fig. 3 has at the bottom a tip opening 1 'for liquid passage, to which a thin Suction tube 2 'connects.
  • the suction tube 2 ' opens into a transition cone 3', the other with an extended work area 4 'communicates.
  • the work area 4 ' opens at the top in a Aufsteckkonus 5' with a plug-in opening 6 'for a Kolbenhubpipette with adjustable piston stroke.
  • the liquid rises through the tip opening 1 and the Suction tube 2 and the transition cone 3 always up in the work area 4 up.
  • the pipette tip in the working area 4 inside is approximately circular cylindrical shaped.
  • the transition cone has a clear cone angle, which helps avoid a fountain effect about 7 °.
  • a typical pipette tip has a total length of about 100 mm, of which about 25 mm on the suction tube 2 omitted. The diameter increases from about 0.5 mm in the tip opening 1 to about 3 mm at the beginning of the working area 4 on.

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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 (12)

  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 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: Rabs. = VFlüss. - (VFlüss. + ρ ghPo . Vt - b)1a ou
       Rrel = Rabs./VFlüss. = écart relatif de précision,
    avec:
    Rabs = VFlüss. - VAnz. = écart absolu de précision
    VFlüss. =
    Figure 00180001
    = volume de liquide reçu
    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
  5. Procédé suivant l'une des revendications 1 à 4, 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.
  6. Procédé suivant la revendication 5, caractérisé en ce que la zone de transition est déterminée par un angle le plus faible possible.
  7. Procédé suivant l'une des revendication 1 à 6, caractérisé en ce qu'on utilise une pointe de pipette disposant d'un fin tube d'aspiration adjacent à l'ouverture de pointe.
  8. Procédé suivant l'une des revendication 1 à 7, caractérisé en ce qu'on règle le facteur de correction de pipette.
  9. Procédé suivant l'une des revendication 1 à 8, caractérisé en ce qu'on règle le décalage du point zéro de l'affichage.
  10. Procédé suivant l'une des revendication 1 à 9, caractérisé en ce qu'on règle le volume mort.
  11. Procédé suivant la revendication 10, 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.
  12. Procédé suivant l'une des revendication 1 à 11, 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
DE4209620 1992-03-25
DE4209620A DE4209620C1 (de) 1992-03-25 1992-03-25 Verfahren zur Korrektur des Volumenfehlers ïV bei einem Pipettiersystem

Publications (5)

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

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

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EP1541235A1 (fr) * 2003-11-27 2005-06-15 Gilson S.A.S. Procédé de corriger l'affichage d'une valeur d'un volume d'un échantillon liquide à prélever avec une pipette
RU2353426C2 (ru) * 2003-11-27 2009-04-27 Джилсон С.А.С. Способ и система для точного измерения жидкости в пипетке для взятия проб жидкости
FR2862888B1 (fr) * 2003-11-27 2006-07-07 Gilson Sas Procede d'affichage d'une valeur d'un volume d'un echantillon liquide a prelever avec une pipette, a precision amelioree
FR2867397B1 (fr) * 2004-03-09 2006-06-30 Gilson Sas Procede d'affichage d'une valeur d'un volume d'un echantillon liquide a prelever avec une pipette, a precision amelioree
US7976793B2 (en) * 2003-11-27 2011-07-12 Gilson S.A.S. Electronic pipette
JP2005189014A (ja) * 2003-12-25 2005-07-14 Oval Corp 基準体積管
FI20050340A0 (fi) * 2005-04-01 2005-04-01 Thermo Electron Oy Kalibroitava pipetti
DE102006032859A1 (de) 2006-07-14 2008-01-17 Eppendorf Ag Elektronische Dosiervorrichtung zum Dosieren von Flüssigkeiten
DK1887330T3 (da) * 2006-08-11 2010-03-22 Biohit Oyj Fremgangsmåde til afprøvning af pipetter
DE602006017388D1 (de) * 2006-09-19 2010-11-18 Suisse Electronique Microtech Vorrichtung und Verfahren zum Kalibrieren einer Pipette oder einer Dispenservorrichtung
DE102007010299B4 (de) 2007-03-02 2009-01-29 Eppendorf Ag Handpipettiervorrichtung
CN102527457B (zh) * 2011-12-12 2013-12-11 浙江清华长三角研究院萧山生物工程中心 一种用于多道电子移液器的校正方法
JP7228576B2 (ja) * 2017-09-19 2023-02-24 メトラー－トレド・ライニン，エルエルシー ピペットのクイックセットボリューム調整機構
SG11202103018QA (en) 2018-08-31 2021-04-29 Beckman Coulter Inc High and low volume precision pipettor
CN113252147A (zh) * 2021-05-18 2021-08-13 山东度一测控技术有限公司 一种活塞式流量标准装置
CN119091387B (zh) * 2024-11-07 2025-04-04 大龙兴创实验仪器(北京)股份公司 一种基于计算机视觉的移液器液差补偿监测方法及系统

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Also Published As

Publication number Publication date
EP0562358B8 (fr) 2005-10-26
DE4209620C1 (de) 1993-12-16
EP0562358A2 (fr) 1993-09-29
US5481900A (en) 1996-01-09
EP0562358A3 (en) 1994-05-18
EP0562358B1 (fr) 1997-07-16
DE59306907D1 (de) 1997-08-21

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