EP1507592A1 - Procede et dispositif pour doser de petits volumes de liquides - Google Patents

Procede et dispositif pour doser de petits volumes de liquides

Info

Publication number
EP1507592A1
EP1507592A1 EP03718583A EP03718583A EP1507592A1 EP 1507592 A1 EP1507592 A1 EP 1507592A1 EP 03718583 A EP03718583 A EP 03718583A EP 03718583 A EP03718583 A EP 03718583A EP 1507592 A1 EP1507592 A1 EP 1507592A1
Authority
EP
European Patent Office
Prior art keywords
liquid
volume
dispensed
tip
outlet opening
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.)
Withdrawn
Application number
EP03718583A
Other languages
German (de)
English (en)
Inventor
Ernst BÜRGISSER
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.)
EPR Labautomation AG
Original Assignee
EPR Labautomation AG
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 EPR Labautomation AG filed Critical EPR Labautomation AG
Publication of EP1507592A1 publication Critical patent/EP1507592A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01F—MEASURING VOLUME, VOLUME FLOW, MASS FLOW OR LIQUID LEVEL; METERING BY VOLUME
    • G01F11/00—Apparatus requiring external operation adapted at each repeated and identical operation to measure and separate a predetermined volume of fluid or fluent solid material from a supply or container, without regard to weight, and to deliver it
    • G01F11/02—Apparatus requiring external operation adapted at each repeated and identical operation to measure and separate a predetermined volume of fluid or fluent solid material from a supply or container, without regard to weight, and to deliver it with measuring chambers which expand or contract during measurement
    • G01F11/021—Apparatus requiring external operation adapted at each repeated and identical operation to measure and separate a predetermined volume of fluid or fluent solid material from a supply or container, without regard to weight, and to deliver it with measuring chambers which expand or contract during measurement of the piston type
    • G01F11/029—Apparatus requiring external operation adapted at each repeated and identical operation to measure and separate a predetermined volume of fluid or fluent solid material from a supply or container, without regard to weight, and to deliver it with measuring chambers which expand or contract during measurement of the piston type provided with electric controlling means
    • 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
    • B01L3/022—Capillary pipettes, i.e. having very small bore
    • 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
    • B01L3/0241—Drop counters; Drop formers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01F—MEASURING VOLUME, VOLUME FLOW, MASS FLOW OR LIQUID LEVEL; METERING BY VOLUME
    • G01F11/00—Apparatus requiring external operation adapted at each repeated and identical operation to measure and separate a predetermined volume of fluid or fluent solid material from a supply or container, without regard to weight, and to deliver it
    • G01F11/02—Apparatus requiring external operation adapted at each repeated and identical operation to measure and separate a predetermined volume of fluid or fluent solid material from a supply or container, without regard to weight, and to deliver it with measuring chambers which expand or contract during measurement
    • G01F11/021—Apparatus requiring external operation adapted at each repeated and identical operation to measure and separate a predetermined volume of fluid or fluent solid material from a supply or container, without regard to weight, and to deliver it with measuring chambers which expand or contract during measurement of the piston type
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009—Characterised by arrangements for controlling the aspiration or dispense of liquids
    • G01N35/1016—Control of the volume dispensed or introduced
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/02—Drop detachment mechanisms of single droplets from nozzles or pins
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0622—Valves, specific forms thereof distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0633—Valves, specific forms thereof with moving parts
    • B01L2400/065—Valves, specific forms thereof with moving parts sliding valves
    • 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
    • B01L3/0234—Repeating pipettes, i.e. for dispensing multiple doses from a single charge

Definitions

  • the invention relates to a method for dosing small volumes of liquid according to the preamble of claim 1.
  • the invention further relates to a device for dosing small volumes of liquid according to the preamble of claim 9.
  • piezo pipettes which have a specially designed nozzle and a piezo crystal acting on the nozzle, so that the liquid drop is pressed out of the nozzle by means of a piezoelectric pulse.
  • This allows very small drops of liquid to be dispensed without contact.
  • a disadvantage of this piezo pipette is that it is very expensive, that it is not suitable for highly parallelized arrangements and that it does not allow the use of disposable tips.
  • Patent EP-0876219 discloses a further solution for overcoming the adhesive forces.
  • the pipetting device disclosed therein has a pulse generator which is arranged between the syringe, including the piston with pipette cylinder, and the tip, and which effects a pulse on the liquid column located in the pipetting device.
  • the disadvantage of this pipetting device is that it is less suitable for highly parallelized arrangements, that a large number of pulse generators are required for safe, highly parallelized delivery, that the pipetting device is therefore correspondingly expensive, and that an additional device for air-free filling is required ,
  • the object of the invention is to create a method and a device for dosing small liquid volumes of less than 1 ⁇ l, which is inexpensive, and which in particular also allows a reliable dispensing of liquid volumes of less than 100 nl.
  • the object is achieved in particular with a method for metering a liquid volume to be dispensed of less than 1 ⁇ l by means of a metering pump, in particular a pipette, a tip being filled with a liquid and containing the liquid to be dispensed at least in the area of its outlet opening, and a gas volume exceeding the outlet opening is drawn into the tip, and a volume corresponding to the sum of the liquid volume to be dispensed and the gas volume is then fed to the tip such that the liquid volume is dispensed via the outlet opening without contact.
  • a metering pump in particular a pipette
  • the basic idea of the method according to the invention is essentially to use a syringe with a motor-driven syringe plunger for dosing very small volume units in the range between, for example, 10 nl and 1 ⁇ l, by first drawing up a gas or air volume in a liquid-filled tip, and then the volume Syringe plunger is moved with great acceleration in the opposite direction, the volume displaced over the travel path of the syringe plunger being the sum of the Air volume and the volume of liquid to be dispensed correspond, so that the volume of liquid to be dispensed is pressed out of the syringe or its tip, and the adhesive forces acting on the volume of liquid to be dispensed are overcome.
  • the method according to the invention surprisingly enables very small and precisely metered liquid volumes with a lower limit of up to 10 nm to be reproducibly dispensed with a syringe with a conventionally motor-driven syringe plunger.
  • the volume discharged from the tip or moved by the syringe plunger is composed of two partial volumes, namely the gas volume dispensed and the liquid volume dispensed.
  • the syringe plunger can be accelerated to the required plunger speed, which is required in order to accelerate the liquid volume to be dispensed to the flow or exit speed required to overcome the adhesive force.
  • An electric motor in particular a commercially available, inexpensive electric motor, is particularly suitable as a motor drive, which is not a matter of course, since such electric motors have a certain inherent inertia.
  • the inherent inertia of the electric motor and its limited acceleration capacity are of secondary importance in the method according to the invention, since the motor drive is accelerated during the non-time-critical ejection of the gas volume.
  • the inertia of the liquid-filled syringe is also of minor importance during the expulsion of the gas volume.
  • the distance of the travel of the syringe plunger required to eject the gas volume and also the duration of the ejection can be varied within a wide range by the size of the gas volume and by a suitable choice of the diameter of the tip and / or the syringe plunger, so that it can also be used, for example a relatively slow electric motor is possible to achieve the piston speed required to deliver the liquid volume.
  • An electric motor such as a stepper motor or a servo motor is particularly suitable as a motor drive.
  • another drive for example a hydraulic or pneumatic motor, could also be suitable as a motor drive.
  • the volume dispensed can be metered precisely and that the motor drive accelerates the syringe plunger during the expulsion of the air volume in such a way that the syringe plunger has a sufficiently high speed at the time of dispensing the liquid volume.
  • the syringe plunger must be braked very quickly to a standstill. This braking can be done by the motor drive.
  • a brake for example a disc brake, is also advantageously provided, which brakes the syringe plunger and / or the motor drive additionally or alone.
  • the method according to the invention has the advantage that an inexpensive electric motor can be used to drive the syringe and that a highly parallelized arrangement of, for example, 96 or 384 syringes arranged in parallel can also be driven with a single electric motor.
  • the method according to the invention has the further advantage that no additional device such as a pulse generator is required in order to accelerate the liquid.
  • the sealing element is not attached to the moving syringe plunger but to the stationary pipette cylinder.
  • An annular sealing element is arranged in a stationary manner in the pipette cylinder and effects a seal between the pipette cylinder and the syringe plunger.
  • the sealing ring is thus arranged stationary, the piston acting as a displacer.
  • the syringe plunger slides over the sealing element with little resistance, so that the syringe plunger can be moved easily.
  • the bore of the Pipette cylinders are not designed precisely, so that an inexpensive pipette cylinder can be used. Since only the ring-shaped sealing element constitutes a closing part, the syringe can be operated long and inexpensively.
  • Figure 1 is a schematic representation of a pipetting device
  • FIG. 2 shows a tip filled with liquid
  • Figure 3 shows a tip which is partially filled with air
  • FIG. 4 shows a tip which delivers the liquid volume VF
  • Figure 5 is a speed-time diagram of the piston of the
  • FIG. 6 shows a longitudinal section through a piston pipette
  • FIG. 1 schematically shows a highly parallelized arrangement of, for example, 96 or 384 metering pumps 2 arranged in parallel.
  • the metering pumps are designed as syringes 2, only two of these syringes 2 being shown. However, the arrangement according to the invention could also have only a single syringe 2.
  • Each syringe 2 comprises a pipette cylinder 5, within which a syringe plunger 6 is mounted so as to be displaceable in the direction of movement Z. All syringe pistons 6 are connected to the beam 8 via piston rods 7.
  • the bar 8 is coupled to the electric motor 1 via the spindle la.
  • the syringe plungers 6 are thus moved back and forth by the electric motor 1 in the direction of movement Z.
  • the pipette cylinders 5 are stored in a carrier plate 3a.
  • the carrier plate 3a forms a valve 3 together with the valve plate 3b.
  • the valve plate 3b which is mounted so as to be displaceable in the vertical direction, comprises a channel system 10 with channels 11 and 12.
  • tips 13 with nozzle openings 13a are arranged in the valve plate 3b.
  • the tips 13 are arranged above a plate 14 with cavities 15.
  • the syringe system must be complete with a
  • Liquid 16a for example water, can be filled without any air inclusion.
  • the three-way valve 3 with reagent supply channels 11 and disposal channels 12 arranged between the syringe 2 and the tip 13 serves this purpose.
  • a brake 9 on the spindle 1 a or on the motor 1 in order to bring the rotating motor 1 to a standstill as quickly as possible.
  • the device shown in Figure 1 is operated as follows.
  • the syringe system is completely filled with a liquid 16a, also referred to as a system liquid, water being used as the system liquid, for example.
  • the syringe system is filled in such a way that there are no air pockets.
  • the sample 16 is aspirated from a Storage container is added to the pipette tip 13 so that the tip 13 is at least partially filled with the liquid to be dispensed.
  • an air volume VL is drawn into the tip 13 via the nozzle opening 13a of the tip 13.
  • a certain volume of liquid is dispensed without contact by moving the syringe plunger 6 downward with great acceleration in the opposite direction, displacing the air volume VL and the certain liquid volume.
  • the syringe plunger 6 is then stopped, and the tip 13, as shown in FIG. 2, is completely filled with the liquid 16.
  • FIG. 2 shows a schematic longitudinal section through a tip 13 filled with liquid 16, the liquid volume VF to be dispensed being shown in the region of the nozzle opening 13a.
  • This liquid volume VF forms part of the liquid 16 and is shown in FIG. 2 only for better understanding with different hatching.
  • the air or gas volume VL is drawn into the tip 13.
  • the gas volume VL is advantageously a multiple, for example ten times the volume VF to be dispensed. For example, with a liquid volume VF to be dispensed of 50nl, the gas or air volume VL could be 500nl.
  • the syringe plunger 6 is moved downward by the electric motor 1 with great acceleration, the syringe plunger 6 displacing a volume corresponding to the sum of the air volume VL and the liquid volume VF to be dispensed until it stops, so that, as shown in FIG. 4, the liquid volume VF to be dispensed the tip 13 is pressed.
  • the diameter of the nozzle opening 13a and the exit velocity of the liquid are chosen such that the acting adhesive forces are overcome, and the volume of liquid VF to be dispensed, which for example, can have a volume in the range between 10 nl and 1 ⁇ l, detaches from the tip 13.
  • the process shown in Figures 2, 3 and 4 can be repeated several times in succession.
  • the same amount of liquid volume VF can always be dispensed.
  • the dispensed liquid volume VF can also be varied by actuating the syringe plunger 6 in such a way that the dispensed liquid volume VF is changed.
  • the nozzle opening 13a can, for example, have a diameter of 0.2 mm, and in particular a diameter between 0.1 mm and 0.3 mm.
  • the exit velocity of the liquid from the nozzle opening 13a can be, for example, 5 m / sec, and in particular between 2 m / sec and 20 m / sec.
  • the air volume VL can be, for example, 950 nl, and the liquid volume VF to be dispensed, for example, 50 nl.
  • the electric stepper motor 1 is activated, for example, in such a way that the syringe plunger 6 is conveyed upward in the direction of movement Z during 950 steps of the stepper motor, and that the syringe plunger 6 is then initially moved downwards in 1000 steps in the direction of movement Z and is then braked ,
  • the acceleration path BS resulting in the tip 13 is shown in FIG. 3.
  • FIG. 5 shows, schematically and by way of example, a speed-time diagram of the speed v of the speed v moving downward in the direction of movement Z in FIG. 5
  • Syringe plunger 6 During this movement, the air volume VL and subsequently the liquid volume VF to be dispensed is first ejected via the nozzle opening 13a. The syringe plunger 6 is accelerated to a speed vmax during the ejection of the air volume VL and maintains this speed until the movement of the syringe plunger 6 is braked. During the delivery of the air volume VL, the syringe plunger 6 covers the distance SL, and the hatched line shown during delivery of the liquid volume VF Distance SF.
  • the dispensing of the liquid volume VF already begins during the movement at a constant speed vmax, for example at the time T, or for example only at the time Tl at which the braking process begins.
  • the dispensing of the liquid volume VF preferably begins before the point in time T1, and, as shown in FIG. 5 with the hatched area, lasts from the point in time T until the syringe plunger 6 comes to a standstill the exit velocity of one located in the outlet opening 13a
  • Liquid element is proportional to the speed of the syringe plunger 6, or that the acceleration of this liquid element is proportional to the acceleration of the syringe plunger 6. If, for example, the syringe plunger 6 has an inner diameter of 2 mm and the outlet opening 13a has an inner diameter of 0.2 mm, the speed of the syringe plunger 6 has an inner diameter of 2 mm and the outlet opening 13a has an inner diameter of 0.2 mm, the speed of the
  • the stop ramp SR In order to achieve the greatest possible negative acceleration during braking of the syringe plunger 6, the stop ramp SR must be correspondingly steep. If the electric motor 1 does not generate a sufficiently steep stop ramp SR, the
  • Steepness can be increased by an additional brake 9 acting.
  • the stop ramp SR has such a negative acceleration that the liquid element located in the outlet opening 13a experiences a negative acceleration of approximately 100 m / sec.
  • the air volume VL requires a certain size so that the syringe plunger 6 reaches the speed vmax. If the air volume VL is chosen too small, the syringe plunger 6 has to be reached the speed vmax can already be braked again.
  • the maximum speed vmax is preferably matched to the cross section of the outlet opening 13a and the volume of liquid VF to be dispensed, and in particular also to the viscosity of the liquid to be dispensed.
  • the size of the outlet opening 13a is preferably in relation to the amount of to be dispensed
  • Liquid volume VF selected adjusted, the geometry and the surface properties of the outlet opening 13a are also to be taken into account.
  • the negative acceleration of the stop ramp SR is preferably in a range such that a negative acceleration between 50 m / sec and 200 m / sec 2 is brought about on the liquid element located in the outlet opening (13a).
  • FIG. 6 shows a longitudinal section through a smooth-running piston pipette 2.
  • a sealing ring 19 is firmly connected to the pipette cylinder 5.
  • the syringe plunger 6 is mounted displaceably in the direction of movement Z and displaces a volume within the pipette cylinder 5.
  • FIG. 7 shows a longitudinal section through a tip 13, in which the liquids were received in such a way that an air bubble 18 results between the sample 16 and the system liquid 16a.
  • This air bubble 18 prevents mixing of sample 16 and system liquid 16a.
  • the size of the air bubble 18 is preferably selected depending on the cross section of the tip 13. For example, the air bubble 18 could be dispensed
  • Liquid volume VF of 100 nl have a volume of about 1 ⁇ l. If the liquid volume VF to be dispensed is very low, for example 10 nm preferably a very small air bubble 18 is selected, or the system is filled with liquid such that there is no air bubble 18 between the sample 16 and the system liquid 16a, so that the system has no air or gas inclusions.
  • FIG. 1 also shows a control device 17 for controlling the metering device according to the invention.
  • This control device 17 can be part of the metering device, or can also be designed as an independent device which is connected to an existing metering device.
  • the control device 17 is designed to control the drive device 1 of the metering pump 2 such that the metering pump 2 draws in a liquid volume of system liquid 16a corresponding to the gas volume VL immediately before the delivery of a liquid volume VF to be dispensed, and subsequently a liquid volume of system liquid 16a corresponding to the sum of gas volumes VL and dispenses liquid volume VF to be dispensed, so that a liquid volume VF of the sample 16 is dispensed from the tip 13.

Abstract

L'invention concerne un procédé pour doser un volume de liquide (VF) à évacuer, ce volume étant inférieur à 1 ñl, au moyen d'une pompe de dosage et notamment d'une pipette (2). Selon l'invention, une pointe (13) est entièrement remplie d'un liquide et, au moins dans la zone de son orifice de sortie (13a), contient le liquide (16) à évacuer, un volume de gaz (VL) est aspiré dans la pointe (13) par l'orifice de sortie (13a), puis un volume correspondant à la somme du volume de liquide (VF) à évacuer et du volume de gaz (VL) est amené vers la pointe (13) de manière à ce que le volume de liquide (VF) sorte par l'orifice de sortie (13a) sans contact. Le dispositif utilisé comprend une pluralité de pompes de dosage à actionnement simultané, notamment de pipettes (2) actionnées par un piston d'injection (6), ainsi qu'un moteur (1) entraînant les pistons d'injection (6) et une soupape (3) disposée entre la pompe de dosage et sa pointe, afin de remplir entièrement et sans gaz un système quelconque d'un liquide.
EP03718583A 2002-05-24 2003-05-12 Procede et dispositif pour doser de petits volumes de liquides Withdrawn EP1507592A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CH8672002 2002-05-24
CH8672002 2002-05-24
PCT/CH2003/000300 WO2003099442A1 (fr) 2002-05-24 2003-05-12 Procede et dispositif pour doser de petits volumes de liquides

Publications (1)

Publication Number Publication Date
EP1507592A1 true EP1507592A1 (fr) 2005-02-23

Family

ID=29555527

Family Applications (1)

Application Number Title Priority Date Filing Date
EP03718583A Withdrawn EP1507592A1 (fr) 2002-05-24 2003-05-12 Procede et dispositif pour doser de petits volumes de liquides

Country Status (4)

Country Link
US (1) US20050214172A1 (fr)
EP (1) EP1507592A1 (fr)
AU (1) AU2003222704A1 (fr)
WO (1) WO2003099442A1 (fr)

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