WO2005082535A1 - Microfluides actionnes par la flottabilite - Google Patents

Microfluides actionnes par la flottabilite Download PDF

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Publication number
WO2005082535A1
WO2005082535A1 PCT/IE2005/000019 IE2005000019W WO2005082535A1 WO 2005082535 A1 WO2005082535 A1 WO 2005082535A1 IE 2005000019 W IE2005000019 W IE 2005000019W WO 2005082535 A1 WO2005082535 A1 WO 2005082535A1
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WO
WIPO (PCT)
Prior art keywords
channel
fluid
control
residency
zone
Prior art date
Application number
PCT/IE2005/000019
Other languages
English (en)
Inventor
Mark Davies
Tara Dalton
Edmond Walsh
Ronan Grimes
Original Assignee
University Of Limerick
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 University Of Limerick filed Critical University Of Limerick
Publication of WO2005082535A1 publication Critical patent/WO2005082535A1/fr

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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
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • 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
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • 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
    • B01L7/54—Heating or cooling apparatus; Heat insulating devices using spatial temperature gradients

Definitions

  • the invention relates to microfiuidics such as for the pumping, control and manipulation of samples in a micro total analysis system ( ⁇ TAS) device to detect the population of rare mutated cells.
  • ⁇ TAS micro total analysis system
  • the primary method to identify rare cells in a sample is to probe the sample using known genetic markers, the markers being specific to the type of mutation being sought, and then amplify the same sample. If the mutations are present then the amplification can be detected, usually using optical techniques. It is also possible, depending on the amplification used, to detect the number of mutated cells in the original sample; a number extremely important as firstly, it can be linked to the progress of the cancer and secondly, it provides a quantitative measure with which to diagnose remission.
  • PCR Polymerase Chain Reaction
  • the invention is therefore directed towards achieving improved methods of control, manipulation and pumping in a microfluidic system for applications such as the above.
  • a microfluidic system comprising: a microfluidic device comprising a channel forming a circuit; means for delivering a carrier fluid and sample reactors into the channel; a support for supporting the microfluidic device in a non-horizontal plane; and a controller for applying different temperatures to the carrier fluid at different positions of the channel according to a control scheme to control carrier fluid and reactor flow by buoyancy and fluid properties.
  • the channel is differentially heated by heating the channel walls, which in turn heat the carrier fluid, and which in turn heat the reactors.
  • the channel comprises a plurality of branches, and flow is directed into the branches according to heating control to provide a multi-port valve.
  • channel temperature is dynamically controlled so that at any time one or more selected branches are at a common lower temperature on a gravity- assisted side or at a higher temperature on a gravity-opposed side.
  • the channel comprises a residency zone and at least one control zone forming a loop with the residency zone, and the controller varies temperature of the control zone to vary fluid residency time is the residency zone.
  • the residency zone receives gravity-assisted flow, and the controller increases control zone temperature to decrease residency time, and vice versa.
  • the residency zone receives gravity-opposed flow, and the controller decreases control zone temperature to decrease residency time and vice versa.
  • the microfluidic device comprises a plurality of residency zones, each independently controlled.
  • control zones for the residency zone.
  • control zones there are two control zones, and they are on opposed sides of the residency zone.
  • system further comprises means for applying a standing wave in the channel to maintain the reactors substantially centrally in the channel.
  • said means comprises a piezo electric device.
  • the controller varies the plane of the channel to vary flow in the channel according to varying gravitational effects.
  • the controller directs thermal fluid into the channel to heat or cool the carrier fluid.
  • the thermal fluid enters and exits from opposed sides of the channel.
  • the controller controls thermal fluid flowrate to contribute to control of the flowrate of the carrier fluid and reactors.
  • the invention also provides an analysis system comprising any microfluidic system as described above, means for delivering samples as the reactors into the carrier fluid, means in the controller for controlling the carrier fluid temperature according to required reaction temperatures in addition to flow control criteria, and a detector for monitoring the reactors.
  • the invention provides a microfluidic system comprising: a microfluidic device comprising a channel forming a circuit; means for delivering a fluid into the channel; a support for supporting the microfluidic device in a non-horizontal plane; a controller for applying different temperatures to the fluid at different positions of the channel according to a control scheme to control fluid and reactor flow by buoyancy and fluid properties, and wherein the channel comprises a plurality of parallel branches, and flow is directed into the branches according to temperature control of fluid in the branches to provide a multi-port valve.
  • channel temperature is dynamically controlled so that at any time one or more selected branches are at a lower temperature on a gravity-assisted side or at a higher temperature on a gravity-opposed side.
  • the invention provides a microfluidic system comprising: a microfluidic device comprising a channel forming a circuit; means for delivering a fluid into the channel; a support for supporting the microfluidic device in a non-horizontal plane; a controller for applying different temperatures to the fluid at different positions of the channel according to a control scheme to control carrier fluid and reactor flow by buoyancy and fluid properties, and wherein the channel comprises a residency zone and at least one parallel control zone forming a loop with the residency zone, and the controller varies temperature of the control zone to vary fluid residency time is the residency zone.
  • FIG. 1 is a diagram of a ⁇ -TAS system incorporating microfluidic devices of the invention
  • Fig. 2 is a plan view and a perspective view of a microfluidic device
  • Fig. 3 shows a simple microfluidic channel for conveying sample reactors in a carrier fluid in which pumping control is achieved without moving parts;
  • Fig. 4 shows two representations of a microfluidic circuit incorporating a multi-port valve, again without need for moving parts;
  • Fig. 5 shows two representations of a microfluidic circuit having three PCR stages, with control of residency time without need for moving parts;
  • Fig. 6 is a representation of a simple microfluidic circuit used for tests, the results of which are shown in Fig. 6;
  • Fig. 7 is a velocity and temperature plot of flows
  • Fig. 8 is a set of diagrams illustrating velocity vectors for flow in the circuit of Fig. 6;
  • Fig. 9 is a set of diagrams showing an apparatus used to experimentally demonstrate the pumping, manipulation and control of microfluidic flows without moving parts using buoyancy.
  • Fig. 10 is a diagram showing piezo control of the reactors in a conduit.
  • a ⁇ -TAS system 1 comprises an electronic controller 2 and: a mixing/sipping stage 3, a microfluidic device 4 providing a PCR reactor, and an optical detection system 5.
  • This diagram also illustrates systems for interfacing 10 with a patient to obtain a relevant sample, sample preparation 11, and bio-informatics 12.
  • the microfluidic device 4 has a circular enclosed path 20 for droplets (also "bubbles” or “reactors”) of reagent enveloped by a carrier fluid.
  • droplets also "bubbles” or “reactors”
  • reactor is used because each bubble or droplet is an individual micro reactor in its own environment, within which the chemical processes of amplification take place under thermal conditions imposed by the carrier fluid.
  • thermal zones each comprising an outer channel 21 and an inner channel 22 the inlets and outlets of which are below the plane of the path 20.
  • the channels 21 and 22 direct thermal fluid at selected temperatures.
  • the carrier fluid which circulates with the reactors and the thermal fluid which enters and exits at the three thermal zones prevent contact between the reactors and the device surfaces.
  • the thermal fluid also controls the residency time within the thermal zones by virtue of viscous drag applied to the carrier fluid, and heating or cooling of the fluids. Pumping of the carrier fluid arises because the path 20 is in a non-horizontal plane in the gravitational field and there are temperature differences between the thermal zones.
  • flow-rates and temperatures of the reagents are controlled by: (a) flowrate of the carrier fluid; and/or (b) flowrates of the thermal fluids (individually controlled in each thermal zone); and/or (c) temperatures of the thermal fluids (individually controlled in each thermal zone); and/or (d) orientation of the path 20 from horizontal to vertical with infinite adjustment within this range, and/or (e) Rotation about the central axis of the device
  • the factors (c), (d), and (e) above contribute to buoyancy of the reactors conveyed in the carrier fluid.
  • flow control is achieved without moving parts through using buoyancy in different geometrical arrangements.
  • buoyancy is used to control flow in terms of flowrate, routing through channels, and residency times in which the buoyancy is controlled by temperature variations between channel walls which heat or cool the carrier fluid. There is therefore no need for thermal fluids in the following embodiments. Also, the plane of the microfluidic circuit is not varied in these embodiments, sufficient control being achieved by virtue of the temperature control while maintaining the circuit in a vertical plane.
  • the resultant equation for a circular cross section channels is; ⁇ P P M X2 L c 2 where, g - gravity; h - vertical height, p - density, ⁇ - thermal expansion coefficient of fluid, T - temperature, ⁇ - dynamic viscosity, u - velocity, 1 - channel length, c - channel diameter.
  • Fluid properties, geometry and temperature difference can all vary to obtain better designs depending on the application. Fluid property variations with temperature are also important.
  • Figs. 3 to 6 numerical solutions were used to demonstrate the flow, and hence reactor, pumping, manipulation and control without moving parts. There is a valve for inlet and outlet of fluid and reactors, but this is closed during use. The black circles represent the reactors. Arrows represent the direction of the fluid in the channel to the left, with the velocity of the fluid being proportional to the length of the arrow. Temperature values represent the temperature in the channel to the left. These numerical solutions were done for the properties of water in a channel of 1mm diameter and height of 38mm and are shown to scale. Wall temperatures are applied as the boundary conditions.
  • Fig. 3 shows a buoyancy-driven pump in microfluidic device 30 having a single closed loop channel with a right leg 31 and a left leg 32.
  • the reactors are immersed into an immiscible bio-compatible carrier fluid.
  • Pumping, control, and manipulation of the carrier fluid results in pumping, control and manipulation of the reactors without moving parts.
  • the carrier fluid is heated through the walls, which in turn heats the reactors efficiently due to their small size, typically between 100 and 300 microns in diameter.
  • the reactors never come in contact with each other or a solid wall, thus avoiding possible contamination problems.
  • the fluid moves with a velocity so as to satisfy Equation 1, carrying with it the reactors.
  • With control of the carrier fluid there is continuous pumping of many samples in the same device without the need for moving parts, while also removing contamination risk, as is necessary in cell culturing and PCR for example.
  • the temperatures are 368K and 340K.
  • Fig. 4 shows a circuit 40 with buoyancy-driven pumping and manipulation in parallel channels.
  • the representations are velocity (left) and temperature (right).
  • the right channel is 41, and there are five parallel channels 42-46 on the left.
  • a convection loop is created as in Fig. 3.
  • the fluid is controlled and manipulated, without moving parts, to flow through any single or combination of the five parallel channels by varying wall temperatures in these channels.
  • all the fluid is controlled to flow down channel number 42, while channels 44, 45, and 46 have almost zero velocity fluid, and channel 43 has reversed fluid which further accelerates the fluid velocity in channel 42.
  • the velocity in channel 42 has a magnitude which is approximately the sum of the magnitudes of the velocities for channels 41 and 43. Less or more than five channels can be used, thereby allowing the system to meet high throughput requirements.
  • each sample can go through a different process depending on what channel they are diverted into (the channel currently at lowest temperature), and each channel could contain a PCR reaction with different temperatures and residency times. Effectively, this creates a 5 -port valve system without moving parts using buoyancy and temperature-dependent properties of the carrier fluid for use in cell sorting, sample preparation and detection for example.lt will be understood that two or more of the channels 42-46 may be at the same (lower) temperature for approximately equally divided flow through them. Of course, if the multi-port valve were on the right side then the temperature control would be in the opposite sense as the flow would become gravity-opposed..
  • the stage 52 has a residency zone 53 and a pair of control zones 54.
  • the stage 60 has a residency zone 61 and a pair of control zones 62, while the stage 70 has a residency zone 71 and a pair of control zones 72.
  • Each residency zone represents one of the thermal zones of a PCR cycle.
  • the embodiment gives the flexibility to vary the temperatures and times depending on sample to be processed.
  • the embodiment shown in Fig. 5 uses two control zones for each stage, to vary the residency time of samples in each residency zone using buoyancy and fluid properties. On the gravity-assisted side, by setting the temperature of the control zones below that of the residency zone, the flow is predominately diverted through the control zones and thus the flow velocity in the residency zone is decreased. The opposite is the case on the gravity opposed side.
  • the control zones effectively behave as a controlled micro valve or regulator.
  • a device such as the device 50 may be in any of the parallel channels of a larger device such as shown in Fig. 4.
  • Fig. 6 shows the boundary conditions used in a numerical model which was used to quantify the control of residency times using buoyancy as discussed in the preceding section.
  • the overall circuit has the numeral 100, and it comprises a main channel 101 and a stage having a residency zone 102 and control zones 103.
  • the geometry is 2D and was meshed using a structured grid of 17,004 cells.
  • the channel width is 3.5mm for all channels.
  • the centreline to centreline width of the main loop was 42mm and of the control loop is 21mm.
  • the centreline to centreline height of the main loop is 129.5mm and of the control loop is 42mm.
  • the walls of the main loop are set to constant temperatures, representative of a PCR thermal cycle. The wall temperatures were varied, which in turn varied the fluid temperatures in as per Fig. 7.
  • Fig. 8 shows velocity vectors showing control wall temperatures of: (a) 340K (b) 380K (c) 435K.
  • Fig. 8(d) shows the overall geometry of the CFD model indicating the area of interest shown in (a), (b) and (c).
  • Fig. 9 shows an apparatus used to experimentally demonstrate the flow regimes.
  • the arrows illustrate heat addition and removal, Ql and Q2 respectively from the channels for each of the flow regimes.
  • the channels are initially in an isothermal state.
  • the time taken for dye to travel through the residency zone was recorded in Table 1 below. It is seen that there is a large change in velocity representing the regulation and switching effect.
  • the central channel residency times for the regimes of Fig. 9 are: Table 1 Regime 1 , 20s, Regime 2, 5 s, Regime 3, 60s.
  • the system of the invention may comprise a piezo-electric device mounted to apply a standing wave in the channel to maintain the reactors substantially centrally in the channel.
  • piezoelectric transducers 51 are excited using an AC voltage source, this makes them oscillate in the z plane.
  • a standing wave is created within the channel. This frequency is given by; where n is the mode, c and c ' the velocity of sound in the carrier fluid and channel walls, L the distance between the two transducers. This creates a force towards the node plane as shown in Fig.
  • Integrated pumping, manipulation, control and thermal cycling of the reactors without moving parts, and without cross-contamination Very high throughputs as measured by: processing time for one sample. number of samples processed in series. number of samples processed in parallel. Low cost and high reliability, due to the absence of moving parts. Facility to independently control and vary all PCR parameters for process optimisation.
  • a multi-port valve such as shown in Fig. 4 is particularly suitable for the sample preparation 11, mixing and sipping, and PCR reactor stages.
  • the device of Fig. 5 is particularly suitable for the PCR stage.
  • the reactors may be cells which are cultured in the carrier fluid giving the advantage that they do not come in contact with a surface and are easily nurtured for screening of these cells.
  • the reactors may be slugs which are conveyed by the carrier fluid without contacting each other but in contact with the channel walls.
  • the channel walls may in this embodiment have a repellent coating to avoid contamination.
  • the multi-port valve and residency time aspects of the invention may be applied to applications other than analysis systems with reactors. For example, they may control microfluidic flow of fluid for chemical reactions, without conveying reactors.

Abstract

L'invention porte sur un système microfluidique (40) qui parvient à un contrôle et une manipulation de flux de fluides sans bouger des éléments. Ce système comprend un canal situé dans un circuit dans un plan vertical pourvu d'une jambe opposée par gravité (41) et d'un côté assisté par gravité pourvu de cinq branches (42-46). Un fluide porteur transportant de petits échantillons de réacteurs circule dans le circuit. Le débit total du circuit est contrôlé par variation du différentiel de température entre le côté gauche et droit afin de modifier la flottabilité. Le contrôle sélectif des températures dans les branches (42-46) permet d'acheminer le flux dans une ou plusieurs branches le cas échéant. Par exemple, si la température est inférieure dans une branche (42), le flux sera dévié à travers celle-ci. Cet arrangement fournit une soupape à ports multiples. Le circuit peut aussi comprendre une zone de résidence (61) et le débit (par conséquent une durée de résidence) dans cette zone est contrôlé par variation de la température d'une paire de zones de contrôle (62) sur n'importe quel côté. Le dispositif microfluidique permet d'obtenir un contrôle de flux total sans bouger des éléments au moyen du contrôle de température et des propriétés de flottabilité et des propriétés fluidiques.
PCT/IE2005/000019 2004-02-27 2005-02-25 Microfluides actionnes par la flottabilite WO2005082535A1 (fr)

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US54810804P 2004-02-27 2004-02-27
US60/548,108 2004-02-27

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7641860B2 (en) 2006-06-01 2010-01-05 Nanotek, Llc Modular and reconfigurable multi-stage microreactor cartridge apparatus
US7797988B2 (en) 2007-03-23 2010-09-21 Advion Biosystems, Inc. Liquid chromatography-mass spectrometry
WO2010136336A2 (fr) * 2009-05-27 2010-12-02 Siemens Aktiengesellschaft Réacteur microfluidique à chambre de réaction annulaire
US7854902B2 (en) 2006-08-23 2010-12-21 Nanotek, Llc Modular and reconfigurable multi-stage high temperature microreactor cartridge apparatus and system for using same
US7998418B1 (en) 2006-06-01 2011-08-16 Nanotek, Llc Evaporator and concentrator in reactor and loading system
CN103381376A (zh) * 2012-05-02 2013-11-06 李木 一种无人值守型数字式微流控系统及其控制方法

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WO1999041015A1 (fr) * 1998-02-11 1999-08-19 Institut für Physikalische Hochtechnologie e.V. Reacteur a flux miniaturise comportant differentes zones de temperatures
US20020037499A1 (en) * 2000-06-05 2002-03-28 California Institute Of Technology Integrated active flux microfluidic devices and methods

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Publication number Priority date Publication date Assignee Title
US5270183A (en) * 1991-02-08 1993-12-14 Beckman Research Institute Of The City Of Hope Device and method for the automated cycling of solutions between two or more temperatures
WO1999041015A1 (fr) * 1998-02-11 1999-08-19 Institut für Physikalische Hochtechnologie e.V. Reacteur a flux miniaturise comportant differentes zones de temperatures
US20020037499A1 (en) * 2000-06-05 2002-03-28 California Institute Of Technology Integrated active flux microfluidic devices and methods

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Title
AGRAWAL, N., UGAZ, V.: "Pumping and Thermocycling in Closed-loop convective flow systems", March 2004 (2004-03-01), ADS ABSTRACT SERVICE, XP002330395, Retrieved from the Internet <URL:http://adsabs.harvard.edu/cgi-bin/nph-bib_query?bibcode=2004APS..MAR.R1099A&db_key=PHY> *
AGRAWAL, N., UGAZ, V.M.: "Rapid DNA Amplification in Buoyancy Driven Closed Loop Microfluidic Systems", May 2005 (2005-05-01), NSTI CONFERENCE NANOTECH 2005, XP002330397, Retrieved from the Internet <URL:http://www.nsti.org/Nanotech2005/showabstract.html?absno=876> [retrieved on 20050602] *
ANALYTICAL CHEMISTRY, vol. 76, 1 July 2004 (2004-07-01), THERMOSIPHON-BASED PCR REACTOR, pages 3707 - 3715, XP002330396 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7641860B2 (en) 2006-06-01 2010-01-05 Nanotek, Llc Modular and reconfigurable multi-stage microreactor cartridge apparatus
US7790124B2 (en) 2006-06-01 2010-09-07 Nanotek, Llc Modular and reconfigurable multi-stage microreactor cartridge apparatus
US7998418B1 (en) 2006-06-01 2011-08-16 Nanotek, Llc Evaporator and concentrator in reactor and loading system
US7854902B2 (en) 2006-08-23 2010-12-21 Nanotek, Llc Modular and reconfigurable multi-stage high temperature microreactor cartridge apparatus and system for using same
US7797988B2 (en) 2007-03-23 2010-09-21 Advion Biosystems, Inc. Liquid chromatography-mass spectrometry
WO2010136336A2 (fr) * 2009-05-27 2010-12-02 Siemens Aktiengesellschaft Réacteur microfluidique à chambre de réaction annulaire
WO2010136336A3 (fr) * 2009-05-27 2011-09-22 Siemens Aktiengesellschaft Réacteur microfluidique à chambre de réaction annulaire
CN103381376A (zh) * 2012-05-02 2013-11-06 李木 一种无人值守型数字式微流控系统及其控制方法

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