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
• • 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
  • B01L2300/00—Additional constructional details
  • B01L2300/08—Geometry, shape and general structure
  • B01L2300/0809—Geometry, shape and general structure rectangular shaped
  • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• • 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
  • B01L2300/08—Geometry, shape and general structure
  • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
  • B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
• • 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
  • B01L2300/08—Geometry, shape and general structure
  • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
  • B01L2300/0883—Serpentine channels
• • 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
  • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• • 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
  • B01L2300/1838—Means for temperature control using fluid heat transfer medium
• • 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
  • B01L2300/1883—Means for temperature control using thermal insulation
• • 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
  • B01L2400/04—Moving fluids with specific forces or mechanical means
  • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
  • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
• • 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
  • B01L3/502707—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 manufacture of the container or its components

Definitions

• the present invention relates to a chemical and / or biological analysis device, equipped with an analysis support which may be of the single-use type.
• the invention finds applications in the fields of chemistry and biology.
• the device can be used in chemical amplification or PCR polymerase reaction (Polymerase Chain Reaction) methods for the analysis of genetic material (DNA).
• Macroscopic systems of chemical or biological analysis are currently known using titration plates. These plates have cuvettes in which samples and reagents are mixed by pipetting (with a pipette). To allow chemical or biological reactions, the plates are successively heated to set temperatures by successive stoving, then are cooled. With such systems, the supply of reagents is a long and complex operation, in particular because of a successive supply of each reagent. In addition, the thermal inertia of heating and cooling of the titration plates proves to be too high and lengthens the analysis time.
• the equipment also has a high manufacturing cost.
• the object of the invention is to propose a biological and / or chemical analysis device which does not have the limitations mentioned above.
• Another aim is to reduce the heating and cooling times and to allow precise and selective temperature control of the components to be analyzed during the various reaction phases.
• Another object of the invention is to propose such a device which can be quickly adapted to different types of products to analyze and not requiring complex connection operations.
• the invention also aims to provide a device with an analysis support, very low cost, which can be for single use, which can be discarded and replaced after each use, or after a limited number of uses. For example, consider a thousand sequential analyzes with a device before discarding it.
• the invention more specifically relates to a chemical and / or biological analysis device comprising an analysis support with at least one inlet cuvette for collecting a sample, at least one outlet cuvette for delivering said sample, at least one internal channel passing through the support for connecting the inlet cuvette to the outlet cuvette, and at least one reagent tank, connected to each channel respectively between an inlet cuvette and an outlet cuvette, in which the inlet cuvette, the outlet cuvette and the reservoir open onto a first face of the analysis support.
• the device may in particular comprise a plurality of inlet bowls and a corresponding plurality of outlet bowls, each inlet bowl being respectively connected to an associated outlet bowl, by means of a channel.
• the placing of the liquids to be analyzed in the inlet cuvettes and / or of the reagents in the corresponding tanks can take place by micropipetting (with a micropipette).
• the placement of liquids to be analyzed in the cuvettes inlet and / or reagents in the corresponding tanks can take place by sealed means for supplying fluids such as a plug deposited on the tank or the bowl and connected to a syringe or to a pressurized tank.
• the supply of liquids to be analyzed and / or reagents can be automated by means of a high resolution dispensing robot (exemption).
• the sequential analyzes presupposing the replacement over time of at least one reagent by another can be automated by the sequential introduction into the corresponding reservoir of several distinct reagents.
• a neutral buffer liquid may or may not be introduced into the reservoir.
• the internal channel or channels can be provided to extend near at least one second face of the analysis support so as not to be separated from said second face only by a thin wall.
• the thin wall may have a thickness of less than 100 ⁇ .
• the wall is chosen to be thin enough to allow heat exchange with thermal sources external to the analysis support.
• the wall separating the channels from the second face can be chosen to be thinner than a wall separating the channels from one another or from bowls.
• the face of the channels opposite to the thin wall may have a thermal barrier, this being able to be produced by a layer of material with low thermal conductivity and / or a structuring of the substrate making it possible to locate on the channels a cavity filled with air or a gas with little heat transfer.
• the device can also comprise a thermal support independent of the analysis support, the thermal support having an exchange face. thermal with at least one thermal source, and said thermal support can be removably attached to the analysis support in order to bring the heat exchange face into contact with the second face of the analysis support.
• this support can be of the single-use or multi-use type, that is to say be discarded after one or more uses. By use is meant the sequential realization of a number of analyzes close to, for example, 1000.
• the heat exchange face may include one or more thermostatically controlled zones, each equipped with at least one thermal source. Thermostated zones coincide with at least one zone of the analysis support located downstream of a connection between a reagent tank and a channel.
• thermo support By associating a thermostated zone of the thermal support with a corresponding zone of the analysis support situated respectively in the vicinity, for example downstream, of each reagent reservoir, it is possible to selectively control and adapt the temperature of the liquid to be analyzed by function of each reagent used.
• the downstream term used here is understood in relation to a direction of flow of the liquids to be analyzed from the inlet basins to the outlet basins.
• the thermal sources may include one or more thermostatically controlled electric heating resistors.
• the thermal sources may also include one or more channels through which a heat transfer fluid passes. This fluid can be used to locally heat or cool the analysis support.
• this may include a first substrate having through openings which respectively form the cuvettes and reservoirs, and a second substrate, bonded to the first substrate, the second substrate having grooves, covered by the first substrate to form channels, and coinciding respectively with the through openings.
• the first substrate may have two thermally non-conductive layers, for example a few microns thick.
• the first substrate may have at least one non-through opening so as to create at least one thermal insulation cavity.
• the invention also relates to a method of implementing an analysis device as described above, according to which the analysis support is brought into contact with the thermal support during an analysis phase of determined duration, at least one sample to be analyzed and at least one reagent being introduced into the analysis support prior to the analysis phase or during the analysis phase, then, after the analysis phase, the analysis support is removed thermal support. At the end of the analysis, the analysis support can be reused.
• FIG. 1A is a simplified schematic section of an analysis support according to the invention.
• Figure 1B shows the analysis support of Figure 1A equipped with means for filling reagent tanks.
• FIG. 2 is an exploded perspective showing more precisely the structure of the analysis support.
• FIG. 3 is a simplified perspective view of an analysis support according to the figure
• FIG. 4 is a schematic longitudinal section of the analysis support placed on the thermal support.
• FIG. 5 is a plan view of part of an analysis support according to the invention, constituting a variant with respect to Figures 1 to 4.
• FIG. 6 is a simplified cross-section of an analysis support including a part in accordance with Figure 5.
• FIG. 7 is a simplified cross section of an analysis support including a part in accordance with FIG. 5 and constituting a variant with respect to FIG. 6.
• FIG. 12 is a cross section of an analysis support and a thermal support according to one invention and illustrating a particular embodiment of the thermal support.
• FIG. 1 is a section of an analysis support 100 according to the invention.
• an inlet bowl 102 formed essentially by a through opening made in a substrate 100a of the support, in the vicinity of one of its ends.
• an outlet bowl 104 is formed in the vicinity of a second end. The bowls 102,
• An internal channel 108 connects the inlet and outlet bowls.
• the channel 108 is in the form of a groove etched in a second substrate 100b bonded to the first substrate so that the latter covers the groove. It is observed that the depth of the groove is practically equal to the thickness of the second substrate
• the support 100 is of generally parallelepiped shape and the first and second faces are the main opposite and parallel faces.
• the figure also shows in section the reagent reservoirs 120a, 120b, 120c arranged between the inlet 102 and outlet 104 cuvettes.
• the reservoirs also open on the first face 106 of the analysis support 100. Fittings or passages 122a are provided to connect each of the tanks to channel 108.
• passages 122a are shown in the plane of the figure, so that the reservoirs are not distinguished from the inlet and outlet bowls in FIG. 1.
• the liquid to be analyzed can be introduced into the inlet cuvettes by means of a pipette.
• the reagent reservoirs can be filled in the same way.
• FIG. 1B shows an analysis support in accordance with FIG. 1A, the reservoirs 120a, 120b and 120c are respectively associated with fluid supply means 150a, 150b and 150c.
• These means comprise plugs, or supply caps 152a, 152b, 152c applied in leaktight manner above the reservoirs and connected to syringe pumps 154a, 154b and 154c which contain reagents.
• the caps can be glued to the surface of the test stand or tightened against the surface, using a gasket.
• the references 156a, 156b and 156c respectively designate pressure sensors formed on pipes connecting the syringe pumps to the caps 152a, 152b and 152c, so as to control the pressure and / or the flow rate of the reagents.
• a similar feeding system can also be fitted to the inlet bowls.
• the inlet basins and the tanks are subjected to atmospheric pressure, or to a pressure fixed by the supply system, while a vacuum line 124 is applied to the outlet basins .
• a first spontaneous filling of the analysis support can be carried out with a polar solvent (such as alcohol) followed by a nominal solvent in order to avoid the formation of bubbles.
• a polar solvent such as alcohol
• a nominal solvent such as water
• Figure 2 shows more precisely and separately the two substrates 100a and 100b which form the analysis support.
• the analysis support comprises a plurality of inlet cuvettes 102 and a plurality of outlet cuvettes 104.
• the bowls have the form of through openings made in the first substrate 100a. These openings have a flared V shape, forming a funnel.
• each inlet bowl 102 is individually connected to an outlet bowl 104 by a channel 108.
• the analysis support includes three reagent reservoirs 120a, 120b, 120c.
• each reservoir is common to several channels 108 to which it is connected by means of fittings 122a, 122b.
• the reference 122a more precisely designates holes in the first substrate 110a connecting a reservoir to corresponding branches 122b, formed in the second substrate 110b and connected respectively to the channels. (Of course, reservoirs can also be individualized for the different channels).
• the quantities of liquids (liquids to be analyzed, and reagents) which mix at the intersection of the branches 122b and the channels 108 depend on the respective size of these branches and the channels 108.
• FIG. 3 shows an analysis support 100, in accordance with that of FIG. 2, the substrates 100a and 100b of which are permanently bonded. The analysis support is shown above a corresponding thermal support 200.
• the thermal support 200 has a heat exchange face 212 facing the second face 112 of the analysis support 100, in the vicinity of which the channels are located.
• the heat exchange face 212 of the thermal support 200 and the second face 112 of the analysis support are intended to be brought into contact.
• the heat exchange face 212 has three thermostatically controlled zones 220a, 220b, 220c each equipped with one or more thermal sources (not shown).
• the three thermostatically controlled zones 220a, 220b, 220c are arranged so as to coincide with portions of channels of the analysis support located in the vicinity respectively of the reservoirs 120a, 120b, 120c, or more precisely of the branches supplying the reagents.
• the fluid in the channel 122b can pass through each thermal zone only once or several times thanks to suitable channel patterns as shown in FIG. 5 described later.
• FIG. 4 is a diagrammatic section of the analysis support transferred to the thermal support making it possible to represent in more detail the thermostatically controlled zones.
• the thermostat zones can have several thermal sources. This is the case of the thermostatically controlled area 220a.
• This comprises a first thermal source 230 formed of electrical resistors, such as for example microresistors in platinum. It also includes two sources 232 and 234 in the form of channels crossed by heat transfer fluids.
• the electrical resistances of the first source 230 can be brought to a temperature of 94 ° C, the heat transfer fluid of the second heat source 232 to a temperature of 55 ° C and the heat transfer fluid from the third heat source 234 at a temperature of 72 ° C.
• FIG. 5 is a top view of part of a first substrate 100a of an analysis support and shows an alternative embodiment of a channel 108.
• the channel 108 is folded in a repeated geometric pattern.
• FIG. 6 and 7 show two alternative embodiments of the device making it possible to improve the uniformity of the temperature in the channels by isolating their upper face, that is to say the face opposite to said second face 112 of the support. analysis.
• a second solution represented in FIG. 7, consists in placing between the upper and lower parts 100a, 100b of the analysis support a layer 100c of weakly conductive material of the heat. It is also possible to use an upper substrate equipped with a layer 100c of a thermal insulating material.
• FIG. 8 shows an example of a method of producing an analysis support as described above.
• a first substrate plate 100a for example made of silicon
• openings are made, as shown in FIG. 8.
• These openings constitute the bowls or tanks 102, 104, 120a, 120b, 120c.
• the chemically etched openings are made with inclined sides by anisotropic chemical etching for example (KOH) so as to give them a flared shape.
• the location of the openings is defined by an etching mask (not shown) coincident with the pattern of the grooves.
• the drilling of the layer 100c of thermal insulating material for example Si0 2 in the case of the variant proposed in FIG. 7, can be carried out, for example, by etching CHF 3 by dry process, the dimension of the perforation being defined by an etching mask or by using the walls of the chemically created hole as a mask.
• FIG. 9 shows the etching of grooves, forming the channels 108, in a second substrate 100b, for example of silicon.
• the etching is carried out through an etching mask (not shown) having a pattern corresponding to the desired channels. This is, for example, chemical etching (KOH).
• KOH chemical etching
• the depth of the grooves is for example of the order of 100 ⁇ m for a substrate 100b with a thickness of 250 to 450 ⁇ m.
• a third step shown in FIG. 10 comprises the sealing of the first and second substrates 100a and 100b, so as to put the cuvettes or reservoirs 102, 104, 120a, 120b, 120c into communication with the corresponding channels (grooves) 108. Sealing takes place, for example, by direct (molecular) bonding of the two substrates.
• the grooves 108 of the second substrate 100b are covered by the first substrate 100a to form the channels.
• a last step, represented in FIG. 11, comprises the thinning of the second substrate 100b so as to preserve between the channel 108 and the external surface 112 only a thin wall 110.
• This wall 110 has a thickness of
• Thinning is achieved by etching and / or mechanochemical polishing.
• a plurality of analysis supports in accordance with the invention can be produced simultaneously and collectively according to the above method in two silicon wafers (corresponding to the first and second substrates).
• the process is completed by cutting the slices with a saw to individualize the analysis supports.
• FIG. 12 shows a particular embodiment of the thermal support 200 of an analysis device according to the invention.
• the thermal support 200 essentially comprises a base 202 on which one or more thermostatic bars are arranged.
• the thermal support comprises three thermostatic bars 320a, 320b, 320c which respectively form three thermostatically controlled zones. All or part of the bars can be embedded in a thermal insulating material.
• two bars 320b and 320c are surrounded by a solid thermal insulating material, while the first bar 320a is left in free contact with the ambient air on its lateral faces.
• Each bar is equipped with heating and / or cooling means.
• the first bar 320a is equipped with a channel 322a which runs through it and which makes it possible to thermostate it by circulation of a heat transfer fluid.
• the other bars 320b and 320c are also equipped with such channels 322b and 322c.
• the channels are respectively connected to thermostatic baths with pumping systems (not shown) for circulating the heat transfer fluid.
• connection between the baths and the bars can take place by means of hydraulic connections, not shown.
• the channels can be of the circular type, as shown in the figures, but can also be provided with fin systems to optimize the heat exchanges. Additional heating elements can be integrated into the bars.
• the third bar 320c is equipped with an electrical resistance 330. The electrical resistance is used here as “hot source” while the heat transfer fluid is used as “cold source”.
• the second bar 320b comprises an element 340 for measuring the temperature, such as a resistor, used to control, for example, the temperature of the associated thermostated bath.
• the reference 100 generally designates a removable analysis support disposed on the thermal support so as to be in contact with the thermostatic bars. The detailed description of such a support is not repeated here. On this subject, reference can be made to the explanations given with reference to the preceding figures.
• the analysis support 100 can be simply placed on the thermal support 200. It can also be pressed against the thermal support by means of a flange or a suction system not shown.
• the means for temperature control that is to say in particular the baths thermostates and thermostats bars, are integral with the thermal support or in fluid connection with the thermal support, and as the analysis support is removable, it is possible to produce the latter in a simple and inexpensive manner.

Landscapes

• Chemical & Material Sciences (AREA)
• Health & Medical Sciences (AREA)
• Clinical Laboratory Science (AREA)
• Chemical Kinetics & Catalysis (AREA)
• General Health & Medical Sciences (AREA)
• Dispersion Chemistry (AREA)
• Molecular Biology (AREA)
• Biochemistry (AREA)
• Life Sciences & Earth Sciences (AREA)
• Analytical Chemistry (AREA)
• Hematology (AREA)
• Automatic Analysis And Handling Materials Therefor (AREA)
• Investigating Or Analysing Biological Materials (AREA)
• Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
• Apparatus Associated With Microorganisms And Enzymes (AREA)
• Sampling And Sample Adjustment (AREA)

Applications Claiming Priority (3)

Publications (2)

Family

ID=9531670

Family Applications (1)

Country Status (6)

Families Citing this family (20)

Family Cites Families (10)

• 1999
  • 1999-10-14 DE DE69913721T patent/DE69913721T2/de not_active Expired - Lifetime
  • 1999-10-14 WO PCT/FR1999/002499 patent/WO2000023190A1/fr active IP Right Grant
  • 1999-10-14 AT AT99947579T patent/ATE256501T1/de not_active IP Right Cessation
  • 1999-10-14 US US09/806,515 patent/US6680193B1/en not_active Expired - Fee Related
  • 1999-10-14 EP EP99947579A patent/EP1121199B1/de not_active Expired - Lifetime
  • 1999-10-14 JP JP2000576958A patent/JP4398096B2/ja not_active Expired - Fee Related

Non-Patent Citations (1)

Also Published As

Similar Documents

Legal Events