WO2001088527A2 - Procede et dispositif pour l'injection d'un echantillon dans un capillaire d'electrophorese - Google Patents
Procede et dispositif pour l'injection d'un echantillon dans un capillaire d'electrophorese Download PDFInfo
- Publication number
- WO2001088527A2 WO2001088527A2 PCT/FR2001/001429 FR0101429W WO0188527A2 WO 2001088527 A2 WO2001088527 A2 WO 2001088527A2 FR 0101429 W FR0101429 W FR 0101429W WO 0188527 A2 WO0188527 A2 WO 0188527A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- capillary
- sample
- channel
- plate
- well
- 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.)
- Ceased
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44743—Introducing samples
Definitions
- the present invention relates to the injection of a sample into an electrophoresis capillary.
- the electrophoresis by capillary (s) has the advantage of allowing a higher flow rate and a higher automation than the previous electrophoresis techniques.
- the injection techniques mainly used to date indeed require preparations of large quantities of sample volume (10 to 20 ⁇ l of DNA sample for example), of which they use only a small part.
- An object of the invention is to overcome this drawback.
- Another object of the invention is to propose a technique which makes it possible to carry out, in a capillary or a set of capillaries, a reproducible and effective electrokinetic injection.
- Hydrodynamic injection consists in using a pressure difference to bring a certain quantity of sample into a capillary. This technique is often unsatisfactory, insofar as it requires large areas for introducing samples and therefore does not allow good resolution, where it can only be used for low viscosity separation matrices and where it is difficult to implement in parallel on a plurality of capillaries.
- electrokinetic injection which consists in applying, for a short time, a potential difference between the ends of the capillaries to cause the molecules to migrate inside them.
- the quantity of sample introduced by this method can be increased, either by accentuating the difference in potential, either by extending the loading time, without completely controlling the quantity injected.
- the resolution is better than for hydrodynamic injection.
- the invention proposes a method for injecting a sample into an electrophoresis capillary, according to which the capillary is immersed in the sample and an electric field is applied between the ends of the capillary to cause the sample to migrate.
- the capillary characterized in that the sample is introduced beforehand into a part having a channel whose dimensions, perpendicular to the direction in which said channel extends, are less than about four times the outside diameter of the capillary, and in that to immerse the capillary in the sample, said capillary is introduced into said channel.
- It also provides a device for injecting a sample into an electrophoresis capillary comprising a hollow part in which the sample is received and in which the capillary is intended to be immersed, as well as means for applying between the ends.
- an electric field from the capillary for migrating the sample into the capillary characterized in that said hollow part is a part having a channel whose dimensions, perpendicular to the direction in which said channel extends, are less than about four times the outside diameter of the capillary and which is capable of receiving a capillary and in that it further comprises means for introducing the sample into said channel.
- the volume of sample which it is necessary to prepare is considerably restricted: in fact a volume of sample is sufficient to fill the channel over a few millimeters; the entire prepared sample is then used.
- such a channel makes it possible to perfectly control the position of the capillary and in particular to position it with great precision relative to the electrode and thus to create in an reproducible manner an electric field intended to migrate the sample.
- it allows great homogeneity on the field at the inlet of the capillary, and therefore the same migration speed on all of the molecules in the channel.
- the injection device proposed by the invention is advantageously supplemented by the following different characteristics taken alone or in all their possible combinations, which also contribute to achieving the aims of the invention:
- the well-forming element and the part which presents the channel are able to fit into each other, the sample rising by capillarity or pressure difference in the channel when the part which presents the channel is introduced into the part forming a well;
- the well element has a flared shape facilitating the guiding of the part which has a channel; - the channel is extended by a flared opening which facilitates the introduction of the capillary;
- This opening is substantially conical in shape
- the device comprises a plate which has a plurality of elements forming wells intended to cooperate with a plurality of complementary parts each having a channel;
- the device comprises at least one receptacle capable of receiving such a plate and means capable of automatically transporting said receptacle from one work station to another;
- the device further comprises a cover capable of closing the receptacle when such a plate is in place therein;
- the device comprises means forming a seal ensuring the seal between the receptacle and the cover;
- the complementary parts, each having a channel, are carried by a plate and the device comprises means for automatically positioning said plate and the plate which has a plurality of well-forming elements relative to each other.
- the invention also relates to an electrophoresis device comprising such an injection device.
- FIG. 1 is a schematic sectional representation of a device according to a possible embodiment of the invention.
- FIG. 2 is a schematic exploded perspective representation of a handling palette of a device according to an embodiment of the invention
- FIG. 3 is a schematic perspective representation of a storage enclosure of a device according to a possible embodiment of the invention and automatic handling means.
- FIG. 1 shows a system allowing the injection of samples into a capillary 1.
- This system comprises a well 2, the bottom of which is of substantially conical shape and receives a droplet of a sample to be injected into the capillary. It further comprises a part 3 intended to be introduced into this well 2 and which is of a shape allowing it to fit into it.
- This part 3 has a channel 4, in this case tubular, which extends axially over its entire length and in which the capillary 1 is received.
- this channel 4 is defined by a tube 5 made of glass or of a similar material which is of very small internal diameter and in which the capillary 1 is received.
- the part 3 carries, at its end by which the sample is intended to be introduced into the tube 5, then into the capillary 1, an electrode
- the supply of this electrode 6 is carried out by means of a metal wire 7 which extends in the height of the part 3 between said electrode
- the part 3 is fixed in an orifice 9 which passes through the plate 8, for example by screwing.
- This part 3 ends at said orifice 9 by a metal portion 3a (electrical conductor), which is crossed by an axial opening 3b of conical shape which makes it possible to guide the capillary to place it in the tube 5 (the walls of this opening are for example in Teflon).
- a metal portion 3a electrical conductor
- an axial opening 3b of conical shape which makes it possible to guide the capillary to place it in the tube 5 (the walls of this opening are for example in Teflon).
- the capillary 1 is in turn carried by a plate 10 forming a support.
- a drop G of sample is placed at the bottom of the well 2 (that is to say a few ⁇ l, for example from 1 to 5), then the part 3 is introduced into the well 2.
- the drop G then rises by capillary action or pressure in the tube 5.
- a drop of a few micro-liters of sample is enough to thus create in the tube 5 a column of several mm of sample.
- the capillary 1 is then introduced into the part 3 by bringing the plate 10 forming a support onto the plate 8.
- the capillary 1 then bathes in the sample, being located at a well-determined and reproducible distance from the electrode 6.
- the capillary 1 is for example 35 cm long, with an internal diameter of 75 ⁇ m and an external diameter of 200 ⁇ m.
- the part 3 and the plate 10 are for example made of Plexiglas.
- the well 2, the part 3 and the tube 5 are for example of a height of 20 to 25 mm.
- Plate 8 is for example made of stainless steel.
- the tube 5 has for example an internal diameter of 650 ⁇ m and an external diameter of 1600 ⁇ .
- the internal diameter of the conical hole presented by the Teflon portion 3 varies from 1000 ⁇ m to 400 ⁇ m.
- the electrode 6 is for example made of Platinum and is in the form of a disc with an internal diameter of 400 ⁇ m and an external diameter of 700 ⁇ m, attached to the end edge of the part 3.
- the electrode 6 could be carried by the bottom of the well 2. It could consist of a wire, for example made of Platinum, which descends from the plate 8 to the bottom of the well.
- the tube 5 can be made of Platinum and play the role of electrode.
- the part 2 could be a metal part constituting the electrode 6.
- the electrode In all cases, it is advantageous to give the electrode a geometry allowing an electric field as homogeneous as possible in the channel 4.
- the well 2 is advantageously carried by a plate 11, with a plurality of other injection wells.
- the plate 11 can carry 96 wells distributed in an 8 * 12 matrix.
- this plate 11 and the wells 2 which it carries are advantageously received in a receptacle 12.
- This receptacle 12 makes it possible to handle said plates 11 and in particular to transport them from one work station to another. It includes keying means (pins 17) imposing a single possible position for a plate 11 in said receptacle 12.
- a cover 13 is attached to said receptacle 12 to prevent evaporation and exposure to light of the samples.
- the pallets 14 thus formed by a receptacle 12 closed by a cover 13 and containing a plate 11 allow an automaton to handle the plates 11 in the following manner.
- the wells 2 of the plate 11 having been initially filled, the automaton or an operator positions a cover 13 on the receptacle 12 in which said plate 11 is arranged so as to close the receptacle 12.
- the pallet 14 thus formed is transported to an enclosure 15 (FIG. 3), in which it is possible to store a large number of these pallets 14.
- the cabinet 15 comprises for example a refrigerated compartment permanently maintained at 4 ° C or below, which makes it possible to minimize the degradation of the stored samples.
- the device may also also include a heating compartment making it possible to denature the samples, for example by heating them to 95 ° C., before rapidly cooling them to 4 ° C. or below, the automaton moving the pallets 14 by one compartment to another.
- a heating compartment making it possible to denature the samples, for example by heating them to 95 ° C., before rapidly cooling them to 4 ° C. or below, the automaton moving the pallets 14 by one compartment to another.
- the heating compartment can be integrated into the cabinet 15, or be constituted by an independent work station.
- the automaton will seek in the refrigerated compartment of the cabinet 15 the pallet which corresponds to this plate 11, by means of a handling arm 16.
- the new pallet - constituted by a receptacle 12, a plate 11 and a cover 8 carrying a plurality of parts 3 electrode holders - is transported by the automaton in the sequencer. It is inserted therein in a movement parallel to the axis of the capillaries, which are introduced into the tubes 5 of the parts 3 while being guided by the conical openings of the portions 3a.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Sampling And Sample Adjustment (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
Description
Claims
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2001260403A AU2001260403A1 (en) | 2000-05-15 | 2001-05-11 | Method and device for injecting a sample in an electrophoresis capillary |
| JP2001584872A JP2003533695A (ja) | 2000-05-15 | 2001-05-11 | 電気泳動キャピラリーへの試料注入方法及び装置 |
| EP01934092A EP1285263A2 (fr) | 2000-05-15 | 2001-05-11 | Procede et dispositif pour l'injection d'un echantillon dans un capillaire d'electrophorese |
| US10/276,079 US20040011649A1 (en) | 2000-05-15 | 2001-05-11 | Method and device for injecting a sample in an electrophoresis capillary |
| CA002409034A CA2409034A1 (fr) | 2000-05-15 | 2001-05-11 | Procede et dispositif pour l'injection d'un echantillon dans un capillaire d'electrophorese |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR00/06138 | 2000-05-15 | ||
| FR0006138A FR2808883B1 (fr) | 2000-05-15 | 2000-05-15 | Procede et dispositif pour l'injection d'un echantillon dans un capillaire d'electrophorese |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO2001088527A2 true WO2001088527A2 (fr) | 2001-11-22 |
| WO2001088527A3 WO2001088527A3 (fr) | 2002-04-25 |
Family
ID=8850207
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/FR2001/001429 Ceased WO2001088527A2 (fr) | 2000-05-15 | 2001-05-11 | Procede et dispositif pour l'injection d'un echantillon dans un capillaire d'electrophorese |
Country Status (7)
| Country | Link |
|---|---|
| US (1) | US20040011649A1 (fr) |
| EP (1) | EP1285263A2 (fr) |
| JP (1) | JP2003533695A (fr) |
| AU (1) | AU2001260403A1 (fr) |
| CA (1) | CA2409034A1 (fr) |
| FR (1) | FR2808883B1 (fr) |
| WO (1) | WO2001088527A2 (fr) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2301284A1 (es) * | 2005-07-15 | 2008-06-16 | Consejo Superior Inveti. Cientificas | Dispositivo y procedimiento de mejorado para el analisis cuantitativo en electroforesis capilar. |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0581983B1 (fr) * | 1992-08-04 | 1996-10-23 | Hewlett-Packard GmbH | Appareil pour l'exécution d'électrophorèse capillaire |
| US6132582A (en) * | 1998-09-14 | 2000-10-17 | The Perkin-Elmer Corporation | Sample handling system for a multi-channel capillary electrophoresis device |
-
2000
- 2000-05-15 FR FR0006138A patent/FR2808883B1/fr not_active Expired - Fee Related
-
2001
- 2001-05-11 WO PCT/FR2001/001429 patent/WO2001088527A2/fr not_active Ceased
- 2001-05-11 US US10/276,079 patent/US20040011649A1/en not_active Abandoned
- 2001-05-11 JP JP2001584872A patent/JP2003533695A/ja active Pending
- 2001-05-11 CA CA002409034A patent/CA2409034A1/fr not_active Abandoned
- 2001-05-11 AU AU2001260403A patent/AU2001260403A1/en not_active Abandoned
- 2001-05-11 EP EP01934092A patent/EP1285263A2/fr not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2301284A1 (es) * | 2005-07-15 | 2008-06-16 | Consejo Superior Inveti. Cientificas | Dispositivo y procedimiento de mejorado para el analisis cuantitativo en electroforesis capilar. |
| ES2301284B1 (es) * | 2005-07-15 | 2009-05-20 | Consejo Superior Inveti. Cientificas | Dispositivo y procedimiento de mejora para el analisis cuantitativo en electroforesis capilar. |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2001088527A3 (fr) | 2002-04-25 |
| FR2808883B1 (fr) | 2002-08-09 |
| US20040011649A1 (en) | 2004-01-22 |
| FR2808883A1 (fr) | 2001-11-16 |
| JP2003533695A (ja) | 2003-11-11 |
| EP1285263A2 (fr) | 2003-02-26 |
| CA2409034A1 (fr) | 2001-11-22 |
| AU2001260403A1 (en) | 2001-11-26 |
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