Miniaturized planar columns for use in a liquid phase separation apparatus

• US 6,264,892 B1
• Filed: 01/11/2000
• Issued: 07/24/2001
• Est. Priority Date: 10/19/1994
• Status: Expired due to Term

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Swedberg, Sally A., Mittelstadt, Laurie S., Bek, Fritz, Kaltenbach, Patrick, Witt, Klaus E.
• Primary Examiner(s): Tung, T.
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US09/480,922
• Time in Patent Office: 560 Days
• Field of Search: 204/450, 204/451, 204/452, 204/453, 204/455, 204/600, 204/601, 204/602, 204/603, 204/604, 204/605, 422/100, 422/101, 422/99, 422/103, 422/68.1, 422/70, 422/82.01, 422/82.02, 422/82.05, 422/82.09, 435/288.6, 435/288.7, 435/287.2
• US Class Current: 422/68.1
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/5055 Hinged, e.g. opposable surfaces
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: B29C 65/02 by heating, with or without pressure
• CPC Class Codes: B29C 66/54 Joining several hollow-preforms, e.g. half-shells, to form hollow articles, e.g. for making balls, containers; Joining several hollow-preforms, e.g. half-cylinders, to form tubular articles
• CPC Class Codes: B29C 66/549 said hollow-preforms being interconnected during their moulding process, e.g. by a hinge
• CPC Class Codes: B29L 2031/756 Microarticles, nanoarticles
• CPC Class Codes: G01N 2030/0035 electrical field
• CPC Class Codes: G01N 2030/285 electrically driven carrier
• CPC Class Codes: G01N 2030/3007 : same temperature for whole column
• CPC Class Codes: G01N 2030/3015 : temperature gradients along column
• CPC Class Codes: G01N 2030/303 : using peltier elements
• CPC Class Codes: G01N 27/44704 : Details; Accessories
• CPC Class Codes: G01N 27/44708 : Cooling
• CPC Class Codes: G01N 27/44721 : by optical means
• CPC Class Codes: G01N 27/44773 : Multi-stage electrophoresis, e.g. two-dimensional electrophoresis
• CPC Class Codes: G01N 27/44791 : Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 30/02 : Column chromatography
• CPC Class Codes: G01N 30/60 : Construction of the column
• CPC Class Codes: G01N 30/6047 : with supporting means; Holders
• CPC Class Codes: G01N 30/6052 : body
• CPC Class Codes: G01N 30/606 : with fluid access or exit ports
• CPC Class Codes: G01N 30/6095 : Micromachined or nanomachined, e.g. micro- or nanosize
• CPC Class Codes: Y10T 156/1064 : Partial cutting [e.g., grooving or incising]

Abstract

Miniaturized planar column devices for use in a liquid phase separation apparatus are described. The devices include microstructures that have been fabricated by laser ablation in a variety of novel support substrates. Devices formed according to the methods of the invention include associated laser-ablated features required for function, such as on-device reservoirs or makeup flow compartments, analyte detection means and sample injection means. The miniaturized columns can be used in an apparatus intended for analysis of either small and/or macromolecular solutes in the liquid phase which employs chromatographic, electrophoretic or electrochromatographic separation means. The apparatus can include a variety of optional injection means, manifolds, keeper means, post column collection means, and combinations thereof.

Claims

1. A miniaturized column device comprising:

(a) substrate having a first substantially planar surface, molded from a mold insert formed using a technique selected from the group consisting of laser-ablation and LIGA techniques and having a negative structure of the first substantially planar surface comprising;

(i) a negative structure of a separation channel comprising a microchannel;

(ii) a negative structure of a reservoir means comprising a cavity;

(iii) a negative structure of a fluid conducting means comprising a duct enabling fluid communication between the separation channel and the reservoir means; and

(iv) a negative structure of a bypass microchannel that is distinct from and unconnected to the separation channel and the reservoir means;

(b) a first cover plate arranged over the first planar surface of the substrate that respectively defines a separation compartment in combination with the separation channel in the substrate, a reservoir compartment in combination with the reservoir means in the substrate, a fluid conducting compartment in combination with the fluid conducting means in the substrate, and a volumetric sample compartment in combination with the bypass microchannel in the substrate; and

(c) an inlet port and an outlet port communicating with the separation compartment, said ports enabling passage of fluid from an external source through the separation compartment.

2. The miniaturized column of claim 1, further comprising first detection means in communication with the separation compartment thereby enabling detection of a sample passing through the separation compartment.

3. The miniaturized column of claim 1, wherein the cover plate comprises an orifice enabling the passage of fluid from an external source into the reservoir compartment.

4. The miniaturized column of claim 1, further comprising motive means enabling the displacement of fluid from the reservoir compartment through the fluid conducting compartment and into the separation compartment.

5. A miniaturized column device comprising:

(a) a substrate having a first substantially planar surface, molded from a mold insert formed using a technique selected from the group consisting of laser-ablation and LIGA techniques and having a negative structure of the first substantially planar surface comprising;

(i) a negative structure of a separation channel comprising a microchannel, wherein said separation channel has an upstream and a downstream terminus; and

(ii) a negative structure of a makeup flow channel extending substantially along the length of the separation channel, comprising a duct having an upstream terminus and a downstream terminus, wherein the downstream terminus of the makeup flow channel is in communication with the downstream terminus of the separation channel;

(b) a first cover plate arranged over the first planar surface of the substrate that respectively defines a separation compartment in combination with the separation channel of the substrate, and a makeup flow compartment in combination with the makeup flow channel of the substrate; and

(c) an inlet port in communication with the upstream terminus of the separation channel of the substrate, an outlet port in communication with the downstream terminus of the separation compartment, said ports enabling passage of fluid from an external source through the separation compartment, and a makeup compartment inlet in communication with the upstream terminus of the makeup flow channel of the substrate.

6. The miniaturized column of claim 5, further comprising first detection means in communication with the separation compartment thereby enabling detection of a sample passing through the separation compartment.

7. A liquid phase separation apparatus comprising:

(a) a miniaturized column, comprising

(i) a substrate having a first substantially planar surface and molded from a mold insert formed using a technique selected from the group consisting of laser-ablation and LIGA techniques and having a negative structure of the first substantially planar surface comprising a negative structure of a separation channel formed by a microchannel on the substrate,

(ii) a first cover plate arranged over the first planar surface-of the substrate, whereby said cover plate defines a separation compartment in combination with the separation channel, and

(iii) at least one inlet port and at least one outlet port communicating with the separation compartment, said ports enabling the passage of fluid from an external source through the separation compartment;

(b) associated containment means for housing a sample to be separated;

(c) injection means for introducing the sample into the column from the associated containment means;

(d) means for applying a motive force along the column such that the sample is moved therethrough; and

(c) detecting means for detecting separated analytes of interest contained in the sample.

8. The liquid phase separation apparatus of claim 7, wherein the injection means comprises a manifold having a volumetric sample compartment formed therein, said manifold arranged in divertable fluid communication with the inlet port and the associated containment means.

9. The liquid phase separation apparatus of claim 7, further comprising an associate keeper means having first and second opposable elements and adapted for receiving the miniaturized column device.

10. The liquid phase separation apparatus of claim 9, wherein the keeper means is further adapted to provide a means for facilitating the coupling of a fastener with the inlet port, outlet port or detection means.

11. The liquid phase separation apparatus of claim 10, wherein the keeper means comprises at least one annular boss arranged to encompass an inlet port, outlet port or detection means.

12. The liquid phase separation apparatus of claim 11, wherein the annular boss comprises a cylinder having an interior and an exterior surface.

13. The liquid phase separation apparatus of claim 12, wherein the annular boss further comprises lock means for detachably coupling a fastener with the inlet port, outlet port or detection means.

14. The liquid phase separation apparatus of claim 13, wherein the lock means comprises thread means arranged on the exterior surface of the annular boss for threadably engaging a fastener.

15. The liquid phase separation apparatus of claim 12, wherein the annular boss is configured to facilitate interfacing of an associated lightguide means with the detection means.

16. The liquid phase separation apparatus of claim 10, wherein the keeper means comprises at least one aperture arranged to be in coaxial relation with an inlet port, outlet port or detection means when a miniaturized column device is disposed within the keeper means.

17. The liquid phase separation apparatus of claim 16, wherein the aperture has a greater diameter than that of the coaxially arranged inlet port, outlet port or detection means.

18. The liquid phase separation apparatus of claim 7, wherein the miniaturized column inlet port and outlet port enable the formation of a sample eluent within the separation compartment, said separation apparatus further comprising:

(a) sample eluate delivery means comprising a mixing chamber, wherein the mixing chamber includes a first side in fluid communication with the outlet port, a top end in fluid communication with a mixing chamber inlet means, and a bottom end in fluid communication with an outlet nozzle;

(b) a fluid source in divertable fluid communication with the mixing chamber inlet means; and

(c) a post-column collection device comprising a sample eluate receiving means positioned relative to the outlet nozzle, thereby enabling the collection of the sample eluate.

19. The liquid phase separation apparatus of claim 18, further comprising a means for heating the fluid source.

20. The liquid phase separation apparatus of claim 18, further comprising protection means interposed between the miniaturized column device and the sample eluate receiving means, said protection means comprising an opening arranged in axial alignment with the outlet nozzle.

21. A liquid phase separation apparatus comprising:

(a) a miniaturized column, comprising

(i) a substrate having a first substantially planar surface, wherein said substrate is molded from a mold insert formed using a technique selected from the group consisting of laser-ablation and LIGA techniques and having a negative structure of the first substantially planar surface comprising a negative structure of a separation channel comprising a microchannel in the first planar surface,

(ii) a first cover plate arranged over the first planar surface of the substrate, whereby said cover plate defines a separation compartment in combination with the separation channel, and

(iii) at least one inlet port and at least one outlet port communicating with the separation compartment, said ports enabling the passage of fluid from an external source through the separation compartment; and

(iv) a bypass microchannel in the first planar surface, wherein the first cover plate in combination with the bypass microchannel defines a volumetric sample compartment having first and second ends; and

(b) injection means for introducing the sample into the column, comprising a manifold in fluid communication with the containment means and in divertable fluid communication with the volumetric sample compartment and the inlet port.

22. The liquid phase separation apparatus of claim 21, wherein the manifold is further in fluid communication with a second associated containment means housing a liquid medium.

23. The liquid phase separation apparatus of claim 21, wherein the manifold is movable between a plurality of positions relative to the miniaturized column device.

24. The liquid phase separation apparatus of claim 23, wherein the manifold is movable between a first and a second position, whereby the manifold in said first position enables dynamic fluid communication between the volumetric sample compartment and the associated containment means to allow a sample plug to be formed by the passage of sample through the volumetric sample compartment, said plug corresponding to the dimensions of the sample compartment, and further whereby the manifold in said second position allows movement of the sample plug from the sample compartment to the separation compartment.

25. The liquid phase separation apparatus of claim 23, wherein the manifold comprises a rotor detachably coupled to the miniaturized column device.

26. The liquid phase separation apparatus of claim 23, wherein the manifold comprises an elongated housing detachably coupled to the miniaturized column device.

27. The liquid phase separation apparatus of claim 21, wherein the miniaturized column inlet port and outlet port enable the formation of a sample eluent within the separation compartment, said separation apparatus further comprising:

(a) sample eluate delivery means comprising a mixing chamber, wherein the mixing chamber includes a first side in fluid communication with the outlet port, a top end in fluid communication with a mixing chamber inlet means, and a bottom end in fluid communication with an outlet nozzle;

(b) a fluid source in divertable fluid communication with the mixing chamber inlet means; and

(c) a post-column collection device comprising a sample eluate receiving means positioned relative to the outlet nozzle, thereby enabling the collection of the sample eluate.

28. The liquid phase separation apparatus of claim 27, further comprising a means for heating the fluid source.

29. The liquid phase separation apparatus of claim 27, further comprising protection means interposed between the miniaturized column device and the sample eluate receiving means, said protection means comprising an opening arranged in axial alignment with the outlet nozzle.