Apparatus and methods for sample analysis

• US 6,375,817 B1
• Filed: 04/16/1999
• Issued: 04/23/2002
• Est. Priority Date: 04/16/1999
• Status: Active Grant

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Perseptive Biosystems Incorporated (Ecovyst, Inc.)
• Inventors: Koutny, Lance, Carson, William W., Taylor, Todd A.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US09/293,313
• Time in Patent Office: 1,103 Days
• Field of Search: 210/198.2, 210/101, 210/243, 210/656, 204/600, 204/601, 204/602, 204/603, 204/604, 204/605, 204/450, 204/451, 204/452, 204/453, 204/455, 422/100, 436/180
• US Class Current: 204/453
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

The present invention is directed to apparatus and methods for rapid, automated, microscale sample analysis using pressure differentials. The invention includes an apparatus having intersecting channels for introduction of a sample and separation of that sample into its components. The sample introduction and separation channels preferably are etched in a microfabricated device, such as a microchip, to form a junction. Pressure gradients are applied to the channels to form a sample plug in the separation channel. The separation channel may have disposed within it a medium for separation of the components suspected to be contained in the sample. For example, with the proper medium, a voltage gradient may be applied along the separation channel to separate the components of the sample electrophoretically. The apparatus also may include means for detecting the components of the sample subsequent to separation.

Claims

1. A sample plug formation device comprising:

a housing defining

a separation channel comprising a longitudinal axis, and

an introduction channel which forms a juncture with the separation channel;

a pressure control device independently in communication with the separation channel and the introduction channel wherein

a first pressure differential applied to the introduction channel transports a sample to the juncture, and

a second pressure differential applied to the separation channel transports a portion of the sample in the juncture into the separation channel to form a sample plug; and

a voltage generator in communication with the separation channel, wherein the voltage generator is adapted to apply an electric potential along the longitudinal axis.

2. The sample plug formation device of claim 1 further comprising a separation medium disposed within the separation channel.

3. The sample plug formation device of claim 1 wherein the housing comprises a microfabricated solid.

4. The sample plug formation device of claim 1 wherein the introduction channel and the separation channel independently have a mean diameter within the range of about 0.1 μ

m to about 1000 μ

m.

5. A scientific instrument comprising the sample plug formation device of claim 1.

6. The scientific instrument of claim 5 further comprising a computer in communication with the pressure control device to control the pressure control device.

7. The scientific instrument of claim 5 further comprising a detector spaced apart from the junction and in communication with the separation channel to detect a chemical component.

11. The method of claim 6 further comprising the step of analyzing for a component in the sample plug.

8. A method for forming a sample plug comprising the steps of:

(a) providing a sample plug formation device comprising;

a housing defining

a separation channel comprising a longitudinal axis, and

an introduction channel which forms a juncture with the separation channel; and

a pressure control device in communication with the separation channel;

(b) applying a first pressure differential to the introduction channel to transport a sample in communication with the introduction channel to the juncture;

(c) applying a second pressure differential to the separation channel to transport a portion of the sample in the junction into the separation channel to form a sample plug; and

(d) applying an electric potential along the longitudinal axis of the separation channel.

9. The method of claim 8 wherein the first pressure differential applied to the introduction channel is reduced prior to the application of the second pressure differential to the separation channel.

10. The method of claim 8 further comprising applying an electric potential along the longitudinal axis during step (b).

12. The method of claim 8 further comprising the step of applying positive pressure to the separation channel to move the sample plug along the separation channel.

13. The method of claim 12 wherein applying positive pressure separates components in the sample plug.

14. The method of claim 13 further comprising the step of analyzing for a component in the sample plug.

15. The method of claim 8 wherein applying the second pressure differential to the separation channel is for a specified interval.

16. A scientific instrument comprising:

a microfabricated solid comprising,

a housing defining

a separation channel comprising a longitudinal axis,

an introduction channel which forms a juncture with the separation channel, and

a pressure control device independently in communication with the separation channel and the introduction channel wherein

a first pressure differential applied to the introduction channel transports a sample to the juncture, and

a second pressure differential applied to the separation channel transports a portion of the sample in the juncture into the separation channel to form a sample plug;

a voltage generator in communication with the separation channel to apply an electric potential along the longitudinal axis;

a computer in communication with the pressure control device to control the pressure control device; and

a detector spaced apart from the junction and in communication with the separation channel to detect a chemical component.

17. The scientific instrument of claim 16 wherein the detector is an optical detector.