Method of performing gradient-based assays in a microfluidic device

• US 7,112,444 B2
• Filed: 04/24/2003
• Issued: 09/26/2006
• Est. Priority Date: 04/24/2002
• Status: Expired due to Term

• Current Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Original Assignee: Wisconsin Alumni Research Foundation (University of Wisconsin System)
• Inventors: Beebe, David J., Walker, Glenn M.
• Primary Examiner(s): Le, Ong V.
• Assistant Examiner(s): YANG, NELSON C
• Application Number:
  US10/422,236
• Publication Number:
  US 20040063151A1
• Time in Patent Office: 1,251 Days
• Field of Search: 137/3, 137/93, 156/292, 156/273.5, 422/50, 422/58, 422/100, 422 81- 8208, 422/103, 435 6- 72, 436/518, 436/514, 436/52, 436/53, 436/172, 436/177, 436/180
• US Class Current: 436/52
• CPC Class Codes: G01N 33/6818 Sequencing of polypeptides
• CPC Class Codes: Y10T 436/117497 with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/118339 with formation of a segmented stream
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A method is provided for performing a gradient-based assay in a microfluidic device. The method includes the steps of passing first and second fluids through a channel in the microfluidic device. The first fluid has a predetermined concentration of particles therein such that the particles in the first fluid diffuse into the second fluid so as to cause a gradient of concentration of particles in the second fluid as the second fluid flows through the channel. The second fluid sequentially intersects a series of targets along the channel wall as the second fluid flows through the channel.

Claims

1. A method of performing a gradient-based assay in a microfluidic assay, comprising the steps of:

providing a microfluidic device having a channel therethrough, the channel being partially defined by a channel wall extending along an axis;

providing a plurality of biological targets axially spaced along the channel wall;

introducing a stream of first fluid into the channel so as to flow therethrough, the first fluid stream having a predetermined concentration of particles therein;

introducing a stream of second fluid into the channel so as to flow therethrough in laminar flow with the first fluid stream, the particles in the first fluid stream diffusing into the second fluid stream so as to cause a gradient of concentrations of particles in the second fluid stream such that predetermined concentrations of particles in the second stream intersect corresponding biological targets as the second fluid stream flows therepast;

monitoring the plurality of biological targets for optical changes, the optical change of each biological target corresponding to the predetermined concentration of particles in the second fluid stream acting thereon;

selecting one of the predetermined concentrations in response to the optical change of a user-selected biological target; and

providing a plurality of wells spaced along the channel wall, wherein the plurality of biological targets are located within the wells.

2. The method of claim 1 wherein the channel has first and second fluid sides and first and second ends and wherein the channel wall extends along the second side of the channel.

3. The method of claim 2 wherein the first fluid stream is introduced in the first side of the channel adjacent the first end thereof and wherein the second fluid stream is introduced in the second side of the channel adjacent the first end thereof.

4. The method of claim 1 further comprising the step of generating a visual display in response to the second fluid stream flowing past the biological targets.

5. A method of performing a gradient-based assay in a microfluidic assay, comprising the steps of:

providing a microfluidic device having a channel therethrough, the channel being partially defined by a channel wall extending along an axis;

providing a plurality of biological targets axially spaced along the channel wall;

introducing a stream of first fluid into the channel so as to flow therethrough, the first fluid stream having a predetermined concentration of particles therein;

introducing a stream of second fluid into the channel so as to flow therethrough in laminar flow with the first fluid stream, the particles in the first fluid stream diffusing into the second fluid stream so as to cause a gradient of concentrations of particles in the second fluid stream such that predetermined concentrations of particles in the second stream intersect corresponding biological targets as the second fluid stream flows therepast;

monitoring the plurality of biological targets for optical changes, the optical change of each biological target corresponding to the predetermined concentration of particles in the second fluid stream acting thereon;

selecting one of the predetermined concentrations in response to the optical change of a user-selected biological target; and

providing a plurality of sample channels extending from the channel, wherein portions of only the second fluid stream flow into corresponding sample channels,

wherein the biological targets are stationary with respect to the first and second fluid streams flowing through the channel.

6. The method of claim 5 wherein the channel has first and second fluid sides and first and second ends and wherein the channel wall extends along the second side of the channel.

7. The method of claim 6 wherein the first fluid stream is introduced in the first side of the channel adjacent the first end thereof and wherein the second fluid stream is introduced in the second side of the channel adjacent the first end thereof.

8. The method of claim 5 further comprising the step of generating a visual display in response to the second fluid stream flowing past the biological targets.