Rapid detection of species bands in a separation conduit

• US 6,590,653 B2
• Filed: 10/12/2001
• Issued: 07/08/2003
• Est. Priority Date: 07/16/1996
• Status: Expired due to Term

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Parce, J. Wallace
• Primary Examiner(s): STAFIRA, MICHAEL PATRICK
• Application Number:
  US09/977,056
• Publication Number:
  US 20020097398A1
• Time in Patent Office: 634 Days
• Field of Search: 356/344, 356/314, 204/299.R, 204/182.8, 204/180.1, 250/288, 250/288.A, 250/281, 250/282, 210/198.2, 210/198.3, 210/658, 210/656, 73/615.3, 73/233, 73/614.8, 436/180, 436/52, 436/53, 422/56, 422/58, 422/61, 422/68.1, 422/100, 422/102, 422/81, 422/82, 435/287.3, 435/288.6, 435/288.7
• US Class Current: 356/344
• CPC Class Codes: G01N 27/44721 by optical means
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: Y10S 366/02 Micromixers: segmented laminar flow with boundary mixing orthogonal to the direction of fluid propagation with or without geometry influences from the pathway

Abstract

The present invention provides a microfluidic system for fast, accurate and low cost electrophoretic analysis of materials in the fields of chemistry, biochemistry, biotechnology, molecular biology and numerous other fields. Light from periodically spaced regions along a channel in the microfluidic system are received by a photodetector. The intensity of light received by the photodetector is modulated by the movement of species bands through the channel under electrophoretic forces. By Fourier analysis, the velocity of each species band is determined and the identification of the species is made by its electrophoretic mobility in the channel.

Claims

1. A method of detecting fluorescent species bands in a separation conduit, comprising:

directing an activation energy through a mask at a plurality of discrete regions of the separation conduit, the plurality of discrete regions containing a plurality of fluorescent species bands, the fluorescent species bands fluorescing in response to said activation energy; and

simultaneously detecting fluorescence from said plurality of species bands in said plurality of discrete regions.

2. The method of claim 1, wherein the plurality of fluorescent species bands are electrophoresed through the separation conduit during said directing and simultaneously detecting steps.

3. The method of claim 1, wherein the activation energy is directed at a plurality of discrete regions of the separation conduit.

4. The method of claim 3, further comprising determining a rate at which the plurality of species bands are moving in the separation conduit by monitoring a frequency at which fluorescence is detected in the plurality of regions of the separation conduit.

5. The method of claim 1, wherein the separation conduit comprises a channel in a microfluidic device.

6. The method of claim 1, wherein the fluorescent species bands comprise nucleic acids.

7. A method of detecting fluorescent species bands in a separation conduit, comprising:

directing an activation energy at a plurality of discrete regions of the separation conduit, the plurality of discrete regions containing a plurality of fluorescent species bands, the fluorescent species bands fluorescing in response to said activation energy;

simultaneously detecting fluorescence from said plurality of species bands in said plurality of discrete regions; and

wherein the plurality of discrete regions are each between about 50 and 500 μ

m long along the separation conduit.

8. A method of detecting fluorescent species bands in a separation conduit, comprising:

directing an activation energy at a plurality of regions of the separation conduit, the plurality of regions containing a plurality of fluorescent species bands, the fluorescent species bands fluorescing in response to said activation energy; and

simultaneously detecting fluorescence from said plurality of species bands in said plurality of regions, wherein the fluorescence from the plurality of fluorescent species bands is passed through a plurality of slits before being detected.

9. A system for detecting separated fluorescent species bands, comprising:

a separation conduit for separating species bands;

a fluorescence detection system, comprising;

an activation energy source for directing activation energy at a plurality of regions of the separation conduit at a wavelength effective to activate a fluorescent material on the species bands;

a mask positioned between the activation energy source and the separation conduit for directing the activation energy at a plurality of discrete regions of the separation conduit; and

a detector oriented to simultaneously detect fluorescence from the plurality of regions of the separation conduit.

10. The system of claim 9, wherein the detector comprises a CCD array.