Optical approach for microfluidic DNA electrophoresis detection

• US 9,067,207 B2
• Filed: 03/04/2011
• Issued: 06/30/2015
• Est. Priority Date: 06/04/2009
• Status: Active Grant

• Current Assignee: Leidos Innovations Technology, Inc., Microgem NZ Limited, University of Virginia Patent Foundation (University of Virginia)
• Original Assignee: Lockheed Martin Corporation (Martin Marietta Corporation), University of Virginia Patent Foundation (University of Virginia), Zygem Corporation Ltd.
• Inventors: Bell, H. Randall, Landers, James P, Scott, Orion N, Pettit, John W, Bienvenue, Joan M, Marchiarullo, Daniel J, Root, Brian E
• Primary Examiner(s): MUMMERT, STEPHANIE KANE
• Application Number:
  US13/064,091
• Publication Number:
  US 20110229897A1
• Time in Patent Office: 1,579 Days
• Field of Search: None
• US Class Current: 1/1
• 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 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: G01N 2021/4186 Phase modulation imaging
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 29/32 Arrangements for suppressing undesired influences, e.g. temperature or pressure variations , compensating for signal noise

Abstract

Aspects of the disclosure provides a DNA analyzer to facilitate an integrated single-chip DNA analysis. The DNA analyzer includes an interface for coupling a microfluidic chip to the DNA analyzer. The microfluidic chip includes a first domain configured for polymerase chain reaction (PCR) amplification of DNA fragments, and a second domain fluidically coupled to the first domain to receive the DNA fragments and perform electrophoretic separation of the DNA fragments. The DNA fragments are tagged with fluorescent labels. The DNA analyzer includes a detection module to excite the fluorescent labels to emit fluorescence and detect the emitted fluorescence. The detection module includes a laser source, a set of optical elements, a filter module and a photo-detector.

Claims

1. A DNA analyzer, comprising:

an interface for coupling a microfluidic chip to the DNA analyzer, wherein the microfluidic chip includes;

a first domain configured for polymerase chain reaction (PCR) amplification of DNA fragments, the DNA fragments being tagged with fluorescent labels; and

a second domain fluidically coupled to the first domain to receive the DNA fragments, the second domain having a separation channel for electrophoretic separation of the DNA fragments; and

a detection module configured to be optically coupled with the microfluidic chip that includes;

a laser source configured to generate a laser beam;

a set of optical elements configured to direct the laser beam to the separation channel to excite the fluorescent labels to emit fluorescence while the DNA fragments migrate in the separation channel, and to collect the emitted fluorescence into an optical signal, the set of optical elements including an objective lens configured to be aligned with the separation channel to direct the laser beam to the separation channel and to collect the emitted fluorescence from the separation channel;

a motor configured to align the objective lens to the separation channel;

a filter module configured to filter the optical signal to allow a first portion of the optical signal having a first wavelength to pass;

a photo-detector configured to generate an electrical detection signal in response to the filtered optical signal;

a modulation signal generator coupled to the filter module, the modulation signal generator being configured to generate a modulation signal having a modulation frequency, and a reference signal having the modulation frequency, the modulation signal being used by the filter module to modulate the filtered optical signal; and

a phase-sensitive detector configured to receive the reference signal and the electrical detection signal corresponding to the modulated filtered optical signal, and demodulate the electrical detection signal based on the reference signal to remove noise due to the photo-detector.

2. The DNA analyzer of claim 1, wherein the photo-detector further comprises:

a photo-multiplier tube configured to generate the electrical detection signal in response to the filtered optical signal.

3. The DNA analyzer of claim 1, wherein the filter module further comprises:

an acousto-optic tunable filter (AOTF) configured to filter the optical signal to allow the first portion of the optical signal having the first wavelength to pass based on an electrical tuning signal having a first tuning frequency, the first wavelength satisfying a matching condition of the AOTF with the first tuning frequency.

4. The DNA analyzer of claim 3, further comprising:

a controller configured to generate a control signal indicative of the first tuning frequency; and

a synthesizer configured to generate the electrical tuning signal having the first tuning frequency based on the control signal.

5. The DNA analyzer of claim 4, wherein

the controller adjusts is configured to adjust the control signal to be indicative of a second tuning frequency, and that-the adjusted control signal causes:

the synthesizer generates the to generate an electrical tuning signal having the second tuning frequency based on the control signal; and

the AOTF filters to filter the optical signal to allow a second portion of the optical signal having a second wavelength to pass based on the electrical tuning signal, the second wavelength satisfying the matching condition of the AOTF with the second tuning frequency.

6. The DNA analyzer of claim 1, further comprising:

a pressure module configured to flow liquid in the micro fluidic chip;

a thermal module configured to induce thermal cycling at the first domain of the microfluidic chip for the PCR amplification;

a power module configured to generate voltages to be applied to the second domain of the micro fluidic chip for the electrophoretic separation; and

a controller module configured to control the pressure module, the thermal module, the power module, and the detection module according to a control procedure to act on the microfluidic chip for a single-chip DNA analysis.

7. A DNA analyzer, comprising:

an interface for coupling a micro fluidic chip to the DNA analyzer, wherein the microfluidic chip includes;

a first domain configured for polymerase chain reaction (PCR) amplification of DNA fragments, the DNA fragments being tagged with fluorescent labels; and

a second domain fluidically coupled to the first domain to receive the DNA fragments, the second domain having a separation channel for electrophoretic separation of the DNA fragments;

a detection module configured to be optically coupled with the microfluidic chip that includes;

a laser source configured to generate a laser beam;

a passive optics module including passive units that are pre-configured to receive the laser beam and transmit the laser beam; and

an active optics module including at least an active unit to focus the laser beam to the separation channel to excite the fluorescent labels to emit fluorescence while the DNA fragments migrate in the separation channel, and to collect the emitted fluorescence from the separation channel into an optical signal for return, wherein;

the passive optics module includes;

a filter module configured to filter the optical signal to allow a first portion of the optical signal having a first wavelength to pass; and

a photo-detector configured to generate an electrical detection signal in response to the filtered optical signal;

the active optics module is configured to be calibrated with respect to each microfluidic chip and the passive optics module is configured not to be adjusted for every microfluidic chip;

a modulation signal generator coupled to the filter module, the modulation signal generator being configured to generate a modulation signal having a modulation frequency, and a reference signal having the modulation frequency, the modulation signal being used by the filter module to modulate the filtered optical signal; and

a phase-sensitive detector configured to receive the reference signal and the electrical detection signal corresponding to the modulated filtered optical signal, and demodulate the electrical detection signal based on the reference signal to remove noise due to the photo-detector.

8. The DNA analyzer of claim 7, wherein the photo-detector further comprises:

a photo-multiplier tube configured to generate the electrical detection signal in response to the filtered optical signal.

9. The DNA analyzer of claim 7, wherein the active optics module further comprises:

an objective lens configured to be aligned with the separation channel to direct the laser beam to the separation channel and to collect the emitted fluorescence from the separation channel.

10. The DNA analyzer of claim 9, wherein the active optics module further comprises:

a motor configured to align the objective lens to the separation channel.

11. The DNA analyzer of claim 7, wherein the filter module further comprises:

an acousto-optic tunable filter (AOTF) configured to filter the optical signal to allow the first portion of the optical signal having the first wavelength to pass based on an electrical tuning signal having a first tuning frequency, the first wavelength satisfying a matching condition of the AOTF with the first tuning frequency.

12. The DNA analyzer of claim 11, further comprising:

a controller configured to generate a control signal indicative of the first tuning frequency; and

a synthesizer configured to generate the electrical tuning signal having the first tuning frequency based on the control signal.

13. The DNA analyzer of claim 12, wherein

the controller adjusts is configured to adjust the control signal to be indicative of a second tuning frequency, and the adjusted control signal causes:

the synthesizer generates the to generate an electrical tuning signal having the second tuning frequency; and

the AOTF filters to filter the optical signal to allow a second portion of the optical signal having a second wavelength to pass based on the electrical tuning signal, the second wavelength satisfying the matching condition of the AOTF with the second tuning frequency.

14. The DNA analyzer of claim 7, further comprising:

a pressure module configured to flow liquid in the microfluidic chip;

a thermal module configured to induce thermal cycling at the first domain of the micro fluidic chip for the PCR amplification;

a power module configured to generate voltages to be applied to the second domain of the microfluidic chip for the electrophoretic separation; and

a controller module configured to control the pressure module, the thermal module, the power module, and the detection module according to a control procedure to act on the microfluidic chip for a single-chip DNA analysis.

15. The DNA analyzer of claim 7, wherein the active optics module is configured to align the objective lens in conjunction with an alignment mark on the microfluidic chip.