System and method for dual-detection of a cellular response

• US 20090325211A1
• Filed: 05/05/2008
• Published: 12/31/2009
• Est. Priority Date: 10/06/2007
• Status: Abandoned Application

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Ferrie, Ann M., Lahiri, Joydeep, Fang, Ye, Wu, Qi, Mudaliar, Deepti J.
• Application Number:
  US12/151,179
• Publication Number:
  US 20090325211A1
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 21/7743 the reagent-coated grating coupling light in or out of the waveguide

Abstract

A system and method as defined herein for dual-detection of evanescent-wave label-free light and evanescent-wave excited-fluorescent label-emitted light in an optical biosensor.

Claims

1. A system for evanescent-wave label-free light and evanescent-wave excited-fluorescence light detection, the system comprising:

an optical sensor;

a light source to illuminate the sensor;

an optical detector to collect the evanescent-wave label-free light and the evanescent-wave excited-fluorescence light emitted from the sensor; and

a processor to analyze the collected light.

2. The system of claim 1, wherein the optical sensor comprises an array of waveguide grating coupled sensors on a microplate.

3. The system of claim 1, wherein the optical sensor has at least one live-cell immobilized on the sensor surface.

4. The system of claim 1, wherein the light source comprises a fiber coupled tunable laser system having wavelengths from about 400 to about 900 nanometers.

5. The system of claim 1, wherein the optical detector includes a self-referenced interferometer.

6. The system of claim 1, wherein the optical detector comprises a first beam splitter that adjusts the incident angle of the light source'"'"'s beam and a second beam splitter that selects evanescent-wave label-free reflected light and evanescent-wave excited fluorescent label emitted light.

7. The system of claim 1, wherein the optical detector comprises a first digital camera for collecting evanescent-wave label-free reflected light and a second digital camera for collecting evanescent-wave excited-fluorescent label emitted light.

8. The system of claim 1, wherein the optical detector comprises at least one of:

a collimating lens;

an excitation filter optionally having a bandwidth of ±

1 nm;

an optical shutter;

a polarization controller;

an imaging lense;

a notch filter;

a fluorescence emission filter;

or a combination thereof.

9. A method for characterizing a live-cell, the method comprising:

providing the system of claim 1 having a live-cell immobilized on the sensor'"'"'s surface;

contacting the immobilized cell with a first fluorescent-labeled stimulus;

detecting the effect of the first fluorescent-labeled stimulus contact on a selected cellular target by interrogating the sensor for evanescent-wave label-free light and evanescent-wave excited-fluorescent label-emitted light; and

comparing the sensor'"'"'s evanescent wave label-free light and evanescent wave excited-fluorescent label-emitted light in the presence and absence of a second stimulus.

10. The method of claim 9, wherein the fluorescent-labeled stimulus has an affinity for at least one target associated with the live-cell immobilized on the sensor'"'"'s surface.

11. The method of claim 9, wherein interrogating the sensor excites the fluorescent-labeled stimulus having an association with the basal cell membrane surface of the immobilized live-cell on the surface of the sensor.

12. The method of claim 9, wherein interrogating the sensor provides evanescent-wave label-free light associated with a dynamic mass redistribution event of the immobilized live-cell.

13. The method of claim 9, wherein interrogating the sensor for evanescent-wave fluorescence and evanescent-wave label-free light is accomplished sequentially, simultaneously, or a combination thereof.

14. The method of claim 9, wherein the sensor is a resonant waveguide grating biosensor, a surface plasmon resonance, a photonic crystal biosensor, or a resonant mirror.

15. A method for characterizing a live-cell, the method comprising:

providing the system of claim 1 having a live-cell immobilized on the sensor'"'"'s surface, the live-cell having a fluorescent target;

contacting the immobilized cell with a stimulus;

detecting the stimulus induced changes on the fluorescent target by interrogating the sensor for evanescent-wave fluorescence light; and

detecting the stimulus induced changes in the evanescent-wave label-free light.

16. The method of claim 15, wherein the live-cell having a fluorescent target is accomplished with a gene expression vector which expresses a fluorescent protein.

17. The method of claim 16, wherein the live-cell having a fluorescent target is accomplished with transfection, insertion of a lipid target into the cell surface membrane, or combination thereof.

18. A dual-detection system for evanescent-wave label-free light and evanescent-wave excited-fluorescence light detection, the system comprising:

an optical sensor;

a light source to illuminate the sensor;

a first optical detector to collect the evanescent-wave label-free light from the sensor;

a second detector to collect evanescent-wave excited-fluorescence light from the sensor; and

a processor to analyze the collected light.

19. The dual-detection system of claim 18, wherein the optical sensor comprises a patterned reference region, a sample region having a live-cell or a biomolecule thereon, or a combination thereof.

20. A method to enhance detection of a single resonant wavelength of an evanescent-wave label-free signal and an evanescent-wave excited-fluorescence signal from a single sensor, the method comprising:

measuring the evanescent-wave excited-fluorescence signal of a specific target having a fluorescent label, and measuring the label-free dynamic mass redistribution signal upon stimulation; and

correlating the measured fluorescence signal from the target and the label-free dynamic mass redistribution signal.

21. The method of claim 20, wherein correlating the fluorescence signal and the label-free dynamic mass redistribution signal comprises, at least one of:

comparing the kinetic profiles of both signals;

comparing the modulation profiles of both signals by alteration of signaling cascades;

comparing the impact of a gene alteration on the cellular response;

or a combination thereof.

22. A method for dual-detection of ion-channel activity in a live-cell, the method comprising:

providing a biosensor having at least one live-cell immobilized on the biosensor surface;

furnishing the immobilized cell with a membrane-potential sensitive dye;

contacting the immobilized cell having the membrane-potential sensitive dye with a stimulus; and

detecting the stimulus-induced optical label-free signal and evanescent wave excited fluorescence signal.

23. A method for dual-detection of ion-channel activity in a live-cell, the method comprising:

providing a biosensor having at least one live-cell immobilized on the biosensor surface;

furnishing the immobilized live-cell with a membrane-potential sensitive dye and a fluorescent lipid;

contacting the immobilized cell having the dye and the lipid with a stimulus; and

detecting the stimulus-induced optical label-free signal and evanescent wave excited fluorescence signal, the fluorescence signal changes in relation to a change in fluorescent resonant energy transfer between the dye and the lipid.

24. A method dual-detection of ion-channel activity in a live-cell, the method comprising:

providing a biosensor having at least one live-cell immobilized on the biosensor surface;

furnishing the immobilized cell with a membrane-potential sensitive dye and a quencher lipid;

contacting the immobilized cell having the dye and the quencher lipid with a stimulus; and

detecting the stimulus-induced optical label-free signal and evanescent wave excited fluorescence signal, the detected fluorescence signal changes in relation to a change in distance between the quencher lipid and the membrane-potential sensitive dye.