Single-Cell Label-Free Assay

• US 20090093013A1
• Filed: 05/28/2008
• Published: 04/09/2009
• Est. Priority Date: 10/06/2007
• Status: Active Grant

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Ferrie, Ann M., Fang, Ye, Wu, Qi
• Granted Patent: US 8,703,428 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: G01N 21/211 Ellipsometry optical thickness measurement G01B11/06
• CPC Class Codes: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• CPC Class Codes: G01N 21/554 detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

The disclosure provides a system and method for characterizing a single live-cell response to a stimulus with a biosensor imaging system having cells immobilized on the biosensor at a resolution level of a single cell, as defined herein.

Claims

1. A method for characterizing a single live-cell response to a stimulus, the method comprising:

contacting the single live-cell immobilized on a biosensor with a stimulus for a selected cellular target;

interrogating and detecting the dynamic mass redistribution of the contacted cell with a biosensor imaging system; and

determining the difference effect of the stimulus on the cell-signaling of the cellular target.

2. The method of claim 1 wherein the single live-cell comprises at least one of:

a single-cell per biosensor, a single-cell cluster per biosensor, a sub-single-cell per biosensor, or a combination thereof.

3. The method of claim 1 wherein the difference effect of the stimulus on the cell-signaling of the cellular target comprises comparing the dynamic mass redistribution of the cell-signaling of the cellular target in the presence and in the absence of the stimulus.

4. The method of claim 1 wherein the biosensor imaging system'"'"'s resolution is from about 1 to about 50 pixels per cell.

5. The method of claim 1 wherein the biosensor imaging system'"'"'s resolution is from about 1 to 10 pixels per cell at least in the direction perpendicular to the propagation direction of resonant light within the biosensor.

6. The method of claim 1 wherein the cell immobilized on the biosensor'"'"'s surface has a confluency of from about 0.5% to about 100%.

7. The method of claim 1 wherein the cell immobilized on the biosensor'"'"'s surface has a confluency of from about 80% to about 100%.

8. The method of claim 1 wherein the biosensor imaging system comprises a swept wavelength optical interrogation imaging system for a resonant waveguide grating biosensor, imaging ellipsometry, surface plasmon resonance imaging, or a combination thereof.

9. The method of claim 1 wherein the dynamic mass redistribution signal comprises an optical signal that measures real time kinetics of a stimulus-induced cellular response as a function of time.

10. The method of claim 1 wherein the dynamic mass redistribution signal comprises an optical signal that measures an endpoint or multiple points of a stimulus-induced cellular response over time and throughout a stimulation event.

11. The method of claim 1 wherein the biosensor imaging system provides biosensor output comprising at least one of:

the overall dynamics, the phase, the amplitude and kinetics of the phase, the transition time from one phase to another of the dynamic mass redistribution signal, or a combination thereof.

12. The method of claims 1 wherein the dynamic mass redistribution is the resonant wavelength or the resonant angle of the coupled light at each pixel or location and which wavelength or angle depends on the image interrogation mode of the biosensor system.

13. The method of claim 1 wherein the cellular target comprises a feature on the cell'"'"'s surface comprising at least one of a G protein-coupled receptor (GPCR), an ion channel, a receptor tyrosine kinase, a cytokine receptor, an immuno-receptor, an integrin receptor, an ion transporter, a pathogen recognition target, or a combination thereof.

14. The method of claim 1 wherein the immobilized cells are patterned on the surface of the biosensor.

15. The method of claim 1 wherein the interrogating and detecting comprises a period of time of at least one of:

from about seconds to about minutes, from about minutes to about hours, from about days to about weeks, or a combination thereof.

16. The method of claim 1 wherein the cellular target comprises an intracellular target comprising at least one of:

an enzyme, a kinase, a phosphatase, a monomeric or a dimeric receptor, an homologous or an heterologous receptor complex, or combinations thereof.

17. The method of claim 1 wherein the single live-cell immobilized on the biosensor comprises an adherent cell, a suspension cell, or a combination thereof.