Self-referencing biodetection method and patterned bioassays

1. A self-referencing colorimetric resonant optical biosensor comprising:

(a) one or more liquid-holding vessels comprising a colorimetric resonant optical biosensor as a surface; and

(b) one or more specific binding substances immobilized on a first portion of the colorimetric resonant optical biosenor of each liquid-holding vessel forming a reaction surface, and no specific binding substances immobilized on a second portion of the colorimetric resonant optical biosenor of each liquid-holding vessel forming a reference surface, wherein the biosensor comprises two or more reaction surfaces in each liquid holding vessel and two or more reference surfaces in each liquid holding vessel.

2. The self-referencing colorimetric resonant optical biosensor of claim 1 , wherein when the biosensor is illuminated a resonant grating effect is produced on the reflected radiation spectrum and wherein the depth and period of a grating of the biosensor are less than a wavelength of the resonant grating effect.

3. The self-referencing colorimetric resonant optical biosensor of claim 1 , wherein a narrow band of optical wavelengths is reflected from the biosensor when the biosensor is illuminated with a broad band of optical wavelengths.

4. The self-referencing colorimetric resonant optical biosensor of claim 1 , wherein the liquid-holding vessel is selected from the group consisting of a microtiter plate well, a test tube, a Petri dish and a microfluidic channel.

5. A method of detecting the binding of one or more specific binding substances to their respective binding partners in a self-referencing colorimetric resonant optical biosensor of claim 1 comprising:

(a) applying one or more specific binding partners in a reaction volume to the one or more liquid holding vessels of claim 1 ;

(b) illuminating the reaction surfaces and the reference surfaces with light;

(b) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the reaction surfaces and the reference surfaces; and

(c) comparing the maxima or minima of the one or more reference surfaces to the maxima or minima from the one or more reaction surfaces;

wherein the binding of one or more specific binding substances to their respective binding partners is detected.

6. A method of detecting activity of an enzyme in a self-referencing colorimetric resonant optical biosensor of claim 1 comprising:

(a) applying one or more enzymes in a reaction volume to the one or more liquid holding vessels of claim 1 ;

(b) illuminating the one or more reaction surfaces and the one or more reference surfaces with light;

(c) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the one or more reaction surfaces and the one or more reference surfaces; and

(d) comparing the maxima or minima of the one or more reference surfaces to the maxima or minima from the one or more reaction surfaces;

wherein the activity of an enzyme is detected.

7. A method of detecting inhibition activity of one or more molecules against one or more enzymes or specific binding partners in a self-referencing colorimetric resonant optical biosensor of claim 1 comprising:

(a) applying one or more molecules suspected of having inhibition activity in a reaction volume to the one or more liquid holding vessels of claim 1 ;

(b) applying one or more enzymes or specific binding partners in a reaction volume to the one or more liquid holding vessels;

(c) illuminating the one or more reaction surfaces and the one or more reference surfaces with light;

(d) detecting a maxima in reflected wavelength, or a minima in transmitted wavelength of light from the one or more reaction surfaces and the one or more reference surfaces; and

(e) comparing the maxima or minima of the one or more reference surfaces to the maxima or minima from the one or more reaction surfaces;

wherein the inhibition activity of one or more molecules is detected.