Optical-waveguide sensor chip, method of manufacturing the same, method of measuring substance, substance-measuring kit and optical-waveguide sensor

1. An optical-waveguide sensor chip, comprising:

a plane optical waveguide;

a cap formed on the plane optical waveguide and including a dent for forming a measurement region with the plane optical waveguide;

a first substance which is immobilized on a part of the surface of the plane optical waveguide in the measurement region, and which is specifically reactive with an analyte substance in an analyte sample solution;

an inlet hole opened in the cap so as to communicate with the measurement region, the inlet hole configured to introduce the analyte sample solution and a dispersion of fine particles including a second substance immobilized on the surface thereof into the measurement region, the second substance being specifically reactive with the analyte substance in the analyte sample solution;

an outlet hole opened in the cap so as to communicate with the measurement region; and

an incident-sided grating and an outgoing-sided grating disposed on the plane optical waveguide to interpose the measurement region therebetween, and configured to generate evanescent light on the plane optical waveguide including the measurement region by introducing incident light to the incident-sided grating.

2. The optical-waveguide sensor chip according to claim 1 , wherein the plane optical waveguide is a plate-shaped glass.

3. The optical-waveguide sensor chip according to claim 1 , wherein the plane optical waveguide is an organic resin film having a thickness of 3 to 300 μm.

4. The optical-waveguide sensor chip according to of claim 1 , wherein the fine particles are resin beads.

5. The optical-waveguide sensor chip according to claim 1 , wherein the fine particles are metal colloids.

6. The optical-waveguide sensor chip according to claim 1 , wherein the analyte substance is an antigen and each of the first and second substances, which are specifically reactive with the analyte substance, are antibodies.

7. The optical-waveguide sensor chip according to claim 1 , wherein each of the fine particles has a diameter of 50 nm to 10 μm.

8. A method of measuring a substance, comprising:

preparing an optical-waveguide sensor chip according to claim 1 ;

dropping an analyte sample solution on the surface of a plane optical waveguide in a measurement region of the optical-waveguide sensor chip by introducing the analyte sample solution into the measurement region through an inlet hole to allow specific reaction between a first substance on the surface of the plane optical waveguide and an analyte substance in the analyte sample solution;

introducing a dispersion of fine particles including a second substance, which is immobilized on the surface of each of the particles, on the surface of the plane optical waveguide in the measurement region through the inlet hole to specifically react the second substance with the analyte substance, thereby immobilizing the fine particles on the surface of the plane optical waveguide in the measurement region by binding the analyte substance with the first substance on the surface of the plane optical waveguide and by binding the second substance with the analyte substance;

generating evanescent light on the plane optical waveguide including the measurement region while light is irradiated through an incident-sided grating to the plane optical waveguide including the measurement region to propagate the incident light in the plane optical waveguide including the measurement region and releasing the incident light from an outgoing-sided grating; and

detecting reduction in an intensity of the light released from the outgoing-sided grating to measure said substance, where the reduction is caused in accordance with attenuation of the evanescent light absorbed or scattered by the fine particles immobilized on the surface of the plane optical waveguide in the measurement region.

9. The method according to claim 8 , wherein the analyte substance in the analyte sample solution is an antigen and each of the first and second substances, which are specifically reactive with the analyte substance, are antibodies.

10. A substance-measuring kit comprising, in combination:

an optical-waveguide sensor chip comprising:

a plane optical waveguide,

a cap formed on the plane optical waveguide and including a dent for forming a measurement region with the plane optical waveguide,

a first substance which is immobilized on a part of the surface of the plane optical waveguide in the measurement region, and which is specifically reactive with an analyte substance in an analyte sample solution,

an inlet hole opened in the cap so as to communicate with the measurement region, the inlet hole configured to introduce the analyte sample solution and a dispersion of fine particles including a second substance immobilized on the surface thereof into the measurement region, the second substance being specifically reactive with the analyte substance in the analyte sample solution,

an outlet hole opened in the cap so as to communicate with the measurement region, and

an incident-sided grating and an outgoing-sided grating disposed on the plane optical waveguide to interpose the measurement region therebetween, and configured to generate evanescent light on the plane optical waveguide including the measurement region by introducing incident light to the incident-sided grating; and

a package accommodating the dispersion of fine particles having a second substance immobilized on the surface thereof, the second substance being specifically reactive with the analyte substance in the analyte sample solution.

11. The substance-measuring kit according to claim 10 , wherein the analyte substance is an antigen and each of the first and second substances, which are specifically reactive with the analyte substance, are antibodies.

12. The substance-measuring kit according to claim 10 , wherein the plane optical waveguide is a plate-shaped glass.

13. The substance-measuring kit according to claim 10 , wherein the plane optical waveguide is an organic resin film having a thickness of 3 to 300 μm.

14. The substance-measuring kit according to claim 10 , wherein each of the fine particles has a diameter of 50 nm to 10 μm.

15. The substance-measuring kit according to claim 10 , wherein the fine particles are resin beads.

16. The substance-measuring kit according to claim 10 , wherein the fine particles are metal colloids.

17. A method of measuring a substance by using the substance-measuring kit according to claim 10 , comprising:

dropping an analyte sample solution on the surface of a plane optical waveguide in a measurement region of an optical-waveguide sensor chip by introducing the analyte sample solution into the measurement region through an inlet hole to allow specific reaction between a first substance immobilized on the surface of the plane optical waveguide and an analyte substance in the analyte sample solution;

introducing a dispersion of fine particles in the package on the surface of the plane optical waveguide in the measurement region through the inlet hole to specifically react a second substance on the surface of the fine particles with the analyte substance, thereby immobilizing the fine particles on the surface of the plane optical waveguide in the measurement region by binding the analyte substance with the first substance on the surface of the plane optical waveguide and by binding the second substance with the analyte substance;

generating evanescent light on the plane optical waveguide including the measurement region while light is irradiated through an incident-sided grating to the plane optical waveguide including the measurement region to propagate the incident light in the plane optical waveguide including the measurement region and releasing the incident light from an outgoing-sided grating; and

detecting reduction in an intensity of the light released from the outgoing-sided grating to measure said substance, where the reduction is caused in accordance with attenuation of the evanescent light absorbed or scattered by the fine particles immobilized on the surface of the plane optical waveguide in the measurement region.

18. The method according to claim 17 , wherein the analyte substance in the analyte sample solution is an antigen and each of the first and second substances, which are specifically reactive with the analyte substance, are antibodies.

19. An optical-waveguide sensor, comprising:

an optical-waveguide sensor chip comprising:

a plane optical waveguide,

a cap formed on the plane optical waveguide and including a dent for forming a measurement region with the plane optical waveguide,

a first substance which is immobilized on a part of the surface of the plane optical waveguide in the measurement region, and which is specifically reactive with an analyte substance in an analyte sample solution,

an inlet hole opened in the cap so as to communicate with the measurement region, the inlet hole configured to introduce the analyte sample solution and a dispersion of fine particles including a second substance immobilized on the surface thereof into the measurement region, the second substance being specifically reactive with the analyte substance in the analyte sample solution,

an outlet hole opened in the cap so as to communicate with the measurement region, and

an incident-sided grating and an outgoing-sided grating disposed on the plane optical waveguide to interpose the measurement region therebetween, and configured to generate evanescent light on the plane optical waveguide including the measurement region by introducing incident light to the incident-sided grating;

a light source configured to irradiate a light into the incident-sided grating of the optical-waveguide sensor chip; and

a light-receiving device configured to receive light released from the outgoing-sided grating of the optical-waveguide sensor chip.