Method for enhancing fluorescence

• US 6,118,126 A
• Filed: 11/06/1998
• Issued: 09/12/2000
• Est. Priority Date: 10/31/1997
• Status: Expired due to Fees

• Current Assignee: Sarnoff Corporation (SRI International, Inc.)
• Original Assignee: Sarnoff Corporation (SRI International, Inc.)
• Inventors: Zanzucchi, Peter John
• Primary Examiner(s): Minnifield, Nita
• Assistant Examiner(s): BASKAR, PADMAVATHI
• Application Number:
  US09/187,355
• Time in Patent Office: 676 Days
• Field of Search: 436/172, 436/518, 436/524, 436/531, 422/82.07, 422/82.08, 250/458.1, 250/461.1, 250/461.2
• US Class Current: 250/458.1
• CPC Class Codes: B01L 2300/168 Specific optical properties, e.g. reflective coatings
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/545 Synthetic resin
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

The present invention relates to a method for the enhancement of fluorescence wherein a fluorophor is connected to a textured material. The method can be used in any forensic or medical diagnostic assay, particularly where the absence or presence of a molecule having a concentration of less than about 1 μg/ml is desirably determined.

Claims

1. A device for conducting reactions comprising:

an assay device comprising a detection or reaction chamber which contains a textured material that enhances fluorescence of a fluorophor associated therewith, the association of the textured material with the fluorophor providing at least a 10-fold enhancement in fluorescence yield of the fluorophor, wherein the textured material comprises voids or pores effective to create backscatter of the fluorescence exciting light;

a source of light selected to emit light that is not absorbed by the textured material in an amount effective to diminish the fluorescence enhancement, which light source is aligned with at least one detection or reaction chamber, wherein the light source directs light at an angle sufficiently close to a 90°

angle relative to the textured surface so that the fluorescence yield is at least 75% of that available with a 90°

angle; and

a detector aligned to collect emissions located within a 20°

cone surrounding the axis of reflection of the light emitted by the light source.

2. The device of claim 1, wherein the the textured material is a porous membrane that is formed of nylon, poly(carbonate) or nitrocellulose.

3. The device of claim 1, wherein association of the fluorophor and textured material yield at least a 50-fold enhancement in the fluorescence yield of the fluorophor.

4. The device of claim 1, wherein the light source is a source of coherent light.

5. The device of claim 1, wherein the detector is aligned to collect emissions located within no more than a 10°

cone surrounding the axis of reflection of the light emitted by the light source.

6. The device of claim 1, wherein the detector is aligned to collect emissions located within no more than a 5°

cone surrounding the axis of reflection of the light emitted by the light source.

7. The device of claim 1, wherein the light source directs light at an angle effective to yield at least 85% of the fluorescence yield available using a 90°

angle relative to the textured surface.

8. The device of claim 7, wherein the textured material is a porous membrane that is formed of nylon, poly(carbonate) or nitrocellulose.

9. The device of claim 1, wherein the light source directs light at an angle effective to yield at least 90% of the fluorescence yield available using a 90°

angle relative to the textured surface.

10. The device of claim 1, wherein the light source directs light at an angle effective to yield at least 95% of the fluorescence yield available using a 90°

angle relative to the textured surface.

11. The device of claim 1, wherein the light source directs light at a 90°

angle relative to the textured surface.

13. The device of claim 8, wherein the embossed, textured material is sealed on the opposing side to prevent fluid transit therethrough.

12. A device for conducting a fluorescence measurement comprising an textured material, which textured material enhances the fluorescence of a fluorophor associated therewith providing at least a 10-fold enhancement in the fluorescence yield of the fluorophor when detected in backscatter mode, embossed to define a reaction well or chamber, wherein the textured material comprises voids or pores effective to create backscatter of the fluorescence exciting light.