Fluorescent assays, including immunoassays, with feature of flowing sample
First Claim
1. In a method for performing assays involving measurement of fluorescence excited in a fluid sample by an evanescent wave at a surface region of a totally internally reflecting substrate, the improvement comprising:
- exciting said fluorescence while flowing through an enclosure of fixed dimensions bounded in part by said region, a sufficient volume of said sample to maintain said enclosure filled with flowing sample.
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Abstract
An assay method and apparatus employing total internal reflection of excitation radiation at an activated region on the surface of an optically conductive rod or fiber surrounded by a liquid phase of lower index of refraction, so as to excite into fluorescence fluorophores in the liquid present in a thin layer around the rod surface. The induced fluorescence is then gathered and measured. Excitation occurs while flowing a sample of the liquid phase through an enclosure of fixed dimensions bounded in part by the activated region.
87 Citations
11 Claims
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1. In a method for performing assays involving measurement of fluorescence excited in a fluid sample by an evanescent wave at a surface region of a totally internally reflecting substrate, the improvement comprising:
exciting said fluorescence while flowing through an enclosure of fixed dimensions bounded in part by said region, a sufficient volume of said sample to maintain said enclosure filled with flowing sample. - View Dependent Claims (2, 3)
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4. In apparatus for assaying a fluid sample, said apparatus including a totally internally reflecting substrate transmissive to radiation capable of exciting fluorescence in fluorescent material disposed at least on a portion of the surface of said substrate, said substrate also being transmissive to said fluorescence, and means spaced from at least said portion of said surface of said substrate so as to define an enclosure bounded in part by said portion, the improvement comprising:
means for flowing substantially continuously said sample over at least said portion of said surface and in sufficient volume to fill said enclosure, while exciting said fluorescence. - View Dependent Claims (5, 6, 7, 8, 9, 10, 11)
Specification