Immunoassays at a quartz-liquid interface: Theory, instrumentation and preliminary application to the fluorescent immunoassay of human immunoglobulin G

Sutherland, Ranald M.; Dähne, Claus; Place, John F.; Ringrose, Anthony R.

doi:10.1016/0022-1759(84)90292-8

Cited by 58 publications

(9 citation statements)

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“…The principle of the evanescent field sensing enables the detection of fluorophores exclusively in the close proximity of the optical fiber. 19 The maximum penetration depth of the evanescent field into the surrounding medium is of the order of some hundred nanometers, allowing detection of the affinity partners which are bound to immobilized recognition elements. Fluorophores outside of the evanescent field do not contribute to the emission signal.…”

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confidence: 99%

Fiber-Optic Evanescent Wave Biosensor for the Detection of Oligonucleotides

et al. 1996

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An automated optical biosensor system based on fluorescence excitation and detection in the evanescent field of a quartz fiber was used to detect 16-mer oligonucleotides in DNA hybridization assays. A biotinylated capture probe was immobilized on the fiber surface via avidin or streptavidin. The hybridization with fluorescein-labeled complementary strands was monitored in real time by fluorescence detection. The double strands formed by hybridization could be dissociated by chemical or thermal regeneration, allowing one to perform hundreds of assay cycles with the same fiber. The signal loss during longtime measurements, i.e., consecutive hybridization assays, can be described by a single-exponential function. Over more than 200 cycles, the net signal decreased by 50% with a signal variation of 2.4% after correction for this signal loss. By binding the capture probe with the 5'-end to the optical fiber surface, and by using a 50% (w/w) aqueous urea solution for chemical regeneration, the duration of an assay cycle could be reduced to 3 min. By applying longer assay cycles, the detection limit for the hybridization with a complementary fluorescein-labeled oligonucleotide was 2.0 x 10(-13) M (24 fmol). To detect an unlabeled complementary 16-mer oligonucleotide, competitive hybridization assays were performed, resulting in a detection limit of 1.1 x 10(-9) M (132 pmol). Poly-(acrylic acid) 5100 sodium salt and Tween 20 were used in the hybridization buffer to prevent nonspecific binding caused by ionic or hydrophobic interaction. The amount of nonspecific binding of noncomplementary oligonucleotides was in the range of 1-2%, compared with the specific binding in the different hybridization assays.

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Fiber-Optic Evanescent Wave Biosensor for the Detection of Oligonucleotides

et al. 1996

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“…The parameter d p , also called the penetration depth, is defined as the distance at which the electric field amplitude reduces to 1/e of E 0 . The term d p can be shown to be given as (Sutherland et al, 1984;Boisdé and Harmer, 1996).…”

Section: Total Internal Reflection Fluorescence Microscopymentioning

confidence: 99%

Bioimaging: Principles and Techniques

Prasad¹

2003

Introduction to Biophotonics

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“…Antibody-antigen reactions may be monitored optically by coating the antibody onto a fibre (often in place of part of the cladding, making an intrinsic sensor). Subsequent combination with the antigen changes the transmission properties and may thus be detected as a change in intensity [24].…”

Section: Sensors For Biological Compoundsmentioning

confidence: 99%

Fibreoptic Sensors in Clinical Measurement

Yellowlees

1991

British Journal of Anaesthesia

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Immunoassays at a quartz-liquid interface: Theory, instrumentation and preliminary application to the fluorescent immunoassay of human immunoglobulin G

Cited by 58 publications

References 11 publications

Fiber-Optic Evanescent Wave Biosensor for the Detection of Oligonucleotides

Fiber-Optic Evanescent Wave Biosensor for the Detection of Oligonucleotides

Bioimaging: Principles and Techniques

Fibreoptic Sensors in Clinical Measurement

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