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Novel Diode Laser‐compatible Fluorophores and Their Application to Single Molecule Detection, Protein Labeling and Fluorescence Resonance Energy Transfer Immunoassay
Author(s) -
Oswald Bernhard,
Gruber Michaela,
Böhmer Martin,
Lehmann Frank,
Probst Mario,
Wolfbeis Otto S.
Publication year - 2001
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1562/0031-8655(2001)0740237ndlcfa2.0.co2
Subject(s) - cyanine , förster resonance energy transfer , covalent bond , chemistry , fluorescence , context (archaeology) , photochemistry , immunoassay , resonance fluorescence , resonance (particle physics) , laser , quantum yield , optics , organic chemistry , physics , particle physics , antibody , immunology , biology , paleontology
We describe a series of new long‐wave absorbing and fluorescing cyanine dyes and labels (based on a general logic for the design of such dyes), their spectra, covalent and noncovalent linkage to proteins, their use in single molecule detection (SMD) and as donors and acceptors, respectively, in fluorescence resonance energy transfer studies. The new labels represent water‐soluble and reactive fluorophores whose quantum yields increase substantially if noncovalently or covalently bound to proteins. Due to their strong absorptions between 550 and 700 nm they are excitable by light‐emitting diodes or diode lasers. Their high absorbances (ε around 100 000) and adequate fluorescence quantum yields (φ up to 0.68 if bound to proteins) along with their availability as reactive NHS esters make them viable labels for proteins and oligomers, e.g. in context with SMD or fluorescence energy transfer immunoassay which is demonstrated for the system HSA/anti‐HSA.