Instrumentation for fluorescence-based fiber optic biosensors.

This chapter summarizes the construction principles, operation, and calibration of (single-fiber) fluorescence-based fiber optic sensors. These sensors transduce recognition of a chemical analyte by a transducer such as a protein molecule as a change in fluorescence wavelength or lifetime that can be measured remotely through a length of fiber optic. Examples are given of determination of metal ions in aqueous solution by fluorescence ratio and lifetime.

DsRed as a highly sensitive, selective, and reversible fluorescence-based biosensor for both Cu(+) and Cu(2+) ions.

The wild type form of Red fluorescent protein (DsRed), an intrinsically fluorescent protein found in tropical corals, is found to be highly selective, reversible and sensitive for both Cu(+) and Cu(2+), with a nanomolar detection limit. The selectivity towards these ions is retained even in the presence of other heavy metal ions. The K(d) values for monovalent and divalent copper, based on single binding isotherms, are 450 and 540 nM, respectively.

Excitation ratiometric fluorescent biosensor for zinc ion at picomolar levels.

Zinc is a metal ion of increasing significance in several biomedical fields, including neuroscience, immunology, reproductive biology, and cancer. Fluorescent indicators have added greatly to our understanding of the biology of several metal ions, most notably calcium. Despite substantial efforts, only recently have zinc indicators been developed which are sufficiently selective for use in the complex intra- and extracellular milieus, and which are capable of quantifying the free zinc levels with some degree of reliability.

Fluorescent zinc indicators for neurobiology.

Mounting evidence indicates that zinc has multiple roles in cell biology, viz. as a part of metalloenzyme catalytic sites, as a structural component of gene regulatory proteins, and (like calcium) as a free signal ion, particularly in the cortex of the brain. While most Zn(II) in the brain is tightly bound, such that free Zn(II) levels extracellularly and intracellularly are likely to be picomolar, a subset of glutamatergic neurons possess weakly bound zinc in presynaptic boutons which is released at micromolar levels in response to a variety of stimuli.