Fluorescein sensors

Fluo-3 (1) is a Ca -responsive fluorescein sensor based on the PET mechanism. The fluorescence response of Fluo-3 to Cd " is the same as that observed with Ca " or 7x3 moreover, neither of these metals influences the Cd -induced Huo-3 fluorescence due to the large difference in their binding constants. In practical applications, the probe is loaded into rat thymocytes as the membrane-permeable acetoxymethyl (AM) ester form. Subsequent flow cytometric analysis is characterized by increased intensity of the Fluo-3 fluorescence when the cells are exposed to Cd ", corresponding to the formation of the Cd -Huo-3 complex in the cells [37,38]. 

One of the interesting features in the structure-photophysical property relationship of fluorescein is that the quantum yield of fluorescein increases under the basic condition. Therefore, many of fluorescein derivatives have been used as pH sensors to measure intracellular pH due to their pH-responding photophysical property [53]. Although fluorescein itself is slightly fluorescent in alcoholic solutions, the addition of alkali (pH > 8) to the fluorescein solution produces the very intense fluorescent alkali salt. The salt form of fluorescein... 

L.A. Saari, W.R. Seitz 1982 pH sensor based on immobilized fluorescein... 

The first sensor proposed for detecting gastric and oesophageal pH24, made use of two fluorophores, fluorescein and eosin, immobilised onto fibrous particles of amino-ethyl cellulose, fixed on polyester foils. Only tested in vitro, the sensor reveals a satisfactory response time of around 20 seconds. 

CL sensors based on immobilization of nonenzyme reagents have been extensively studied in recent years. Nakagama et al. [63] developed a CL sensor for monitoring free chlorine in tap water. This sensor consisted of a Pyrex tube, packed with the uranine (fluoresceine disodium) complex immobilized on IRA-93 anion-exchange resin, and a PMT placed close to the Pyrex tube. It was used for monitoring the concentration of free chlorine (as HCIO) in tap water, up to 1 mmol/L, with a detection limit of 2 nmol/L. The coefficient of variation (n =... 

The intrinsic sensors are based on the direct recognition of the chemicals by its intrinsic optical activity, such as absorption or fluorescence in the UV/Vis/IR region. In these cases, no extra chemical is needed to generate the analytical signal. The detection can be a traditional spectrometer or coupled with fiber optics in those regions. Sensors have been developed for the detection of CO, C02 NOx, S02, H2S, NH3, non-saturated hydrocarbons, as well as solvent vapors in air using IR or NIR absorptions, or for the detection of indicator concentrations in the UV/ Vis region and fluorophores such as quinine, fluorescein, etc. 

E. Wang, L. Zhu, L. Ma and H. Patel, Optical sensors for sodium, potassium and ammonium ions based on lipophilic fluorescein anionic dye and neutral carriers, Anal. Chim. Acta, 357 (1997) 85-90. 

Various pH sensors have been built with a fluorescent pH indicator (fluorescein, eosin Y, pyranine, 4-methylumbelliferone, SNARF, carboxy-SNAFL) immobilized at the tip of an optical fiber. The response of a pH sensor corresponds to the titration curve of the indicator, which has a sigmoidal shape with an inflection point for pH = pK , but it should be emphasized that the effective pKa value can be strongly influenced by the physical and chemical properties of the matrix in which the indicator is entrapped (or of the surface on which it is immobilized) without forgetting the dependence on temperature and ionic strength. In solution, the dynamic range is restricted to approximately two pH units, whereas it can be significantly extended (up to four units) when the indicator is immobilized in a microhetero-geneous microenvironment (e.g. a sol-gel matrix). 

Fluorescence resonance energy transfer has also been used for ionic strength measurements.(95) Fluorescein labeled dextran (donor) and polyethyleneimine-Texas Red (acceptor) were placed behind a dialysis membrane. The polymer association is ionic strength dependent and the ratio of intensities (F o/Fw) was used as the measured parameter. Since both the donor and acceptor are fluorescent, this kind of sensor may allow expand the sensitive ionic strength range by shifts in observation wavelength, as was discussed for pH probe Carboxy SNAFL-2 (see Section 10.3). 

Some analytes, such as riboflavin (vitamin B2)16 and polycyclic aromatic compounds (an important class of carcinogens), are naturally fluorescent and can be analyzed directly. Most compounds are not luminescent. However, coupling to a fluorescent moiety provides a route to sensitive analyses. Fluorescein is a strongly fluorescent compound that can be coupled to many molecules for analytical purposes. Fluorescent labeling of fingerprints is a powerful tool in forensic analysis.17 Sensor molecules whose luminescence responds selectively to a variety of simple cations and anions are available.18 Ca2+ can be measured from the fluorescence of a complex it forms with a derivative of fluorescein called calcein. 

Applications of fiber-optic pH sensors in environmental analysis, biomedical research, medical monitoring, and industrial process control have been reviewed by Lin [67]. A multitude of luminescent systems for pH monitoring are commercially available, mostly under special trademarks. Pyrene [68-70], coumarin, bromothymol blue [71] and fluorescein [72-74] derivatives are typical examples that have been used in research in the past two decades. Carboxyfluorescein derivatives have been directly applied to skin tissue samples for the lifetime imaging of pH gradients in the extracellular matrix of the epidermis [75]. Two-photon excitation microscopy became an estab-... 

Ping et al. have fabricated an integrated microsensor array on a silicon wafer for pH imaging [89]. Six different pH-sensitive colorimetric dyes (methyl violet 6B, phenolic red, alizarin complexone, 5-carboxy-fluorescein, alizarin red and methylthymol blue) were used to cover the whole pH range. The dyes were adsorbed on microbeads and placed in etched microwells on the silicon wafer. The indicator array was also used as a cation sensor chip (see Sect. 2.4). 

The PAH polymeric layer played an important role in our fluorescence sensor design. First, its positive charges enabled the deposition of anionic dextran that was labeled with the pH indicator fluorescein on the surface of the nanoparticles. More importantly, the PAH polymeric layer separated between the fluorescein molecules and the metal particle. In fact, the thickness of the polymeric layer was over 10 nm, which is larger than the Forster distance required for efficient energy transfer between the fluorophore and the metallic gold particles. 

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