Fluorescence sensitive detection

Dansyl-amino acid (fluorescence-sensitive detection)... 

Liquid scintillation counting is by far the most common method of detection and quantitation of -emission (12). This technique involves the conversion of the emitted P-radiation into light by a solution of a mixture of fluorescent materials or fluors, called the Hquid scintillation cocktail. The sensitive detection of this light is affected by a pair of matched photomultiplier tubes (see Photodetectors) in the dark chamber. This signal is amplified, measured, and recorded by the Hquid scintillation counter. Efficiencies of detection are typically 25—60% for tritium >90% for and P and... 

For an analyte of molecular weight 5000 and good chromatographic conditions, most photometric detectors can be expected to provide detection limits of 2—5 ng. Improvement into the mid-picogram or lower range normally requires the use of more sensitive detection means such as fluorescence or electrochemical detectors. 

More sensitive detection methods and more objective recording methods (e g the employment of scanners) are constantly been striven for m order to overcome this illusion It IS for this reason too that fluorescent methods have been introduced to an increasing extent on account of their higher detection sensitivity This allows an appreciable reduction in the amount of sample applied, so that possible interfering substances are also present m smaller quantibes This increases the quality of the chromatographic separation and the subsequent m situ analysis... 

The formation of a microphase structure can be sensitively detected by using hydrophobic fluorescent probes. Hydrophobic microdomains tend to solubilize hydrophobic small molecules present together in aqueous solution. For example, diphenylhexatriene (DHT) is hydrophobically bound to the St aggregates in ASt-x in aqueous solution and, as a result, the fluorescence intensity is greatly enhanced. Figure 9 shows the fluorescence intensity of DHT in the presence of ASt-x relative to the intensity in its absence (I/I0) as a function of the ASt-x concentration [29],... 

If the signal decay is a single-exponential curve, equations 16 and 17 result in values for X that are in agreement with each other. Dissimilar values indicate multiexponential decay, which usually means that the sample contains more than one fluorophore. Multiexponential decay can be resolved by using a phase fluorometer with phase sensitive detection. A time-independent, direct-current signal is produced that is proportional to the cosine of the difference between the phase angle of the detector ( D) and the phase angle of the fluorescence ( ) ... 

The simplest fluorescence measurement is that of intensity of emission, and most on-line detectors are restricted to this capability. Fluorescence, however, has been used to measure a number of molecular properties. Shifts in the fluorescence spectrum may indicate changes in the hydrophobicity of the fluorophore environment. The lifetime of a fluorescent state is often related to the mobility of the fluorophore. If a polarized light source is used, the emitted light may retain some degree of polarization. If the molecular rotation is far faster than the lifetime of the excited state, all polarization will be lost. If rotation is slow, however, some polarization may be retained. The polarization can be related to the rate of macromolecular tumbling, which, in turn, is related to the molecular size. Time-resolved and polarized fluorescence detectors require special excitation systems and highly sensitive detection systems and have not been commonly adapted for on-line use. 

Yamauchi, S., Nakai, C., Nimura, N., Kinoshita, T., and Hanai, T., Development of a highly sensitive fluorescence reaction detection system for liquid chromatographic analysis of reducing carbohydrates, Analyst, 118, 773,1993. 

The detection of the migrating sample boundary in CE can be accomplished by UV, fluorescent, electrochemical, radiochemical, conductivity, and mass spectrometry (MS) means. The use of high-sensitivity detection systems is always a key issue in CE applications. The sensitivity of HPCE detectors may be at least 2 to 3 orders of magnitude better than that of HPLC detectors. Since the detection cell volume is very small, the concentration sensitivity... 

Mass spectrometry (MS) is also being used to add another dimension of analysis to achiral-chiral analysis. Recently, an achiral-chiral column-switching LC/LC-MS/MS method was reported for the pindolol enantiomers in human serum (Motoyama et al., 2002) and phenprocoumon metabolites (Kammerer et al., 1998). For analytes that have very poor chromophores or cannot naturally fluoresce, MS detection can be more sensitive for the underivatized form of the analyte. Also, MS detection can be particularly useful when very similar analytes that differ in mass (such as some amino acids and metabolites) cannot be satisfactorily separated chromatographically,... 

The chiral amines (55) were first derivatized by conventional reaction with fluorescene-isothio-cyanate (57) leading to fluorescein-active compounds (58) (Equation (9)) that enable a sensitive detection system, specifically laser-induced fluorescence detection (LIF), to be used.90 Although... 

Goedhart, J., Vermeer, J. E., Adjobo-Hermans, M. J., van Weeren, L. and Gadella, T. W., Jr. (2007). Sensitive detection of p65 homodimers using red-shifted and fluorescent protein-based FRET couples. PLoS ONE 2, elOll. 

When cells are suspended in a biological fluid or culture medium, both serum proteins and cells interact with the surface substrate. Serum protein adsorption behavior on SAMs has been examined with various analytical methods, including SPR [58-61], ellipsometry [13, 62, 63], and quartz QCM [64—66]. These methods allow in situ, highly sensitive detection of protein adsorption without any fluorescence or radioisotope labeling. SPR and QCM are compatible with SAMs that comprise alkanethiols. In our laboratory, we employed SPR to monitor protein adsorption on SAMs. 

Figure 7.21 Dendrimers that are fluorescently labeled as well as biotinylated create enhanced detection reagents for use in (strept)avidin-biotin-based assays. Large complexes containing multiple fluorescent dendrimers can bind to antigens and form a highly sensitive detection system that exceeds the detection capability of fluorescently labeled antibodies.

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