Sensitivity fluorescence methods

LDH (lactate dehydrogenase Membrane integrity Easy to measure. Fluorescent methods sensitive... 

Note that in liquid phase chromatography there are no detectors that are both sensitive and universal, that is, which respond linearly to solute concentration regardless of its chemical nature. In fact, the refractometer detects all solutes but it is not very sensitive its response depends evidently on the difference in refractive indices between solvent and solute whereas absorption and UV fluorescence methods respond only to aromatics, an advantage in numerous applications. Unfortunately, their coefficient of response (in ultraviolet, absorptivity is the term used) is highly variable among individual components. 

X-Ray Fluorescence analysis (XRF) is a well-established instrumental technique for quantitative analysis of the composition of solids. It is basically a bulk evaluation method, its analytical depth being determined by the penetration depth of the impinging X-ray radiation and the escape depth of the characteristic fluorescence quanta. Sensitivities in the ppma range are obtained, and the analysis of the emitted radiation is mosdy performed using crystal spectrometers, i.e., by wavelength-dispersive spectroscopy. XRF is applied to a wide range of materials, among them metals, alloys, minerals, and ceramics. 

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... 

It is instructive to compare the sensitivity which may be achieved by absorption and fluorescence methods. The overall precision with which absorbance can be measured is certainly not better than 0.001 units using a 1 cm cell. Since for most molecules the value of emax is rarely greater than 105, then on the basis of the Beer-Lambert Law the minimum detectable concentration is given by cmin> 10 3/105= 10 8M. 

The sensitivity of the fluorescence methods varies considerably with the instrument used. Advances in modern instrumentation and the power of today s computers allow for a much improved sensitivity. Using commercially available instruments and modern computers equipped with appropriate software, detection limits down to 10 pg of calf thymus DNA can be achieved using ethidium bromide. (We have achieved such levels using several Perkin-Elmer MPF66 Instruments at various locations.)... 

Applications of the oxalate-hydrogen peroxide chemiluminescence-based and fluorescence-based assays with NDA/CN derivatives to the analysis of amino acids and peptides are included. The sensitivity of the chemiluminescence and fluorescence methods is compared for several analytes. In general, peroxyoxalate chemiluminescence-based methods are 10 to 100 times more sensitive than their fluorescence-based counterparts. The chief limitation of chemiluminescence is that chemical excitation of the fluorophore apparently depends on its structure and oxidation potential. 

Franco, C. M., Fente, C. A., Vazquez, B., Cepeda, A., Lallaoui, L., Prognon, P., and Mahuzier, G., Simple and sensitive high-performance liquid chromatography-fluorescence method for the determination of citrinin. Application to the analysis of fungal cultures and cheese extracts, /. Chromatogr. A, 723, 69, 1996. 

The Af-dansylated amino acid (e.g., glycine, leucine, proline) exhibits a yellow fluorescence. The sensitivity of detection for amino acids by this method is about 10 9 mol of amino acid. The advantage of this prodecure in comparison with that using dansyl chloride is the fact that it can be carried out in homogenous aqueous solution without addition of a cosolvent. 191... 

Most studies of the physical binding of hydrocarbon metabolites and metabolite model compounds have measured the effect of DNA binding on hydrocarbon fluorescence intensities, fluorescence lifetimes and UV absorption spectra Radioactive labelling has also been used, but less frequently. Spectroscopic methods are particularly convenient. These methods, especially fluorescence methods, are also very sensitive. All of the hydrocarbons in Figure 1 except the epoxides have high fluorescence quantum yields, which permit routine detection in the 10 -10 7 M concentration range. 

In the determination of carbohydrates, sensitivity can often be increased by using fluorescence rather than absorbance for the final determination. With compounds that are not normally fluorescent, it becomes necessary to find fluorescent derivatives. Hirayama [160] concentrated the carbohydrates in coastal water samples, using electrodialysis and evaporation, and made fluorescent derivatives using anthrone and 5-hydroxyl-1-tetralone, determining pentoses separately from hexoses in the process. While this method does seem to have the extra sensitivity expected from fluorescent methods, the extra manipulations render it unsatisfactory for routine use. 

Over the course of the last three decades, a variety of techniques have been used to characterize the circuitry of serotonergic neurons in the central nervous system. The density of serotonergic innervation in the forebrain was initially underestimated because the original histo-fluorescence method was limited in sensitivity and did not permit the detection of many fine axons and terminals. 

The detection of flu viruses via a fluorescent sandwich immunoassay was reported by Bucher.(10) However, the method sensitivity was too low for direct detection of the virus. A novel sandwich immunoassay was described by Ogcr((lff7 for the detection of Botulinum Toxin A. Antibodies specific for Clostridium botulinum were covalently attached to the surface of a tapered fiber. After the capture of the antigen, a sandwich was formed with a rhodamine-labeled anti-toxin IgG, and the evanescent wave was measured. The assay was highly specific with detection limits near 5 ppb. 

An ideal sensor recognizes analytes in a sensitive, selective, and reversible manner. This recognition is in turn reported as a clear response. In recent years, conducting polymers have emerged as practical and viable transducers for translating analyte-receptor and nonspecific interactions into observable signals. Transduction schemes include electronic sensors using conductometric and potentiometric methods and optical sensors based on colorimetric and fluorescence methods [1]. 

Silver metal and its contents in silver alloys and salts can be measured at trace levels by various instrumental techniques such as flame- and furnace-AA, ICP-AES, ICP/MS and x-ray fluorescence methods. It is solubilized by digestion with nitric acid prior to analysis. The AA measurement may be carried out at the wavelength 328.1 nm and ICP analysis at 328.07 nm. ICP/MS is the most sensitive technique while x-ray fluorescence is relatively less sen-... 

The sensitivity of fluorescence methods is generally 10 to 10 times more than the sensitivity of absorption methods. 

Rapsomanikis, S., M. Wake, A.-M. N. Kitto, and R. M. Harrison, Analysis of Atmospheric Ammonia and Particulate Ammonium by a Sensitive Fluorescence Method, Environ. Sci. Technol., 22, 948-952 (1988). 

We demonstrate the Mil method, which couples the sensitivity of multiphoton excitation on the spectral phase of the laser pulses to probe microscopic chemical environment-induced changes in the multiphoton excitation spectrum of sensitive reporter molecules. We carry out the optimization of the required phase functions in solution and provide theoretical simulations. We show experimental images whereby pH-selective two-photon microscopy is achieved and demonstrate how selective excitation can be used to enhance contrast and, consequently, to achieve functional imaging, using fluorescent probes sensitive to changes in their local environment. 

The main advantage of fluorescence methods is their high sensitivity. abouL one part in It)8, in many determinations both inorganic and organic. This is two or three orders of magnilude belter than absorption melhods. where the sensitivity is limited by the necessity of delecting a very small fractional decrease in the light transmitted by the solution. 

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