Fluorescence staining techniques

During the past two decades, cell-biological and biomedical research has greatly benefited from innovations in fluorescence microscopy. Both the increase in the repertoire of fluorescence staining techniques at the (sub)cellular level and the development of a multitude of novel fluorescence microscopy techniques contributed significantly. 

The search for more rapid and sensitive methods of protein detection after electrophoresis led to the development of fluorescent staining techniques. Two commonly used fluorescent reagents are fluorescamine and anilinonaphthalene sulfonate. New dyes based on silver salts (silver diamine or silver-tungstosilicic acid complex) have been developed for protein staining. They are 10 to 100 times more sensitive than Coomassie Blue (Fig. 4.7). 

From the species of the first triad only D. subobscura is distributed all over Europe. D. guanche is endemic to Canary Islands, D. madeirensis to the island of Madeira. Morphological discrimination is extremmely difficult and intercrosses between D. madeirensis with D. guanche or D. madeirensis with D. subobscura (Krimbas and Loukas, 1984) are possible. By use of molecular, and by specific fluorescent staining techniques of mitotic chromosomes, however, a marked difference with respect to the composition of satellite DNA could be detected. 

Pernis et al. (41) found, by fluorescence staining techniques, that an appreciable fraction of rabbit lymphoid cells (up to 15%) bear IgM on their surfaces but have IgG in their cytoplasm. In a heterozygous rabbit the surface IgM and internal IgG of a single cell are of the same allotype (a and b loci) (42). This supports the conclusion that the surface IgM is synthesized by the cell and is not absorbed from surrounding fluids. The authors concluded that their data are consistent with a switch, within a single cell, from the biosynthesis of IgM to IgG. 

Clauss, M. Springorum, A. C. Hartung, J. Comparison of different fluorescence and non-fluorescence staining techniques for rapid detection of airborne micro-organisms collected on room temperature vulcanizing (RTV) silicones from generated aerosols and from ambient air. Aerosol Sci. Technol. 2012,46, 818-827. 

Tlie morphology of some bacteria, especially those that form spores, is distinctive enough under the light microscope to have value for identification. This means that differential staining techniques, such as the Gram stain or acid-fast stain, and fluorescence microscopy may help to determine the iden-... 

Abstract Fluorescent molecules have been widely used as biomolecular labels, enzyme substrates, environmental indicators, and cellular stains and thus constitute indispensable tools in chemistry, physics, biology, and medicinal sciences. The large variation in the photophysics of the available fluorophores connected with chemical alterations give fluorescent probe techniques an almost unlimited scope for the detection of specific molecules and the investigation of intermolecular interactions on a molecular scale. 

TRITC has been used in numerous applications involving fluorescence detection, including double-staining techniques with fluorescein-labeled probes (Mossberg and Ericsson, 1990), the synthesis of fluorescently labeled DNA probes (Smith et al., 1985), as a label in homogeneous... 

Texas Red hydrazide is a derivative of Texas Red sulfonyl chloride made by reaction with hydrazine (Invitrogen). The result is a sulfonyl hydrazine group on the No. 5 carbon position of the lower-ring structure of sulforhodamine 101. The intense Texas Red fluorophore has a QY that is inherently higher than either the tetramethylrhodamine or Lissamine rhodamine B derivatives of the basic rhodamine molecule. Texas Red s luminescence is shifted maximally into the red region of the spectrum, and its emission peak only minimally overlaps with that of fluorescein. This makes derivatives of this fluorescent probe among the best choices of labels for use in double-staining techniques. 

Precise quantifications are an important quality in molecular biology. There are slight differences in the methods used for global and targeted proteomics. In experiments intended to visualize as many proteins as possible, it is highly desirable to have a parallel quantification method that builds on the display technique. For 2D gel electrophoresis, fluorescent staining methods are under development (Urwin and Jackson, 1993), but they still lack overall sensitivity. Labeling proteins with radioactive isotopes is the most precise method for quantification but is limited to cell cultures, and alternatives are desirable. Recently, a precise method... 

TRITC has been used in numerous applications involving fluorescence detection, including double-staining techniques with fluorescein-labeled probes (Mossberg and... 

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