Screening fluorescence

Du, Y. et al. 2007. High-throughput screening fluorescence polarization assay for tumor-specific Hsp90. J. 

Once phage or scFv which specifically bind the antigen of interest have been identified it may be necessary to cany out further screening tests to assess which antibodies fiom the positive population have the highest affinities. There are a variety of wtq of achieving this including surface plasmon resonance (BlAcore) screening, fluorescence quench measmement, and competition ELISAs. Descriptions of these protocols are beyond the scope of this chapter. 

The basic device is very simple. A tip of refractory metal, such as tungsten, is electrically heat-polished to yield a nearly hemispherical end of about 10" cm radius. A potential of about 10 kV is applied between the tip and a hemispherical fluorescent screen. The field, F, falls off with distance as kr, and if the two radii of curvature are a and b, the total potential difference V is then... 

The final grid is positively charged to accelerate the accepted electrons onto the fluorescent screen. The diffraction pattern may then be photographed. 

Zinc sulfide is used in making luminous dials. X-ray and TV screens, and fluorescent lights. 

The Calcium Halophosphate Phosphors. Early fluorescent lamps used various combinations of naturally occurring fluorescent minerals. The development of the calcium halophosphate phosphor, Ca (P0 2(Cl, F) Sb ", Mn, in the 1940s was a significant breakthrough in fluorescent lighting (7). As is often the case in new phosphor discoveries, this phosphor was found accidentally while searching for phosphors for radar screens. 

Early fluorescent pigments were promoted and adopted for use in screen inks for poster boards and paints for safety applications. These thermoset pigments were not well-suited because of their poor fightfastness. Also, because of their relatively coarse particle size, their use in thinner film applications, such as gravure or flexo, was limited. 

Whereas the eadiest fluorescent-dye pigments would last only 20 days outdoors in a screen-ink film, fade resistance has been improved to such an extent that some modem daylight-fluorescent coated panels stiU have useful color after nine months or mote in Florida sunlight in a 45° exposure tack facing south. The fluorescent layer is usually coated with an acrylic film containing a uv absorber. Indoor-accelerated exposure equipment is, of course, invaluable in the development of such systems. Better dyes and resins very likely will make possible fat mote stable coatings in the future. 

Filtered-Particle Inspection. Solids containing extensive inteiconnected porosity, eg, sintered metallic or fired ceramic bodies formed of particles that ate typically of 0.15-mm (100-mesh) screen size, are not inspectable by normal Hquid penetrant methods. The preferred test medium consists of a suspension of dyed soHd particles, which may be contained in a Hquid vehicle dyed with a different color. Test indications can form wherever suspensions can enter cracks and other discontinuities open to the surface and be absorbed in porous material along interior crack walls. The soHd particles that form test indications ate removed by filtration along the line of the crack at the surface where they form color or fluorescent indications visible under near-ultraviolet light (1,3). 

Direct quantitation of receptor concentrations and dmg—receptor interactions is possible by a variety of techniques, including fluorescence, nmr, and radioligand binding. The last is particularly versatile and has been appHed both to sophisticated receptor quantitation and to dmg screening and discovery protocols (50,51). The use of high specific activity, frequendy pH]- or p lj-labeled, dmgs bound to cmde or purified cellular materials, to whole cells, or to tissue shces, permits the determination not only of dmg—receptor saturation curves, but also of the receptor number, dmg affinity, and association and dissociation kinetics either direcdy or by competition. Complete theoretical and experimental details are available (50,51). 

Includes fluorescent pigments for gravure, flexo, and screen inks. 

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