Fluorochrome

The following guidelines will help in choosing fluorophores. 

Allophycocyanin (APC) 604 (650) 650 (660) Polypeptide with exceptional Fluorescence for cytometry 

3-Aminocouma ri n [1635-31-0] 350 445 Forms fluorescent derivatives via reaction with the 3-NH2 group 

Auramine 0 (basic yellow 2) [2465-27-2] 440 530 Fluorescent stain for bacteria and probe for certain enzymes, e.g. alcohol dehydrogenases 

4-Bromomethyl-7- methoxycoumarin [35231-44-8] 360 (340) 410 (421) Forms fluorescent probes with various compds for TLC/HPLC 

Fluorescamine [38183-12-9] Reacts with various R-NH2 to form fluorescent compds 

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The principle underlying the BAT is that the attachment of the antigen to the IgE present on the surface of the basophil leads to the activation of the basophil and the release of its mediators (histamine, leukotrienes, prostaglandins, etc.) and the expression on its membrane of molecules such as CD63, CD203c or others which are markers of basophil activation. The basophils are identified with monoclonal antibodies marked with fluorochromes and anti-IgE and anti-CD63 receptors [for a complete review, we suggest readers read references 19-22]. 

Immunophenotype The process of identification and quantitation of cellular antigens through fluorochrome-labeled monoclonal antibodies. 

Flame Photometry and Gas Chromatography (CyTerra) -Aerodynamic Particle Size and Shape Analysis (BIRAL) -Flow Cytometry (Luminex, LLNL) -Semiconductor-Based Ultraviolet Light (DARPA) -Polymer Fluorochrome (Echo Technology) -Laser-Induced Breakdown Spectroscopy -Raman Scattering -Infrared Absorption -Terahertz Spectroscopy -UV LIDAR... 

Anderson MT, Baumgarth N, Haugland RP et al (1998) Pairs of violet light excited fluorochromes for flow cytometry analysis. Cytometry 33 435-444... 

The nonfluorescent monofunctional azulenyl squaraine labels NIRQ700 an<3 NIRQ7so with absorption maxima between 600 and 750 nm are capable of quenching a number of NIR fluorochromes. They were specifically developed to extend the spectrum for FRET-based assays [100, 101]. 

Lin Y, Weissleder R, Tung CH (2002) Novel near-infrared cyanine fluorochromes synthesis, properties, and bioconjugation. Bioconjugate Chem 13 605-610... 

Acid-fast staining for Cryptosporidium sp. has recently become important because this parasite is now recognized as a cause of severe diarrhea in immunodeficient patients such as those with AIDS, and it can cause transient diarrhea in immunocompetent individuals. The modified acid-fast stain recommended is similar to that used to stain Nocardia spp. in that it uses milder acid decolorization. A variety of acid-fast and fluorochrome staining procedures have been described for Cryptosporidium spp., and all the procedures appear to work. 

Burns J, Hambridge M, Taylor CR. Intracellular immunoglobulins. A comparative study of three standard tissue processing methods using horseradish peroxidase and fluorochrome conjugates. J. Clin. Pathol. 1974 27 548-557. 

Duijndam, W.A.L., Wiegant, J., Van Duijn, P., and Haaijman, J.J. (1988) A simple method for labeling the carbohydrate moieties of antibodies with fluorochromes. J. Immunol. Meth. 109, 289-290. 

Goding, J.W. (1976) Conjugation of antibodies with fluorochromes Modifications to the standard methods./. Immunol. Meth. 13, 215-226. 

Sippel, T.O. (1981) New fluorochromes for thiols Maleimide and iodoacetamide derivatives of 3-phenyl-coumarin fluorophore./. Histochem. Cytochem. 29, 314. 

The digital data from the flow cytometer are collected in list mode files that contain a record of all of the fluorescent events associated with each individual cell in a sample. For any individual cell, the number of events collected is dependent on the number of fluorochromes utilized in the analysis that bind to the cell, which can be amplified by increasing the variety/number of lasers utilized to excite the fluorochrome. These raw data can then be analyzed on-line or off-line by software provided by the manufacturer of... 

Representative Fluorochromes/Dyes and Applications for Functional Flow Cytometry3... 

Application Fluorochrome/Dye Excitation Wavelength (nm) Emission Wavelength (nm) Comments... 

This chapter offers some select, recent developments regarding fixation, dehydration, and embedding. In addition, some tried-and-true procedures are described for the localizations of cellular and tissue chemicals in stems and roots of young Zea mays seedlings. Also provided are more recently developed fluorochromes for DN A and RNA localizations (18,19). 

DNA Feulgen Acridine orange as a fluorochrome Ethidium bromide as a fluorochrome 4, 6 - diamido-2-phenylindole as a fluorochrome... 

Memhrane-Impermeant Fluorochromes into Living Cells... 

Controls for Membrane Impermeance. Some fluoro-chromes can enter protoplasts via probenecid-sensitive anion channels (19) and control experiments must establish membrane impermeance. Soak the tissues in the fluorochrome at the concentration used for up to 12 h, rinsing and examining the tissues for fluorescence at hourly intervals. Cell viability is assessed by rate of cytoplasmic streaming, or by chloroplast autofluorescence, which declines in dead cells. 

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