Lucifer Yellow CH and

Lucifer Yellow CH and Rhodamine dextran (Molecular Probes, Inc., OR). 

Lil) of 0.05% of Lucifer Yellow CH and Rhodamine dextran (mw 10,000) in PBS at room temperature. The dish is swirled to distribute the solution evenly, and then cells in the culture are scraped off the dish with a cell scraper. Cells are immediately resuspended in complete culture medium, rinsed at least twice by centrifugation to remove unincorporated macromolecules, and replated on a new dish containing a culture of cells which are the potential recipients of the dye via gap junction formation. After 1 h of incubation at 37°C, the dish can be examined under an epifluorescence phase microscope at the appropriate wavelengths. 

This fluorophore has an excitation maximum at 428 nm and an emission maximum at 534nm. The extinction coefficient of the molecule in aqueous solution is about 12,000M 1cm 1. Cascade Blue hydrazide and Lucifer Yellow CH derivatives can be excited simultaneously by light of less than 400 nm, resulting in the possibility for two-color detection at 420 and 534 nm. 

Lucifer Yellow CH is soluble in aqueous solution, and it should be stable for awhile if protected from light. The reagent is available as three different salts of the sulfonate groups. The ammonium salt of the fluorophore is soluble to a level of 9 percent in water, while the lithium and potassium salts have a solubility of 5 and 1 percent, respectively. A concentrated stock solution of the fluorophore may be prepared in water and an aliquot added to a buffered reaction medium to facilitate the transfer of small quantities. For aqueous reactions, a pH range of 5-9 will result in efficient hydrazone formation with aldehyde or ketone residues. 

Spiegel, S., Wilchek, M., and Fishman, P.H. (1983) Fluorescent labeling of cell surface glycoconjugates with Lucifer Yellow CH. Biochem. Biophys. Res. Comm. 112, 872-877. 

Lucifer Yellow CH is commonly used as a neuronal tracer by staining cells and then... 

Small water-soluble molecules can also be introduced into cells via stimulation of fluid-phase uptake by the described protocol. Uptake of Lucifer Yellow (m.w. 457.2 Lucifer Yellow CH, dilithium salt from Sigma Chemicals, Rehovot, Israel) has been stimulated by 2.2-fold and 2.7-fold in COS 5-7 and HaCaT cell lines, respectively, as compared with the constitutive uptake. 

Usually, a 4-10% solution of Lucifer Yellow CH in lithium chloride is iontophoreticaUy injected through glass capillaries by passing current (1-3 nA) pulses (50-500 nsec) at 10 or 1 Hz, respectively (Miller and Goodenough, 1985 Schuetze and Good-enough, 1982). Transfer of the dye into the surrounding cells can be observed under a fluorescence microscope about 10 min after the injection. If required, cells can be fixed after dye injection by incubation for at least 20 min in 4% paraformaldehyde in PBS (Ren et al., 1990). 

Klein M, Martinoia E, Weissenbock G (1997) Transport of lucifer yellow CH into plant vacuoles - evidence for direct energization of a sulphonated substance and implications for the design of new molecular probes. FEBS Lett 420 86-92... 

Electrodes were made from two-barrel borosilicate glass tubing with filaments (WPI). One barrel was filled with 3 M KCl (2-10 MU) for stimulation and recording. For microejections, the other barrel was filled with 5% Lucifer Yellow (Lucifer Yellow CH, lithium salt. Molecular Probes) in 150 mM LiCl or with lOmM Alexa fluor 488 (hydrazide, sodium salt. Molecular Probes) in 200 mM KCl. When the double-barrel microelectrode was inserted into a cell, dye ejection was accomplished with 100-msec negative current pulses (1-50 nA) delivered through the dye-filled barrel and applied for 1-5 min at 100-msec intervals. After dye ejection the tissue was fixed. 

Oparka, K. J. Prior, D. A. M. Movement of Lucifer Yellow CH in potato tuber storage tissues a comparison of symplastic and apoplastic transport. 

Reardon PM, Gochoco CH, Audus KL, Wilson G, Smith PL (1993) In vitro nasal transport across ovine mucosa effects of ammonium glycyrrhizinate on electrical properties and permeability of growth hormone releasing peptide, mannitol, and lucifer yellow. Pharm Res 10 553-561. 

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