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This time I want to ask another question about the dye, sodium
fluorescein. This dye can be used in the in vivo experiment to trace the
diffusion of the cerebrospinal fluid. But before we start any in vivo
experiment, we test the excitation and emission peak in spectrofluorameter
(Pekin Elmer LS50B). We found a very different excitation but the same
emission wavelength in compare with the other literatures. (The others use
around 490nm, Sigma suggestion is 460nm and our result is 390!) The
emission wavelength is the same: 520nm. I found this difference is very
suprise. Can anyone give me a reason why I get a different excitation
peak. (The spectrofluorameter lamp is xenon lamp.)
The second questions is whether there is anyone have experience
using this dye. Will this dye go into the cell? The fluorescein part, in
principle, should not go into the cell because of its hydrophobic
structure. But when this part is with sodium together, this will make this
fluorescein part to be hydrophilic. From literature search, some say they
will go into the cell but say not. From our hand, we get a little signal
when the dye is incubated with the cells. Can anyone give me some suggestion?
Thank you very much.
Raymond, Chuen-Chung CHANG
(DAAD Research Fellow)
Institute for Surgical Research,
Klinikum Grosshadern,
University of Munich,
D-81366 Munich,
Germany
Fax:+49-89-7095-8897
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