Dear Colleagues William Smith raised an important point which others are probably better qualified to address than myself. Dr Smith asked 'Are all FITCs the same ?' and then quoted diffences in observed spectral overlap when using FITC conjugated to different molecules viz antibodies, dextran ... In our paper: Flow cytometric quantification of electroporation mediated uptake of macromolecules into plant protoplasts, Protoplasma 186:50-56, 1995, we were able to distinguish between FITC and dextran-conjugated FITC based on yellow/orange discrimination. The spectral properties of fluorescein are dependent upon the miroenvironment, principally pH. It is therefore not surprising, in a simplistic way, that different FITC-labelled probes have characteristic properties. Perhaps some of our more enlightened colleagues could explain Dr Smith's observations. Yours sincerely, Nigel Blackhall, University of Nottingham, UK
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