ÀÎ DISTRIBUTION AND FLUORESCENCE SPECTRA IN MYOBLASTS AND SINGLE MUSCLE FIBRES
T. N. Belyaeva, S. A. Krolenko, E. A. Leontieva, T. P. Mozhenok, A. V. Salova, M. D. Faddejeva
Institute of Cytology RAS, St. Petersburg;
e-mail: tbel@mail.cytspb.rssi.ru
Using spectral scanning regime of Leica TCS SL confocal microscope, acridine orange (ÀÎ) fluorescence spectra in nuclei and cytoplasms of living myoblasts
L6J1 and frog single muscle fibres have been studied. ÀÎ fluorescence spectra in salt solutions dependent on free ÀÎ concentrations and in ÀÎ complexes with
DNA have also been obtained for comparison. Myoblasts nuclei fluoresced in green spectral region with maximum at ~530 nm (corresponding ÀÎ monomers
fluorescence), nucleoli fluoresced most brightly. Nuclear chromatin fluoresced not uniformly in these cells. We saw similar to myoblasts ÀÎ emission in nucleoli
and nuclei of frog single muscle fibres. The uniformed weak green fluorescence was observed for myoblast cytoplasm. As to the muscle fibres sarcoplasm, we saw
also ÀÎ green fluorescence in A-discs. In myoblasts and muscle fibre cytoplasm we saw the fluorescent red, yellow and green granules which were acidic organelles.
The comparison of ÀÎ fluorescence spectra in living cells with fluorescence spectra of different ÀÎ concentrations and complexes of ÀÎ with DNA in buffer solutions
allows estimation of ÀÎ concentration in acidic granules which is of interest in the investigation of cellular organelles functions in the processes of intracellular
transport, adaptation, apoptosis and a number of pathological conditions.
Key words: acridine orange, acid organelles, confocal microscopy, fluorescence spectra
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