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Fluorescence microscopy

LM László Mózsik
MH Mirthe Hoekzema
NK Niels A. W. de Kok
RB Roel A. L. Bovenberg
YN Yvonne Nygård
AD Arnold J. M. Driessen
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For visualization of DsRed-SKL fluorescent protein, liquid shake-flask cultures were cultivated for 5 days in SMP, and mycelium was collected and re-suspended in phosphate-buffered saline (58 mM Na2HPO4; 17 mM NaH2PO4; 68 mM NaCl, pH 7.3). Confocal imaging was performed on a Carl Zeiss LSM800 confocal microscope using 20 × objective and ZEN 2009 software (Carl Zeiss, Oberkochen, Germany). The DsRed signal was visualized by excitation with a 543 nm helium neon laser (Lasos Lasertechnik, Jena, Germany), and emission was detected using a 565 to 615 nm band-pass emission filter60.

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