Hbo 100 microscope illuminating system

Aspergillus fumigatus spores (1 × 10⁵) were inoculated onto plastic coverslips in 24 well plates and grown in the presence of analogs overnight at 37°C in RPMI buffered with MOPS pH 7.0. After 24 h, plastic coverslips were washed once in PBS and stained. Calcofluor (1 mg/ml) and 10% (w/v) potassium hydroxide were added drop-wise to coverslips, washed in PBS, mounted, and viewed. Concanavalin A (Con A) and FUN-1 were added at 50 μg/ml in 1 ml PBS and incubated at 37°C for 30 min. All imaging was carried out on a Zeiss LSM5 confocal microscope. Confocal images were recorded under a bright field lens using ×20 objective magnification. Filter cubes facilitating fluorescent imaging were used to record images for Calcofluor at Abs_405nm, Con A at Abs_488nm and FUN-1 at Abs_543nm. All images were captured using the Zeiss HBO-100 microscope illuminating system, processed using the Zen AIM application imaging program and converted to JPG using Axiovision 40 Ver. 4.6.3.0.

BY4741 wild-type or cdc55Δ strain expressing Msn2p-GFP was pre-cultured in YPD medium overnight at 30 °C, inoculated into YPD20 medium at a final OD₆₀₀ of 0.1, and further incubated at 30 °C without shaking for 24 h. Collected cells were immediately observed under a fluorescent microscope Axiovert 200 M (Carl Zeiss) for GFP signal. Images were captured with a HBO 100 Microscope Illuminating System (Carl Zeiss) digital camera.