Figure 3. Confocal analysis of C. albicans OPC 84 and C. glabrata 90030 after incubation with rhBD-2. C. albicans OPC 84 or C. glabrata 90030 yeast cells were respectively incubated with rhBD-2, at 37°C, for 48 hrs. Cells were then incubated with a staining solution containing Alexa Fluor 488 conjugated Concanavalin A (ConA) (50 µg/mL) (Molecular Probes, Eugene, OR, USA) and FUN^® 1 (20 µM) (Molecular Probes) in PBS, 37°C, 30 min. Stained cells were observed with the use of a dual scanning confocal microscopy system (LSM 510, Zeiss, Oberkochen, Germany). (A) Normal C. albicans cells showing uniform membrane thickness (green boundaries) and normal metabolic activity (red inside yeast cells); (B,C) C. albicans after incubation with rhBD-2, showing a lack of FUN 1 staining within the cells, indicating severe metabolic deficiencies, and evidence of thinning of the cell walls (B, arrows); (C) shows leakage of cytoplasmic material (arrow); (D) C. glabrata 90030 showing uniform membrane thickness (green boundaries) and normal metabolic activity (red inside yeast cells); (E) C. glabrata after incubation with rhBD-2 does not show appreciable differences in either envelope thickness, metabolic activity, or cytoplasmic debris when compared with untreated cells (D).