Live cell imaging was performed to monitor phagosomal maturation by by using a Zeiss Axio Observer inverted microscope (Carl Zeiss Inc., Thornwood, NY) equipped with a confocal laser system housed in the Duke University Light Microscope Core Facility (LMCF) or by using an UltraVIEW Vox live cell imaging system equipped with a CSU-1 spinning disk scanner, housed in the Institute of Medical Sciences at the University of Aberdeen. Images obtained were analyzed and processed with the Zeiss LSM 510 version 4.2 (Carl Zeiss Inc., Thornwood, NY) or with the Volocity software (PerkinElmer), repectively. Phagosome maturation was monitored by measuring cathepsin B activity by employing the Magic Red Cathepsin B assay kit (ImmunoChemistry) following the manufacturer’s instructions. Hoechst 33342 was used to stain macrophage nuclei.
Time lapse analyses for hyphal growth of wild-type and cnaBΔ mutants were conducted by using a Zeiss Axio Observer Z1 microscope system (Carl Zeiss Inc., Thornwood, NY) with an Opto-electronically motorized XY stage, Pecon XL S1 incubator, and Coolsnap ES2 high-resolution CCD camera, housed in the Duke University Light Microscopy Core Facility (LMCF).
Transmission electron microscopy for spores, wild-type hyphae, wild-type yeast, and cnbRΔ yeast-locked mutants, was performed as described previously (Li et al., 2011 (link)).
Time lapse analyses for hyphal growth of wild-type and cnaBΔ mutants were conducted by using a Zeiss Axio Observer Z1 microscope system (Carl Zeiss Inc., Thornwood, NY) with an Opto-electronically motorized XY stage, Pecon XL S1 incubator, and Coolsnap ES2 high-resolution CCD camera, housed in the Duke University Light Microscopy Core Facility (LMCF).
Transmission electron microscopy for spores, wild-type hyphae, wild-type yeast, and cnbRΔ yeast-locked mutants, was performed as described previously (Li et al., 2011 (link)).