Metamorph v6

Log phase yeast culture cells were spotted on a glass slide and imaged with an Olympus BX-51 microscope equipped with Infinity software v.5.0.3 (Lumenera, Ottawa, Canada) for image acquisition. Alternatively, live cells were imaged using a laser scanning confocal microscope (Zeiss LSM410). MetaMorph v6.1 software (Universal Imaging Corporation; Downington, PA) was used to construct calibrated overlays of z-stacked images. Worms were mounted on 2% agarose pads spotted with 20 μl of 10 mM Na-azide. Confocal microscopy was performed using a Zeiss LSM410 confocal laser scanning microscope. Z-stacks were acquired and processed using the Zeiss LSM software package. Alternatively, imaging was performed using an Applied Precision DeltaVision microscope (GE). Image processing was performed using Softworx software (G.E./Applied Precision).

Yeast cells were spotted on a glass slide and imaged with an Olympus BX-51 microscope equipped with Infinity software v.5.0.3 (Lumenera, Ottawa, Canada) for image acquisition. Alternatively, mounted cells were imaged using a laser scanning confocal microscope (Zeiss LSM 510). MetaMorph v6.1 software (Universal Imaging Corporation; Downington, PA) was used to reconstruct 3D images from the calibrated overlays of the z-stacks. Emission spectra were obtained by scanning sample fluorescence intensity over an emission wavelength of 400-700 nm while applying a constant excitation wavelength of 495 nm (Preston et al. 1989 (link)). To image old cells, calcofluor-white, which stains chitin in bud scars was used (Bartholomew and Mittwer 1953 (link); Cabib and Bowers 1971 (link); Smeal et al. 1996 (link)).