Preparation of Yeast Cells for Confocal Microscopy

Confocal scanning microscopy has been successfully used for immunofluorescence work in yeast. The major axis of a Saccharomyces cerevisiae haploid cell is approx 4 μm. Optical sections of approximately 2� 0.4 μm thickness from fluorescently-labeled yeast cells can be obtained using the laser scanning confocal microscope (1 ). This means that it is possible to look at optical sections corresponding to about one tenth of a yeast cell. For example, confocal scanning microscopy has been used to examine the distribution of actin in fixed cells prepared from reverting protoplasts, and to show that a monoclonal antibody raised to rat liver nuclear proteins recognized two protein components of the yeast nuclear pore complex, p95 and p110 (2 ,3 ).