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).