Media-Agarose Slides: A Mounting Method for Observation of Living Cells

Extract

Often during light and fluorescent microscopy, one wishes to observe samples of cells (e.g., yeast) grown in liquid culture. This is sometimes difficult as the cells can move around under the coverslip and a wet mount can dry out, killing the cells. Protocols in which the cells are resuspended in molten agarose results in cells that are in many different focal planes and often stressed by the high temperature of the molten agarose. This protocol describes a method of depositing small pads of growth media containing agarose onto slides, which eliminates these problems. Choice of growth media depends upon both the growth requirements of the cells and the requirements of the microscopy method. For example, yeast synthetic media works better for fluorescent microscopy than yeast rich media, because the yeast extract in rich media is autofluorescent in the green (fluorescein/GFP) channel.