Methods for studying the yeast vacuole.

摘要: Publisher Summary This chapter describes techniques used in the study of vacuole and focuses on procedures laboratories. The yeast is a large, dynamic organelle that can be detected by fight microscopy. It most prominent when cells are viewed using differential interference-contrast optics. Reliable detection vacuoles vital staining with fluorescent dyes conjunction fluorescence microscopy described chapter. Several methods available for purification vacuole, all which take advantage low buoyant density vacuole. method choice modified version procedure involves enzymatic removal cell walls, osmotic lysis spheroplasts, followed flotation discontinuous Ficoll gradient. Other use polybase induced spheroplasts under isotonic conditions gradient centrifugation. follows has consistently given 10–20% yield contamination other organelles.