Abstract
Four basic approaches have been utilized for isolation of higher plant vacuoles and one specific procedure for each of these is offered here as a broadly tested representative of each of the basic approaches. Criteria for quality of isolated vacuoles may vary according to the intended use of the organelles. Guidelines and contamination markers for assessing purity of preparations are discussed. The remarkable stability-extensibility of tonoplast has made it possible not only to isolate intact vacuoles (largest isolated vacuoles are about 150 μm in diameter yet tonoplast is about 100 Å thick) but also to manipulate them. Exploitation of isolated vacuoles and tonoplast for fundamental studies, especially transport studies, has just begun.
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