Abstract
Yeast cells are well suited to visualizing organelles by 4D confocal microscopy. Typically, one or more cellular compartments are labeled with a fluorescent protein or dye, and a stack of confocal sections spanning the entire cell volume is captured every few seconds. Under appropriate conditions, organelle dynamics can be observed for many minutes with only limited photobleaching. Images are captured at a relatively low signal-to-noise ratio and are subsequently processed to generate movies that can be analyzed and quantified. Here, we describe methods for acquiring and processing 4D data using conventional scanning confocal microscopy.
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1 Electronic Supplementary Material
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4D movie of yeast Golgi compartments before deconvolution. See the legend to Fig. 1 (MOV 1139 kb)
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Day, K.J., Papanikou, E., Glick, B.S. (2016). 4D Confocal Imaging of Yeast Organelles. In: Brown, W. (eds) The Golgi Complex. Methods in Molecular Biology, vol 1496. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6463-5_1
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DOI: https://doi.org/10.1007/978-1-4939-6463-5_1
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6461-1
Online ISBN: 978-1-4939-6463-5
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