Computational protocol: Convoluted Plasma Membrane Domains in the Green Alga Chara are Depleted of Microtubules and Actin Filaments

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Protocol publication

[…] The confocal laser scanning microscopes used in this study were a Leica TCS SP5 coupled to a DMI 6000B inverted microscope and a Zeiss LSM 510 coupled to a Zeiss Axiovert inverted microscope. All images included in this study are single optical sections and are positioned with vertical sides parallel to the long axes of the cells. For comparison of the distribution of microtubules or actin filaments with that of charasomes, Z-stacks of the cortical cytoplasm were taken using a ×40 water immersion objective with a numerical aperture of 1.2 or a ×63 oil immersion objective with a numerical aperture of 1.4. Usually, single optical sections with a thickness of 1.2 μm were sufficient for the analysis of the cytoskeleton at the cytoplasmic surface of the charasomes and the adjacent smooth plasma membrane areas. If charasomes were thicker than 1.2 μm, a projection of two optical sections was necessary to visualize all cytoskeletal elements in the peripheral cell cortex. Statistical analysis was performed using Image J (http://imagej.nih.gov/ij) and SigmaPlot (Systat Software). […]

Pipeline specifications

Software tools ImageJ, SigmaPlot
Applications Miscellaneous, Laser scanning microscopy, Microscopic phenotype analysis
Chemicals Carbon, Hydrogen, Cytochalasin D, Paclitaxel