Visualization of apical membrane domains in Aspergillus nidulans by Photoactivated Localization Microscopy (PALM)

Norio Takeshita, Yuji Ishitsuka, Yiming Li, Ulrich Nienhaus, Reinhard Fischer

Author address: 

Dept. of Microbiology, Karlsruhe Institute of Technology, Karlsruhe, Germany

Abstract: 

Apical sterol-rich plasma membrane domains (SRDs), which can be viewed using the sterol-binding fluorescent dye filipin, are gaining attention for their important roles in polarized growth of filamentous fungi. The size of SRDs is around a few mm, whereas the size of lipid rafts ranges in general between 10-200 nm. In recent years, super-resolution microscope techniques have been improving and breaking the diffraction limit of conventional light microscopy whose resolution limit is 250 nm. In this method, a lateral image resolution as high as 20 nm will be a powerful tool to investigate membrane microdomains. To investigate deeply the relation of lipid membrane domains and protein localization, the distribution of microdomains in SRDs were analyzed by super-resolution microscope technique, Photoactivated Localization Microscopy (PALM). Membrane domains were visualized by each marker protein tagged with photoconvertible fluorescent protein mEosFP for PALM. Size, number, distribution and dynamics of membrane domains, and dynamics of single molecules were investigated. Time-laps analysis revealed the dynamic behavior of exocytosis.
2013

abstract No: 

N/A

Full conference title: 

27th Fungal Genetics Conference
    • Fungal Genetics Conference 27th (2013)