Molecular mapping of neuronal architecture using STORM microscopy and new fluorescent probes for SMLM imaging

Molecular mapping of neuronal architecture using STORM microscopy and new fluorescent probes for SMLM imaging

Tools and methodologies that enable precise imaging of neuronal architecture are in great demand as scientists seek to effectively analyze the structure of synapses and their associated proteins.  Recently the Danglot group reported on new software in combination with membrane labeling probes to detect pre and postsynaptic proteins.  The statistical object distance analysis (SODA) software effectively analyzes cellular shape and spot density and was able to detect distances between two synapse-associated proteins synapsin and PSD95.  Importantly, the SODA software was even used to perform 3D analysis, making it suitable for use with thick 3D volume STORM images.  The software's primary limitation is the need to properly identify the cell boundary; thus, the group used SODA in combination with the fluorgenic membrane probe called Membright.  The group highlighted several important features of the probe that make it an excellent tool for live and fixed cell labeling and even highlighted important conditions and factors to optimize membrane labeling with this tool.  They described specific uses for the membright probe, including extracellular vesicle labeling, and important points to consider when using the probe for this application.  Finally, information about the probe’s use in single-molecule localization microscopy was discussed.  Overall, they’ve identified a great combination of tools to investigate a nanometric molecular map of proteins that can be used in neuronal cells as well as other cell models.  Cytoskeleton Inc.’s MemGlow™ 590 (Cat. # MG03) is the Cy3.5-Mem bright probe that was highlighted throughout these studies.