Development in the STORM

Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address: .
Developmental Cell (Impact Factor: 9.71). 12/2012; 23(6):1103-10. DOI: 10.1016/j.devcel.2012.10.003
Source: PubMed


The recent invention of superresolution microscopy has brought up much excitement in the biological research community. Here, we focus on stochastic optical reconstruction microscopy/photoactivated localization microscopy (STORM/PALM) to discuss the challenges in applying superresolution microscopy to the study of developmental biology, including tissue imaging, sample preparation artifacts, and image interpretation. We also summarize new opportunities that superresolution microscopy could bring to the field of developmental biology.

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    • "The first involves patterned light illumination, allowing superresolution acquisitions by two fundamentally different methods, stimulated emission depletion (STED; Hell, 2007) and structured illumination microscopy (SIM; Gustafsson, 2000). The second interrogates the precision of fluorophore localization and includes stochastic optical reconstruction microscopy (STORM; Kamiyama and Huang, 2012) and photoactivation localization microscopy (PALM; Sengupta et al., 2012). The above regimes differ in translational and axial resolution, and their temporal efficiency depends on the size of the imaged area. "
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    • "These clusters appear at a size that is in the order of the diffraction limit of light microscopy ($200 nm in the imaging plane), which restricts the characterization of size and protein numbers. In order to overcome the resolution limit, we use here single-molecule localization-based super-resolution microscopy, which typically achieves a lateral resolution of $20–30 nm (Kamiyama and Huang, 2012). Individual protein clusters which might overlap in a conventional microscopy image can thereby be discerned (van de Linde et al., 2008). "
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