Development in the STORM
ABSTRACT 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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ABSTRACT: The structure of dendritic spines determines synaptic efficacy, a plastic process that mediates information processing in the vertebrate nervous system. Aberrant spine morphology, including alterations in shape, size, and number, are common in different brain diseases. Because of this, accurate and unbiased characterization of dendritic spine structure is vital to our ability to explore and understand their involvement in neuronal development, synaptic plasticity, and synaptic failure in neurological diseases. Investigators have attempted to elucidate the precise structure and function of dendritic spines for more than a hundred years, but their fundamental role in synaptic plasticity and neurological diseases remains elusive. Limitations and ambiguities in imaging techniques have exacerbated the challenges of acquiring accurate information about spines and spine features. However, recent advancements in molecular biology, protein engineering, immuno-labelling techniques, and the use of super-resolution nano-microscopy along with powerful image analysis software have provided a better understanding of dendritic spine architecture. Here we describe the pros and cons of the classical staining techniques used to study spine morphology, and the alteration of dendritic spines in various neuropathological conditions. Finally, we highlight recent advances in super-resolved nanoscale microscopy, and their potentials and pitfalls when used to explore dendritic spine dynamics. Copyright © 2015. Published by Elsevier Ireland Ltd.Neuroscience Research 02/2015; DOI:10.1016/j.neures.2015.02.007 · 2.15 Impact Factor
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ABSTRACT: A discussion is presented of the process of looking into science, particularly microscopy, whilst being concerned with its representation and its public understanding and dissemination in the context of curating exhibitions. Historical examples explored include the studies of the fruit fly, Drosophila, and the works of noted scientists including Mendel, Grew, Leeuwenhoek, Hooke, Baker, Lister and Quekett. How the development of modern equipment and techniques such as scanning electron microscopy led to advances in histology and physiology is examined. The current importance of medical imaging techniques such as positron emission tomography (PET) and computerized tomography (CT) scanning is discussed. A particular mention is made of the artistic works of Kessler based on electron microscopy.Interdisciplinary Science Reviews 06/2014; 39(2):187-199. DOI:10.1179/0308018814Z.00000000083 · 0.38 Impact Factor
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ABSTRACT: Single molecule localization microscopy (SMLM) techniques allow for sub-diffraction imaging with spatial resolutions better than 10 nm reported. Much has been discussed relating to different variations of SMLM and all-inclusive microscopes can now be purchased, removing the need for in-house software or hardware development. However, little discussion has occurred examining the reliability and quality of the images being produced, as well as the potential for overlooked preparative artifacts. As a result of the up to an order-of-magnitude improvement in spatial resolution, substantially more detail is observed, including changes in distribution and ultrastructure caused by the many steps required to fix, permeabilize, and stain a sample. Here we systematically investigate many of these steps including different fixatives, fixative concentration, permeabilization concentration and timing, antibody concentration, and buffering. We present three well-optimized fixation protocols for staining microtubules, mitochondria and actin in a mammalian cell line and then discuss various artifacts in relation to images obtained from samples prepared using the protocols. The potential for such errors to go undetected in SMLM images and the complications in defining a 'good' image using previous parameters applied to confocal microscopy are also discussed.Scientific Reports 01/2015; 5:7924. DOI:10.1038/srep07924 · 5.08 Impact Factor