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    Mapping sub-cellular protein aggregates and lipid inclusions using synchrotron ATR-FTIR microspectroscopy

    Access Status
    Fulltext not available
    Authors
    Hartnell, David
    Hollings, Ashley
    Ranieri, Anna Maria
    Lamichhane, Hum Bahadur
    Becker, Thomas
    Sylvain, Nicole J.
    Hou, Huishu
    Pushie, M.J.
    Watkin, Elizabeth
    Bambery, K.R.
    Tobin, M.J.
    Kelly, Michael E.
    Massi, Max
    Vongsvivut, J.
    Hackett, Mark
    Date
    2021
    Type
    Journal Article
    
    Metadata
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    Citation
    Hartnell, D. and Hollings, A. and Ranieri, A.M. and Lamichhane, H.B. and Becker, T. and Sylvain, N.J. and Hou, H. et al. 2021. Mapping sub-cellular protein aggregates and lipid inclusions using synchrotron ATR-FTIR microspectroscopy. Analyst. 146 (11): pp. 3516-3525.
    Source Title
    Analyst
    DOI
    10.1039/d1an00136a
    ISSN
    0003-2654
    Faculty
    Faculty of Health Sciences
    Faculty of Science and Engineering
    School
    Curtin Medical School
    School of Molecular and Life Sciences (MLS)
    Funding and Sponsorship
    http://purl.org/au-research/grants/arc/FT190100017
    URI
    http://hdl.handle.net/20.500.11937/90112
    Collection
    • Curtin Research Publications
    Abstract

    Visualising direct biochemical markers of cell physiology and disease pathology at the sub-cellular level is an ongoing challenge in the biological sciences. A suite of microscopies exists to either visualise sub-cellular architecture or to indirectly view biochemical markers (e.g. histochemistry), but further technique developments and innovations are required to increase the range of biochemical parameters that can be imaged directly, in situ, within cells and tissue. Here, we report our continued advancements in the application of synchrotron radiation attenuated total reflectance Fourier transform infrared (SR-ATR-FTIR) microspectroscopy to study sub-cellular biochemistry. Our recent applications demonstrate the much needed capability to map or image directly sub-cellular protein aggregates within degenerating neurons as well as lipid inclusions within bacterial cells. We also characterise the effect of spectral acquisition parameters on speed of data collection and the associated trade-offs between a realistic experimental time frame and spectral/image quality. Specifically, the study highlights that the choice of 8 cm-1 spectral resolutions provide a suitable trade-off between spectral quality and collection time, enabling identification of important spectroscopic markers, while increasing image acquisition by ∼30% (relative to 4 cm-1 spectral resolution). Further, this study explores coupling a focal plane array detector with SR-ATR-FTIR, revealing a modest time improvement in image acquisition time (factor of 2.8). Such information continues to lay the foundation for these spectroscopic methods to be readily available for, and adopted by, the biological science community to facilitate new interdisciplinary endeavours to unravel complex biochemical questions and expand emerging areas of study.

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