TY - JOUR
T1 - Assaying three-dimensional cellular architecture using X-ray tomographic and correlated imaging approaches
AU - Bayguinov, Peter O.
AU - Fisher, Max R.
AU - Fitzpatrick, James A.J.
N1 - Funding Information:
Acknowledgments—This work utilized the resources of the Washington University Diabetes Research Center (supported by National Institutes of Health (NIH) Grant P30 DK020579, the Washington University Rheumatic Diseases Research Resource-based Center (NIH Grant P30 AR073752), the Washington University Musculoskeletal Research Center (NIH Grant P30 AR074992), and the Site-man Cancer Center of Barnes-Jewish Hospital and Washington University School of Medicine (NIH Grant P30 CA091842). The three-dimensional X-ray phase-contrast microscope for sub- micron quantitative biological imaging was acquired with the support of NIH Grant S10 OD021694.
Funding Information:
This work was supported by Children?s Discovery Institute of Washington University and St. Louis Children?s Hospital Grants CDI-CORE-2015-505 and CDI-CORE-2019-813 (to J. A. J. F.), and Foundation for Barnes-Jewish Hospital Grants 3770 and 4642 (to J. A. J. F.).
Funding Information:
Funding and additional information—This work was supported by Children’s Discovery Institute of Washington University and St. Louis Children’s Hospital Grants CDI-CORE-2015-505 and CDI-CORE-2019-813 (to J. A. J. F.), and Foundation for Barnes-Jewish Hospital Grants 3770 and 4642 (to J. A. J. F.).
Publisher Copyright:
© 2020 Bayguinov et al.
PY - 2020/11/13
Y1 - 2020/11/13
N2 - Much of our understanding of the spatial organization of and interactions between cellular organelles and macromolecular complexes has been the result of imaging studies utilizing either light- or electron-based microscopic analyses. These classical approaches, while insightful, are nonetheless limited either by restrictions in resolution or by the sheer complexity of generating multidimensional data. Recent advances in the use and application of X-rays to acquire micro- and nanotomographic data sets offer an alternative methodology to visualize cellular architecture at the nanoscale. These new approaches allow for the subcellular analyses of unstained vitrified cells and three-dimensional localization of specific protein targets and have served as an essential tool in bridging light and electron correlative microscopy experiments. Here, we review the theory, instrumentation details, acquisition principles, and applications of both soft X-ray tomography and X-ray microscopy and how the use of these techniques offers a succinct means of analyzing three-dimensional cellular architecture. We discuss some of the recent work that has taken advantage of these approaches and detail how they have become integral in correlative microscopy workflows.
AB - Much of our understanding of the spatial organization of and interactions between cellular organelles and macromolecular complexes has been the result of imaging studies utilizing either light- or electron-based microscopic analyses. These classical approaches, while insightful, are nonetheless limited either by restrictions in resolution or by the sheer complexity of generating multidimensional data. Recent advances in the use and application of X-rays to acquire micro- and nanotomographic data sets offer an alternative methodology to visualize cellular architecture at the nanoscale. These new approaches allow for the subcellular analyses of unstained vitrified cells and three-dimensional localization of specific protein targets and have served as an essential tool in bridging light and electron correlative microscopy experiments. Here, we review the theory, instrumentation details, acquisition principles, and applications of both soft X-ray tomography and X-ray microscopy and how the use of these techniques offers a succinct means of analyzing three-dimensional cellular architecture. We discuss some of the recent work that has taken advantage of these approaches and detail how they have become integral in correlative microscopy workflows.
UR - http://www.scopus.com/inward/record.url?scp=85096202734&partnerID=8YFLogxK
U2 - 10.1074/jbc.REV120.009633
DO - 10.1074/jbc.REV120.009633
M3 - Review article
C2 - 32938716
AN - SCOPUS:85096202734
VL - 295
SP - 15782
EP - 15793
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 46
ER -