Nanoscale soft x-ray tomography is a powerful new imaging technology for biomedical and translational research. This technique produces high spatial resolution, high-fidelity 3-Dimensional tomographic reconstructions of the specimen, and does so with unprecedented sample throughput. The work of this proposal will further enhance the applicability, usability and fidelity of soft x-ray tomography as a biomedical imaging technique, make it applicable to a greater range of specimen types, significantly increase spatial resolution, and extend the tools and options available for protein localization. We will continue development of our pioneering work on correlated cryogenic fluorescence microscopy. These efforts, and the planned instrumental and technological developments, will allow multi-modal imaging to be carried out on specimens containing endogenous or exogenous fluorescent labels. The end result will be a suite of imaging options that allow protein localization data to be optimally positioned within a 3-Dimensional reconstruction of the specimen. In soft x-ray tomography the image contrast is derived from the biochemical composition of the specimen. Apart from eliminating the need to fix and stain the specimen this results in each organelle having a signature linear x-ray absorption coefficient. This measurement can be used to identify and quantifiably characterize organelles within a specimen and between different specimens. This type of information, together with the location of specific molecules in the specimen is key to understanding the cellular effects of disease, identifying and validating drug targets, and determining modes of action of molecules with therapeutic potential.

Public Health Relevance

Technologies are being developed to visualize the effects of disease on cell structure and organization. This is important information in the identification and validation of targets for drug design, and for determining the mode of action of molecules with therapeutic potential. This technology can be applied to understanding the cellular effects of cancer, or infection, whether it be microbial, viral or parasitic in origin.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Biotechnology Resource Grants (P41)
Project #
5P41GM103445-09
Application #
8643771
Study Section
Special Emphasis Panel (ZRG1-BST-K (40))
Program Officer
Swain, Amy L
Project Start
2004-05-06
Project End
2015-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
9
Fiscal Year
2013
Total Cost
$838,887
Indirect Cost
$304,280
Name
University of California San Francisco
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
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Le Gros, Mark A; Clowney, E Josephine; Magklara, Angeliki et al. (2016) Soft X-Ray Tomography Reveals Gradual Chromatin Compaction and Reorganization during Neurogenesis In Vivo. Cell Rep 17:2125-2136
Tjong, Harianto; Li, Wenyuan; Kalhor, Reza et al. (2016) Population-based 3D genome structure analysis reveals driving forces in spatial genome organization. Proc Natl Acad Sci U S A 113:E1663-72
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Do, Myan; Isaacson, Samuel A; McDermott, Gerry et al. (2015) Imaging and characterizing cells using tomography. Arch Biochem Biophys 581:111-21
Elgass, Kirstin D; Smith, Elizabeth A; LeGros, Mark A et al. (2015) Analysis of ER-mitochondria contacts using correlative fluorescence microscopy and soft X-ray tomography of mammalian cells. J Cell Sci 128:2795-804
Ugarte, Fernando; Sousae, Rebekah; Cinquin, Bertrand et al. (2015) Progressive Chromatin Condensation and H3K9 Methylation Regulate the Differentiation of Embryonic and Hematopoietic Stem Cells. Stem Cell Reports 5:728-40
Le Gros, Mark A; McDermott, Gerry; Cinquin, Bertrand P et al. (2014) Biological soft X-ray tomography on beamline 2.1 at the Advanced Light Source. J Synchrotron Radiat 21:1370-7

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