title: Three-dimensional imaging of human stem cells using soft X-ray tomography.
abstract: Three-dimensional imaging of human stem cells using transmission soft X-ray tomography (SXT) is presented for the first time. Major organelle types--nuclei, nucleoli, mitochondria, lysosomes and vesicles--were discriminated at approximately 50 nm spatial resolution without the use of contrast agents, on the basis of measured linear X-ray absorption coefficients and comparison of the size and shape of structures to transmission electron microscopy (TEM) images. In addition, SXT was used to visualize the distribution of a cell surface protein using gold-labelled antibody staining. We present the strengths of SXT, which include excellent spatial resolution (intermediate between that of TEM and light microscopy), the lack of the requirement for fixative or contrast agent that might perturb cellular morphology or produce imaging artefacts, and the ability to produce three-dimensional images of cells without microtome sectioning. Possible applications to studying the differentiation of human stem cells are discussed.
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria 3800, Australia The Florey Institute of Neuroscience and Mental Health, Melbourne University, Parkville, Victoria 3052, Australia.
authors.forename: J C
authors.initials: JC
authors.lastname: Niclis
authors.affiliation: The Ritchie Centre, Monash Institute of Medical Research, Monash University, Clayton, Victoria 3800, Australia Wake Forest Baptist Medical Center, Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC, USA.
authors.forename: S V
authors.initials: SV
authors.lastname: Murphy
authors.affiliation: Advanced Light Source, Lawrence Berkeley National Laboratory, US Department of Energy, Berkeley, CA, USA.
authors.forename: D Y
authors.initials: DY
authors.lastname: Parkinson
authors.affiliation: Advanced Light Source, Lawrence Berkeley National Laboratory, US Department of Energy, Berkeley, CA, USA.
authors.forename: A
authors.initials: A
authors.lastname: Zedan
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: A H
authors.initials: AH
authors.lastname: Sathananthan
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: D S
authors.initials: DS
authors.lastname: Cram
authors.affiliation: Department of Anatomy and Developmental Biology, Monash University, Clayton, Victoria 3800, Australia Centre for Biospectroscopy, School of Chemistry, Monash University, Melbourne, Victoria, Australia phil.heraud@monash.edu.
authors.forename: P
authors.initials: P
authors.lastname: Heraud
mesh_terms: D000906:Antibodies / Q000737:chemistry; D002466:Cell Nucleolus / Q000648:ultrastructure*; D055032:Electron Microscope Tomography; D006046:Gold / Q000737:chemistry; D006801:Humans; D021621:Imaging, Three-Dimensional; D008247:Lysosomes / Q000648:ultrastructure*; D008928:Mitochondria / Q000648:ultrastructure*; D013194:Staining and Labeling; D013234:Stem Cells / Q000648:ultrastructure*; D014056:Tomography, X-Ray
chemical_list: D000906:Antibodies; D006046:Gold
keywords: human; stem cells; three-dimensional; transmission soft X-ray microscopy