title: Towards an atlas of mammalian cell ultrastructure by cryo soft X-ray tomography.
abstract: We provide a catalog of 3D cryo soft X-ray tomography (cryo-SXT) images obtained from ∼6 to 12μm thick mouse adenocarcinoma cells. Included are multiple representative images of nuclei, nucleoli, nuclear membrane, nuclear membrane channels, mitochondria, lysosomes, endoplasmic reticulum, filaments and plasma membrane, plus three structures not previously described by cryo-SXT, namely Golgi, microvilli and nuclear-membrane blebs. Sections from the 3D cryo-SXT tomograms for all the preceding structures closely resemble those seen by thin-section transmission electron microscopy (TEM). Some structures such as nuclear-membrane channels and nuclear-membrane blebs are more easily detected by cryo-SXT than TEM most likely due to their better contrast and cellular preservation in cryo-SXT combined with the ability to rapidly locate these structures within a full 3D image. We identify and discuss two current limitations in cryo-SXT: variability in image quality and difficulties in detecting weaker contrast structures such as chromatin and various nuclear bodies. Progress on these points is likely to come from the solution of several technical problems in image acquisition, plus the implementation of advanced cryo soft X-ray microscopy approaches such as phase contrast or optical sectioning.
authors.affiliation: Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, 41 Library Drive, Bethesda, MD 20892, USA.
authors.forename: Waltraud G
authors.initials: WG
authors.lastname: Müller
authors.forename: J Bernard
authors.initials: JB
authors.lastname: Heymann
authors.forename: Kunio
authors.initials: K
authors.lastname: Nagashima
authors.forename: Peter
authors.initials: P
authors.lastname: Guttmann
authors.forename: Stephan
authors.initials: S
authors.lastname: Werner
authors.forename: Stefan
authors.initials: S
authors.lastname: Rehbein
authors.forename: Gerd
authors.initials: G
authors.lastname: Schneider
authors.forename: James G
authors.initials: JG
authors.lastname: McNally
mesh_terms: D000818:Animals; D001271:Atlases as Topic; D002462:Cell Membrane / Q000648:ultrastructure*; D022003:Cell Nucleus Structures / Q000648:ultrastructure; D015925:Cryopreservation; D021621:Imaging, Three-Dimensional; D051379:Mice; D008954:Models, Biological; D015388:Organelles / Q000648:ultrastructure*; D015203:Reproducibility of Results; D014056:Tomography, X-Ray / Q000379:methods*; D014407:Tumor Cells, Cultured