title: Correlative cryogenic tomography of cells using light and soft x-rays.
abstract: Correlated imaging is the process of imaging a specimen with two complementary modalities, and then combining the two data sets to create a highly informative, composite view. A recent implementation of this concept has been the combination of soft x-ray tomography (SXT) with fluorescence cryogenic microscopy (FCM). SXT-FCM is used to visualize cells that are held in a near-native, cryopreserved. The resultant images are, therefore, highly representative of both the cellular architecture and molecular organization in vivo. SXT quantitatively visualizes the cell and sub-cellular structures; FCM images the spatial distribution of fluorescently labeled molecules. Here, we review the characteristics of SXT-FCM, and briefly discuss how this method compares with existing correlative imaging techniques. We also describe how the incorporation of a cryo-rotation stage into a cryogenic fluorescence microscope allows acquisition of fluorescence cryogenic tomography (FCT) data. FCT is optimally suited for correlation with SXT, since both techniques image the specimen in 3-D, potentially with similar, isotropic spatial resolution.
authors.affiliation: Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Smith
authors.affiliation: Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States.
authors.forename: Bertrand P
authors.initials: BP
authors.lastname: Cinquin
authors.affiliation: Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States.
authors.forename: Myan
authors.initials: M
authors.lastname: Do
authors.affiliation: Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States.
authors.forename: Gerry
authors.initials: G
authors.lastname: McDermott
authors.affiliation: Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; Physical BioSciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States. Electronic address: MALegros@lbl.gov.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Le Gros
authors.affiliation: Department of Anatomy, School of Medicine, University of California San Francisco, San Francisco, CA, United States; Physical BioSciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States; National Center for X-ray Tomography, Advanced Light Source, Berkeley, CA, United States. Electronic address: carolyn.larabell@ucsf.edu.
authors.forename: Carolyn A
authors.initials: CA
authors.lastname: Larabell
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods; D008027:Light; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D014057:Tomography, X-Ray Computed / Q000379:methods*; D014965:X-Rays
keywords: Cell structure; Imaging; Molecular localization