Format

Send to

Choose Destination
Ultramicroscopy. 2014 Aug;143:33-40. doi: 10.1016/j.ultramic.2013.10.013. Epub 2013 Nov 7.

Correlative cryogenic tomography of cells using light and soft x-rays.

Author information

1
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.
2
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.
3
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.

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.

KEYWORDS:

Cell structure; Imaging; Molecular localization

PMID:
24355261
PMCID:
PMC4013260
DOI:
10.1016/j.ultramic.2013.10.013
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center