title: Targeted nano analysis of water and ions using cryocorrelative light and scanning transmission electron microscopy.
abstract: Cryo fluorescence imaging coupled with the cryo-EM technique (cryo-CLEM) avoids chemical fixation and embedding in plastic, and is the gold standard for correlated imaging in a close to native state. This multi-modal approach has not previously included elementary nano analysis or evaluation of water content. We developed a new approach allowing analysis of targeted in situ intracellular ions and water measurements at the nanoscale (EDXS and STEM dark field imaging) within domains identified by examination of specific GFP-tagged proteins. This method allows both water and ions- fundamental to cell biology- to be located and quantified at the subcellular level. We illustrate the potential of this approach by investigating changes in water and ion content in nuclear domains identified by GFP-tagged proteins in cells stressed by Actinomycin D treatment and controls. The resolution of our approach was sufficient to distinguish clumps of condensed chromatin from surrounding nucleoplasm by fluorescence imaging and to perform nano analysis in this targeted compartment.
authors.affiliation: Laboratoire de Recherche en Nanosciences EA 4682, UFR Sciences Exactes et Naturelles, Université de Reims Champagne Ardenne, 21 rue Clément ADER, Reims, France.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Nolin
authors.forename: Dominique
authors.initials: D
authors.lastname: Ploton
authors.forename: Laurence
authors.initials: L
authors.lastname: Wortham
authors.forename: Pavel
authors.initials: P
authors.lastname: Tchelidze
authors.forename: Gérard
authors.initials: G
authors.lastname: Balossier
authors.forename: Vincent
authors.initials: V
authors.lastname: Banchet
authors.forename: Hélène
authors.initials: H
authors.lastname: Bobichon
authors.forename: Nathalie
authors.initials: N
authors.lastname: Lalun
authors.forename: Christine
authors.initials: C
authors.lastname: Terryn
authors.forename: Jean
authors.initials: J
authors.lastname: Michel
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D007477:Ions / Q000737:chemistry*; D017348:Microscopy, Electron, Scanning Transmission / Q000379:methods*; D014867:Water / Q000737:chemistry*
chemical_list: D007477:Ions; D014867:Water