Correlative transmission electron microscopy and high-resolution hard X-ray fluorescence microscopy of cell sections to measure trace element concentrations at the organelle level.
J Struct Biol
; 213(3): 107766, 2021 09.
Article
in En
| MEDLINE
| ID: mdl-34216761
ABSTRACT
Metals are essential for life and their concentration and distribution in organisms are tightly regulated. Indeed, in their free form, most transition metal ions are toxic. Therefore, an excess of physiologic metal ions or the uptake of non-physiologic metal ions can be highly detrimental to the organism. It is thus fundamental to understand metal distribution under physiological, pathological or environmental conditions, for instance in metal-related pathologies or upon environmental exposure to metals. Elemental imaging techniques can serve this purpose, by allowing the visualization and the quantification of metal species in tissues down to the level of cell organelles. Synchrotron radiation-based X-ray fluorescence (SR-XRF) microscopy is one of the most sensitive techniques to date, and great progress was made to reach nanoscale spatial resolution. Here we propose a correlative method to couple SR-XRF to electron microscopy (EM), with the possibility to quantify selected elemental contents in a specific organelle of interest with 50 × 50 nm2 raster scan resolution. We performed EM and SR-XRF on the same section of hepatocytes exposed to silver nanoparticles, in order to identify mitochondria through EM and visualize Ag co-localized with these organelles through SR-XRF. We demonstrate the accumulation of silver in mitochondria, which can reach a 10-fold higher silver concentration compared to the surrounding cytosol. The sample preparation and experimental setup can be adapted to other scientific questions, making the correlative use of SR-XRF and EM suitable to address a large panel of biological questions related to metal homeostasis.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Trace Elements
/
Metal Nanoparticles
Language:
En
Journal:
J Struct Biol
Journal subject:
BIOLOGIA MOLECULAR
Year:
2021
Document type:
Article
Affiliation country:
France