Correlative 3D cryo X-ray imaging reveals intracellular location and effect of designed antifibrotic protein-nanomaterial hybrids

DOI: 10.1039/D1SC04183E

Authors: Johannes Groen (ALBA Synchrotron Light Source) , Ana R. Palanca (Instituto de Biomedicina y Biotecnologia de Cantabria (IBBTEC), University of Cantabria) , Antonio Aires (Center for Cooperative Research in Biomaterials (CIC biomaGUNE), Basque Research and Technology Alliance (BRTA)) , Javi Conesa (ALBA Synchrotron Light Source; National Center for Biotechnology CSIC (CNB-CSIC)) , David Maestro (Instituto de Biomedicina y Biotecnologia de Cantabria (IBBTEC), University of Cantabria) , Stefan Rehbein (Bessy II) , Maria Harkiolaki (Diamond Light Source) , Ana Villar (Instituto de Biomedicina y Biotecnologia de Cantabria (IBBTEC), University of Cantabria) , Aitziber L. Cortajarena (Center for Cooperative Research in Biomaterials (CIC biomaGUNE), Basque Research and Technology Alliance (BRTA); Ikerbasque, Basque Foundation for Science) , Eva Pereiro (ALBA Synchrotron Light Source)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Chemical Science

State: Published (Approved)
Published: October 2021
Diamond Proposal Number(s): 25162

Abstract: Revealing the intracellular location of novel therapeutic agents is paramount for the understanding of their effect at the cell ultrastructure level. Here, we apply a novel correlative cryo 3D imaging approach to determine the intracellular fate of a designed protein-nanomaterial hybrid with antifibrotic properties that shows great promise in mitigating myocardial fibrosis. Cryo 3D structured illumination microscopy (cryo-3D-SIM) pinpoints the location and cryo soft X-ray tomography (cryo-SXT) reveals the ultrastructural environment and subcellular localization of this nanomaterial with spatial correlation accuracy down to 70 nm in whole cells. This novel high resolution 3D cryo correlative approach unambiguously locates the nanomaterial after overnight treatment within multivesicular bodies which have been associated with endosomal trafficking events by confocal microscopy. Moreover, this approach allows assessing the cellular response towards the treatment by evaluating the morphological changes induced. This is especially relevant for the future usage of nanoformulations in clinical practices. This correlative super-resolution and X-ray imaging strategy joins high specificity, by the use of fluorescence, with high spatial resolution at 30 nm (half pitch) provided by cryo-SXT in whole cells, without the need of staining or fixation, and can be of particular benefit to locate specific molecules in the native cellular environment in bio-nanomedicine.

Subject Areas: Biology and Bio-materials


Instruments: B24-Cryo Soft X-ray Tomography

Other Facilities: Mistral at ALBA

Added On: 28/10/2021 09:58

Documents:
d1sc04183e.pdf

Discipline Tags:

Non-Communicable Diseases Health & Wellbeing Structural biology Life Sciences & Biotech

Technical Tags:

Imaging Tomography Cryo-soft X-ray Tomography (Cryo-SXT)