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

• DOI: 10.1039/d1sc04183e
• Publication Date: 2021-10-19T12:26:40Z
• Authors: J. Groen; A. Palanca; A. Aires; J. J. Conesa; D. Maestro; S. Rehbein; M. Harkiolaki; A. V. Villar; A. L. Cortajarena; E. Pereiro
• Keywords: Correlative; STED microscopy; Nanomaterials

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