Multiscale and Multimodal Microscopy of physiological and pathological tissues with crystals

Using advanced optical, X-ray and scanning electron microscopy techniques in vivo or under cryogenic conditions, we investigate physiological and pathological tissues from the millimeter to the nanometer scale, in two, three and four dimensions, preserving them hydrated, as close as possible to the native conditions. In biomineralization, we reconstruct pathways of skeleton formation in unicellular organisms such as the green alga Phacotus lenticularis (Noy Shaked, PhD research) and in multicellular organisms such as sea urchins [1], from ion concentration to mineral deposition. We use sequential procedures involving in vivo confocal microscopy, soft X-ray cryo-microscopy, micro-spectroscopy, and scanning electron microscopy techniques in 2D and in 3D. In the investigation of atherosclerotic lesions, we built a sequence involving micro-CT, followed by cryo-SEM, cathodoluminescence imaging, and cryo-FIB-SEM. We observe cholesterol crystal formation, accumulation and dissolution in rabbit and human lesions, which allows us to reconstruct some of the events occurring in pathological plaques [2]. These observations were made possible by using combinations of imaging in vivo or in cryo-fixed, unembedded and minimally processed tissues, and can be advantageous in multiscale imaging of other pathological or functional tissues.

[1] K Kahil, N Varsano, A Sorrentino, E Pereiro, P Rez, S Weiner, L Addadi. Cellular pathways of calcium transport and concentration toward mineral formation in sea urchin larvae. PNAS 2020

[2] Capua-Shenkar J, Varsano N, Itzhak NR, Kaplan-Ashiri I, Rechav K, Jin X, Niimi M, Fan J, Kruth H, Addadi L. Examining atherosclerotic lesions in three dimensions at the nanometer scale with cryo-FIB-SEM. PNAS, 2022