Carbon speciation in organic fossils using 2D to 3D x-ray Raman multispectral imaging. Georgiou Rafaella,Gueriau Pierre,Sahle Christoph J,Bernard Sylvain,Mirone Alessandro,Garrouste Romain,Bergmann Uwe,Rueff Jean-Pascal,Bertrand Loïc Science advances The in situ two-dimensional (2D) and 3D imaging of the chemical speciation of organic fossils is an unsolved problem in paleontology and cultural heritage. Here, we use x-ray Raman scattering (XRS)-based imaging at the carbon K-edge to form 2D and 3D images of the carbon chemistry in two exceptionally preserved specimens, a fossil plant dating back from the Carboniferous and an ancient insect entrapped in 53-million-year-old amber. The 2D XRS imaging of the plant fossil reveals a homogeneous chemical composition with micrometric "pockets" of preservation, likely inherited from its geological history. The 3D XRS imaging of the insect cuticle displays an exceptionally well preserved remaining chemical signature typical of polysaccharides such as chitin around a largely hollowed-out inclusion. Our results open up new perspectives for in situ chemical speciation imaging of fossilized organic materials, with the potential to enhance our understanding of organic specimens and their paleobiology. 10.1126/sciadv.aaw5019