Fig. 2

Application of the CLEM holder for bone study. A Schematics of the CLEM holder applied for LM observation. In the case of the UTS specimen, RLM is effective due to interference colors providing high contrast by reflective light. In addition, the light absorption plate was used together to avoid light reflecting back after passing through the specimen. B The configuration for STEM-in-SEM EDS using the CLEM holder and its components. C A RLM micrograph of a femoral transverse section from an 8-week-old rat. D A STEM-in-SEM micrograph obtained at the location shown in the marked region of (C). E An EDS spectrum acquired from the bone matrix of (D). The spectrum shows Ca, P, C, and O as the main composing elements of bone. The weak Cu, Al, and Si signals are generated from the TEM grid, holder, and STEM detector, respectively. (EDS: Energy-dispersive spectroscopy. LM: Light microscopy. RLM: Reflected light microscopy. STEM-in-SEM: Scanning transmission electron microscopy-in-scanning electron microscopy. UTS: Ultra-thin sections)