The use of dual beam ESEM FIB to reveal the internal ultrastructure of hydroxyapatite nanoparticle-sugar-glass composites.

Microparticles (MP) spray dried from hydroxyapatite (HA) nanoparticle (NP) sugar suspensions are currently under development as a prolonged release vaccine vehicle. Those with a significant sugar component cannot be sectioned by ultramicrotomy as resins are excluded by the sugar. Focused ion beam (FIB) milling is the only method to prepare thin sections that enables the inspection of the MPs ultrastructure by transmission electron microscopy (TEM). Several methods have been explored and we have found it is simplest to encapsulate MPs in silver dag, sandwiched between gold foils for FIB-milling to enable multiple MPs to be sectioned simultaneously. Spray dried MPs containing 80% sugar have an inter-nanoparticle separation that is comparable with NP size (approximately 50 nm). MPs spray dried with 50% sugar or no sugar are more tightly packed. Nano-porosity in the order of 10 nm exists between NPs. MPs spray dried in the absence of sugar and sectioned by ultramicrotomy or by FIB-milling have comparable nanoscale morphologies. Selected area electron diffraction (SAED) demonstrates that the HA remains (substantially) crystalline following FIB-milling.

Electron microscopic study of novel threadlike structures on the surfaces of mammalian organs.

The ultrastructures of novel threadlike structures (NTSs) and corpuscles on the surfaces of internal organs of rats were investigated using electron microscopy. The samples were studied in situ by using a stereomicroscope and were taken for further morphological analysis. Scanning electron microscope (SEM) images revealed a bundle structure of threadlike tissue, which was composed of several 10-micro m-thick subducts. The surfaces of the corpuscles were rather coarse and fenestrated. The corpuscles had cucumber-like shapes with an average length of about 2 mm and a thickness of about 400 micro m. Transmission electron microscope (TEM) images disclosed disordered collagen fibers, which formed the extracellular matrix of the threadlike tissue, and immune-function cells, like macrophages, mast cells, and eosinophils. Sinuses of various diameters, which were thought to be cross-sections of the lumens of the subducts, were observed in the TEM, cryo-SEM and focused-ion-beam SEM images. These SEM images were obtained for the first time to reveal the detailed structure of the NTSs that were only recently discovered.