While electron microscopes and AFMs are capable of high resolution imaging to molecular levels, there is an ongoing
problem in integrating these results into the larger scale structure and functions of tissue and organs within a complex
organism. Imaging biological samples with optical microscopy is predominantly done with histology and
immunohistochemistry, which can take up to a several weeks to prepare, are artifact prone and only available as
individual 2D images. At the nano resolution scale, the higher resolution electron microscopy and AFM are used, but
again these require destructive sample preparation and data are in 2D. To bridge this gap, we describe a rapid non
invasive hierarchical bioimaging technique using a novel lab based x-ray computed tomography to characterize complex
biological organism in multiscale- from whole organ (mesoscale) to calcified and soft tissue (microscale), to subcellular
structures, nanomaterials and cellular-scaffold interaction (nanoscale). While MicroCT (micro x-ray computed
tomography) is gaining in popularity for non invasive bones and tissue imaging, contrast and resolution are still vastly
inadequate compared to histology. In this study we will present multiscale results from a novel microCT and nanoCT
(nano x-ray tomography system). The novel MicroCT can image large specimen and tissue sample at histology
resolution of submicron voxel resolution, often without contrast agents, while the nanoCT using x-ray optics similar to
those used in synchrotron radiation facilities, has 20nm voxel resolution, suitable for studying cellular, subcellular
morphology and nanomaterials. Multiscale examples involving both calcified and soft tissue will be illustrated, which
include imaging a rat tibia to the individual channels of osteocyte canaliculli and lacunae and an unstained whole murine
lung to its alveoli. The role of the novel CT will also be discussed as a possible means for rapid virtual histology using a
biopsy of a human regenerated bone sample done without contrast agents and that of other soft tissues with contrast
agents. Comparison between histology, SEM and MRI will be given.
The most commonly employed tools for wafer thickness and topography metrology are based on capacitance method, which due to physical size of probes, and may not be suitable for direct measurement of multi-layer non-conductive wafers or Micro Electromechanical Systems (MEMS) structures. Recently developed that low coherence interferometry provides solution, which overcomes limitations of these methods. Selected MEMS applications including characterization of deep (high aspect) trenches and membrane structures have been also developed. The above listed applications were limited to measurements of relative distance between two optical interfaces in material transparent at the wavelength of probing radiation. Absolute distance gauging by fiber optic low coherence interferometer is difficult due to large thermal drift (of the order of 0.04 mm/K). We demonstrate that this drift is a result of thermal changes of refractive index of fiber optic glass. We present solution eliminating this drift is based on introduction of the additional reference plane in the signal arm of the Michelson interferometer. Use of this reference plane eliminates influence of changes of refractive index of glass fibers on result of measurement and improves thermal stability of low coherence interferometer by three orders of magnitude.
Novel nondestructive method based on low coherence optical interferometry for measurement of deep etched trenches in MEMs structures is presented. The proposed technique proves to provide very reproducible results and can be easily extended to metrology of other materials such as metals and dielectrics. We present results in real life semiconductor structures and discuss practical and fundamental limits of this technique
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