0000000000223062
AUTHOR
Francesca Cella Zanacchi
Light Sheet Fluorescence Microscopy (LSFM) for Two-Photon Excitation Imaging of Thick Samples.
Over the last decades, fluorescence microscopy techniques have been developed in order to provide a deeper, faster and higher resolution imaging of three-dimensional biological samples. Within this framework, Light Sheet Fluorescence Microscopy (LSFM) became an increasingly useful and popular imaging technique able to answer several biological questions in the field of developmental biology [1]. Thanks to the spatial confinement of the excitation process within a thin sheet in the focal plane, it provides an intrinsic optical sectioning and a reduced phototoxicity. On the other side, Two-Photon Excitation (2PE), thanks to the use of IR wavelengths, has become an invaluable tool to improve i…
4D (x-y-z-t) imaging of thick biological samples by means of Two-Photon inverted Selective Plane Illumination Microscopy (2PE-iSPIM)
AbstractIn the last decade light sheet fluorescence microscopy techniques, such as selective plane illumination microscopy (SPIM), has become a well established method for developmental biology. However, conventional SPIM architectures hardly permit imaging of certain tissues since the common sample mounting procedure, based on gel embedding, could interfere with the sample morphology. In this work we propose an inverted selective plane microscopy system (iSPIM), based on non-linear excitation, suitable for 3D tissue imaging. First, the iSPIM architecture provides flexibility on the sample mounting, getting rid of the gel-based mounting typical of conventional SPIM, permitting 3D imaging of…
Biocompatibility and biodistribution of functionalized carbon nano-onions (f-CNOs) in a vertebrate model
AbstractFunctionalized carbon nano-onions (f-CNOs) are of great interest as platforms for imaging, diagnostic and therapeutic applications due to their high cellular uptake and low cytotoxicity. To date, the toxicological effects of f-CNOs on vertebrates have not been reported. In this study, the possible biological impact of f-CNOs on zebrafish during development is investigated, evaluating different toxicity end-points such as the survival rate, hatching rate, and heart beat rate. Furthermore, a bio-distribution study of boron dipyrromethene (BODIPY) functionalized CNOs in zebrafish larvae is performed by utilizing inverted selective plane illumination microscopy (iSPIM), due to its intri…
Influence of Nanoparticle Exposure on Nervous System Development in Zebrafish Studied by Means of Light Sheet Fluorescence Microscopy
Zebrafish has a remarkable similarity in the molecular signaling processes, cellular structure, anatomy and physiology to other higher order vertebrates, making it an excellent vertebrate model organism (1). Recently, zebrafish has been used for neurotoxicity screening of numerous nanomaterials with a focus on the developmental effects due to the possibility of in vivo visualization of specific neurons and axon tracts by injecting dyes in live animals as well in fixed ones (2). Here we propose Light Sheet Fluorescence Microscopy (LSFM) (3) (4) to perform neurotoxicity studies, in order to study the nervous system architecture and to image 3D structures in the brain of live larvae during the…