Some fluorescence microscopy techniques, such as confocal laser scanning microscopy (CLSM), have a limited penetration depth. Consequently, the visualization and imaging of three-dimensional (3-D) cell cultures, such as spheroids, using these methods can be a significant challenge. Therefore, to improve the imaging of 3-D tissues, optical clearing methods have been optimized to render transparency to the opaque spheroids. The influence of the polyethylene glycol (PEG) molecular weight (MW) used in the ClearT2 method for the imaging of propidium iodide (PI)-stained spheroids was investigated. The results demonstrated that the ClearT2 clearing method contributes to spheroids transparency and to the preservation of PI fluorescence intensity for all the PEG MW used (4000, 8000, and 10,000 Da). Furthermore, the ClearT2 method performed using PEG 4000 Da allowed a better PI signal penetration depth and cross-section depth. Overall, the optimization of PEG MW can improve the imaging of intact spheroids by CLSM. Furthermore, this work may also contribute to increase the application of 3-D cell culture models by the pharmaceutical industry for the high-throughput screening of therapeutics.
Underwater survey, compared to land archaeology, needs some specific techniques, because the application of some active 3D sensor, such as laser scanner, is obviously impossible. The necessity to produce three-dimensional survey, offering the same accuracy of classical terrestrial laserscanning or photogrammetric methods, combined with the request of low costs and rapid solutions, led the researchers to test and apply oftentimes image-based techniques.<p> </p> In the last two years the Ca' Foscari University and University IUAV of Venice are conducting a research on the application of integrated techniques to support underwater metric documentation, comparing them to the manual traditional one. The gained experience (and confirmed by other recently published papers) shows that the actual multiimage digital photogrammetry is a good solution for the underwater archaeology. This approach is useful both from a metric and from a recording point of view, because it achieves high quality results, such as accurate 3D models or 2D representations, offering a complete documentation of underwater sites.<p> </p> But photogrammetry has to be supported by a topographical survey (to acquire ground control points - GCP) to georeference all the finds in the same reference system. This paper presents the integrated survey of two roman shipwrecks, approaching differently in the GCP’s acquisition just for the different morphological characteristic of the sites. The wrecks’ cargos are huge marble blocks, presenting differences in quantities, layout and depths. Those characteristics determine the choice of the topographic survey. <p> </p>The results of the survey are two 3D polygonal textured models of the sites, which can be easily used for different analyses and reconstructive hypothesis, opening new possibilities of documentation with both specialists and the wider public. Furthermore, 3D models are the geometric base for 2D orthophoto and cross section extraction. <p> </p>The paper will illustrate all the phases regarding the survey’s design, acquisition and realization and the data processing to obtain 2D and 3D final representation.