We have developed a randomized grating condenser zone plate (GCZP) that provides a µm-scale probe for use in x-ray ptychography. This delivers a significantly better x-ray throughput than probes defined by pinhole apertures, while providing a clearly-defined level of phase diversity to the illumination on the sample, and helping to reduce the dynamic range of the detected signal by spreading the zero-order light over an extended area of the detector. The first use of this novel x-ray optical element has been demonstrated successfully for both amplitude and phase contrast imaging using soft x-rays on the TwinMic beamline at the Elettra synchrotron.
Advanced imaging is useful for understanding the three-dimensional (3D) growth of cells. X-ray tomography serves as a powerful noninvasive, nondestructive technique that can fulfill these purposes by providing information about cell growth within 3D platforms. There are a limited number of studies taking advantage of synchrotron X-rays, which provides a large field of view and suitable resolution to image cells within specific biomaterials. In this study, X-ray synchrotron radiation microtomography at Diamond Light Source and advanced image processing were used to investigate cellular infiltration of HeLa cells within poly L-lactide (PLLA) scaffolds. This study demonstrates that synchrotron X-rays using phase contrast is a useful method to understand the 3D growth of cells in PLLA electrospun scaffolds. Two different fiber diameter (2 and 4 µm) scaffolds with different pore sizes, grown over 2, 5 and 8 days in vitro, were examined for infiltration and cell connectivity. After performing visualization by segmentation of the cells from the fibers, the results clearly show deeper cell growth and higher cellular interconnectivity in the 4 µm fiber diameter scaffold. This indicates the potential for using such 3D technology to study cell–scaffold interactions for future medical use.
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