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Contact X‐ray microscopy of living cells by using LiF crystal as imaging detector
Authors:L. REALE  F. BONFIGLI  A. LAI  F. FLORA  P. ALBERTANO  M. L. DI GIORGIO  L. MEZI  R.M. MONTEREALI  A. FAENOV  T. PIKUZ  S. ALMAVIVA  M. FRANCUCCI  P. GAUDIO  S. MARTELLUCCI  M. RICHETTA  A. POMA
Affiliation:1. Campus Biomedico, Via álvaro del Portillo 21, Rome, Italy;2. ENEA, C.R. Frascati, UTAPRAD, Technical Unit Development and Applications of Radiation, Frascati, Rome, Italy;3. Università di Roma “Tor Vergata,” Laboratorio di Biologia delle Alghe, Dipartimento di Biologia, via della Ricerca scientifica, Rome, Italy;4. University of L'Aquila, Department of Life, Health and Environmental Sciences, Coppito, L'Aquila, Italy;5. Institute for Academic Initiatives, Osaka University, Japan and Joint Institute for High Temperatures Russian Academy of Sciences, Moscow, Russia;6. Graduate School of Engineering, Osaka University, Japan and Joint Institute for High Temperatures Russian Academy of Sciences, Moscow, Russia;7. University of Rome “Tor Vergata,”, Department of Engineering, Rome, Italy
Abstract:In this paper, the use of lithium fluoride (LiF) as imaging radiation detector to analyse living cells by single‐shot soft X‐ray contact microscopy is presented. High resolved X‐ray images on LiF of cyanobacterium Leptolyngbya VRUC135, two unicellular microalgae of the genus Chlamydomonas and mouse macrophage cells (line RAW 264.7) have been obtained utilizing X‐ray radiation in the water window energy range from a laser plasma source. The used method is based on loading of the samples, the cell suspension, in a special holder where they are in close contact with a LiF crystal solid‐state X‐ray imaging detector. After exposure and sample removal, the images stored in LiF by the soft X‐ray contact microscopy technique are read by an optical microscope in fluorescence mode. The clear image of the mucilaginous sheath the structure of the filamentous Leptolyngbya and the visible nucleolus in the macrophage cells image, are noteworthiness results. The peculiarities of the used X‐ray radiation and of the LiF imaging detector allow obtaining images in absorption contrast revealing the internal structures of the investigated samples at high spatial resolution. Moreover, the wide dynamic range of the LiF imaging detector contributes to obtain high‐quality images. In particular, we demonstrate that this peculiar characteristic of LiF detector allows enhancing the contrast and reveal details even when they were obscured by a nonuniform stray light.
Keywords:Contact X‐ray microscopy  lithium fluoride detectors  algae  Chlamydomonas  Leptolyngbya  Macrophage RAW 264.7
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