Contact X-ray microscopy of living cells by using LiF crystal as imaging detector

J Microsc. 2015 May;258(2):127-39. doi: 10.1111/jmi.12226. Epub 2015 Feb 2.

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: Chlamydomonas; Contact X-ray microscopy; Leptolyngbya; Macrophage RAW 264.7; algae; lithium fluoride detectors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chlamydomonas / ultrastructure
  • Cyanobacteria / ultrastructure
  • Fluorides*
  • Lasers
  • Lithium Compounds*
  • Macrophages / ultrastructure
  • Mice
  • Microscopy / methods*
  • RAW 264.7 Cells
  • X-Rays

Substances

  • Lithium Compounds
  • lithium fluoride
  • Fluorides