AI Article Synopsis
- Most halophilic Archaea, like those from the Halobacteriaceae family, need high concentrations of sodium chloride (NaCl) for growth, causing challenges in structural studies using electron microscopy due to low contrast.
- Researchers focused on Halobacterium salinarum and found that they could adapt these microbes to survive at lower NaCl levels through dialysis, maintaining vital cells at 3 M salt.
- The study ultimately showed that with careful adjustments to vitrification and cell thinning techniques, structural studies of these microorganisms can be successfully performed in situ, even under high salt conditions.
Article Abstract
Most halophilic Archaea of the class Halobacteriaceae depend on the presence of several molar sodium chloride for growth and cell integrity. This poses problems for structural studies, particularly for electron microscopy, where the high salt concentration results in diminished contrast. Since cryo-electron microscopy of intact cells provides new insights into the cellular and molecular organization under close-to-live conditions, we evaluated strategies and conditions to make halophilic microbes available for investigations in situ. Halobacterium salinarum, the test organism for this study, usually grows at 4.3 M NaCl. Adaptation to lower concentrations and subsequent NaCl reduction via dialysis led to still vital cells at 3 M salt. A comprehensive evaluation of vitrification parameters, thinning of frozen cells by focused-ion-beam micromachining, and cryo-electron microscopy revealed that structural studies under high salt conditions are possible in situ.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5329092 | PMC |
| http://dx.doi.org/10.1007/s00792-016-0912-0 | DOI Listing |
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