Phosphorus detection in vitrified bacteria by cryo-STEM annular dark-field analysis

Sharon Grayer Wolf, Peter Rez, Michael Elbaum

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Bacterial cells often contain dense granules. Among these, polyphosphate bodies (PPBs) store inorganic phosphate for a variety of essential functions. Identification of PPBs has until now been accomplished by analytical methods that required drying or chemically fixing the cells. These methods entail large electron doses that are incompatible with low-dose imaging of cryogenic specimens. We show here that Scanning Transmission Electron Microscopy (STEM) of fully hydrated, intact, vitrified bacteria provides a simple means for mapping of phosphorus-containing dense granules based on quantitative sensitivity of the electron scattering to atomic number. A coarse resolution of the scattering angles distinguishes phosphorus from the abundant lighter atoms: carbon, nitrogen and oxygen. The theoretical basis is similar to Z contrast of materials science. EDX provides a positive identification of phosphorus, but importantly, the method need not involve a more severe electron dose than that required for imaging. The approach should prove useful in general for mapping of heavy elements in cryopreserved specimens when the element identity is known from the biological context.

Original languageEnglish (US)
Pages (from-to)227-233
Number of pages7
JournalJournal of Microscopy
Volume260
Issue number2
DOIs
Publication statusPublished - Nov 1 2015

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Keywords

  • Agrobacterium
  • Analytical microscopy
  • Cryo-microscopy
  • EDS
  • EFTEM
  • Scanning transmission electron microscopy

ASJC Scopus subject areas

  • Histology
  • Pathology and Forensic Medicine

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