X-ray microtomographic visualization of Escherichia coli by metalloprotein overexpression

J Synchrotron Radiat. 2013 Jul;20(Pt 4):581-6. doi: 10.1107/S0909049513008467. Epub 2013 May 1.

Abstract

This paper reports X-ray microtomographic visualization of the microorganism Escherichia coli overexpressing a metalloprotein ferritin. The three-dimensional distribution of linear absorption coefficients determined using a synchrotron radiation microtomograph with a simple projection geometry revealed that the X-ray absorption was homogeneously distributed, suggesting that every E. coli cell was labeled with the ferritin. The ferritin-expressing E. coli exhibited linear absorption coefficients comparable with those of phosphotungstic acid stained cells. The submicrometer structure of the ferritin-expressing E. coli cells was visualized by Zernike phase contrast using an imaging microtomograph equipped with a Fresnel zone plate. The obtained images revealed curved columnar or bunching oval structures corresponding to the E. coli cells. These results indicate that the metalloprotein overexpression facilitates X-ray visualization of three-dimensional cellular structures of biological objects.

Keywords: bacteria; ferritin; high-Z element; micro-CT; three-dimensional structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / metabolism*
  • Base Sequence
  • DNA Primers
  • Electron Probe Microanalysis
  • Escherichia coli / metabolism*
  • Metalloproteins / metabolism*
  • X-Ray Microtomography

Substances

  • Bacterial Proteins
  • DNA Primers
  • Metalloproteins