Quantitative X-ray phase contrast waveguide imaging of bacterial endospores

J Appl Crystallogr. 2015 Mar 24;48(Pt 2):464-476. doi: 10.1107/S1600576715003593. eCollection 2015 Apr 1.

Abstract

Quantitative waveguide-based X-ray phase contrast imaging has been carried out on the level of single, unstained, unsliced and freeze-dried bacterial cells of Bacillus thuringiensis and Bacillus subtilis using hard X-rays of 7.9 keV photon energy. The cells have been prepared in the metabolically dormant state of an endospore. The quantitative phase maps obtained by iterative phase retrieval using a modified hybrid input-output algorithm allow for mass and mass density determinations on the level of single individual endospores but include also large field of view investigations. Additionally, a direct reconstruction based on the contrast transfer function is investigated, and the two approaches are compared. Depending on the field of view and method, a resolution down to 65 nm was achieved at a maximum applied dose of below 5 × 105 Gy. Masses in the range of about ∼110-190 (20) fg for isolated endospores have been obtained.

Keywords: Bacillus subtilis; Bacillus thuringiensis; contrast transfer function; endospores; holotomography; phase retrieval; waveguide imaging.