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High‐resolution X‐ray imaging of Plasmodium falciparum ‐infected red blood cells
Author(s) -
Williams Garth J.,
Hanssen Eric,
Peele Andrew G.,
Pfeifer Mark A.,
Clark Jesse,
Abbey Brian,
Cadenazzi Guido,
de Jonge Martin D.,
Vogt Stefan,
Tilley Leann,
Nugent Keith A.
Publication year - 2008
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20616
Subject(s) - microscopy , resolution (logic) , optics , plasmodium falciparum , coherent diffraction imaging , fluorescence lifetime imaging microscopy , fluorescence , physics , pathology , computer science , phase retrieval , malaria , artificial intelligence , fourier transform , medicine , quantum mechanics
Abstract Methods for imaging cellular architecture and ultimately macromolecular complexes and individual proteins, within a cellular environment, are an important goal for cell and molecular biology. Coherent diffractive imaging (CDI) is a method of lensless imaging that can be applied to any individual finite object. A diffraction pattern from a single biological structure is recorded and an iterative Fourier transform between real space and reciprocal space is used to reconstruct information about the architecture of the sample to high resolution. As a test system for cellular imaging, we have applied CDI to an important human pathogen, the malaria parasite, Plasmodium falciparum . We have employed a novel CDI approach, known as Fresnel CDI, which uses illumination with a curved incident wavefront, to image red blood cells infected with malaria parasites. We have examined the intrinsic X‐ray absorption contrast of these cells and compared them with cells contrasted with heavy metal stains or immunogold labeling. We compare CDI images with data obtained from the same cells using scanning electron microscopy, light microscopy, and scanning X‐ray fluorescence microscopy. We show that CDI can offer new information both within and at the surface of complex biological specimens at a spatial resolution of better than 40 nm. and we demonstrate an imaging modality that conveniently combines scanning X‐ray fluorescence microscopy with CDI. The data provide independent confirmation of the validity of the coherent diffractive image and demonstrate that CDI offers the potential to become an important and reliable new high‐resolution imaging modality for cell biology. CDI can detect features at high resolution within unsectioned cells. © 2008 International Society for Advancement of Cytometry