High-resolution X-ray imaging of Plasmodium falciparum-infected red blood cells.
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.
Williams
Garth J
GJ
ARC Centre of Excellence for Coherent X-Ray Science & School of Physics, The University of Melbourne, Vic., 3010 Australia.
Hanssen
Eric
E
Peele
Andrew G
AG
Pfeifer
Mark A
MA
Clark
Jesse
J
Abbey
Brian
B
Cadenazzi
Guido
G
de Jonge
Martin D
MD
Vogt
Stefan
S
Tilley
Leann
L
Nugent
Keith A
KA
D000818:Animals; D004912:Erythrocytes / Q000000981:diagnostic imaging* / Q000469:parasitology* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D017348:Microscopy, Electron, Scanning Transmission; D008856:Microscopy, Fluorescence; D010963:Plasmodium falciparum / Q000302:isolation & purification*; D011859:Radiography; D014162:Transfection; D014961:X-Ray Diffraction / Q000379:methods*
D049452:Green Fluorescent Proteins