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Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae.


ABSTRACT: Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both cellular structure and trace element distribution in frozen-hydrated cells at cryogenic temperatures, with high structural and chemical fidelity. Ptychographic reconstruction algorithms deliver phase and absorption contrast images at a resolution beyond that of the illuminating lens or beam size. Using 5.2-keV X-rays, we have obtained sub-30-nm resolution structural images and ?90-nm-resolution fluorescence images of several elements in frozen-hydrated green algae. This combined approach offers a way to study the role of trace elements in their structural context.

SUBMITTER: Deng J 

PROVIDER: S-EPMC4345580 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae.

Deng Junjing J   Vine David J DJ   Chen Si S   Nashed Youssef S G YS   Jin Qiaoling Q   Phillips Nicholas W NW   Peterka Tom T   Ross Rob R   Vogt Stefan S   Jacobsen Chris J CJ  

Proceedings of the National Academy of Sciences of the United States of America 20150209 8


Trace metals play important roles in normal and in disease-causing biological functions. X-ray fluorescence microscopy reveals trace elements with no dependence on binding affinities (unlike with visible light fluorophores) and with improved sensitivity relative to electron probes. However, X-ray fluorescence is not very sensitive for showing the light elements that comprise the majority of cellular material. Here we show that X-ray ptychography can be combined with fluorescence to image both ce  ...[more]

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