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Title: 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. Finally, this combined approach offers a way to study the role of trace elements in their structural context.

Authors:
 [1];  [2];  [2];  [2];  [1];  [3];  [2];  [2];  [2];  [1]
  1. Northwestern Univ., Evanston, IL (United States)
  2. Argonne National Lab. (ANL), Argonne, IL (United States)
  3. La Trobe Univ., Melbourne, VIC (Australia); Commonwealth Scientific and Industrial Research Organization Manufacturing Flagship, Parkville, VIC (Australia)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1235500
Alternate Identifier(s):
OSTI ID: 1215636; OSTI ID: 1346725
Grant/Contract Number:  
AC02-06CH11357; 1R01GM104530; 1S10RR029272-01
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 112; Journal Issue: 8; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ptychography; X-ray fluorescence microscopy; cryogenic biological samples

Citation Formats

Deng, Junjing, Vine, David J., Chen, Si, Nashed, Youssef S. G., Jin, Qiaoling, Phillips, Nicholas W., Peterka, Tom, Ross, Rob, Vogt, Stefan, and Jacobsen, Chris J. Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae. United States: N. p., 2015. Web. doi:10.1073/pnas.1413003112.
Deng, Junjing, Vine, David J., Chen, Si, Nashed, Youssef S. G., Jin, Qiaoling, Phillips, Nicholas W., Peterka, Tom, Ross, Rob, Vogt, Stefan, & Jacobsen, Chris J. Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae. United States. doi:10.1073/pnas.1413003112.
Deng, Junjing, Vine, David J., Chen, Si, Nashed, Youssef S. G., Jin, Qiaoling, Phillips, Nicholas W., Peterka, Tom, Ross, Rob, Vogt, Stefan, and Jacobsen, Chris J. Tue . "Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae". United States. doi:10.1073/pnas.1413003112.
@article{osti_1235500,
title = {Simultaneous cryo X-ray ptychographic and fluorescence microscopy of green algae},
author = {Deng, Junjing and Vine, David J. and Chen, Si and Nashed, Youssef S. G. and Jin, Qiaoling and Phillips, Nicholas W. and Peterka, Tom and Ross, Rob and Vogt, Stefan and Jacobsen, Chris J.},
abstractNote = {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. Finally, this combined approach offers a way to study the role of trace elements in their structural context.},
doi = {10.1073/pnas.1413003112},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 8,
volume = 112,
place = {United States},
year = {2015},
month = {2}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1073/pnas.1413003112

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Cited by: 42 works
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