TY - GEN
T1 - Sub-100-nm 3D-elemental mapping of frozen-hydrated cells using the Bionanoprobe
AU - Chen, Si
AU - Yuan, Ye
AU - Deng, Junjing
AU - Mak, Rachel
AU - Jin, Qiaoling
AU - Paunesku, Tatjana
AU - Gleber, Sophie C.
AU - Vine, David
AU - Flachenecker, Claus
AU - Hornberger, Benjamin
AU - Shu, Deming
AU - Lai, Barry
AU - Maser, Jörg
AU - Finney, Lydia
AU - Roehrig, Christian
AU - VonOsinski, Jay
AU - Bolbat, Michael
AU - Brister, Keith
AU - Jacobsen, Chris
AU - Woloschak, Gayle
AU - Vogt, Stefan
PY - 2013
Y1 - 2013
N2 - Hard X-ray fluorescence microscopy is one of the most sensitive techniques to perform trace elemental analysis of unsectioned biological samples, such as cells and tissues. As the spatial resolution increases beyond sub-micron scale, conventional sample preparation method, which involves dehydration, may not be sufficient for preserving subcellular structures in the context of radiation-induced artifacts. Imaging of frozen-hydrated samples under cryogenic conditions is the only reliable way to fully preserve the three dimensional structures of the samples while minimizing the loss of diffusible ions. To allow imaging under this hydrated natural-state condition, we have developed the Bionanoprobe (BNP), a hard X-ray fluorescence nanoprobe with cryogenic capabilities, dedicated to studying trace elements in frozen-hydrated biological systems. The BNP is installed at an undulator beamline at Life Sciences Collaboration Access Team at the Advanced Photon Source. It provides a spatial resolution of 30 nm for fluorescence imaging by using Fresnel zone plates as nanofocusing optics. Differential phase contrast imaging is carried out in parallel to fluorescence imaging by using a quadrant photodiode mounted downstream of the sample. By employing a liquid-nitrogen-cooled sample stage and cryo specimen transfer mechanism, the samples are well maintained below 110 K during both transfer and X-ray imaging. The BNP is capable for automated tomographic dataset collection, which enables visualization of internal structures and composition of samples in a nondestructive manner. In this presentation, we will describe the instrument design principles, quantify instrument performance, and report the early results that were obtained from frozen-hydrated whole cells.
AB - Hard X-ray fluorescence microscopy is one of the most sensitive techniques to perform trace elemental analysis of unsectioned biological samples, such as cells and tissues. As the spatial resolution increases beyond sub-micron scale, conventional sample preparation method, which involves dehydration, may not be sufficient for preserving subcellular structures in the context of radiation-induced artifacts. Imaging of frozen-hydrated samples under cryogenic conditions is the only reliable way to fully preserve the three dimensional structures of the samples while minimizing the loss of diffusible ions. To allow imaging under this hydrated natural-state condition, we have developed the Bionanoprobe (BNP), a hard X-ray fluorescence nanoprobe with cryogenic capabilities, dedicated to studying trace elements in frozen-hydrated biological systems. The BNP is installed at an undulator beamline at Life Sciences Collaboration Access Team at the Advanced Photon Source. It provides a spatial resolution of 30 nm for fluorescence imaging by using Fresnel zone plates as nanofocusing optics. Differential phase contrast imaging is carried out in parallel to fluorescence imaging by using a quadrant photodiode mounted downstream of the sample. By employing a liquid-nitrogen-cooled sample stage and cryo specimen transfer mechanism, the samples are well maintained below 110 K during both transfer and X-ray imaging. The BNP is capable for automated tomographic dataset collection, which enables visualization of internal structures and composition of samples in a nondestructive manner. In this presentation, we will describe the instrument design principles, quantify instrument performance, and report the early results that were obtained from frozen-hydrated whole cells.
KW - Cryogenic capabilities
KW - Hard X-ray fluorescence microscopy
KW - The Bionanoprobe
KW - Tomography
UR - http://www.scopus.com/inward/record.url?scp=84888167149&partnerID=8YFLogxK
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U2 - 10.1117/12.2025169
DO - 10.1117/12.2025169
M3 - Conference contribution
AN - SCOPUS:84888167149
SN - 9780819497017
T3 - Proceedings of SPIE - The International Society for Optical Engineering
BT - X-Ray Nanoimaging
PB - SPIE
T2 - X-Ray Nanoimaging: Instruments and Methods
Y2 - 28 August 2013 through 29 August 2013
ER -