Abstract
Cryo-electron tomography (cryo-ET) has enabled high resolution three-dimensional(3D) structural analysis of virus and host cell interactions and many cell signaling events; these studies, however, have largely been limited to very thin, peripheral regions of eukaryotic cells or to small prokaryotic cells. Recent efforts to make thin, vitreous sections using cryo-ultramicrotomy have been successful, however,this method is technically very challenging and with many artifacts. Here, we report a simple and robust method for creating in situ, frozen-hydrated cell lamellas using a focused ion beam at cryogenic temperature (cryo-FIB), allowing access to any interior cellular regions of interest. We demonstrate the utility of cryo-FIB with high resolution 3D cellular structures from both bacterial cells and large mammalian cells. The method will not only facilitate high-throughput 3D structural analysis of biological specimens, but is also broadly applicable to sample preparation of thin films and surface materials without the need for FIB " lift-out"
| Original language | English (US) |
|---|---|
| Pages (from-to) | 318-326 |
| Number of pages | 9 |
| Journal | Journal of Structural Biology |
| Volume | 180 |
| Issue number | 2 |
| DOIs | |
| State | Published - Nov 1 2012 |
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All Science Journal Classification (ASJC) codes
- Structural Biology
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3D structure determination of native mammalian cells using cryo-FIB and cryo-electron tomography. / Wang, Ke; Strunk, Korrinn; Zhao, Gongpu; Gray, Jennifer L.; Zhang, Peijun.
In: Journal of Structural Biology, Vol. 180, No. 2, 01.11.2012, p. 318-326.Research output: Contribution to journal › Article
TY - JOUR
T1 - 3D structure determination of native mammalian cells using cryo-FIB and cryo-electron tomography
AU - Wang, Ke
AU - Strunk, Korrinn
AU - Zhao, Gongpu
AU - Gray, Jennifer L.
AU - Zhang, Peijun
PY - 2012/11/1
Y1 - 2012/11/1
N2 - Cryo-electron tomography (cryo-ET) has enabled high resolution three-dimensional(3D) structural analysis of virus and host cell interactions and many cell signaling events; these studies, however, have largely been limited to very thin, peripheral regions of eukaryotic cells or to small prokaryotic cells. Recent efforts to make thin, vitreous sections using cryo-ultramicrotomy have been successful, however,this method is technically very challenging and with many artifacts. Here, we report a simple and robust method for creating in situ, frozen-hydrated cell lamellas using a focused ion beam at cryogenic temperature (cryo-FIB), allowing access to any interior cellular regions of interest. We demonstrate the utility of cryo-FIB with high resolution 3D cellular structures from both bacterial cells and large mammalian cells. The method will not only facilitate high-throughput 3D structural analysis of biological specimens, but is also broadly applicable to sample preparation of thin films and surface materials without the need for FIB " lift-out"
AB - Cryo-electron tomography (cryo-ET) has enabled high resolution three-dimensional(3D) structural analysis of virus and host cell interactions and many cell signaling events; these studies, however, have largely been limited to very thin, peripheral regions of eukaryotic cells or to small prokaryotic cells. Recent efforts to make thin, vitreous sections using cryo-ultramicrotomy have been successful, however,this method is technically very challenging and with many artifacts. Here, we report a simple and robust method for creating in situ, frozen-hydrated cell lamellas using a focused ion beam at cryogenic temperature (cryo-FIB), allowing access to any interior cellular regions of interest. We demonstrate the utility of cryo-FIB with high resolution 3D cellular structures from both bacterial cells and large mammalian cells. The method will not only facilitate high-throughput 3D structural analysis of biological specimens, but is also broadly applicable to sample preparation of thin films and surface materials without the need for FIB " lift-out"
UR - http://www.scopus.com/inward/record.url?scp=84867910247&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84867910247&partnerID=8YFLogxK
U2 - 10.1016/j.jsb.2012.07.003
DO - 10.1016/j.jsb.2012.07.003
M3 - Article
C2 - 22796867
AN - SCOPUS:84867910247
VL - 180
SP - 318
EP - 326
JO - Journal of Structural Biology
JF - Journal of Structural Biology
SN - 1047-8477
IS - 2
ER -