TY - JOUR
T1 - 3D Imaging of mammalian cells with ion-abrasion scanning electron microscopy
AU - Heymann, Jurgen A.W.
AU - Shi, Dan
AU - Kim, Sang
AU - Bliss, Donald
AU - Milne, Jacqueline L.S.
AU - Subramaniam, Sriram
PY - 2009/4
Y1 - 2009/4
N2 - Understanding the hierarchical organization of molecules and organelles within the interior of large eukaryotic cells is a challenge of fundamental interest in cell biology. We are using ion-abrasion scanning electron microscopy (IA-SEM) to visualize this hierarchical organization in an approach that combines focused ion-beam milling with scanning electron microscopy. Here, we extend our previous studies on imaging yeast cells to image subcellular architecture in human melanoma cells and melanocytes at resolutions as high as ∼6 and ∼20 nm in the directions parallel and perpendicular, respectively, to the direction of ion-beam milling. The 3D images demonstrate the striking spatial relationships between specific organelles such as mitochondria and membranes of the endoplasmic reticulum, and the distribution of unique cellular components such as melanosomes. We also show that 10 nm-sized gold particles and quantum dot particles with 7 nm-sized cores can be detected in single cross-sectional images. IA-SEM is thus a useful tool for imaging large mammalian cells in their entirety at resolutions in the nanometer range.
AB - Understanding the hierarchical organization of molecules and organelles within the interior of large eukaryotic cells is a challenge of fundamental interest in cell biology. We are using ion-abrasion scanning electron microscopy (IA-SEM) to visualize this hierarchical organization in an approach that combines focused ion-beam milling with scanning electron microscopy. Here, we extend our previous studies on imaging yeast cells to image subcellular architecture in human melanoma cells and melanocytes at resolutions as high as ∼6 and ∼20 nm in the directions parallel and perpendicular, respectively, to the direction of ion-beam milling. The 3D images demonstrate the striking spatial relationships between specific organelles such as mitochondria and membranes of the endoplasmic reticulum, and the distribution of unique cellular components such as melanosomes. We also show that 10 nm-sized gold particles and quantum dot particles with 7 nm-sized cores can be detected in single cross-sectional images. IA-SEM is thus a useful tool for imaging large mammalian cells in their entirety at resolutions in the nanometer range.
KW - Automated 3D imaging
KW - Cancer imaging
KW - Dual beam microscopy
KW - Melanoma detection
KW - Mitochondrial architecture
UR - http://www.scopus.com/inward/record.url?scp=60649115825&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=60649115825&partnerID=8YFLogxK
U2 - 10.1016/j.jsb.2008.11.005
DO - 10.1016/j.jsb.2008.11.005
M3 - Article
C2 - 19116171
AN - SCOPUS:60649115825
SN - 1047-8477
VL - 166
SP - 1
EP - 7
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 1
ER -