| RRC ID |
11173
|
| Author |
Engelhardt P, Meriläinen J, Zhao F, Uchiyama S, Fukui K, Lehto VP.
|
| Title |
Whole-mount immunoelectron tomography of chromosomes and cells.
|
| Journal |
Methods Mol Biol
|
| Abstract |
Standard immunogold-labeling methods in transmission electron microscopy (TEM) are unable to locate immunogold particles in the depth direction. This inability does not only concern bulky whole mounts, but also sections. A partial solution to the problem is stereo inspection. However, three-dimensional reconstruction of immunogold-labeled structures, that is, immuno-electron tomography (IET), is a correct solution for this inconsistency. Striking improvement in resolution is achieved: the 1.4-nm immunogold particles are shown in IET that are not detected in the original tilt series. IET is not restricted to laboratories with advanced medium- or high-voltage TEM and super-computing facilities; the methods we have developed for whole-mounted chromosomes and also for whole-mounted cytoskeleton of fibroblasts work remarkably well with ordinary 80-kV TEMs equipped with a goniometer to collect tilt series for IET on film. In addition, free programs are available to produce three-dimensional reconstructions even without high-performance computers. These improvements make it possible to many laboratories without modern facilities to perform IET reconstruction with standard TEM apparatus.
|
| Volume |
369
|
| Pages |
387-405
|
| Published |
2007-1-1
|
| DOI |
10.1007/978-1-59745-294-6_19
|
| PII |
1-59745-294-7:387
|
| PMID |
17656761
|
| MeSH |
Animals
Cell Line
Cells / ultrastructure
Chick Embryo
Chromosomes / ultrastructure*
Chromosomes, Human / ultrastructure
Gold Colloid
HeLa Cells
Humans
Imaging, Three-Dimensional / methods*
Microscopy, Immunoelectron / methods*
Tomography / methods
|
| Resource |
| Human and Animal Cells |
HeLa(RCB0007)
HeLa.S3(RCB0191)
K562(RCB0027)
BALL-1(RCB0256) |
