Serial section electron tomography: a method for three-dimensional reconstruction of large structures.
We present a method for combining single axis tomography and serial sectioning techniques to derive a three-dimensional reconstruction of large structures at electron microscopic resolution. This serial-tomography method allows the use of sufficiently thin sections to achieve adequate resolution with electron tomography, yet enables the generation of large reconstructions with considerably fewer sections than would be required using a serial thin section reconstruction technique. Serial thick sections (1-2 microns) are cut through the structure of interest, tomographic volume reconstructions are obtained for each section from a single axis tilt series, and the resulting series of volumes are then aligned and combined to form a single large volume. The serial-tomography method is illustrated with several samples, including red blood cells, the Golgi apparatus, and a spiny dendrite of a cortical pyramidal neuron. In some of these samples, the reconstruction is compared to correlated light microscopic views. The resulting large volume reconstructions appear to represent accurately the size and shape of objects such as red blood cells and spiny dendrites. The continuity of complex, tortuous structures such as the Golgi apparatus is also maintained across serial volumes. These examples demonstrate that it is possible to align and link a series of tomographic volumes accurately and that serial-tomography is a useful method for reconstructing relatively large structures without resorting to large numbers of serial thin sections.
Soto
G E
GE
San Diego Microscopy and Imaging Resource, University of California, La Jolla 92093-0608, USA.
Young
S J
SJ
Martone
M E
ME
Deerinck
T J
TJ
Lamont
S
S
Carragher
B O
BO
Hama
K
K
Ellisman
M H
MH
D000818:Animals; D048429:Cell Size / Q000502:physiology; D003199:Computer Systems; D003712:Dendrites / Q000648:ultrastructure; D004912:Erythrocytes / Q000648:ultrastructure; D005727:Ganglia, Spinal / Q000033:anatomy & histology; D006056:Golgi Apparatus / Q000648:ultrastructure; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation*; D008854:Microscopy, Electron / Q000295:instrumentation*; D009474:Neurons / Q000648:ultrastructure; D017966:Pyramidal Cells / Q000648:ultrastructure; D011892:Rana catesbeiana; D051381:Rats; D012984:Software; D014054:Tomography / Q000295:instrumentation*