Stereology meets electron tomography: towards quantitative 3D electron microscopy.
Stereological tools are the gold standard for accurate (i.e., unbiased) and precise quantification of any microscopic sample. The past decades have provided a broad spectrum of tools to estimate a variety of parameters such as volumes, surfaces, lengths, and numbers. Some of them require pairs of parallel sections that can be produced by either physical or optical sectioning, with optical sectioning being much more efficient when applicable. Unfortunately, transmission electron microscopy could not fully profit from these riches, mainly because of the large depth of field. Hence, optical sectioning was a long-time desire for electron microscopists. This desire was fulfilled with the development of electron tomography that yield stacks of slices from electron microscopic sections. Now, parallel optical slices of a previously unimagined small thickness (2-5 nm axial resolution) can be produced. These optical slices minimize problems related to overprojection effects, and allow for direct stereological analysis, e.g., volume estimation with the Cavalieri principle and number estimation with the optical disector method. Here, we demonstrate that the symbiosis of stereology and electron tomography is an easy and efficient way for quantitative analysis at the electron microscopic level. We call this approach quantitative 3D electron microscopy.
Vanhecke
Dimitri
D
University of Bern, Institute of Anatomy, Experimental Morphology, Baltzerstrasse 2, 3000 Bern 9, Switzerland. vanhecke@ana.unibe.ch
Studer
Daniel
D
Ochs
Matthias
M
D006801:Humans; D021621:Imaging, Three-Dimensional; D008168:Lung / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D013238:Stereotaxic Techniques