A new fractionator principle with varying sampling fractions: exemplified by estimation of synapse number using electron microscopy.
The quantification of ultrastructure has been permanently improved by the application of new stereological principles. Both precision and efficiency have been enhanced. Here we report for the first time a fractionator method that can be applied at the electron microscopy level. This new design incorporates a varying sampling fraction paradigm. The method allows for systematic random sampling from blocks of variable slab thickness, thereby eliminating the need for exhaustive serial sectioning through an entire containing space. This novel approach acknowledges the inaccuracy inherent in estimating the total object number using section sampling fractions based on the average thickness of sections of variable thicknesses. As an alternative, this approach estimates the correct particle section sampling probability based on an estimator of the Horvitz-Thompson type, resulting in a theoretically more satisfying and accurate estimate of the expected number of particles for the defined containing space.
Witgen
B M
BM
Stereology and Electron Microscopy Research Laboratory and MIND Center, Aarhus University, Worms Allé 1185, DK 8000 Aarhus C., Denmark. brent.witgen@ki.au.dk
Grady
M S
MS
Nyengaard
J R
JR
Gundersen
H J G
HJ
D000818:Animals; D006624:Hippocampus / Q000166:cytology / Q000648:ultrastructure*; D051379:Mice; D008854:Microscopy, Electron / Q000379:methods*; D011336:Probability; D013238:Stereotaxic Techniques; D013569:Synapses