Unbiased Estimation of Cell Number Using the Automatic Optical Fractionator

Authors

Peter R. Moulton, University of South Florida College of Medicine
Hady Ahmady Phoulady Ph.D., University of Southern Maine
Dmitry Goldgof, University of South Florida College of Medicine
Lawrence O. Hall, University of South Florida College of Medicine
Marcia Gordon, University of South Florida College of Medicine
David Morgan, University of South Florida College of Medicine

Document Type

Article

Publication Date

12-14-2016

Publication Title

Journal of Chemical Neuroanatomy

Abstract

A novel stereology approach, the automatic optical fractionator, is presented for obtaining unbiased and efficient estimates of the number of cells in tissue sections. Used in combination with existing segmentation algorithms and ordinary immunostaining methods, automatic estimates of cell number are obtainable from extended depth of field images built from three-dimensional volumes of tissue (disector stacks). The automatic optical fractionator is more accurate, 100% objective and 8-10 times faster than the manual optical fractionator. An example of the automatic fractionator is provided for counts of immunostained neurons in neocortex of a genetically modified mouse model of neurodegeneration. Evidence is presented for the often overlooked prerequisite that accurate counting by the optical fractionator requires a thin focal plane generated by a high optical resolution lens.

Comments

doi:10.1016/j.jchemneu.2016.12.002

Recommended Citation

Mouton PR, Ahmady Phoulady H, Goldgof DB, Hall LO, Gordon M, Morgan D. 2016. "Unbiased Estimation of Cell Number Using the Automatic Optical Fractionator.”, Journal of Chemical Neuroanatomy. 80: A1-A8.