New stereological methods for counting neurons

The issue of whether or not neurons die with age is of fundamental importance for understanding senescent decline in motor, sensory, and cognitive functions. Yet after over 100 years of attempts to quantify age-related neuron loss, this fundamental issue is "swirling in controversy" and the results of numerous studies addressing this issue "rife with confusion and seemingly contradictory data" (7). This is in large part due to deficiencies in the way we have thought about what constitutes evidence for neuron loss and deficiencies in the methods that have been available for counting neurons. Over the past several years, a new generation of stereological principles have been developed that have made it possible to identify and eliminate these deficiencies. Although there are now a number of review articles that briefly describe these and other new stereological principles (4,9,13,14,15,18,23), for the most part detailed descriptions of the new principles for counting neurons are found in specialized literature and are accompanied by new terms and concepts that have overwhelmed potential users and inhibited their adoption. In this essay, I describe the principles and concepts that are involved in the new stereological approach to neuron counting and discuss why the data obtained from their application represent improvements over those obtained with previously available techniques. A practical guide to the application of the principles to counting neurons or any other cell, organelle, or object present in sectioned material has been included as an appendix.

[1]  E. Weibel.  Stereological Methods. Practical methods for biological morphometry , 1979 .

[2]  H J Gundersen,et al.  The efficiency of systematic sampling in stereology and its prediction * , 1987, Journal of microscopy.

[3]  H. Uylings,et al.  Comments on review by coleman and flood ‘neuron numbers and dendritic extent in normal aging and alzheimer's disease.’ density measures: Parameters to avoid , 1987, Neurobiology of Aging.

[4]  H. Gundersen,et al.  Unbiased stereological estimation of the number of neurons in the human hippocampus , 1990, The Journal of comparative neurology.

[5]  E R Weibel,et al.  Recent stereological methods for cell biology: a brief survey. , 1990, The American journal of physiology.

[6]  H. Gundersen,et al.  Unbiased stereological estimation of the total number of neurons in the subdivisions of the rat hippocampus using the optical fractionator , 1991, The Anatomical record.

[7]  Luis M. Cruz-Orive,et al.  On the empirical variance of a fractionator estimate , 1990 .

[8]  Mark J. West,et al.  Regionally specific loss of neurons in the aging human hippocampus , 1993, Neurobiology of Aging.

[9]  E. Weibel,et al.  A principle for counting tissue structures on random sections. , 1962, Journal of applied physiology.

[10]  T M Mayhew,et al.  A review of recent advances in stereology for quantifying neural structure , 1992, Journal of neurocytology.

[11]  Paul D. Coleman,et al.  Neuron numbers and dendritic extent in normal aging and Alzheimer's disease , 1987, Neurobiology of Aging.

[12]  L M Cruz-Orive,et al.  Estimation of surface area from vertical sections , 1986, Journal of microscopy.

[13]  H J Gundersen,et al.  Stereology: the fast lane between neuroanatomy and brain function ‐ or still only a tightrope? , 1992, Acta neurologica Scandinavica. Supplementum.

[14]  H J Gundersen,et al.  The absolute number of nerve cells in substantia nigra in normal subjects and in patients with Parkinson's disease estimated with an unbiased stereological method. , 1991, Journal of neurology, neurosurgery, and psychiatry.

[15]  T F Bendtsen,et al.  Unbiased estimation of particle number using sections—an historical perspective with special reference to the stereology of glomeruli , 1989, Journal of microscopy.

[16]  L M Cruz-Orive,et al.  Application of the Cavalieri principle and vertical sections method to lung: estimation of volume and pleural surface area , 1988, Journal of microscopy.

[17]  Richard E. Coggeshall,et al.  A consideration of neural counting methods , 1992, Trends in Neurosciences.

[18]  Jean-Pierre Royet,et al.  Stereology: A method for analyzing images , 1991, Progress in Neurobiology.

[19]  Bonaventura Cavalieri,et al.  Geometria degli indivisibili , 1966 .

[20]  H. J. G. GUNDERSEN,et al.  Some new, simple and efficient stereological methods and their use in pathological research and diagnosis , 1988, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[21]  M. Abercrombie.  Estimation of nuclear population from microtome sections , 1946, The Anatomical record.

[22]  H. Gundersen.  Stereology of arbitrary particles * , 1986, Journal of microscopy.

[23]  H. J. G. Gundersen,et al.  The impact of recent stereological advances on quantitative studies of the nervous system , 1986, Journal of Neuroscience Methods.

[24]  H. J. G. Gundersen,et al.  The new stereological tools: Disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis , 1988, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[25]  D. C. Sterio.  The unbiased estimation of number and sizes of arbitrary particles using the disector , 1984, Journal of microscopy.

[26]  H. Gundersen,et al.  Notes on the estimation of the numerical density of arbitrary profiles: the edge effect , 1977 .

[27]  H J Gundersen,et al.  Total number of neurons and glial cells in human brain nuclei estimated by the disector and the fractionator , 1988, Journal of microscopy.

[28]  D. Rohrlich,et al.  Counting sectioned cells via mathematical reconstruction , 1987, The Journal of comparative neurology.