TY - JOUR
T1 - A novel approach to non-biased systematic random sampling
T2 - A stereologic estimate of Purkinje cells in the human cerebellum
AU - Agashiwala, Rajiv M.
AU - Louis, Elan D.
AU - Hof, Patrick R.
AU - Perl, Daniel P.
N1 - Funding Information:
We gratefully acknowledge the excellent assistance of Dr. Dushyant P. Purohit, Ms. Amy Hsu and Ms. Sandy Harry in carrying out this work. We thank Ms. Vina Ayers for her assistance with Fig. 1 . This study was supported by funds from NIH grants NS042859 (Dr. Louis), AD02219, AG05138 and MH058911 (Drs. Perl, Hof).
PY - 2008/10/21
Y1 - 2008/10/21
N2 - Non-biased systematic sampling using the principles of stereology provides accurate quantitative estimates of objects within neuroanatomic structures. However, the basic principles of stereology are not optimally suited for counting objects that selectively exist within a limited but complex and convoluted portion of the sample, such as occurs when counting cerebellar Purkinje cells. In an effort to quantify Purkinje cells in association with certain neurodegenerative disorders, we developed a new method for stereologic sampling of the cerebellar cortex, involving calculating the volume of the cerebellar tissues, identifying and isolating the Purkinje cell layer and using this information to extrapolate non-biased systematic sampling data to estimate the total number of Purkinje cells in the tissues. Using this approach, we counted Purkinje cells in the right cerebella of four human male control specimens, aged 41, 67, 70 and 84 years, and estimated the total Purkinje cell number for the four entire cerebella to be 27.03, 19.74, 20.44 and 22.03 million cells, respectively. The precision of the method is seen when comparing the density of the cells within the tissue: 266,274, 173,166, 167,603 and 183,575 cells/cm3, respectively. Prior literature documents Purkinje cell counts ranging from 14.8 to 30.5 million cells. These data demonstrate the accuracy of our approach. Our novel approach, which offers an improvement over previous methodologies, is of value for quantitative work of this nature. This approach could be applied to morphometric studies of other similarly complex tissues as well.
AB - Non-biased systematic sampling using the principles of stereology provides accurate quantitative estimates of objects within neuroanatomic structures. However, the basic principles of stereology are not optimally suited for counting objects that selectively exist within a limited but complex and convoluted portion of the sample, such as occurs when counting cerebellar Purkinje cells. In an effort to quantify Purkinje cells in association with certain neurodegenerative disorders, we developed a new method for stereologic sampling of the cerebellar cortex, involving calculating the volume of the cerebellar tissues, identifying and isolating the Purkinje cell layer and using this information to extrapolate non-biased systematic sampling data to estimate the total number of Purkinje cells in the tissues. Using this approach, we counted Purkinje cells in the right cerebella of four human male control specimens, aged 41, 67, 70 and 84 years, and estimated the total Purkinje cell number for the four entire cerebella to be 27.03, 19.74, 20.44 and 22.03 million cells, respectively. The precision of the method is seen when comparing the density of the cells within the tissue: 266,274, 173,166, 167,603 and 183,575 cells/cm3, respectively. Prior literature documents Purkinje cell counts ranging from 14.8 to 30.5 million cells. These data demonstrate the accuracy of our approach. Our novel approach, which offers an improvement over previous methodologies, is of value for quantitative work of this nature. This approach could be applied to morphometric studies of other similarly complex tissues as well.
KW - Brain
KW - Cerebellum
KW - Essential tremor
KW - Methodology
KW - Stereology
UR - http://www.scopus.com/inward/record.url?scp=53249130651&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=53249130651&partnerID=8YFLogxK
U2 - 10.1016/j.brainres.2008.07.119
DO - 10.1016/j.brainres.2008.07.119
M3 - Article
C2 - 18725208
AN - SCOPUS:53249130651
SN - 0006-8993
VL - 1236
SP - 73
EP - 78
JO - Brain Research
JF - Brain Research
ER -