An empirical analysis of the precision of estimating the numbers of neurons and glia in human neocortex using a fractionator-design with sub-sampling

TY - JOUR

T1 - An empirical analysis of the precision of estimating the numbers of neurons and glia in human neocortex using a fractionator-design with sub-sampling

AU - Lyck, L.

AU - Santamaria, I.D.

AU - Pakkenberg, B.

AU - Chemnitz, J.

AU - Schroder, H.D.

AU - Finsen, B.

AU - Gundersen, H.J.G.

N1 - Times Cited: 0ArticleEnglishFinsen, BUniv So Denmark, Ctr Med Biotechnol, Inst Med Biol, JB Winslowsvej 25,2nd Floor, DK-5000 Odense C, DenmarkCited References Count: 54491MAELSEVIER SCIENCE BVPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSAMSTERDAM

PY - 2009

Y1 - 2009

N2 - Improving histomorphometric analysis of the human neocortex by combining stereological cell counting with immunchistochemical visualisation of specific neuronal and glial cell populations is a methodological challenge. To enable standardized immunohistochemical staining, the amount of brain tissue to be stained and analysed by cell counting was efficiently reduced using a fractionator protocol involving several steps of sub-sampling. Since no mathematical or statistical tools exist to predict the variance originating from repeated sampling in complex structures like the human neocortex, the variance at each level of sampling was determined empirically. The methodology was tested in three brains analysing the contribution of the multi-step sampling procedure to the precision on the estimated total numbers of immunohistochemically defined NeuN expressing (NeuN(+)) neurons and CD45(+) microglia. The results showed that it was possible, but not straight forward, to combine immunohistochemistry and the optical fractionator for estimation of specific subpopulations of brain cells in human neocortex. (C) 2009 Elsevier B.V. All rights reserved Udgivelsesdato: 2009/9/15

AB - Improving histomorphometric analysis of the human neocortex by combining stereological cell counting with immunchistochemical visualisation of specific neuronal and glial cell populations is a methodological challenge. To enable standardized immunohistochemical staining, the amount of brain tissue to be stained and analysed by cell counting was efficiently reduced using a fractionator protocol involving several steps of sub-sampling. Since no mathematical or statistical tools exist to predict the variance originating from repeated sampling in complex structures like the human neocortex, the variance at each level of sampling was determined empirically. The methodology was tested in three brains analysing the contribution of the multi-step sampling procedure to the precision on the estimated total numbers of immunohistochemically defined NeuN expressing (NeuN(+)) neurons and CD45(+) microglia. The results showed that it was possible, but not straight forward, to combine immunohistochemistry and the optical fractionator for estimation of specific subpopulations of brain cells in human neocortex. (C) 2009 Elsevier B.V. All rights reserved Udgivelsesdato: 2009/9/15

M3 - Tidsskriftartikel

SN - 0165-0270

VL - 182

SP - 143

EP - 156

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

IS - 2

ER -