Spatial distribution of neurons in tissue culture wells: implications for sampling methods to estimate population size.

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Many laboratory procedures require the counting of cells in culture. While many cultured cells may be counted by automated methods, neuronal cultures often require manual cell counting methods that are prohibitively time-consuming. This paper examines methods of sampling from tissue culture wells for estimating total cell counts. Performance of sampling and estimation schemes will depend in part on how the cells distribute themselves within a well. Spatial statistical analysis techniques are applied to the known total number and distribution of neurons in two wells counted in a grid scheme to demonstrate some important features of the neuron distributional patterns. Based on these two wells and simulated realizations from other point processes, a new sampling and estimation technique using open wedge-shaped sampling regions radiating from the centre of the well is proposed. This method is shown to result in more accurate estimates of the total number of neurons in the well than standard methods.