Abstrakt
Three-dimensional (3-D) objects such as fibroblasts distribute in a 3-D volumes such as tissues or artificial scaffolds. Current methods for cell number determination in 2-D (e.g., DNA/protein quantification in cells on plastic, cell counting on surface of 3D objects) suffer from lack of accuracy and descriptions of the spatial distribution of cells.
Further, 2-D sampling of 3-D objects suffer from biases associated with cellular size, shape and orientation1,2. These obstacles can be addressed with the application of unbiased (accurate) methods for acquisition of cell numbers and spatial distribution in a defined region of interest.
The aim of this project is to develop a method for accurate cell number determination and spatial distribution using freely available ImageJ (FIJI) software and stereological principles for cell sampling and quantification. Here we outline an approach that can be broadly applied to quantify cells and their distribution in a 3-D tissue or scaffold volume under normal or experimental conditions.