Three-Dimensional Digital Image Analysis of Immunostained Neurons in Thick Tissue Sections

Detection and three dimensional reconstruction of cell structures from brightfield microscopy video clips using digital image processing algorithms is presented. While the confocal microscopy offers an efficient technique for three dimensional measurements, extensive and repeated measurements are still often better to be performed using permanent staining and brightfield microscopy. By processing of brightfield microscopy videos using automated and efficient digital image processing algorithms, the tedious task of manual analysis can be avoided. Our two-stage algorithm is applied for 1) cell soma detection and 2) identification of the 3D structure of entire neurons. To verify the results, we present 3D reconstructions of the detected cells.