Automated measurement of nerve fiber density using line intensity scan analysis

► We developed an automated method to quantify fluorescently labeled nerve fibers. ► Our method makes unique use of line intensity scan, NIH ImageJ plugin and built-in peak detection algorithms of MATLAB. ► Our method enables efficient and reliable nerve fiber density measurement in large tissue areas from low magnification epifluorescence images. ► We measured cholinergic nerve fiber density in cortical and hippocampal regions and our results are accurate and highly reproducible. ► Our method is highly adaptable and can be broadly useful in both research and clinical investigation.