Objective Morphological Quantification of Microscopic Images Using a Fast Fourier Transform (FFT) Analysis

Quantification of immunohistochemistry (IHC) and immunofluorescence (IF) using image intensity depends on a number of variables. These variables add a subjective complexity in keeping a standard within and between laboratories. Fast Fourier Transformation (FFT) algorithms, however, allow for a rapid and objective quantification (via statistical analysis) using cell morphologies when the microscopic structures are oriented or aligned. Quantification of alignment is given in terms of a ratio of FFT intensity to the intensity of an orthogonal angle, giving a numerical value of the alignment of the microscopic structures. This allows for a more objective analysis than alternative approaches, which rely upon relative intensities. Curr. Protoc. Essential Lab. Tech . 7:9.5.1‐9.5.12. © 2013 by John Wiley & Sons, Inc.