Quantitative Analysis of Photoreceptor Intensity–Response Function in Fly Visual Neurons

In this protocol, we illustrate how to process images acquired during functional imaging of fly visual neurons and how to analyze and quantify visually evoked activities. We use ImageJ/Fiji for the initial imaging processing. All images acquired previously should be registered to compensate for tissue movement. Next, we extract fluorescence signals specifically from neurons that respond to the light by marking the regions of interest (ROIs). The data are further analyzed in a data-analysis program, such as MATLAB, to plot response traces against time. Finally, we obtain different parameters to reveal the neuron's physiological properties by fitting the data with a Naka–Rushton function.