Measurement Errors in Fluorescence Microscopy Image Registration

Image registration is an important processing step in fluorescence microscopy, for example in tracking or super-resolution methods. Precision localization of single fluorescent molecules from a quantum limited photon detection process, subject to Gaussian readout noise, is key to the use of single molecule microscopy. It is therefore important to know the effect that registration has on the accuracy of localizing a single molecule. Here we demonstrate a suitable approach to image registration that accounts for point-wise errors in localizing the control points typically used in fluorescence microscopy. This allows expressions for the localization errors caused by the registration process to be derived, showing dependence on the number of control points and their associated photon counts.