Image sharpness reconstruction for 3D localization and tracking

A novel structure of multiplane image using combination of quadratically distorted grating and lens were proposed by Professor Alan Greenaway 1. Particle’s z position(longitude) can be determined using the sharpness metric and MLE (maximum likelihood estimation) algorithm. Professor Alan Greenaway and Paul have already given a good talk introducing the system and method. This talk will give more detail about how image sharpness and MLE works. The important thing is that the sharpness value calculated should independent with particle’s brightness (total signal intensity) and background. The preview method will subtract the background before calculating sharpness, this may lead some problem. A new method was developed without background subtraction but reconstruct sharpness after using original intensity calculating it. The orininal intensity sharpness is simply a scaling of “true” sharpness, plus a constant (baseline). Hence subtract the baseline, then rescale it will acquire the “true” sharpness.

1. Blanchard, P. M. and A. H. Greenaway (1999). "Simultaneous multiplane imaging with a distorted diffraction grating." Applied Optics 38(32): 6692-6699.