Simultaneous Recovery Of Both Bright And Dim Structures From Noisy Fluorescence Microscopy Images Using A Modified TV Constraint

The quality and information content of biological images can be significantly enhanced by post-acquisition processing using deconvolution and denoising.However,when imaging complex biological samples,such as neurons,stained with fluorescence labels,the signal level of different structures can differ by several orders of magnitude.This imposes a challenge as current image reconstruction algorithms are focused on recovering low signals and generally have sample-dependent performance,requiring tedious manual tuning.This is one of the main hurdles for their wide adoption by non-specialists.In this work we modify the general constrained reconstruction method(in our case utilizing a total variation constraint)so that both bright and dim structures can drive the deconvolution with equal force.In this way,we can simultaneously obtain high quality reconstruction across a wide range of signals within a single image or image sequence.The algorithm is tested on both simulated and experimental data.When compared with current state of art algorithms,our algorithm outperforms others in terms of maintaining the resolution in the high-signal areas and reducing artifacts in the low-signal areas.The algorithm was also tested on image sequences where one set of parameters are used to reconstruct all images,with blind evaluation by a group of biologists demonstrating a marked preference for the images produced by our method.This means that our method is suitable for batch processing of image sequences obtained from either spatial or temporal scanning.In structured illumination microscopy,deconvolution is used to improve resolution and remove noise.There is also a need to reconstruct low and high signals simultaneously.Using the modified total variation constraint,we reformulated the optimization and provided a solution.Using simulated data,we demonstrated the effectiveness of the modified constraint in recovering both bright and dim structures.