Optics Express · 2020 · 15 citations · 19 references
Natural SciencesBioengineeringBiomedical EngineeringBiomedical ImagingBiomedical OpticsBiophotonicsCellular ImagingFluorescence ImagingFluorescence MicroscopyImage QuantificationSuper-resolution MicroscopyThreshold EstimationSignal ThresholdingSuper-resolutionSuper-resolution Fluorescence ImagingSuper-resolution Fluorescence MicroscopySuper-resolution Image AnalysisSuper-resolution ImagingSuper-resolution ProcessingSuper-resolution Reconstruction
Multiple signal classification algorithm (MUSICAL) exploits temporal fluctuations in fluorescence intensity to perform super-resolution microscopy by computing the value of a super-resolving indicator function across a fine sample grid. A key step in the algorithm is the separation of the measurements into signal and noise subspaces, based on a single user-specified parameter called the threshold. The resulting image is strongly sensitive to this parameter and the subjectivity arising from multiple practical factors makes it difficult to determine the right rule of selection. We address this issue by proposing soft thresholding schemes derived from a new generalized framework for indicator function design. We show that the new schemes significantly alleviate the subjectivity and sensitivity of hard thresholding while retaining the super-resolution ability. We also evaluate the trade-off between resolution and contrast and the out-of-focus light rejection using the various indicator functions. Through this, we create significant new insights into the use and further optimization of MUSICAL for a wide range of practical scenarios.
19
Imaging Intracellular Fluorescent Proteins at Nanometer Resolution
Eric Betzig, George H. Patterson, Rachid Sougrat et al. · Science · 2006 · 8.9K citations