3.2.1 SRRF
NanoPyx's Super-Resolution Radial Fluctuations (SRRF) method generates super-resolved images by exploiting the temporal fluctuations of fluorophores in a sequence of microscopy images. Unlike traditional super-resolution techniques that require specialized equipment or complex protocols, SRRF processes standard fluorescence image sequences to enhance resolution beyond the diffraction limit. It works by analyzing the radial symmetry and intensity of fluctuating fluorescent signals to pinpoint the locations of emitters with high precision. These localized emitters are then used to reconstruct a high-resolution image, revealing fine structural details that are not visible in conventional microscopy.
Reference: Gustafsson, N. et al 2016 Nat Commun 7, 12471
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Image Stack: The image stack to be processed, required to have shape: (time, rows, columns).
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Frames Per Time Point: How many frames are used to calculated individual timepoints. For example, given an input image with 500 frames, if using 100 frames per timepoint, SRRF will generate an image stack with 5 super-resolved frames. Defaults to using every frame to calculate a single timepoint.
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Magnification: Desired magnification for the generated radiality image.
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Ring Radius: Radius of the ring used to calculate the radiality (in pixels).
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Radiality Positivity Constraint: Enable radiality positivity constraint
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Apply Intensity Weighting: Whether to calculate intensity values based on the original image fluorescence intensity as opposed to just outputting the radiality values.
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Reconstruction: Type of temporal reconstruction.