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Evaluation of sted super-resolution image quality by image correlation spectroscopy (QuICS). | LitMetric

AI Article Synopsis

  • An unbiased assessment of imaging performance in super-resolution microscopy is essential, and an algorithm using image correlation spectroscopy (ICS) can effectively evaluate the quality of super-resolution images.
  • The algorithm calculates three important parameters: the width of the autocorrelation function for spatial resolution, brightness for image contrast, and relative noise variance for signal-to-noise ratio.
  • The study also explores the effects of STED imaging conditions on DNA replication foci in U937 cells and finds that different methods for separating photons by lifetime tuning (SPLIT) can enhance resolution and contrast in STED microscopy.

Article Abstract

Quantifying the imaging performances in an unbiased way is of outmost importance in super-resolution microscopy. Here, we describe an algorithm based on image correlation spectroscopy (ICS) that can be used to assess the quality of super-resolution images. The algorithm is based on the calculation of an autocorrelation function and provides three different parameters: the width of the autocorrelation function, related to the spatial resolution; the brightness, related to the image contrast; the relative noise variance, related to the signal-to-noise ratio of the image. We use this algorithm to evaluate the quality of stimulated emission depletion (STED) images of DNA replication foci in U937 cells acquired under different imaging conditions. Increasing the STED depletion power improves the resolution but may reduce the image contrast. Increasing the number of line averages improves the signal-to-noise ratio but facilitates the onset of photobleaching and subsequent reduction of the image contrast. Finally, we evaluate the performances of two different separation of photons by lifetime tuning (SPLIT) approaches: the method of tunable STED depletion power and the commercially available Leica Tau-STED. We find that SPLIT provides an efficient way to improve the resolution and contrast in STED microscopy.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8531054PMC
http://dx.doi.org/10.1038/s41598-021-00301-xDOI Listing

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