Super Resolution Drift Correction with Intermittently Observable Markers

Microscope drift correction necessary for super-resolution microscopy over extended time periods generally requires either significant modification of the microscope setup or the introduction of large fiduciary markers into the sample, which may perturb the cell. Further, existing drift correction methods expect the fiduciary markers to be continually observable, ruling out most quantum dots due to blinking. Similarly, single fluorophores have insufficient signal to make ideal markers, and as such, large particles or fluorescent beads are common fiduciary markers. Here, a drift correction method is developed allowing for intermittent observations, allowing quantum dots, which are less likely to perturb cells, to serve as fiduciary markers. A statistical comparison of intuitive methods of drift correction with the method developed here reveals that naively applying typical methods to intermittent particles introduces significant amounts of additional uncertainty, sufficient to significantly reduce the effective localization accuracy over the course of super-resolution experiments. The method developed here works equally well whether particle observations are intermittent or continual, as long as a comparable number of total particle observations are available.