september, 2024
Event Details
Host: Jörg Enderlein (U GOE), MBExC Local organizer: Alexander Egner (IFNANO) Speaker: Carlas Smith, TU Delft How to shatter the glass ceiling
Event Details
Host: Jörg Enderlein (U GOE), MBExC
Local organizer: Alexander Egner (IFNANO)
Speaker: Carlas Smith, TU Delft
How to shatter the glass ceiling of localization precision using modulated illumination?tbd
Abstract:
“This resolution game is not about lenses anymore.”, Stefan Hell remarked during his Nobel Lecture in 2014. Through replacing the lens maker with the statistician, single-molecule localization microscopy (SMLM) is able to circumvent the diffraction limit, by sparse activation and subsequent localization of single fluorescent probes. This leads to the well-known ‘glass ceiling’ that the localization precision of SMLM improves with the square root of the amount of recorded signal photons.
But why stop there? In this talk, we will show how to improve over the glass ceiling of localization precision, by combining modulated illumination with SMLM. Using illumination patterns, additional statistical information is added to the measurements, which encodes the relative position between an emitter and the illumination pattern.
Our method called SIMFLUX encodes this information using sinusoidal illumination patterns, leading to an improvement of at most 2.4 over SMLM. Furthermore, by projecting donut-shaped illumination patterns through a spinning disk, our SpinFlux methodology leads to a maximum improvement of 3.5 times.
After establishing this, we go a step further and show how to completely shatter the glass ceiling of localization precision. By iteratively repositioning illumination patterns based on previous measurements, such as in iterative MINFLUX, we can zoom in on individual molecules and break the scaling law of SMLM. Our research, awarded the prestigious Biophysical Journal 2022 Paper of the Year-Early Career Investigator Award, uncovers the inherent limits and trade-offs on the localization precision, propelling the frontiers of single-molecule microscopy.
Our findings reveal that, in the absence of background noise and with perfect modulation, the information content of signal photons exhibits exponential growth with each iteration. Nevertheless, the rate of information increase deviates from exponential behavior when confronted with non-zero background noise or imperfect modulation. Through advancing our understanding of super-resolution imaging, our work sets the stage for future breakthroughs in this field.
Organizer
MBExC