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Antonio Martinez-Sanchez
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Hauke Hillen
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march
Event Details
Host: Jörg Enderlein (U GOE), MBExC Local organizer: Alexander Egner (IFNANO) Speaker: Carlas S. Smith, Quantitative Nanoscopy, Delft Center for Systems
Event Details
Host: Jörg Enderlein (U GOE), MBExC
Local organizer: Alexander Egner (IFNANO)
Speaker: Carlas S. Smith, Quantitative Nanoscopy, Delft Center for Systems and Control, Delft University of Technology, The Netherlands
How to shatter the glass ceiling of localization precision using modulated illumination?
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.
Please find here the PDF of the announcement:
Organizer
MBExC
april
Event Details
Host: Jörg Enderlein (U GOE), MBExC Local organizer: Alexander Egner (IFNANO) Speaker: Prof. Guillermo Pedro Acuna, Photonic
Event Details
Host: Jörg Enderlein (U GOE), MBExC
Local organizer: Alexander Egner (IFNANO)
Speaker: Prof. Guillermo Pedro Acuna, Photonic Nanosystems, Department of Physics, University of Fribourg
Abstract:
Developments in low-cost microscopy have accelerated greatly in recent years due to the technological advances of modern smartphones. Among different features, these devices have image sensors with more pixels, better quantum efficiencies, better optics design for light collection, and larger focal distances in different lenses of multi-camera smartphones. Distinct aspects of smartphones, i.e., portability and compactness, have also pushed forward the development of specific smartphone-based setups useful in Point-Of-Care (POC) applications like clinical diagnostics, quantification of immunoassays, detection of bacteria, cancer cytology, fresh tissue imaging, lead and microplastics quantification. While most of these applications used optical setups designed for fluorescence imaging, only a few of them focused on the detection of single molecule fluorescence.
Here, we developed a portable and inexpensive smartphone-based fluorescence microscope that detects direct emission from single molecules. We tested its performance by analyzing single-molecule intensity traces with three smartphones. We also demonstrated that it can be used for super-resolution microscopy with a Single-Molecule Localization Microscopy (SMLM), DNA-PAINT. The smartphone-based microscope we present is low-cost, portable, easy to use, and can virtually be used with any smartphone, making an impact on a truly broad audience.
Please find here the PDF of the announcement:
Organizer
MBExC
june
Event Details
Host: Jörg Enderlein (U GOE), MBExC Local organizer: Alexander Egner (IFNANO) Speaker: Maciej Wojtkowski, International Center for Translational Eye Research (ICTER)
Event Details
Host: Jörg Enderlein (U GOE), MBExC
Local organizer: Alexander Egner (IFNANO)
Speaker: Maciej Wojtkowski, International Center for Translational Eye Research (ICTER) and Institute of Physical Chemistry, Polish Academy of Sciences, Warsaw, Poland
Spatio-temporal optical imaging – new method for in vivo imaging
Abstract:
In vivo optical microscopy with quality comparable to fixed samples remains challenging. We developed Spatio-Temporal Optical Coherence (STOC) Imaging, which dynamically adjusts spatial phase relationships in illuminating light to improve imaging quality. Applied to 3-D imaging as Spatio-Temporal Optical Coherence Tomography (STOC-T), this method uses hundreds of uncorrelated spectral interferograms to amplify ballistic photon signals while attenuating scattered photons. STOC-T provides fast, high-contrast imaging, maintaining high resolution at significant depths without repeated measurements. When applied to eye imaging, STOC-T supports functional imaging techniques like Optoretinography (ORG), which measures photoreceptor responses to light stimuli. We introduced Flicker Optoretinography (f-ORG) for measuring rapid optical path length changes under photopic conditions, achieving reproducible, single-nanometer sensitivity in light-adapted eyes. This approach enhances understanding of retinal responses and photopigment photoactivation processes.
Please find here the PDF of the announcement:
Organizer
MBExC
