Super-resolution fluorescence microscopy is a powerful method to image molecular processes that occur beyond the diffraction limit of light. Förster resonance energy transfer (FRET) is widely used to ...

Genetically-encoded fluorescent biosensors have revolutionized our understanding of complex systems by permitting the in situ observation of chemical activities. However, only a comparatively small se...

We present Rho-tag and SiR-tag, engineered protein tags derived from bacterial multidrug-resistance proteins that bind unsubstituted (silicon-) rhodamines with nanomolar affinity, enabling fast, rever...

Université de Bordeaux

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Nature Photonics - The authors present a review of super-resolution optical fluctuation imaging (SOFI), including its core working principle, recent advances and remaining challenges.

PhD Project - Single molecule biology of bacterial cell wall remodelling at University of Warwick, listed on FindAPhD.com

Potassium ion (K+) is the most abundant metal ion in cells and plays an indispensable role in practically all biological systems. Although there have been reports of both synthetic and genetically encoded fluorescent K+ indicators, there remains a need for an indicator that is genetically targetable, has high specificity for K+ versus Na+, and has a high fluorescent response in the red to far-red wavelength range. Here, we introduce a series of chemigenetic K+ indicators, designated as the HaloKbp1 series, based on the bacterial K+-binding protein (Kbp) inserted into HaloTag7 self-labeled with environmentally sensitive rhodamine derivatives. This series of high-performance indicators features high brightness in the red to far-red region, large intensiometric fluorescence changes, and a range of Kd values. We demonstrate that they are suitable for the detection of physiologically relevant K+ concentration changes such as those that result from the Ca2+-dependent activation of the BK potassium channel.

In this study, we develop a general analytical model of the photochromism of fluorescent proteins and apply it to spectroscopic measurements performed on six different labels. Our approach provides quantitative explanations for phenomena such as the existence of positive and negative switching, limitations i

We present the TriScan, a compact and inexpensive fluorescence microscope that combines the speed of widefield microscopy with the 3D-sectioning capabilities of confocal microscopy. The optical layout is based on an add-on module that combines line-scan confocal imaging with a sensitive camera detector, realized using a simple optical layout that permits the use of arbitrarily fast scanning mirrors. The resulting design is theoretically capable of full field-of-view acquisition rates in the kilohertz regime combined with a diffraction-limited resolution and single-molecule sensitivity. Overall, the TriScan microscope provides the ease-of-use and speed of widefield imaging combined with the optical sectioning of one-photon confocal imaging, in a simple and inexpensive design suitable for a broad variety of settings ranging from research to diagnostic applications and screening. This bioRxiv manuscript describes an ongoing research project and associated preliminary data acquired using an early prototype of the instrument. We welcome and appreciate your enquiries, suggestions, and feedback. Updated versions of this manuscript will be deposited as the project progresses. The author list reflects the core team and points of contact working on this project, but does not reflect all of the contributions made to this research thus far. We are particularly grateful to Damla Temel (formerly KU Leuven) for assistance in the construction of the initial prototype, Lydia Danglot (IPNP Paris) and Hugo Vankelecom (KU Leuven) for providing samples, and Marcel Leutenegger (Max Planck Institute for Biophysical Research) for initial discussions regarding the practical implementation. We also thank the Research Foundation-Flanders (FWO) and the European Research Council for financial support. ### Competing Interest Statement The authors have applied for IPR protection on the TriScan design.