scholarly journals Conventional BODIPY Conjugates for Live-Cell Super-Resolution Microscopy and Single-Molecule Tracking

10.3791/60950 ◽  
2020 ◽  
Author(s):  
Santosh Adhikari ◽  
Chiranjib Banerjee ◽  
Joe Moscatelli ◽  
Elias M. Puchner
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Live Cell ◽  
Single Molecule Tracking ◽  
Super Resolution Microscopy

scholarly journals Live cell single molecule-guided Bayesian localization super resolution microscopy

Cell Research ◽  
2016 ◽  
Vol 27 (5) ◽  
pp. 713-716 ◽  
Author(s):  
Fan Xu ◽  
Mingshu Zhang ◽  
Wenting He ◽  
Renmin Han ◽  
Fudong Xue ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Live Cell ◽  
Super Resolution Microscopy

scholarly journals Live cell single molecule-guided Bayesian localization super resolution microscopy

Cell Research ◽  
2016 ◽  
Author(s):  
Fan Xu ◽  
Mingshu Zhang ◽  
Wenting He ◽  
Renmin Han ◽  
Fudong Xue ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Live Cell ◽  
Super Resolution Microscopy

scholarly journals Systematic Tuning of Rhodamine Spirocyclization for Super-Resolution Microscopy

2021 ◽  
Author(s):  
Nicolas Lardon ◽  
Lu Wang ◽  
Aline Tschanz ◽  
Philipp Hoess ◽  
Mai Tran ◽  
...  
Keyword(s):  
Single Molecule ◽  
Large Range ◽  
Dynamic Equilibrium ◽  
Super Resolution ◽  
Live Cell ◽  
Stimulated Emission ◽  
Important Class ◽  
Localization Microscopy ◽  
Super Resolution Microscopy ◽  
Single Molecule Localization Microscopy

Rhodamines are the most important class of fluorophores for applications in live-cell fluorescence microscopy. This is mainly because rhodamines exist in a dynamic equilibrium between a fluorescent zwitterion and a non-fluorescent but cell-permeable spirocyclic form. Different imaging applications require different positions of this dynamic equilibrium, which poses a challenge for the design of suitable probes. We describe here how the conversion of the ortho-carboxy moiety of a given rhodamine into substituted acyl benzenesulfonamides and alkylamides permits the systematic tuning of the equilibrium of spirocyclization with unprecedented accuracy and over a large range. This allows to transform the same rhodamine into either a highly fluorogenic and cell-permeable probe for live-cell stimulated emission depletion (STED) microscopy, or into a spontaneously blinking dye for single molecule localization microscopy (SMLM). We used this approach to generate differently colored probes optimized for different labeling systems and imaging applications.

Nanoparticles for super-resolution microscopy and single-molecule tracking

2018 ◽  
Vol 15 (6) ◽  
pp. 415-423 ◽  
Author(s):  
Dayong Jin ◽  
Peng Xi ◽  
Baoming Wang ◽  
Le Zhang ◽  
Jörg Enderlein ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Single Molecule Tracking ◽  
Super Resolution Microscopy

scholarly journals Photoactivation of silicon rhodamines via a light-induced protonation

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Michelle S. Frei ◽  
Philipp Hoess ◽  
Marko Lampe ◽  
Bianca Nijmeijer ◽  
Moritz Kueblbeck ◽  
...  
Keyword(s):  
Single Molecule ◽  
Particle Tracking ◽  
Single Particle ◽  
Super Resolution ◽  
Spectroscopic Properties ◽  
Single Particle Tracking ◽  
Live Cell ◽  
Localization Microscopy ◽  
Super Resolution Microscopy ◽  
Single Molecule Localization Microscopy

Abstract Photoactivatable fluorophores are important for single-particle tracking and super-resolution microscopy. Here we present a photoactivatable fluorophore that forms a bright silicon rhodamine derivative through a light-dependent protonation. In contrast to other photoactivatable fluorophores, no caging groups are required, nor are there any undesired side-products released. Using this photoactivatable fluorophore, we create probes for HaloTag and actin for live-cell single-molecule localization microscopy and single-particle tracking experiments. The unusual mechanism of photoactivation and the fluorophore’s outstanding spectroscopic properties make it a powerful tool for live-cell super-resolution microscopy.

scholarly journals Super-Beacons: open-source probes with spontaneous tuneable blinking compatible with live-cell super-resolution microscopy

2020 ◽  
Author(s):  
Pedro M. Pereira ◽  
Nils Gustafsson ◽  
Mark Marsh ◽  
Musa M. Mhlanga ◽  
Ricardo Henriques
Keyword(s):  
Open Source ◽  
Single Molecule ◽  
High Intensity ◽  
Transmembrane Proteins ◽  
Super Resolution ◽  
Live Cell ◽  
New Class ◽  
High Illumination ◽  
Super Resolution Microscopy

Localization based super-resolution microscopy relies on the detection of individual molecules cycling between fluorescent and non-fluorescent states. These transitions are commonly regulated by high-intensity illumination, imposing constrains to imaging hardware and producing sample photodamage. Here, we propose single-molecule self-quenching as a mechanism to generate spontaneous photoswitching independent of illumination. To demonstrate this principle, we developed a new class of DNA-based open-source Super-Resolution probes named Super-Beacons, with photoswitching kinetics that can be tuned structurally, thermally and chemically. The potential of these probes for live-cell friendly Super-Resolution Microscopy without high-illumination or toxic imaging buffers is revealed by imaging Interferon Inducible Transmembrane proteins (IFITMs) at sub-100nm resolutions.

scholarly journals Photoactivation of silicon rhodamines via a light-induced protonation

2019 ◽  
Author(s):  
Michelle S. Frei ◽  
Philipp Hoess ◽  
Marko Lampe ◽  
Bianca Nijmeijer ◽  
Moritz Kueblbeck ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Spectroscopic Properties ◽  
Single Particle Tracking ◽  
Live Cell ◽  
Localization Microscopy ◽  
Rhodamine Derivative ◽  
New Type ◽  
Super Resolution Microscopy ◽  
Single Molecule Localization Microscopy

AbstractWe present a new type of photoactivatable fluorophore that forms a bright silicon rhodamine derivative through a light-dependent isomerization followed by protonation. In contrast to other photoactivatable fluorophores, no caging groups are required, nor are there any undesired side-products released. Using this photoactivatable fluorophore, we created probes for HaloTag and actin for live-cell single-molecule localization microscopy and single-particle tracking experiments. The unusual mechanism of photoactivation and the fluorophore’s outstanding spectroscopic properties make it a powerful tool for live-cell super-resolution microscopy.

scholarly journals Optimized localization analysis for single-molecule tracking and super-resolution microscopy

2010 ◽  
Vol 7 (5) ◽  
pp. 377-381 ◽  
Author(s):  
Kim I Mortensen ◽  
L Stirling Churchman ◽  
James A Spudich ◽  
Henrik Flyvbjerg
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Single Molecule Tracking ◽  
Localization Analysis ◽  
Super Resolution Microscopy

scholarly journals Deep-Learning Super-Resolution Microscopy Reveals Nanometer-Scale Intracellular Dynamics at the Millisecond Temporal Resolution

2021 ◽  
Author(s):  
Rong Chen ◽  
Xiao Tang ◽  
Zeyu Shen ◽  
Yusheng Shen ◽  
Tiantian Li ◽  
...  
Keyword(s):  
Deep Learning ◽  
Spatial Resolution ◽  
Temporal Resolution ◽  
Single Molecule ◽  
Super Resolution ◽  
Live Cell ◽  
Acquisition Time ◽  
Microtubule Network ◽  
Single Frame ◽  
Super Resolution Microscopy

AbstractSingle-molecule localization microscopy (SMLM) can be used to resolve subcellular structures and achieve a tenfold improvement in spatial resolution compared to that obtained by conventional fluorescence microscopy. However, the separation of single-molecule fluorescence events in thousands of frames dramatically increases the image acquisition time and phototoxicity, impeding the observation of instantaneous intracellular dynamics. Based on deep learning networks, we develop a single-frame super-resolution microscopy (SFSRM) approach that reconstructs a super-resolution image from a single frame of a diffraction-limited image to support live-cell super-resolution imaging at a ∼20 nm spatial resolution and a temporal resolution of up to 10 ms over thousands of time points. We demonstrate that our SFSRM method enables the visualization of the dynamics of vesicle transport at a millisecond temporal resolution in the dense and vibrant microtubule network in live cells. Moreover, the well-trained network model can be used with different live-cell imaging systems, such as confocal and light-sheet microscopes, making super-resolution microscopy accessible to nonexperts.

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