Dynamic host–guest interaction enables autonomous single molecule blinking and super-resolution imaging

2019 ◽  
Vol 55 (96) ◽  
pp. 14430-14433 ◽  
Author(s):  
Ranjan Sasmal ◽  
Nilanjana Das Saha ◽  
Florian Schueder ◽  
Divyesh Joshi ◽  
Vasu Sheeba ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Dynamic Interaction ◽  
Two Dimensional ◽  
Resolution Imaging ◽  
Fluorescence Blinking ◽  
2D And 3D ◽  
In Cells

Specific yet dynamic interaction in cucurbit[7]uril (CB[7]) system was used to obtain programmable fluorescence blinking with necessary brightness and frequency to enable two-dimensional (2D) and 3D super-resolution imaging of proteins in cells.

scholarly journals Dynamic Supramolecular Interaction in Cucurbit[7]uril Host-Guest Complex Enables Autonomous Single Molecule Blinking and Super-Resolution Imaging in Cells and Tissues

2018 ◽  
Author(s):  
Ranjan Sasmal ◽  
Nilanjana Das Saha ◽  
Florian Schueder ◽  
Divyesh Joshi ◽  
Vasu Sheeba ◽  
...  
Keyword(s):  
Single Molecule ◽  
Guest Molecule ◽  
Noncovalent Interactions ◽  
Super Resolution ◽  
Imaging Method ◽  
Supramolecular Interaction ◽  
Guest Molecules ◽  
Host Guest Complex ◽  
Resolution Imaging ◽  
2D And 3D

Synthetic supramolecular host-guest complexes are inherently dynamic as they employ weak and reversible noncovalent interactions for their recognition process. This dynamic behavior allows host-guest chemistry to be employed for various state of the art applications. Herein, we demonstrate the use of the dynamic supramolecular interaction to enable nanoscopic imaging inside cells and tissues. This imaging method exploits repetitive and transient binding of fluorescently labeled hexamethylenediamine (HMD) guest molecule to complementary cucurbit[7]uril (CB[7]) host to obtain stochastic switching between fluorescence ON- and OFF-states. Through connecting CB[7] hosts to targeting ligands (e.g., antibodies and small molecules), we show that this autonomous blinking enables two-dimensional (2D) and 3D super-resolution imaging of proteins in fixed cells and tissues. Finally, we exploited the capability of host-guest molecules to maintain their interaction specificity in the complexity of the live intracellular environment to obtain super-resolution actin imaging in living HeLa cell.

scholarly journals 4polar-STORM polarized super-resolution imaging of actin filament organization in cells

2021 ◽  
Author(s):  
Caio Vaz Rimoli ◽  
Cesar Augusto Valades Cruz ◽  
Valentina Curcio ◽  
Manos Mavrakis ◽  
Sophie Brasselet
Keyword(s):  
Actin Filament ◽  
Single Molecule ◽  
Protein Function ◽  
Super Resolution ◽  
New Method ◽  
Nanometer Scale ◽  
Protein Assemblies ◽  
Cellular Context ◽  
Resolution Imaging ◽  
In Cells

Advances in single-molecule localization microscopy are providing unprecedented insights into the nanometer-scale organization of protein assemblies in cells and thus a powerful means for interrogating biological function. However, localization imaging alone does not contain information on protein conformation and orientation, which constitute additional key signatures of protein function. Here, we present a new microscopy method which combines for the first time Stochastic Optical Reconstruction Microscopy (STORM) super-resolution imaging with single molecule orientation and wobbling measurements using a four polarization-resolved image splitting scheme. This new method, called 4polar-STORM, allows us to determine both single molecule localization and orientation in 2D and to infer their 3D orientation, and is compatible with high labelling densities and thus ideally placed for the determination of the organization of dense protein assemblies in cells. We demonstrate the potential of this new method by studying the nanometer-scale organization of dense actin filament assemblies driving cell adhesion and motility, and reveal bimodal distributions of actin filament orientations in the lamellipodium, which were previously only observed in electron microscopy studies. 4polar-STORM is fully compatible with 3D localization schemes and amenable to live-cell observations, and thus promises to provide new functional readouts by enabling nanometer-scale studies of orientational dynamics in a cellular context.

scholarly journals Dynamic Supramolecular Interaction in Cucurbit[7]uril Host-Guest Complex Enables Autonomous Single Molecule Blinking and Super-Resolution Imaging in Cells and Tissues

2018 ◽  
Author(s):  
Ranjan Sasmal ◽  
Nilanjana Das Saha ◽  
Florian Schueder ◽  
Divyesh Joshi ◽  
Vasu Sheeba ◽  
...  
Keyword(s):  
Single Molecule ◽  
Guest Molecule ◽  
Noncovalent Interactions ◽  
Super Resolution ◽  
Imaging Method ◽  
Supramolecular Interaction ◽  
Guest Molecules ◽  
Host Guest Complex ◽  
Resolution Imaging ◽  
2D And 3D

Synthetic supramolecular host-guest complexes are inherently dynamic as they employ weak and reversible noncovalent interactions for their recognition process. This dynamic behavior allows host-guest chemistry to be employed for various state of the art applications. Herein, we demonstrate the use of the dynamic supramolecular interaction to enable nanoscopic imaging inside cells and tissues. This imaging method exploits repetitive and transient binding of fluorescently labeled hexamethylenediamine (HMD) guest molecule to complementary cucurbit[7]uril (CB[7]) host to obtain stochastic switching between fluorescence ON- and OFF-states. Through connecting CB[7] hosts to targeting ligands (e.g., antibodies and small molecules), we show that this autonomous blinking enables two-dimensional (2D) and 3D super-resolution imaging of proteins in fixed cells and tissues. Finally, we exploited the capability of host-guest molecules to maintain their interaction specificity in the complexity of the live intracellular environment to obtain super-resolution actin imaging in living HeLa cell.

scholarly journals Comparing lifeact and phalloidin for super-resolution imaging of actin in fixed cells

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0246138
Author(s):  
Hanieh Mazloom-Farsibaf ◽  
Farzin Farzam ◽  
Mohamadreza Fazel ◽  
Michael J. Wester ◽  
Marjolein B. M. Meddens ◽  
...  
Keyword(s):  
Single Molecule ◽  
Cell Biology ◽  
Cell Movement ◽  
Super Resolution ◽  
Actin Binding ◽  
Thin Filaments ◽  
Reversible Binding ◽  
Multiple Regions ◽  
Resolution Imaging ◽  
Division Cell

Visualizing actin filaments in fixed cells is of great interest for a variety of topics in cell biology such as cell division, cell movement, and cell signaling. We investigated the possibility of replacing phalloidin, the standard reagent for super-resolution imaging of F-actin in fixed cells, with the actin binding peptide ‘lifeact’. We compared the labels for use in single molecule based super-resolution microscopy, where AlexaFluor 647 labeled phalloidin was used in a dSTORM modality and Atto 655 labeled lifeact was used in a single molecule imaging, reversible binding modality. We found that imaging with lifeact had a comparable resolution in reconstructed images and provided several advantages over phalloidin including lower costs, the ability to image multiple regions of interest on a coverslip without degradation, simplified sequential super-resolution imaging, and more continuous labeling of thin filaments.

Single molecule Coordinate and Height super-resolution Imaging with Dithering and Orientation (CHIDO)

2020 ◽  
Author(s):  
Luis A. Alemán-Castañeda ◽  
Valentina Curcio ◽  
Thomas G. Brown ◽  
Sophie Brasselet ◽  
Miguel A. Alonso
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Resolution Imaging

scholarly journals Super-resolution fight club: assessment of 2D and 3D single-molecule localization microscopy software

2019 ◽  
Vol 16 (5) ◽  
pp. 387-395 ◽  
Author(s):  
Daniel Sage ◽  
Thanh-An Pham ◽  
Hazen Babcock ◽  
Tomas Lukes ◽  
Thomas Pengo ◽  
...  
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Localization Microscopy ◽  
Fight Club ◽  
2D And 3D ◽  
Single Molecule Localization Microscopy

scholarly journals Fast, Simultaneous Multiple Fluorophore Fitting in Single Molecule Super-Resolution Imaging

2011 ◽  
Vol 100 (3) ◽  
pp. 349a
Author(s):  
Fang Huang ◽  
Samantha L. Schwartz ◽  
Jason M. Byars ◽  
Keith A. Lidke
Keyword(s):  
Single Molecule ◽  
Super Resolution ◽  
Resolution Imaging ◽  
Fitting In
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