Dynamics and Traffic for Transfecting Exogenous MicroRNA as Studied by Live-Cell Microscopy

2021 ◽  
Vol 17 (8) ◽  
pp. 1647-1653
Author(s):  
Ke Yang ◽  
Yuanyuan Wang ◽  
Bo Sun ◽  
Tian Tian ◽  
Zhu Dai ◽  
...  
Keyword(s):  
Dynamic Properties ◽  
Mean Square Displacement ◽  
Small Region ◽  
Single Particle Tracking ◽  
Live Cell ◽  
Calculation Results ◽  
Long Time ◽  
Live Cell Microscopy ◽  
Cell Microscopy

MicroRNA (miRNA) has emerged as an important gene-regulator that shows great potential in gene therapy because of its unique roles in gene-regulation. However, the knowledge on their function and transportation in vivo is still lacking, and there are limited obvious evidences to define intracellular transportation of miRNA. In this study, the dynamics of exogenous miR-21 transfected into HeLa cells was traced by live-cell microscopy. Their transportation at key time points was recorded and dynamic properties were analyzed by single particle tracking (SPT) and mean square displacement (MSD) calculation. Results showed that the exogenous miRNAs bounded to cells quickly and went through lysosome into cytosol, where they were subsequently recruited into p-body. They finally were degraded, otherwise went back to cytosol in some way. Long time observation and analysis of motion mode showed that the miRNAs were confined in a small region and their motion modes were flexible in different intracellular microenvironment after entering the cells.

scholarly journals Bright photoactivatable fluorophores for single-molecule imaging

2016 ◽  
Author(s):  
Jonathan B. Grimm ◽  
Brian P. English ◽  
Anand K. Muthusamy ◽  
Brian P. Mehl ◽  
Peng Dong ◽  
...  
Keyword(s):  
Single Molecule ◽  
Single Particle Tracking ◽  
Live Cell ◽  
Cell Labeling ◽  
Advanced Imaging ◽  
Live Cell Microscopy ◽  
Protein Tags ◽  
Live Cell Labeling ◽  
Cell Microscopy ◽  
General Method

ABSTRACTSmall molecule fluorophores are important tools for advanced imaging experiments. The development of self-labeling protein tags such as the HaloTag and SNAP-tag has expanded the utility of chemical dyes in live-cell microscopy. We recently described a general method for improving the brightness and photostability of small, cell-permeable fluorophores, resulting in the azetidine-containing “Janelia Fluor” (JF) dyes. Here, we refine and extend the utility of the JF dyes by synthesizing photoactivatable derivatives that are compatible with established live-cell labeling strategies. These compounds retain the superior brightness of the JF dyes but their facile photoactivation enables improved single-particle tracking and localization microscopy experiments.

scholarly journals Cooperative 4Pi Excitation and Detection Yields Sevenfold Sharper Optical Sections in Live-Cell Microscopy

2004 ◽  
Vol 87 (6) ◽  
pp. 4146-4152 ◽  
Author(s):  
Hilmar Gugel ◽  
Jörg Bewersdorf ◽  
Stefan Jakobs ◽  
Johann Engelhardt ◽  
Rafael Storz ◽  
...  
Keyword(s):  
Live Cell ◽  
Live Cell Microscopy ◽  
Cell Microscopy

scholarly journals Live-cell microscopy - tips and tools

2009 ◽  
Vol 122 (6) ◽  
pp. 753-767 ◽  
Author(s):  
M. M. Frigault ◽  
J. Lacoste ◽  
J. L. Swift ◽  
C. M. Brown
Keyword(s):  
Live Cell ◽  
Live Cell Microscopy ◽  
Cell Microscopy

scholarly journals Improved Chemical-Genetic Fluorescent Markers for Live Cell Microscopy

Biochemistry ◽  
2018 ◽  
Vol 57 (39) ◽  
pp. 5648-5653 ◽  
Author(s):  
Alison G. Tebo ◽  
Frederico M. Pimenta ◽  
Yu Zhang ◽  
Arnaud Gautier
Keyword(s):  
Live Cell ◽  
Chemical Genetic ◽  
Fluorescent Markers ◽  
Live Cell Microscopy ◽  
Cell Microscopy

Structured illumination for live cell microscopy

2018 ◽  
Author(s):  
Herbert Schneckenburger ◽  
Verena Richter ◽  
Michael Wagner ◽  
Mathis Piper
Keyword(s):  
Live Cell ◽  
Structured Illumination ◽  
Live Cell Microscopy ◽  
Cell Microscopy

scholarly journals Initiation and disassembly of filopodia tip complexes containing VASP and lamellipodin

2020 ◽  
Vol 31 (18) ◽  
pp. 2021-2034 ◽  
Author(s):  
Karen W. Cheng ◽  
R. Dyche Mullins
Keyword(s):  
Actin Filaments ◽  
Leading Edge ◽  
Live Cell ◽  
Size Dependent ◽  
Live Cell Microscopy ◽  
Cell Microscopy

Clustering of the actin polymerase VASP (vasodilator-stimulated phosphoprotein) reorganizes leading-edge actin filaments into filopodia bundles in crawling cells. Here, we use live-cell microscopy to image the earliest events in VASP clustering and find that initiation depends on interactions with lamellipodin and barbed ends, while disassembly is driven by size-dependent instability.

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