BODIPY based realtime, reversible and targeted fluorescent probes for biothiol imaging in living cells

2020 ◽  
Vol 56 (93) ◽  
pp. 14717-14720
Author(s):  
Rongkun He ◽  
Yichuan Zhang ◽  
Suresh Madhu ◽  
Quan Gao ◽  
Qianjin Lian ◽  
...  
Keyword(s):  
Real Time ◽  
Fluorescent Probes ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Living Cells ◽  
Live Cell

Real-time live cell imaging and quantification of biothiol dynamics are important for understanding pathophysiological processes.

scholarly journals “Probe, Sample, and Instrument (PSI)”: The Hat-Trick for Fluorescence Live Cell Imaging

Chemosensors ◽  
2018 ◽  
Vol 6 (3) ◽  
pp. 40 ◽  
Author(s):  
Ludovic Galas ◽  
Thibault Gallavardin ◽  
Magalie Bénard ◽  
Arnaud Lehner ◽  
Damien Schapman ◽  
...  
Keyword(s):  
Organic Synthesis ◽  
Fluorescent Probes ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Cell Labeling ◽  
Key Points ◽  
Research Infrastructures ◽  
New Strategies ◽  
Fluorescence Live Cell Imaging

Cell Imaging Platforms (CIPs) are research infrastructures offering support to a number of scientific projects including the choice of adapted fluorescent probes for live cell imaging. What to detect in what type of sample and for how long is a major issue with fluorescent probes and, for this, the “hat-trick” “Probe–Sample–Instrument” (PSI) has to be considered. We propose here to deal with key points usually discussed in CIPs including the properties of fluorescent organic probes, the modality of cell labeling, and the best equipment to obtain appropriate spectral, spatial, and temporal resolution. New strategies in organic synthesis and click chemistry for accessing probes with enhanced photophysical characteristics and targeting abilities will also be addressed. Finally, methods for image processing will be described to optimize exploitation of fluorescence signals.

Use of rhodamine-allyl Schiff base in chemodosimetric processes for total palladium estimation and application in live cell imaging

2018 ◽  
Vol 42 (21) ◽  
pp. 17351-17358 ◽  
Author(s):  
Anup Kumar Bhanja ◽  
Snehasis Mishra ◽  
Ketaki Kar ◽  
Kaushik Naskar ◽  
Suvendu Maity ◽  
...  
Keyword(s):  
Schiff Base ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Living Cells ◽  
Live Cell ◽  
Raw 264.7 ◽  
Macrophage Cells ◽  
Raw 264.7 Macrophage ◽  
Raw 264.7 Macrophage Cells

An allyl-rhodamine Schiff base shows excellent palladium sensitivity (LOD, 95 nM) irrespective of Pd(0,ii,iv) and practical applicability is judged in living cells of RAW 264.7 (macrophage) cells.

Synthesis of photoactivatable azido-acyl caged oxazine fluorophores for live-cell imaging

2016 ◽  
Vol 52 (60) ◽  
pp. 9442-9445 ◽  
Author(s):  
Andrew V. Anzalone ◽  
Zhixing Chen ◽  
Virginia W. Cornish
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Living Cells ◽  
Live Cell

A new cell-permeable caged oxazine fluorophore was synthesized for protein specific labeling and photoactivation in living cells.

scholarly journals Illuminating the Sites of Enterovirus Replication in Living Cells by Using a Split-GFP-Tagged Viral Protein

mSphere ◽  
2016 ◽  
Vol 1 (4) ◽  
Author(s):  
H. M. van der Schaar ◽  
C. E. Melia ◽  
J. A. C. van Bruggen ◽  
J. R. P. M. Strating ◽  
M. E. D. van Geenen ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Lipid Composition ◽  
Viral Protein ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Living Cells ◽  
Live Cell ◽  
Genome Replication ◽  
Fluorescent Reporter ◽  
Unique Protein

ABSTRACT Enteroviruses induce the formation of membranous structures (replication organelles [ROs]) with a unique protein and lipid composition specialized for genome replication. Electron microscopy has revealed the morphology of enterovirus ROs, and immunofluorescence studies have been conducted to investigate their origin and formation. Yet, immunofluorescence analysis of fixed cells results in a rather static view of RO formation, and the results may be compromised by immunolabeling artifacts. While live-cell imaging of ROs would be preferred, enteroviruses encoding a membrane-anchored viral protein fused to a large fluorescent reporter have thus far not been described. Here, we tackled this constraint by introducing a small tag from a split-GFP system into an RO-resident enterovirus protein. This new tool bridges a methodological gap by circumventing the need for immunolabeling fixed cells and allows the study of the dynamics and formation of enterovirus ROs in living cells. Like all other positive-strand RNA viruses, enteroviruses generate new organelles (replication organelles [ROs]) with a unique protein and lipid composition on which they multiply their viral genome. Suitable tools for live-cell imaging of enterovirus ROs are currently unavailable, as recombinant enteroviruses that carry genes that encode RO-anchored viral proteins tagged with fluorescent reporters have not been reported thus far. To overcome this limitation, we used a split green fluorescent protein (split-GFP) system, comprising a large fragment [strands 1 to 10; GFP(S1-10)] and a small fragment [strand 11; GFP(S11)] of only 16 residues. The GFP(S11) (GFP with S11 fragment) fragment was inserted into the 3A protein of the enterovirus coxsackievirus B3 (CVB3), while the large fragment was supplied by transient or stable expression in cells. The introduction of GFP(S11) did not affect the known functions of 3A when expressed in isolation. Using correlative light electron microscopy (CLEM), we showed that GFP fluorescence was detected at ROs, whose morphologies are essentially identical to those previously observed for wild-type CVB3, indicating that GFP(S11)-tagged 3A proteins assemble with GFP(S1-10) to form GFP for illumination of bona fide ROs. It is well established that enterovirus infection leads to Golgi disintegration. Through live-cell imaging of infected cells expressing an mCherry-tagged Golgi marker, we monitored RO development and revealed the dynamics of Golgi disassembly in real time. Having demonstrated the suitability of this virus for imaging ROs, we constructed a CVB3 encoding GFP(S1-10) and GFP(S11)-tagged 3A to bypass the need to express GFP(S1-10) prior to infection. These tools will have multiple applications in future studies on the origin, location, and function of enterovirus ROs. IMPORTANCE Enteroviruses induce the formation of membranous structures (replication organelles [ROs]) with a unique protein and lipid composition specialized for genome replication. Electron microscopy has revealed the morphology of enterovirus ROs, and immunofluorescence studies have been conducted to investigate their origin and formation. Yet, immunofluorescence analysis of fixed cells results in a rather static view of RO formation, and the results may be compromised by immunolabeling artifacts. While live-cell imaging of ROs would be preferred, enteroviruses encoding a membrane-anchored viral protein fused to a large fluorescent reporter have thus far not been described. Here, we tackled this constraint by introducing a small tag from a split-GFP system into an RO-resident enterovirus protein. This new tool bridges a methodological gap by circumventing the need for immunolabeling fixed cells and allows the study of the dynamics and formation of enterovirus ROs in living cells.

scholarly journals Keeping Life in Focus New Systems Prevent Z-axis Drift in Time Lapse Studies

2006 ◽  
Vol 14 (4) ◽  
pp. 42-46 ◽  
Author(s):  
Edward Lachica
Keyword(s):  
Fluorescent Probes ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Time Lapse ◽  
Living Cells ◽  
Dynamic Processes ◽  
Biological Structure ◽  
Camera Lucida ◽  
Developmental Anatomy ◽  
Organ Level

Just two decades ago, life scientists studied biological structure, developmental anatomy and intracellular processes by describing individual snapshots of kinetic events. Today, with so much bioscience research focusing on dynamic processes that occur on the molecular, cellular and whole organ level, it is important to record events as they happen, over seconds, minutes or hours, in living cells. Photographs and camera lucida drawings of fixed, stained cells have given way to live cell imaging using fluorescent probes, warming trays to promote cell viability and cinemicrography as a method of recording events.

Real-time and quantitative fluorescent live-cell imaging with quadruplex-specific red-edge probe (G4-REP)

2017 ◽  
Vol 1861 (5) ◽  
pp. 1312-1320 ◽  
Author(s):  
Sunny Y. Yang ◽  
Souheila Amor ◽  
Aurélien Laguerre ◽  
Judy M.Y. Wong ◽  
David Monchaud
Keyword(s):  
Real Time ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Live Cell ◽  
Red Edge ◽  
Edge Probe
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