scholarly journals Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging

2019 ◽  
Author(s):  
Mayeul Collot ◽  
Emmanuel Boutant ◽  
Kyong Tkhe Fam ◽  
Lydia Danglot ◽  
Andrey S. Klymchenko
Keyword(s):  
Plasma Membrane ◽  
Molecular Design ◽  
Photophysical Properties ◽  
Tissue Imaging ◽  
Biological Processes ◽  
Fluorescence Staining ◽  
Styryl Dyes ◽  
Mammalian Cell Lines ◽  
Styryl Dye ◽  
Membrane Probes

ABSTRACTThe plasma membrane (PM) plays a major role in many biological processes; therefore its proper fluorescence staining is required in bioimaging. Among the commercially available PM probes, styryl dye FM1-43 is one of the most widely used. In this work, we demonstrated that fine chemical modifications of FM1-43 can dramatically improve the PM staining. The newly developed probes, SP-468 and SQ-535 were found to display enhanced photophysical properties (reduced crosstalk, higher brightness, improved photostability) and unlike FM1-43, provided excellent and immediate PM staining in 5 different mammalian cell lines including neurons (primary culture and tissue imaging). Additionally, we showed that the new probes displayed differences in their internalization pathways compared to their parent FM1-43. Finally, we demonstrated that the modifications made to FM1-43 did not impair the ability of the new probes to stain the PM of plant cells. Overall, this work presents new useful probes for PM imaging in cells and tissues and provides insights on the molecular design of new PM targeting molecules.

scholarly journals Molecular Tuning of Styryl Dyes Leads to Versatile and Efficient Plasma Membrane Probes for Cell and Tissue Imaging

2020 ◽  
Vol 31 (3) ◽  
pp. 875-883 ◽  
Author(s):  
Mayeul Collot ◽  
Emmanuel Boutant ◽  
Kyong Tkhe Fam ◽  
Lydia Danglot ◽  
Andrey S. Klymchenko
Keyword(s):  
Plasma Membrane ◽  
Tissue Imaging ◽  
Styryl Dyes ◽  
Membrane Probes

Two-Dimensional Near-Infrared and Shortwave Infrared Molecular Aggregates: Taking Kasha’s Model to the Next Dimension

2019 ◽  
Author(s):  
Arundhati Deshmukh ◽  
Danielle Koppel ◽  
Chern Chuang ◽  
Danielle Cadena ◽  
Jianshu Cao ◽  
...  
Keyword(s):  
Near Infrared ◽  
Photophysical Properties ◽  
Satellite Telemetry ◽  
Transition Dipole Moment ◽  
Short Wave ◽  
Tissue Imaging ◽  
Transition Dipole ◽  
Deep Tissue Imaging ◽  
Excitonic States ◽  
Shortwave Infrared

Technologies which utilize near-infrared (700 – 1000 nm) and short-wave infrared (1000 – 2000 nm) electromagnetic radiation have applications in deep-tissue imaging, telecommunications and satellite telemetry due to low scattering and decreased background signal in this spectral region. However, there are few molecular species, which absorb efficiently beyond 1000 nm. Transition dipole moment coupling (e.g. J-aggregation) allows for redshifted excitonic states and provides a pathway to highly absorptive electronic states in the infrared. We present aggregates of two cyanine dyes whose absorption peaks redshift dramatically upon aggregation in water from ~ 800 nm to 1000 nm and 1050 nm with sheet-like morphologies and high molar absorptivities (e ~ 10<sup>5 </sup>M<sup>-1</sup>cm<sup>-1</sup>). To describe this phenomenology, we extend Kasha’s model for J- and H-aggregation to describe the excitonic states of <i> 2-dimensional aggregates</i> whose slip is controlled by steric hindrance in the assembled structure. A consequence of the increased dimensionality is the phenomenon of an <i>intermediate </i>“I-aggregate”, one which redshifts yet displays spectral signatures of band-edge dark states akin to an H-aggregate. We distinguish between H-, I- and J-aggregates by showing the relative position of the bright (absorptive) state within the density of states using temperature dependent spectroscopy. Our results can be used to better design chromophores with predictable and tunable aggregation with new photophysical properties.

scholarly journals A family of pathogen-induced cysteine-rich transmembrane proteins is involved in plant disease resistance

Planta ◽  
2021 ◽  
Vol 253 (5) ◽  
Author(s):  
Marciel Pereira Mendes ◽  
Richard Hickman ◽  
Marcel C. Van Verk ◽  
Nicole M. Nieuwendijk ◽  
Anja Reinstädler ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Plasma Membrane ◽  
Disease Resistance ◽  
Plant Disease Resistance ◽  
Transmembrane Proteins ◽  
Series Data ◽  
Immune Gene ◽  
Biological Processes ◽  
Hypocotyl Growth ◽  
Enhanced Resistance

Abstract Main conclusion Overexpression of pathogen-induced cysteine-rich transmembrane proteins (PCMs) in Arabidopsis thaliana enhances resistance against biotrophic pathogens and stimulates hypocotyl growth, suggesting a potential role for PCMs in connecting both biological processes. Abstract Plants possess a sophisticated immune system to protect themselves against pathogen attack. The defense hormone salicylic acid (SA) is an important player in the plant immune gene regulatory network. Using RNA-seq time series data of Arabidopsis thaliana leaves treated with SA, we identified a largely uncharacterized SA-responsive gene family of eight members that are all activated in response to various pathogens or their immune elicitors and encode small proteins with cysteine-rich transmembrane domains. Based on their nucleotide similarity and chromosomal position, the designated Pathogen-induced Cysteine-rich transMembrane protein (PCM) genes were subdivided into three subgroups consisting of PCM1-3 (subgroup I), PCM4-6 (subgroup II), and PCM7-8 (subgroup III). Of the PCM genes, only PCM4 (also known as PCC1) has previously been implicated in plant immunity. Transient expression assays in Nicotiana benthamiana indicated that most PCM proteins localize to the plasma membrane. Ectopic overexpression of the PCMs in Arabidopsis thaliana resulted in all eight cases in enhanced resistance against the biotrophic oomycete pathogen Hyaloperonospora arabidopsidis Noco2. Additionally, overexpression of PCM subgroup I genes conferred enhanced resistance to the hemi-biotrophic bacterial pathogen Pseudomonas syringae pv. tomato DC3000. The PCM-overexpression lines were found to be also affected in the expression of genes related to light signaling and development, and accordingly, PCM-overexpressing seedlings displayed elongated hypocotyl growth. These results point to a function of PCMs in both disease resistance and photomorphogenesis, connecting both biological processes, possibly via effects on membrane structure or activity of interacting proteins at the plasma membrane.

scholarly journals A method for imaging single molecules at the plasma membrane of live cells within tissue slices

2020 ◽  
Vol 153 (1) ◽  
Author(s):  
Gregory I. Mashanov ◽  
Tatiana A. Nenasheva ◽  
Tatiana Mashanova ◽  
Catherine Maclachlan ◽  
Nigel J.M. Birdsall ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Lines ◽  
Single Molecule ◽  
Cardiac Tissue ◽  
Single Molecules ◽  
Live Cells ◽  
Biological Macromolecules ◽  
Tissue Slices ◽  
Tissue Samples ◽  
Mammalian Cell Lines

Recent advances in light microscopy allow individual biological macromolecules to be visualized in the plasma membrane and cytosol of live cells with nanometer precision and ∼10-ms time resolution. This allows new discoveries to be made because the location and kinetics of molecular interactions can be directly observed in situ without the inherent averaging of bulk measurements. To date, the majority of single-molecule imaging studies have been performed in either unicellular organisms or cultured, and often chemically fixed, mammalian cell lines. However, primary cell cultures and cell lines derived from multi-cellular organisms might exhibit different properties from cells in their native tissue environment, in particular regarding the structure and organization of the plasma membrane. Here, we describe a simple approach to image, localize, and track single fluorescently tagged membrane proteins in freshly prepared live tissue slices and demonstrate how this method can give information about the movement and localization of a G protein–coupled receptor in cardiac tissue slices. In principle, this experimental approach can be used to image the dynamics of single molecules at the plasma membrane of many different soft tissue samples and may be combined with other experimental techniques.

Synthesis of novel viscosity sensitive pyrrolo-quinaldine based styryl dyes: Photophysical properties, electrochemical and DFT study

2020 ◽  
Vol 397 ◽  
pp. 112557
Author(s):  
Mahesh Jachak ◽  
Sushil Khopkar ◽  
Ankur Chaturvedi ◽  
Amruta Joglekar ◽  
Ganapati Shankarling
Keyword(s):  
Photophysical Properties ◽  
Dft Study ◽  
Styryl Dyes

Photophysical properties of aqueous solutions of a styryl dye in the presence of cucurbit[n]uril (n = 5, 6, 8)

2016 ◽  
Vol 50 (1) ◽  
pp. 21-26 ◽  
Author(s):  
A. D. Svirida ◽  
D. A. Ivanov ◽  
N. Kh. Petrov ◽  
A. V. Vedernikov ◽  
S. P. Gromov ◽  
...  
Keyword(s):  
Aqueous Solutions ◽  
Photophysical Properties ◽  
Styryl Dye

D-A structured high efficiency solid luminogens with tunable emissions: Molecular design and photophysical properties

2017 ◽  
Vol 28 (11) ◽  
pp. 2133-2138 ◽  
Author(s):  
Yunzhong Wang ◽  
Zihan He ◽  
Gan Chen ◽  
Tong Shan ◽  
Wangzhang Yuan ◽  
...  
Keyword(s):  
High Efficiency ◽  
Molecular Design ◽  
Photophysical Properties

Molecular design, photoisomerization and complexation of crown ether styryl dyes

1991 ◽  
Vol 185 (5-6) ◽  
pp. 455-460 ◽  
Author(s):  
M.V. Alfimov ◽  
S.P. Gromov ◽  
I.K. Lednev
Keyword(s):  
Crown Ether ◽  
Molecular Design ◽  
Styryl Dyes ◽  
Crown Ether Styryl Dyes

scholarly journals Effects of ATP9A on Extracellular Vesicle Release and Exosomal Lipid Composition

2020 ◽  
Vol 2020 ◽  
pp. 1-17
Author(s):  
Xiao Xu ◽  
Limei Xu ◽  
Peng Zhang ◽  
Kan Ouyang ◽  
Yin Xiao ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Extracellular Vesicles ◽  
Lipid Composition ◽  
Extracellular Vesicle ◽  
Biological Processes ◽  
Vesicle Release ◽  
Endosomal Recycling ◽  
Intracellular Compartments ◽  
Intercellular Communications

Numerous biological processes are regulated by the intercellular communications arising from extracellular vesicles (EVs) released from cells. However, the mechanisms that regulate the quantity of EV discharged have yet to be understood. While it is known that ATP9A, a P4-ATPase, is involved in endosomal recycling, it is not clear whether it also contributes to the release of EVs and the makeup of exosomal lipids. This study is aimed at exploring the role of human ATP9A in the process of EV release and, further, to analyze the profiles of EV lipids regulated by ATP9A. Our results demonstrate that ATP9A is located in both the intracellular compartments and the plasma membrane. The percentage of ceramides and sphingosine was found to be significantly greater in the control cells than in the ATP9A overexpression and ATP9A knockout groups. However, EV release was greater in ATP9A knockout cells, indicating that ATP9A inhibits the release of EVs. This study revealed the effects of ATP9A on the release of EVs and the lipid composition of exosomes.

Design of AIEgens for Near-Infrared IIb Imaging Through Structural Modulation at Molecular and Morphological Levels

2019 ◽  
Author(s):  
yuanyuan li ◽  
Zhaochong Cai ◽  
shunjie liu ◽  
Haoke Zhang ◽  
sherman Wong ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Fluorescence Imaging ◽  
Near Infrared ◽  
Molecular Design ◽  
Organic Dyes ◽  
Tissue Imaging ◽  
Deep Penetration ◽  
Structural Modulation ◽  
Organic Fluorophore

<p>Fluorescence imaging in near-infrared IIb (NIR-IIb, 1500-1700 nm) spectrum holds a considerable promise for tissue imaging with deep penetration and high spatial resolution owing to the minimized autofluorescence and suppressed photon scattering. While few inorganic NIR-IIb fluorescent probes have been reported, their organic counterparts are still underdeveloped, possibly due to the lack of efficient materials with long emission wavelength. Herein, we propose a new molecular design philosophy to develop organic NIR-IIb fluorophores with high quantum yield (QY) by manipulation of the effects of twisted intramolecular charge transfer and aggregation-induced emission at the molecular and morphological levels. A pure organic fluorescent dye emitting up to 1600 nm with a QY of 14.2% in the NIR-II region (1000-1600 nm) is developed. For the first time, NIR-IIb fluorescence imaging of blood vessels and deeply-located intestinal tract of live mice based on organic dyes is achieved. The results show that organic fluorophore performs superb imaging ability in both superficial blood vessels and internal organs with high resolution and enhanced signal-to-background ratio in NIR-IIb region. We hope this groundbreakingly study will inspire further research on the evolution of pure organic NIR-IIb probes for in vivo imaging.</p>

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