Bright fluorogenic squaraines with tuned cell entry for selective imaging of plasma membrane vs. endoplasmic reticulum

Collot, Mayeul; Kreder, Rémy; Tatarets, Anatoliy L.; Patsenker, L. D.; Mély, Yves; Klymchenko, Andrey S.

doi:10.1039/c5cc06094j

Cited by 79 publications

(91 citation statements)

References 31 publications

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“…From these two observations, numerous efforts have been made to develop fluorescent PM probes 4,5,6,7 including from our group. [8][9][10][11][12][13][14][15] Among them, FM dyes (named after Fei Mao 16 ), which are styryl dyes introduced by Betz and colleagues in the early 90s' as synaptic vesicle markers, 16 rapidly became among the most famous and used commercially available PM probes. 17,18 These amphiphilic dyes were further developed by the group of Loew to give rise to PM voltage sensitive probes 19,20 and membrane lipid domains probes.…”

Section: Introductionmentioning

confidence: 99%

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

Collot

Boutant

Fam

et al. 2019

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The 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.

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Section: Introductionmentioning

confidence: 99%

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

Collot

Boutant

Fam

et al. 2019

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“…This mechanism is in line with the earlier data obtained with membrane probes based on Nile red 20 and squaraine. 32 In order to determine how fast the aggregates of MemBright disassemble in the presence of LUVs, probes were added to an excess of LUVs and their fluorescence emission intensity was followed over the time (Fig. S8).…”

Section: Resultsmentioning

confidence: 99%

“…30–31 We recently proposed an efficient far-red emitting PM fluorescent marker called dSQ12S based on a squaraine dye with two zwitterionic amphiphilic anchors, exhibiting superior staining compared to commonly used hydrophobic cyanines, such as DiD, PKH family, etc. 32 Application of this unique design concept to cyanine family, highly popular in bioimaging applications, 33 could open numerous opportunities. First, cyanines are among the brightest dyes that can be obtained in any desired emission color spanning from the visible (orange) to the near-infrared.…”

Section: Introductionmentioning

confidence: 99%

MemBright: a Family of Fluorescent Membrane Probes for Advanced Cellular Imaging and Neuroscience

Collot

Ashokkumar

Anton

et al. 2018

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The proper staining of the plasma membrane (PM) is critical in bioimaging as it 16 delimits the cell. Herein, we developed MemBright: a family of six cyanine--based fluorescent 17 turn--on PM probes that emit from orange to near--infrared when reaching the PM, and 18 enable homogeneous and selective PM staining with excellent contrast in mono and two--19 photon microscopy. These probes are compatible with long--term live cell imaging and 20 immunostaining. Moreover, MemBright label neurons in a brighter manner than 21 surrounding cells allowing identification of neurons in acute brain tissue section and 22 neuromuscular--junctions without any use of transfection or transgenic animals. At last, 23MemBright were used in super--resolution imaging to unravel the dendritic spines' neck. 3D 24 multicolor dSTORM in combination with immunostaining revealed en--passant synapse 25 displaying endogenous glutamate receptors clustered at the axonal--dendritic contact site. 26MemBright probes thus constitute a universal toolkit for cell biology and neuroscience 27 biomembrane imaging with a variety of microscopy techniques. 29 30Visualizing plasma membrane is particularly important in neuroscience, because it enables 1 visualization neuronal organization and membrane trafficking involved in the synaptic 2 transmission. 3 Recent years have seen a tremendous expansion of fluorescence imaging 3 techniques and tools for cellular research. In addition to genetically encoded fluorescent 4 proteins, a number of molecular probes for monitoring cellular events have been 5 developed. 4, 5 The key challenge in cellular imaging is to stain cell compartments with high 6 specificity and persistence. Although numerous efficient molecular probes have been 7 designed to selectively stain specific cellular structures including mitochondria, 6 lipid 8 droplets, 7 endoplasmic reticulum, 8 nucleus 9 and lysosomes, 10--11 there is still a demand for 9 efficient and bright fluorescent PM markers. 12 Fluorescently labelled lectins notably wheat 10 germ agglutinin (WGA) and concanavalin A 9 are popular fluorescent membrane probes. 11Despite their efficiency and ease of use, these probes are expensive and much larger than 12 molecular probes. The small size of the latter, comparable to lipids, allows their precise 13 location in the lipid bilayer, which is of key importance for studies of the lateral lipid 14 organization of biomembranes (lipid rafts), 12, 13 FRET with membrane proteins 14 and super--15 resolution imaging. 15--16 Therefore, molecular probes are an interesting alternative to 16 protein--based membrane markers. Although highly hydrophobic fluorophores are efficient 17 markers of membrane models such as liposomes, they generally fail in staining the cell PM of 18 live cells as they tend to precipitate before reaching the membrane and to quickly cross the 19 PM to stain inner membranes. 12, 17 Recent efforts have been made to develop specific and 20 efficient PM probes for cell imaging with various fluorophores including chromone, 18--19 ...

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“…In addition, it is brighter and can therefore be used at reduced concentrations, minimizing again the risk of false positive results. The key difference of MemBright from PKH is the presence of amphiphilic groups, which favor efficient transfer of the fluorophore from aqueous media to lipid membranes (Collot et al, 2015;Kucherak et al, 2010). Therefore, MemBright can be used to confidently track EV dispersion and uptake.…”

Section: Discussionmentioning

confidence: 99%

Studying the fate of tumor extracellular vesicles at high spatio-temporal resolution using the zebrafish embryo

Hyenne

Ghoroghi

Collot³

et al. 2018

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Highlights• MemBright, a new family of membrane probes, allows for bright and specific staining of EVs • Zebrafish melanoma EVs are very similar to human and mouse melanoma EVs in morphology and protein content • The zebrafish embryo is an adapted model to precisely track tumor EVs dynamics and fate in a living organism from light to electron microscopy • Circulating tumor EVs are rapidly uptaken by endothelial cells and patrolling macrophages • Correlated light and electron microscopy can be used in zebrafish to identify cells and compartments uptaking tumor EVs Blurb Dispersion of tumor extracellular vesicles (EVs) throughout the body promotes tumor progression. However the behavior of tumor EVs in body fluids remains mysterious due to their small size and the absence of adapted animal model. Here we show that the zebrafish embryo can be used to track circulating tumor EVs in vivo and provide the first high-resolution description of their dissemination and uptake.

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Bright fluorogenic squaraines with tuned cell entry for selective imaging of plasma membrane vs. endoplasmic reticulum

Cited by 79 publications

References 31 publications

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

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

MemBright: a Family of Fluorescent Membrane Probes for Advanced Cellular Imaging and Neuroscience

Studying the fate of tumor extracellular vesicles at high spatio-temporal resolution using the zebrafish embryo

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