Rapid FLIM Measurement of Membrane Tension Probe Flipper-TR

Pandžić, Elvis; Whan, Renée; Macmillan, Alexander

doi:10.1007/978-1-0716-1843-1_20

Cited by 7 publications

(4 citation statements)

References 9 publications

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“…FLIM imaging was performed using a Picoquant Microtime 200 STED confocal microscope as described previously. [ 37 ] Excitation was performed using a pulsed 485 nm laser Diode (PicoQuant, LDH‐D‐C‐485) operating at 20 MHz, and emission signal was collected through a bandpass 600/50 nm filter using a gated PMA hybrid 40 detector and a TimeHarp 260 NANO TCSPC board (PicoQuant). SymPhoTime 64 software (PicoQuant) was then used to process and analyze the data.…”

Section: Methodsmentioning

confidence: 99%

Hydrogel Microtumor Arrays to Evaluate Nanotherapeutics

Liu

Nemec

Kopecky

et al. 2022

Adv Healthcare Materials

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Nanoparticle drug formulations have many advantages for cancer therapy due to benefits in targeting selectivity, lack of systemic toxicity, and increased drug concentration in the tumor microenvironment after delivery. However, the promise of nanomedicine is limited by preclinical models that fail to accurately assess new drugs before entering human trials. In this work a new approach to testing nanomedicine using a microtumor array formed through hydrogel micropatterning is demonstrated. This technique allows partitioning of heterogeneous cell states within a geometric pattern—where boundary regions of curvature prime the stem cell‐like fraction—allowing to simultaneously probe drug uptake and efficacy in different cancer cell fractions with high reproducibility. Using melanoma cells of different metastatic potential, a relationship between stem fraction and nanoparticle uptake is discovered. Deformation cytometry reveals that the stem cell‐like population exhibits a more mechanically deformable cell membrane. Since the stem fraction in a tumor is implicated in drug resistance, recurrence, and metastasis, the findings suggest that nanoparticle drug formulations are well suited for targeting this dangerous cell population in cancer therapy.

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Section: Methodsmentioning

confidence: 99%

Hydrogel Microtumor Arrays to Evaluate Nanotherapeutics

Liu

Nemec

Kopecky

et al. 2022

Adv Healthcare Materials

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“…Small‐molecule fluorescent membrane probes are chemistry tools made to enable the fluorescence microscopy imaging of the respective biological functions by simple addition of the probes, without the need of genetic engineering [1–18] . Fluorescent flippers [19] have been developed from 2012 [20] –2016 [21,22] as bioinspired [23] probes to image membrane tension [24–26] in living systems, and have enabled mechanobiology research ever since [19,27,28] . Demand implied that fluorescent flippers satisfy a need in the community and thus encouraged efforts to further improve the original tools ( Figure 1).…”

Section: Introductionmentioning

confidence: 99%

“…[1 -18] Fluorescent flippers [19] have been developed from 2012 [20] -2016 [21,22] as bioinspired [23] probes to image membrane tension [24][25][26] in living systems, and have enabled mechanobiology research ever since. [19,27,28] Demand implied that fluorescent flippers satisfy a need in the community and thus encouraged efforts to further improve the original tools (Figure 1). Today, a rich flipper collection is available to target essentially any membrane of interest within cells, [19] using either empirical trackers [19,29,30] or cellular engineering [31,32] together with controlled release by chemical stimulation [31] or with light.…”

Section: Introductionmentioning

confidence: 99%

Fluorescent Membrane Probes Obey the Israelachvili Rules

Bayard,

Matile

2024

Helvetica Chimica Acta

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When developing fluorescent membrane probes, we naturally tend to focus on the fluorophore itself. In this study, we show that sometimes it can be beneficial to shift attention from the center to the periphery, to maximize multiple interfacing with complex changing environments. Palmitylation for hydrophobic interfacing and glutamate dendrons for hydrophilic interfacing are combined to improve the performance of fluorescent flipper probes. We show that to increase performance in membranes, solubility in water is important, and to increase solubility in water, we increase the hydrophobicity of the flipper probe. These seemingly paradoxical measures are taken to satisfy the Israelachvili rules. They state that only inverted cone amphiphiles form soluble micelles in water, while inverted micelles from cone shaped amphiphiles precipitate into hexagonal 1 supramolecular polymers, and the intermediate cylindrical amphiphiles show intermediate behavior dominated by bilayers and lamellar precipitates. The normal micelles obtained from inverted cone flipper amphiphiles prevent precipitation and prepare for efficient transfer into the lipid bilayer membranes. The results are flippers that break all records set by the 2016 original with regard to effective brightness, responsiveness to membrane order, anchoring in disordered membranes, partitioning into membranes of high order, and stable labeling of membranes of interest.

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“…Fluorescent flippers like 3 without triple bonds in the core, i.e., 0Y-Flippers in the context of this study, have been introduced as small-molecule fluorescent probes for the imaging of membrane tension, i.e., a physical force, in living systems. [41][42][43] These bioinspired [44][45][46] planarizable push-pull probes [46] are composed around an electron-rich and an electron-poor dithienothiophene. The two are twisted out of co-planarity by electrostatic repulsion around the rotatable central bond.…”

Section: Introductionmentioning

confidence: 99%

Core‐Alkynylated Fluorescent Flippers: Altered Ultrafast Photophysics to Track Thick Membranes

Pamungkas,

Fureraj,

Assies

et al. 2024

Angewandte Chemie

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Fluorescent flippers have been introduced as small‐molecule probes to image membrane tension in living systems. This study describes the design, synthesis, spectroscopic and imaging properties of flippers that are elongated by one and two alkynes inserted between the push and the pull dithienothiophene domains. The resulting mechanophores combine characteristics of flippers, reporting on physical compression in the ground state, and molecular rotors, reporting on torsional motion in the excited state, to take their photophysics to new level of sophistication. Intensity ratios in broadened excitation bands from differently twisted conformers of core‐alkynylated flippers thus report on mechanical compression. Lifetime boosts from ultrafast excited‐state planarization and lifetime drops from competitive intersystem crossing into triplet states report on viscosity. In standard lipid bilayer membranes, core‐alkynylated flippers are too long for one leaflet and tilt or extend into disordered interleaflet space, which preserves rotor‐like torsional disorder and thus weak, blue‐shifted fluorescence. Flipper‐like planarization occurs only in highly ordered membranes of matching leaflet thickness, where they light up and selectively report on these thick membranes with red‐shifted, sharpened excitation maxima, high intensity and long lifetime.

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Rapid FLIM Measurement of Membrane Tension Probe Flipper-TR

Cited by 7 publications

References 9 publications

Hydrogel Microtumor Arrays to Evaluate Nanotherapeutics

Hydrogel Microtumor Arrays to Evaluate Nanotherapeutics

Fluorescent Membrane Probes Obey the Israelachvili Rules

Core‐Alkynylated Fluorescent Flippers: Altered Ultrafast Photophysics to Track Thick Membranes

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