bc8b00828_si_003.avi (11.78 MB)
BODIPY with Tuned Amphiphilicity as a Fluorogenic Plasma Membrane Probe
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posted on 2018-12-18, 00:00 authored by Mayeul Collot, Emmanuel Boutant, Maxime Lehmann, Andrey S. KlymchenkoStaining
of the plasma membrane (PM) is essential in bioimaging,
as it delimits the cell surface and provides various information regarding
the cell morphology and status. Herein, the lipophilicity of a green
emitting BODIPY fluorophore was tuned by gradual functionalization
with anchors composed of zwitterionic and aliphatic groups, thus yielding
three different amphiphilic dyes. We found that BODIPY bearing one
or three anchors failed in efficiently staining the PM: the derivative
with one anchor showed low affinity to PM and exhibited strong fluorescence
in water due to high solubility, whereas BODIPY with three anchors
aggregated strongly in media and precipitated before binding to the
PM. In sharp contrast, the BODIPY bearing two anchors (B-2AZ, MemBright-488)
formed virtually nonfluorescent soluble aggregates in aqueous medium
that quickly deaggregated in the presence of PM, leading to a bright
soluble molecular form (quantum yield of 0.92). This fluorogenic response
allowed for efficient probing of the PM at low concentration (20 nM)
with high signal to background ratio images in mono- as well as two-photon
excitation microscopy. B-2AZ proved to selectively stain the PM in
a more homogeneous manner than the commercially available fluorescently
labeled lectin WGA. Finally, it was successfully used in 3D-imaging
to reveal fine intercellular tunneling nanotubes in KB cells and to
stain the PM in glioblastoma cells in spheroids.
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Tuned Amphiphilicityamphiphilic dyesintercellular tunneling nanotubescell morphologyBODIPY fluorophoretwo-photon excitation microscopyPMglioblastoma cells3 D-imagingKB cellsplasma membranebackground ratio imagesfluorogenic responseb -2AZcell surfacealiphatic groupsFluorogenic Plasma Membrane Probeanchors aggregatedlectin WGA
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