Naturally
occurring
fluorescent
proteins
(FPs)
are
the
most
widely
used
tools
for
tracking
cellular
and
sensing
events.
Here,
we
chemically
evolved
self-labeling
SNAP-tag
into
a
palette
of
mimics
(SmFPs)
that
possess
bright,
rapidly
inducible
fluorescence
ranging
from
cyan
to
infrared.
SmFPs
integral
chemical-genetic
entities
based
on
same
fluorogenic
principle
as
FPs,
i.e.,
induction
non-emitting
molecular
rotors
by
conformational
locking.
We
demonstrate
usefulness
these
in
real-time
protein
expression,
degradation,
binding
interactions,
trafficking,
assembly,
show
optimally
designed
outperform
FPs
like
GFP
many
important
ways.
further
circularly
permuted
is
sensitive
changes
their
fusion
partners,
partners
can
be
development
single
SmFP-based
genetically
encoded
calcium
sensors
live
cell
imaging.
Angewandte Chemie International Edition,
Год журнала:
2020,
Номер
60(30), С. 16294 - 16308
Опубликована: Авг. 12, 2020
The
development
of
fluorophores
for
the
second
near-infrared
window
(NIR-II,
1000-1700
nm)
represents
an
emerging,
significant,
and
vibrant
field
in
analytic
chemistry,
chemical
biology,
biomedical
engineering.
wavelength,
brightness,
stability
are
three
crucial
factors
that
determine
performance
NIR-II
fluorophore.
Up
to
now,
significant
progress
has
been
made
fluorescence
molecular
probes,
including
synthesis
D-A-D
D-π-A
with
improved
imaging
construction
off-on
probes
ratiometric
via
energy
transfer
or
structure
modification.
In
this
review,
we
summarize
most
recent
advances
engineering
design
strategies
then
highlight
a
selection
bioimaging
biosensing
applications.
We
also
provide
perspectives
on
potential
challenges
opportunities
emerging
field.
ACS Central Science,
Год журнала:
2019,
Номер
5(9), С. 1602 - 1613
Опубликована: Сен. 5, 2019
Rhodamine
dyes
exist
in
equilibrium
between
a
fluorescent
zwitterion
and
nonfluorescent
lactone.
Tuning
this
toward
the
lactone
form
can
improve
cell-permeability
allow
creation
of
"fluorogenic"
compounds-ligands
that
shift
to
upon
binding
biomolecular
target.
An
archetype
fluorogenic
dye
is
far-red
tetramethyl-Si-rhodamine
(SiR),
which
has
been
used
create
exceptionally
useful
labels
for
advanced
microscopy.
Here,
we
develop
quantitative
framework
development
new
dyes,
determining
lactone-zwitterion
constant
(KL-Z)
sufficient
predict
fluorogenicity.
This
rubric
emerged
from
our
analysis
known
fluorophores
yielded
with
improved
performance
cellular
imaging
experiments.
We
then
designed
novel
fluorophore-Janelia
Fluor
526
(JF526)-with
SiR-like
properties
but
shorter
fluorescence
excitation
emission
wavelengths.
JF526
versatile
scaffold
probes
including
ligands
self-labeling
tags,
stains
endogenous
structures,
spontaneously
blinking
super-resolution
immunofluorescence.
constitutes
label
microscopy
experiments,
will
enable
rational
design
other
bioimaging.
Organic & Biomolecular Chemistry,
Год журнала:
2020,
Номер
18(30), С. 5747 - 5763
Опубликована: Янв. 1, 2020
Guidelines
based
on
photophysical
tuning,
reactivity,
isomerization,
and
energy
transfer
for
rational
design
of
synthetic
fluorescent
probes
biological
systems.
Journal of the American Chemical Society,
Год журнала:
2020,
Номер
142(42), С. 17828 - 17844
Опубликована: Окт. 9, 2020
Single-molecule
super-resolution
microscopy
has
developed
from
a
specialized
technique
into
one
of
the
most
versatile
and
powerful
imaging
methods
nanoscale
over
past
two
decades.
In
this
perspective,
we
provide
brief
overview
historical
development
field,
fundamental
concepts,
methodology
required
to
obtain
maximum
quantitative
information,
current
state
art.
Then,
will
discuss
emerging
perspectives
areas
where
innovation
further
improvement
are
needed.
Despite
tremendous
progress,
full
potential
single-molecule
is
yet
be
realized,
which
enabled
by
research
ahead
us.
Journal of Nanobiotechnology,
Год журнала:
2021,
Номер
19(1)
Опубликована: Май 10, 2021
Abstract
Molecular
imaging
technology
enables
us
to
observe
the
physiological
or
pathological
processes
in
living
tissue
at
molecular
level
accurately
diagnose
diseases
an
early
stage.
Optical
can
be
employed
achieve
dynamic
monitoring
of
and
has
promising
applications
biomedicine.
The
traditional
first
near-infrared
(NIR-I)
window
(NIR-I,
range
from
700
900
nm)
technique
been
available
for
more
than
two
decades
extensively
utilized
clinical
diagnosis,
treatment
scientific
research.
Compared
with
NIR-I,
second
NIR
optical
(NIR-II,
1000
1700
low
autofluorescence,
a
high
signal-to-noise
ratio,
penetration
depth
large
Stokes
shift.
Recently,
this
attracted
significant
attention
also
become
heavily
researched
topic
In
study,
characteristics
different
fluorescence
nanoprobes
latest
reports
regarding
application
NIR-II
biological
tissues
will
described.
Furthermore,
existing
problems
future
perspectives
probes
discussed.
ACS Central Science,
Год журнала:
2021,
Номер
7(7), С. 1144 - 1155
Опубликована: Июнь 3, 2021
The
light-promoted
conversion
of
extensively
used
cyanine
dyes
to
blue-shifted
emissive
products
has
been
observed
in
various
contexts.
However,
both
the
underlying
mechanism
and
species
involved
this
photoconversion
reaction
have
remained
elusive.
Here
we
report
that
irradiation
heptamethine
cyanines
provides
pentamethine
cyanines,
which,
turn,
are
photoconverted
trimethine
cyanines.
We
detail
an
examination
substrate
scope
remarkable
two-carbon
phototruncation
reaction.
Supported
by
computational
analysis,
propose
involves
a
singlet
oxygen-initiated
multistep
sequence
involving
key
hydroperoxycyclobutanol
intermediate.
Building
on
mechanistic
framework,
identify
conditions
improve
yield
over
order
magnitude.
then
demonstrate
can
be
applied
super-resolution
single-molecule
localization
microscopy,
leading
improved
spatial
resolution
with
shorter
imaging
times.
anticipate
these
insights
will
help
transform
common,
but
previously
mechanistically
ill-defined,
chemical
transformation
into
valuable
optical
tool.
Angewandte Chemie International Edition,
Год журнала:
2022,
Номер
62(1)
Опубликована: Сен. 30, 2022
Decoding
cellular
processes
requires
visualization
of
the
spatial
distribution
and
dynamic
interactions
biomolecules.
It
is
therefore
not
surprising
that
innovations
in
imaging
technologies
have
facilitated
advances
biomedical
research.
The
advent
super-resolution
has
empowered
researchers
with
ability
to
answer
long-standing
questions
about
at
an
entirely
new
level.
Fluorescent
probes
greatly
enhance
specificity
resolution
experiments.
Here,
we
introduce
key
technologies,
a
brief
discussion
on
single-molecule
localization
microscopy
(SMLM).
We
evaluate
chemistry
photochemical
mechanisms
fluorescent
employed
SMLM.
This
Review
provides
guidance
identification
adoption
single
molecule
inspire
design
next-generation
amenable
imaging.
