Signaling
dynamics
are
crucial
in
biological
systems,
and
biosensor-based
real-time
imaging
has
revolutionized
their
analysis.
Fluorescence
lifetime
microscopy
(FLIM)
excels
over
the
widely
used
fluorescence
intensity
by
allowing
measurement
of
absolute
signal
levels,
independent
sensor
concentration.
This
capability
enables
comparison
signaling
across
different
animals,
body
regions,
timeframes.
However,
FLIM’s
advantage
can
be
compromised
factors
like
autofluorescence
experiments.
To
address
this,
we
introduce
FLiSimBA,
a
flexible
computational
framework
for
realistic
F
luorescence
Li
fetime
Sim
ulation
B
iological
A
pplications.
Through
simulations,
analyze
signal-to-noise
ratios
data,
determining
uncertainty
providing
necessary
error
bars
measurements.
Furthermore,
challenge
belief
that
is
unaffected
expression
establish
quantitative
limits
to
this
insensitivity
applications.
Additionally,
propose
innovations,
notably
multiplexed
dynamic
combines
innovation
transform
number
signals
simultaneously
monitored,
thereby
enabling
systems
approach
studying
dynamics.
Thus,
incorporating
diverse
into
our
simulation
framework,
uncover
surprises,
identify
limitations,
advancements
biology.
supports
rigorous
experimental
design,
facilitates
accurate
data
interpretation,
paves
way
technological
imaging.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 12, 2025
The
concentrations
of
extracellular
and
intracellular
signaling
molecules,
such
as
dopamine
cAMP,
change
over
both
fast
slow
timescales
impact
downstream
pathways
in
a
cell-type
specific
manner.
Fluorescence
sensors
currently
used
to
monitor
signals
vivo
are
typically
optimized
detect
fast,
relative
changes
concentration
the
target
molecule.
They
less
well
suited
slowly-changing
rarely
provide
absolute
measurements
either
components.
Here,
we
developed
system
for
fluorescence
lifetime
photometry
at
high
temporal
resolution
(FLIPR)
that
utilizes
frequency-domain
analog
processing
measure
genetically-encoded
speed
but
with
long-term
stability
picosecond
precision
freely
moving
mice.
We
applied
FLIPR
investigate
two
functionally
distinct
regions
striatum,
nucleus
accumbens
core
(NAC)
tail
striatum
(TOS).
observed
higher
tonic
levels
baseline
TOS
compared
NAC
detected
differential
dynamic
responses
phasic
appetitive
aversive
stimuli.
Thus,
enables
simple
monitoring
time-scale
neuronal
units,
revealing
previously
unappreciated
spatial
variation
even
well-studied
systems.
Nature Methods,
Journal Year:
2024,
Volume and Issue:
21(10), P. 1916 - 1925
Published: Sept. 20, 2024
Abstract
Genetically
encoded
fluorescent
calcium
indicators
allow
cellular-resolution
recording
of
physiology.
However,
bright,
genetically
targetable
that
can
be
multiplexed
with
existing
tools
in
vivo
are
needed
for
simultaneous
imaging
multiple
signals.
Here
we
describe
WHaloCaMP,
a
modular
chemigenetic
indicator
built
from
bright
dye-ligands
and
protein
sensor
domains.
Fluorescence
change
WHaloCaMP
results
reversible
quenching
the
bound
dye
via
strategically
placed
tryptophan.
is
compatible
rhodamine
fluoresce
green
to
near-infrared,
including
several
efficiently
label
brain
animals.
When
near-infrared
dye-ligand,
shows
7×
increase
fluorescence
intensity
2.1-ns
lifetime
upon
binding.
We
use
WHaloCaMP1a
image
Ca
2+
responses
flies
mice,
perform
three-color
functional
hundreds
neurons
astrocytes
zebrafish
larvae
quantify
concentration
using
microscopy
(FLIM).
Scientific Reports,
Journal Year:
2024,
Volume and Issue:
14(1)
Published: Feb. 6, 2024
Intracellular
signaling
dynamics
play
a
crucial
role
in
cell
function.
Protein
kinase
A
(PKA)
is
key
molecule
that
has
diverse
functions,
from
regulating
metabolism
and
brain
activity
to
guiding
development
cancer
progression.
We
previously
developed
an
optical
reporter,
FLIM-AKAR,
allows
for
quantitative
imaging
of
PKA
via
fluorescence
lifetime
microscopy
photometry.
However,
using
viral
infection
or
electroporation
the
delivery
FLIM-AKAR
invasive
results
variable
expression.
Here,
we
reporter
mouse,
FL-AK,
which
expresses
Cre-dependent
manner
ROSA26
locus.
FL-AK
provides
robust
consistent
expression
over
time.
Functionally,
mouse
line
reports
increase
response
activation
both
G
Signaling
dynamics
are
crucial
in
biological
systems,
and
biosensor-based
real-time
imaging
has
revolutionized
their
analysis.
Fluorescence
lifetime
microscopy
(FLIM)
excels
over
the
widely
used
fluorescence
intensity
by
allowing
measurement
of
absolute
signal
levels,
independent
sensor
concentration.
This
capability
enables
comparison
signaling
across
different
animals,
body
regions,
timeframes.
However,
FLIM’s
advantage
can
be
compromised
factors
like
autofluorescence
experiments.
To
address
this,
we
introduce
FLiSimBA,
a
flexible
computational
framework
for
realistic
F
luorescence
Li
fetime
Sim
ulation
B
iological
A
pplications.
Through
simulations,
analyze
signal-to-noise
ratios
data,
determining
uncertainty
providing
necessary
error
bars
measurements.
Furthermore,
challenge
belief
that
is
unaffected
expression
establish
quantitative
limits
to
this
insensitivity
applications.
Additionally,
propose
innovations,
notably
multiplexed
dynamic
combines
innovation
transform
number
signals
simultaneously
monitored,
thereby
enabling
systems
approach
studying
dynamics.
Thus,
incorporating
diverse
into
our
simulation
framework,
uncover
surprises,
identify
limitations,
advancements
biology.
supports
rigorous
experimental
design,
facilitates
accurate
data
interpretation,
paves
way
technological
imaging.
Journal of Agricultural and Food Chemistry,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 11, 2024
As
the
proportion
of
aging
population
globally
is
surging
year
by
year,
age-associated
diseases,
including
neurodegenerative,
metabolic,
and
cardiovascular
have
recently
attracted
widespread
attention
food
scientists
nutritionists.
Polymethoxyflavonoids
(PMFs),
a
type
dietary
flavonoids,
emerged
as
potential
antiaging
candidates
owing
to
their
diverse
bioactivities,
encompassing
antioxidant,
anti-inflammatory,
neuroprotective,
metabolic
regulatory
effects.
Herein,
this
comprehensive
updated
review
has
summarized
discussed
effects
PMFs
on
aging,
possible
mechanisms
that
link
PMFs-mediated
modulation
prevention
or
treatment
various
aging-related
diseases
been
elaborated
in
detail.
Furthermore,
biological
fate
elaborately
from
absorption,
distribution,
metabolism,
excretion
vivo.
Special
given
bioavailability-bioactivity
relationship
PMFs,
PMF's
activity
significantly
hampered
poor
bioavailability.
Overall,
all
these
conclusions
may
help
providing
perspective
for
further
study
aging.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2023,
Volume and Issue:
unknown
Published: Dec. 21, 2023
Abstract
Signaling
dynamics
are
crucial
in
biological
systems,
and
biosensor-based
real-time
imaging
has
revolutionized
their
analysis.
Fluorescence
lifetime
microscopy
(FLIM)
excels
over
the
widely
used
fluorescence
intensity
by
allowing
measurement
of
absolute
signal
levels,
independent
sensor
concentration.
This
capability
enables
comparison
signaling
across
different
animals,
body
regions,
timeframes.
However,
FLIM’s
advantage
can
be
compromised
factors
like
autofluorescence
experiments.
To
address
this,
we
introduce
FLiSimBA,
a
flexible
computational
framework
for
realistic
F
luorescence
Li
fetime
Sim
ulation
B
iological
A
pplications.
Through
simulations,
analyze
signal-to-noise
ratios
data,
determining
uncertainty
providing
necessary
error
bars
measurements.
Furthermore,
challenge
belief
that
is
unaffected
expression
establish
quantitative
limits
to
this
insensitivity
applications.
Additionally,
propose
innovations,
notably
multiplexed
dynamic
combines
innovation
transform
number
signals
simultaneously
monitored,
thereby
enabling
systems
approach
studying
dynamics.
Thus,
incorporating
diverse
into
our
simulation
framework,
uncover
surprises,
identify
limitations,
advancements
biology.
supports
rigorous
experimental
design,
facilitates
accurate
data
interpretation,
paves
way
technological
imaging.
