International Journal of Molecular Sciences,
Год журнала:
2024,
Номер
25(23), С. 12493 - 12493
Опубликована: Ноя. 21, 2024
The
calcium
cation
is
a
crucial
signaling
molecule
involved
in
numerous
cellular
pathways.
Beyond
its
role
as
messenger
or
modulator
intracellular
cascades,
calcium's
function
excitable
cells,
including
nerve
impulse
transmission,
remarkable.
central
of
nervous
activity
has
driven
the
rapid
development
fluorescent
techniques
for
monitoring
this
living
cells.
Specifically,
genetically
encoded
indicators
(GECIs)
are
most
in-demand
molecular
tools
their
class.
In
work,
we
address
two
issues
imaging
by
designing
based
on
successful
GCaMP6
backbone
and
protein
BrUSLEE.
first
indicator
variant
(GCaMP6s-BrUS),
with
reduced,
calcium-insensitive
fluorescence
lifetime,
potential
dynamics
high
temporal
resolution
combination
advanced
microscopy
techniques,
such
light
beads
microscopy,
where
lifetime
limits
acquisition
speed.
Conversely,
second
(GCaMP6s-BrUS-145),
flexible,
calcium-sensitive
relevant
static
measurements,
particularly
determining
absolute
concentration
values
using
(FLIM).
To
identify
structural
determinants
sensitivity
these
variants,
determine
spatial
structures.
A
comparative
analysis
allowed
optimization
GCaMP6s-BrUS
construct,
resulting
an
combining
behavior
time
domain
enhanced
brightness.
Our
data
may
serve
starting
point
further
engineering
efforts
towards
improved
GECI
variants
fine-tuned
lifetimes.
Frontiers in Oncology,
Год журнала:
2025,
Номер
14
Опубликована: Янв. 8, 2025
Photodynamic
diagnosis
(PDD)
and
autofluorescence
imaging
(AFI)
are
emerging
cancer
diagnostic
technologies
that
offer
significant
advantages
over
traditional
white-light
endoscopy
in
detecting
precancerous
lesions
early-stage
cancers;
moreover,
they
hold
promising
potential
fluorescence-guided
surgery
(FGS)
for
tumors.
However,
their
shortcomings
have
somewhat
hindered
the
clinical
application
of
PDD
AFI.
Therefore,
it
is
imperative
to
enhance
efficacy
AFI,
thereby
maximizing
practical
use.
This
article
reviews
principles,
characteristics,
current
research
status,
advancements
focusing
on
analyzing
discussing
optimization
strategies
AFI
tumor
FGS
scenarios.
Considering
technical
feasibility,
optimizing
may
result
an
effective
real-time
tool
guide
clinicians
surgical
guidance
achieve
best
results.
Biosensors,
Год журнала:
2025,
Номер
15(1), С. 43 - 43
Опубликована: Янв. 13, 2025
Fluorescence
lifetime
imaging
(FLIM)
has
established
itself
as
a
pivotal
tool
for
investigating
biological
processes
within
living
cells.
However,
the
extensive
duration
necessary
to
accumulate
sufficient
photons
accurate
fluorescence
calculations
poses
significant
obstacle
achieving
high-resolution
monitoring
of
cellular
dynamics.
In
this
study,
we
introduce
an
image
reconstruction
method
based
on
edge-preserving
interpolation
(EPIM),
which
transforms
rapidly
acquired
low-resolution
FLIM
data
into
high-pixel
images,
thereby
eliminating
need
extended
acquisition
times.
Specifically,
decouple
grayscale
and
matrix
perform
individual
each.
Following
intensity
image,
apply
wavelet
transformation
adjust
coefficients
according
gradients.
After
inverse
transformation,
original
is
obtained
subjected
noise
reduction
complete
process.
Subsequently,
each
pixel
pseudo-color-coded
its
lifetime,
preserving
both
structural
temporal
information.
We
evaluated
performance
bicubic
our
approach
microspheres
fixed-cell
samples,
demonstrating
their
effectiveness
in
enhancing
quality
images.
By
applying
these
techniques
live-cell
imaging,
can
successfully
obtain
images
at
shortened
intervals,
facilitating
capture
rapid
events.
Sperm
metabolic
pathways
that
generate
energy
for
motility
are
compartmentalized
within
the
flagellum.
Dysfunctions
in
compartments,
namely
mitochondrial
respiration
and
glycolysis,
can
compromise
male
fertility.
Studying
these
compartments
is
thus
required
fertility
treatment.
However,
it
very
challenging
to
capture
images
of
motile
spermatozoa
because
fast
beating
flagellum
introduces
motion
blur.
Therefore,
most
approaches
immobilize
prior
imaging.
Our
findings
indicate
immobilizing
sperm
alters
their
profile,
highlighting
necessity
measuring
metabolism
during
movement.
We
achieved
this
by
encapsulating
mouse
epididymis
a
hydrogel
followed
two-photon
fluorescence
lifetime
imaging
microscopy
situ.
The
autofluorescence
endogenous
metabolites—FAD,
NADH,
NADPH—enabled
us
visualize
without
staining.
trained
machine
learning
automated
image
segmentation
generated
fingerprints
using
object-based
phasor
analysis.
show
compartment
(1)
distinguish
individual
males
genetic
background,
age,
or
fecundity
status,
(2)
correlate
with
fertility,
(3)
change
age
likely
due
increased
oxidative
metabolism.
approach
eliminates
need
immobilization
labeling
captures
native
state
This
technique
could
be
adapted
metabolism-based
selection
assisted
reproduction.
Cells,
Год журнала:
2025,
Номер
14(10), С. 732 - 732
Опубликована: Май 17, 2025
Drug
delivery
to
solid
tumors
is
challenged
by
multiple
physiological
barriers
arising
from
the
tumor
microenvironment,
including
dense
extracellular
matrix,
cellular
heterogeneity,
hypoxic
gradients,
and
elevated
interstitial
fluid
pressure.
These
features
hinder
uniform
distribution
accumulation
of
therapeutics,
reducing
treatment
efficacy.
Despite
their
widespread
use,
conventional
two-dimensional
monolayer
cultures
fail
reproduce
these
complexities,
contributing
poor
translational
predictability
many
preclinical
candidates.
Three-dimensional
multicellular
spheroids
have
emerged
as
more
representative
in
vitro
models
that
capture
essential
architecture,
stromal
interactions,
microenvironmental
resistance
mechanisms.
Spheroids
exhibit
spatially
organized
regions
proliferation,
quiescence,
hypoxia,
can
incorporate
non-tumor
cells
mimic
tumor–stroma
crosstalk.
Advances
spheroid
analysis
now
enable
detailed
evaluation
drug
penetration,
migration,
cytotoxic
response,
molecular
gradients
using
techniques
such
optical
confocal
imaging,
large-particle
flow
cytometry,
biochemical
viability
assays,
microfluidic
integration.
By
combining
relevance
with
analytical
accessibility,
support
mechanistic
studies
transport
efficacy
under
tumor-like
conditions.
Their
adoption
into
routine
workflows
has
potential
improve
accuracy
while
reliance
on
animal
models.
Journal of Neurochemistry,
Год журнала:
2025,
Номер
169(6)
Опубликована: Июнь 1, 2025
ABSTRACT
Neural
networks
are
responsible
for
processing
sensory
stimuli
and
driving
the
synaptic
activity
required
brain
function
behavior.
This
computational
capacity
is
expensive
requires
a
steady
supply
of
energy
building
blocks
to
operate.
Importantly,
neural
composed
different
cell
populations,
whose
metabolic
profiles
differ
between
each
other,
thus
endowing
them
with
capacities,
such
as,
example,
ability
synthesize
specific
precursors
or
variable
proficiency
manage
their
waste.
These
marked
differences
likely
prompted
emergence
diverse
intercellular
interactions,
in
which
shuttling
cycling
metabolites
cells
allows
separation
workload
efficient
control
demand
within
central
nervous
system.
Nevertheless,
our
knowledge
about
bioenergetics
adaptations
still
warrants
further
studies.
In
this
review,
originated
from
Fourth
International
Society
Neurochemistry
(ISN)
Journal
(JNC)
Flagship
School
held
Schmerlenbach,
Germany
(2022),
we
describe
discuss
cells,
exchanges
these
how
bioenergetic
activities
shape
Furthermore,
potential
role
faulty
etiology
progression
Alzheimer's
disease,
Parkinson
Amyotrophic
lateral
sclerosis.
We
foresee
that
deeper
understanding
metabolism
will
provide
crucial
insights
into
higher‐order
functions
emerge
reveal
roots
neuropathological
conditions
hallmarks
include
impaired
function.
image
