Label Free Assessment of Key Biological Autofluorophores: Material Characteristics and Opportunities for Clinical Applications
Advanced Materials,
Год журнала:
2024,
Номер
unknown
Опубликована: Май 22, 2024
Autofluorophores
are
endogenous
fluorescent
compounds
that
naturally
occur
in
the
intra
and
extracellular
spaces
of
all
tissues
organs.
Most
have
vital
biological
functions
-
like
metabolic
cofactors
NAD(P)H
FAD
Язык: Английский
Autofluorescence lifetime flow cytometry with time‐correlated single photon counting
Cytometry Part A,
Год журнала:
2024,
Номер
105(8), С. 607 - 620
Опубликована: Июнь 28, 2024
Autofluorescence
lifetime
imaging
microscopy
(FLIM)
is
sensitive
to
metabolic
changes
in
single
cells
based
on
the
protein-binding
activities
of
co-enzymes
NAD(P)H.
However,
FLIM
typically
relies
time-correlated
single-photon
counting
(TCSPC)
detection
electronics
laser-scanning
microscopes,
which
are
expensive,
low-throughput,
and
require
substantial
post-processing
time
for
cell
segmentation
analysis.
Here,
we
present
a
fluorescence
lifetime-sensitive
flow
cytometer
that
offers
same
TCSPC
temporal
resolution
geometry,
with
low-cost
excitation
sources,
throughput
tens
per
second,
real-time
single-cell
The
system
uses
375
nm
picosecond-pulsed
diode
laser
operating
at
50
MHz,
alkali
photomultiplier
tubes,
an
FPGA-based
tagger,
can
provide
phasor-based
classification
(i.e.,
gating)
flowing
cells.
A
CMOS
camera
produces
simultaneous
brightfield
images
using
far-red
illumination.
second
PMT
provides
two-color
Cells
injected
into
microfluidic
channel
syringe
pump
2-5
mm/s
nearly
5
ms
integration
cell,
resulting
light
dose
2.65
J/cm
Язык: Английский
Accelerating biopharmaceutical cell line selection with label-free multimodal nonlinear optical microscopy and machine learning
Communications Biology,
Год журнала:
2025,
Номер
8(1)
Опубликована: Фев. 3, 2025
The
selection
of
high-performing
cell
lines
is
crucial
for
biopharmaceutical
production
but
often
time-consuming
and
labor-intensive.
We
investigated
label-free
multimodal
nonlinear
optical
microscopy
non-perturbative
profiling
based
on
their
intrinsic
molecular
contrast.
Employing
simultaneous
autofluorescence
multiharmonic
(SLAM)
with
fluorescence
lifetime
imaging
(FLIM),
we
characterized
Chinese
hamster
ovary
(CHO)
at
early
passages
(0–2).
A
machine
learning
(ML)-assisted
analysis
pipeline
leveraged
high-dimensional
information
to
classify
single
cells
into
respective
lines.
Remarkably,
the
monoclonal
line
classifiers
achieved
balanced
accuracies
exceeding
96.8%
as
passage
2.
Correlation
features
FLIM
modality
played
pivotal
roles
in
classification.
This
integrated
bioimaging
approach
presents
a
promising
solution
expedite
process
while
ensuring
identification
techniques
have
potential
broader
single-cell
characterization
applications
stem
research,
immunology,
cancer
biology
beyond.
Label-free
enable
early,
classification
lines,
accelerating
processes
opening
avenues
studies.
Язык: Английский
CHARACTERIZATION OF AUTOFLUORESCENCE AS AN INDICATOR OF ACTIVATION STATE IN NEURAL STEM CELLS
Journal of Stem Cell Research and Tissue Engineering,
Год журнала:
2024,
Номер
8(1), С. 37 - 42
Опубликована: Май 28, 2024
Recent
advancements
in
stem
cell
research
have
uncovered
a
novel
autofluorescence
marker
pivotal
for
investigating
the
dormant
state
of
cells.
This
presents
groundbreaking
opportunity
to
monitor
transition
cells
from
quiescent
an
active
state,
facilitating
identification
entering
cycle.
The
primary
objective
this
is
comprehensively
review
marker's
efficacy
with
aim
developing
therapeutic
strategies
generating
human
nerve
A
systematic
literature
search
initially
yielded
2297
articles
on
characterization
as
indicator
activation
neural
(NSCs).
However,
only
three
met
stringent
inclusion
criteria,
underscoring
novelty
and
scarcity
domain.
Autofluorescence,
particularly
NSCs,
offers
non-invasive
approach
studying
molecular
processes
discerning
various
states,
obviating
need
external
labels.
technique
not
preserves
intrinsic
properties
but
also
circumvents
biases
inherent
traditional
labeling
methods.
Moreover,
when
coupled
cutting-edge
technologies
such
Optical
Coherence
Tomography
Spectral
Inverse
Analysis
(OCSI),
it
enables
precise,
real-time
monitoring
metabolic
alterations
NSCs
during
their
dormancy
activity.
Язык: Английский
Long-term tracing of individual human neural cells using multiphoton microscopy and photoconvertible polymer capsules
Journal of The Royal Society Interface,
Год журнала:
2024,
Номер
21(219)
Опубликована: Окт. 1, 2024
The
study
of
human
neural
cells,
their
behaviour
and
migration
are
important
areas
research
in
the
biomedical
field,
particularly
for
potential
therapeutic
applications.
safety
using
cells
therapy
is
still
a
concern
due
to
lack
information
on
long-term
changes
that
may
occur.
While
current
methods
cell
tracing
explore
gene
manipulations,
we
elaborate
approaches
marking
with
no
genetic
interference.
In
this
study,
present
novel
method
labelling
tracking
cell-impregnatable
photoconvertible
polyelectrolyte
microcapsules.
These
capsules
demonstrated
low
cytotoxicity
effect
differentiation
ability
maintained
high
level
fluorescent
signal
individual
over
7
days.
modified
rhodamine-
fluorescein-based
dyes
were
undergo
photoconversion
by
both
one-
two-photon
lasers
while
being
internalized
cells.
finding
gives
possibility
select
inside
multicellular
structures
like
spheroids
tissues
alternate
appearance.
Thus,
can
track
paths
complex
systems.
This
new
offers
promising
alternative
studying
cells’
systems
such
as
three-dimensional
cellular
populations.
Язык: Английский
Endogenous mitochondrial NAD(P)H fluorescence can predict lifespan
Communications Biology,
Год журнала:
2024,
Номер
7(1)
Опубликована: Ноя. 21, 2024
Many
aging
clocks
have
recently
been
developed
to
predict
health
outcomes
and
deconvolve
heterogeneity
in
aging.
However,
existing
are
limited
by
technical
constraints,
such
as
low
spatial
resolution,
long
processing
time,
sample
destruction,
a
bias
towards
specific
phenotypes.
Therefore,
here
we
present
non-destructive,
label-free
subcellular
resolution
approach
for
quantifying
through
optically
resolving
age-dependent
changes
the
biophysical
properties
of
NAD(P)H
mitochondria
fluorescence
lifetime
imaging
(FLIM)
endogenous
fluorescence.
We
uncover
mitochondrial
across
tissues
C.
elegans
that
associated
with
decline
physiological
function
construct
cellular
models
prediction
age,
which
refer
"mito-NAD(P)H
age
clocks."
Mito-NAD(P)H
can
resolve
rate
individuals
remaining
lifespan.
Moreover,
spatiotemporally
within
tissues,
revealing
multiple
modes
asynchrony
show
longevity
is
ubiquitous
attenuation
these
changes.
Our
data
high-resolution
view
aging,
providing
insights
broaden
our
understanding
how
change
during
approaches
expand
toolkit
quantify
Endogenous
enables
quantification
lifespan
tracking
mitochondria.
Язык: Английский