View,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 21, 2025
Abstract
Over
the
recent
years,
fluorescence
microscopy
has
seen
substantial
progress,
transitioning
from
primarily
qualitative
analysis
to
a
profoundly
quantitative
methodology,
yet
limitations
in
imaging
depth
and
data
quality
persist
due
tissue
opacity.
Tissue
optical
clearing
(TOC)
methodologies
were
implemented
overcome
this
hurdle,
enabling
of
large
samples
such
as
entire
murine
organs.
Further
improvements,
including
expansion
merge
TOC
benefits
with
improved
resolution,
converting
cells
tissues
into
tissue‒gel
hybrids.
Yet,
its
intricate
protocols
lead
remarkable,
∼10‒20×
volume
expansion,
making
acquisition
even
thin
sections
exceedingly
challenging
terms
time,
labor,
handling.
Here,
we
present
novel
solution,
Improved
Segmentation
by
gEntle
Expansion
(ISEE),
for
both
gentle
(1.6‒1.7×)
significantly
precision
fluorescent
signal
segmentation,
while
maintaining
tolerable
increase
during
everyday
laboratory
practice.
When
applied
thick
∼40‒200
µm
slices,
ISEE
renders
them
transparent
ultimately
expanded
within
minutes,
simple,
one‐step
process
compatible
immunohistochemistry
well
all
major
Frontiers in Neuroscience,
Journal Year:
2022,
Volume and Issue:
16
Published: March 31, 2022
Since
1995,
more
than
100
transgenic
(Tg)
mouse
models
of
Alzheimer's
disease
(AD)
have
been
generated
in
which
mutant
amyloid
precursor
protein
(APP)
or
APP/presenilin
1
(PS1)
cDNA
is
overexpressed
(
Nature Biotechnology,
Journal Year:
2023,
Volume and Issue:
42(4), P. 617 - 627
Published: July 10, 2023
Abstract
Whole-body
imaging
techniques
play
a
vital
role
in
exploring
the
interplay
of
physiological
systems
maintaining
health
and
driving
disease.
We
introduce
wildDISCO,
new
approach
for
whole-body
immunolabeling,
optical
clearing
mice,
circumventing
need
transgenic
reporter
animals
or
nanobody
labeling
so
overcoming
existing
technical
limitations.
identified
heptakis(2,6-di-
O
-methyl)-β-cyclodextrin
as
potent
enhancer
cholesterol
extraction
membrane
permeabilization,
enabling
deep,
homogeneous
penetration
standard
antibodies
without
aggregation.
WildDISCO
facilitates
peripheral
nervous
systems,
lymphatic
vessels
immune
cells
whole
mice
at
cellular
resolution
by
diverse
endogenous
proteins.
Additionally,
we
examined
rare
proliferating
effects
biological
perturbations,
demonstrated
germ-free
mice.
applied
wildDISCO
to
map
tertiary
lymphoid
structures
context
breast
cancer,
considering
both
primary
tumor
metastases
throughout
mouse
body.
An
atlas
high-resolution
images
showcasing
nervous,
vascular
is
accessible
http://discotechnologies.org/wildDISCO/atlas/index.php
.
Proceedings of the National Academy of Sciences,
Journal Year:
2024,
Volume and Issue:
121(6)
Published: Jan. 31, 2024
Neurotransmitter
receptors
are
essential
components
of
synapses
for
communication
between
neurons
in
the
brain.
Because
spatiotemporal
expression
profiles
and
dynamics
neurotransmitter
involved
many
functions
delicately
governed
brain,
vivo
research
tools
with
high
resolution
intact
brains
highly
desirable.
Covalent
labeling
by
chemical
reaction
(chemical
labeling)
proteins
without
genetic
manipulation
is
now
a
powerful
method
analyzing
vitro.
However,
selective
target
receptor
brain
has
not
yet
been
achieved.
This
study
shows
that
ligand-directed
alkoxyacylimidazole
(LDAI)
chemistry
can
be
used
to
selectively
tether
synthetic
probes
endogenous
living
mouse
brains.
The
reactive
LDAI
reagents
negative
charges
were
found
diffuse
well
over
whole
could
label
receptors,
including
AMPAR,
NMDAR,
mGlu1,
GABA
A
R.
simple
robust
protocol
was
then
various
applications:
three-dimensional
spatial
mapping
healthy
disease-model
mice;
multi-color
imaging;
pulse–chase
analysis
postnatal
Here,
results
demonstrated
bioorthogonal
modification
animal
may
provide
innovative
molecular
contribute
in-depth
understanding
complicated
functions.
Nature Biotechnology,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 14, 2025
Efficient
and
accurate
nanocarrier
development
for
targeted
drug
delivery
is
hindered
by
a
lack
of
methods
to
analyze
its
cell-level
biodistribution
across
whole
organisms.
Here
we
present
Single
Cell
Precision
Nanocarrier
Identification
(SCP-Nano),
an
integrated
experimental
deep
learning
pipeline
comprehensively
quantify
the
targeting
nanocarriers
throughout
mouse
body
at
single-cell
resolution.
SCP-Nano
reveals
tissue
distribution
patterns
lipid
nanoparticles
(LNPs)
after
different
injection
routes
doses
as
low
0.0005
mg
kg-1-far
below
detection
limits
conventional
imaging
techniques.
We
demonstrate
that
intramuscularly
injected
LNPs
carrying
SARS-CoV-2
spike
mRNA
reach
heart
tissue,
leading
proteome
changes,
suggesting
immune
activation
blood
vessel
damage.
generalizes
various
types
nanocarriers,
including
liposomes,
polyplexes,
DNA
origami
adeno-associated
viruses
(AAVs),
revealing
AAV2
variant
transduces
adipocytes
body.
enables
comprehensive
three-dimensional
mapping
bodies
with
high
sensitivity
should
accelerate
precise
safe
nanocarrier-based
therapeutics.
