ACS Nano,
Journal Year:
2024,
Volume and Issue:
18(6), P. 5167 - 5179
Published: Feb. 1, 2024
Elucidation
of
biological
phenomena
requires
imaging
microenvironments
in
vivo.
Although
the
seamless
visualization
vivo
hypoxia
from
level
whole-body
to
single-cell
has
great
potential
discover
unknown
and
medical
fields,
no
methodology
for
achieving
it
been
established
thus
far.
Here,
we
report
whole-organ
hypoxia,
an
important
microenvironment,
at
resolution
using
activatable
covalent
fluorescent
probes
compatible
with
tissue
clearing.
We
initially
focused
on
overcoming
incompatibility
dyes
refractive
index
matching
solutions
(RIMSs),
which
greatly
hindered
development
molecular
field
The
RIMS
were
then
incorporated
into
hypoxia.
combined
clearing,
comprehensive
single-cell-resolution
a
whole
mouse
body
organs.
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
.
Communications Biology,
Journal Year:
2023,
Volume and Issue:
6(1)
Published: May 9, 2023
Abstract
Light-sheet
fluorescence
microscopy
has
transformed
our
ability
to
visualize
and
quantitatively
measure
biological
processes
rapidly
over
long
time
periods.
In
this
review,
we
discuss
current
future
developments
in
light-sheet
that
expect
further
expand
its
capabilities.
This
includes
smart
adaptive
imaging
schemes
overcome
traditional
trade-offs,
i.e.,
spatiotemporal
resolution,
field
of
view
sample
health.
microscopy,
a
microscope
will
autonomously
decide
where,
when,
what
how
image.
We
assess
image
restoration
techniques
provide
avenues
these
tradeoffs
“open
top”
microscopes
may
enable
multi-modal
with
high
throughput.
As
such,
predict
fulfill
an
important
role
biomedical
clinical
the
future.
Tissue
clearing
for
whole
organ
cell
profiling
has
revolutionized
biology
and
imaging
exploration
of
organs
in
three-dimensional
space
without
compromising
tissue
architecture.
But
complicated,
laborious
procedures,
or
expensive
equipment,
as
well
the
use
hazardous,
organic
solvents
prevent
widespread
adoption
these
methods.
Here,
we
report
a
simple
rapid
method,
EZ
Clear,
that
can
clear
adult
mouse
48
hr
just
three
steps.
Samples
stay
at
room
temperature
remain
hydrated
throughout
process,
preserving
endogenous
synthetic
fluorescence,
altering
sample
size.
After
wholemount
imaging,
samples
processed
with
Clear
be
subjected
to
downstream
applications,
such
embedding
cryosectioning
followed
by
standard
histology
immunofluorescent
staining
loss
fluorescence
signal
from
reporters.
Furthermore,
demonstrate
brains
successfully
immunolabeled
fluorescent
while
still
retaining
Overall,
simplicity,
speed,
flexibility
make
it
easy
adapt
implement
diverse
modalities
biomedical
research.
Nature Communications,
Journal Year:
2024,
Volume and Issue:
15(1)
Published: June 12, 2024
Despite
widespread
adoption
of
tissue
clearing
techniques
in
recent
years,
poor
access
to
suitable
light-sheet
fluorescence
microscopes
remains
a
major
obstacle
for
biomedical
end-users.
Here,
we
present
descSPIM
(desktop-equipped
SPIM
cleared
specimens),
low-cost
($20,000-50,000),
low-expertise
(one-day
installation
by
non-expert),
yet
practical
do-it-yourself
microscope
as
solution
this
bottleneck.
Even
the
most
fundamental
configuration
enables
multi-color
imaging
whole
mouse
brains
and
cancer
cell
line-derived
xenograft
tumor
mass
visualization
neurocircuitry,
assessment
drug
distribution,
pathological
examination
false-colored
hematoxylin
eosin
staining
three-dimensional
manner.
Academically
open-sourced
(
https://github.com/dbsb-juntendo/descSPIM
),
allows
routine
samples
minutes.
Thus,
dissemination
will
accelerate
discoveries
driven
technologies.
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
