bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2022,
Volume and Issue:
unknown
Published: Dec. 2, 2022
Abstract
Tissue
clearing
and
labeling
techniques
have
revolutionized
brain-wide
imaging
analysis,
yet
their
application
to
clinical
formalin-fixed
paraffin-embedded
(FFPE)
blocks
remains
challenging.
We
introduce
HIF-Clear,
a
novel
method
for
efficiently
centimeter-thick
FFPE
specimens
using
elevated
temperature
concentrated
detergents.
HIF-Clear
with
multi-round
immunolabeling
reveals
neuron
circuitry
regulating
multiple
neurotransmitter
systems
in
whole
mouse
brain,
is
able
be
used
as
the
evaluation
of
disease
treatment
efficiency.
also
supports
expansion
microscopy
can
performed
on
non-sectioned
15-year-old
specimen,
well
3-month
brain.
Thus,
represents
feasible
approach
researching
archived
future
neuroscientific
3D
neuropathological
analyses.
Nature Communications,
Journal Year:
2024,
Volume and Issue:
15(1)
Published: Dec. 30, 2024
Abstract
Biological
systems
are
complex,
encompassing
intertwined
spatial,
molecular
and
functional
features.
However,
methodological
constraints
limit
the
completeness
of
information
that
can
be
extracted.
Here,
we
report
development
INSIHGT,
a
non-destructive,
accessible
three-dimensional
(3D)
spatial
biology
method
utilizing
superchaotropes
host-guest
chemistry
to
achieve
homogeneous,
deep
penetration
macromolecular
probes
up
centimeter
scales,
providing
reliable
semi-quantitative
signals
throughout
tissue
volume.
Diverse
antigens,
mRNAs,
neurotransmitters,
post-translational
modifications
well-preserved
simultaneously
visualized.
INSIHGT
also
allows
multi-round,
highly
multiplexed
3D
probing
is
compatible
with
downstream
traditional
histology
nucleic
acid
sequencing.
With
map
undescribed
podocyte-to-parietal
epithelial
cell
microfilaments
in
mouse
glomeruli
neurofilament-intensive
inclusion
bodies
human
cerebellum,
identify
NPY-proximal
types
defined
by
morpho-proteomics
hypothalamus.
We
anticipate
form
foundations
for
multi-omics
technology
holistic
studies.
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2022,
Volume and Issue:
unknown
Published: Dec. 2, 2022
Abstract
Tissue
clearing
and
labeling
techniques
have
revolutionized
brain-wide
imaging
analysis,
yet
their
application
to
clinical
formalin-fixed
paraffin-embedded
(FFPE)
blocks
remains
challenging.
We
introduce
HIF-Clear,
a
novel
method
for
efficiently
centimeter-thick
FFPE
specimens
using
elevated
temperature
concentrated
detergents.
HIF-Clear
with
multi-round
immunolabeling
reveals
neuron
circuitry
regulating
multiple
neurotransmitter
systems
in
whole
mouse
brain,
is
able
be
used
as
the
evaluation
of
disease
treatment
efficiency.
also
supports
expansion
microscopy
can
performed
on
non-sectioned
15-year-old
specimen,
well
3-month
brain.
Thus,
represents
feasible
approach
researching
archived
future
neuroscientific
3D
neuropathological
analyses.
