Deep Tissue Second–Harmonics of Collagen Fibers in a Transparent Rat Heart from a Myocardial Infarction Model
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2025,
Volume and Issue:
unknown
Published: Jan. 12, 2025
Abstract
The
combination
of
tissue-clearing
techniques
with
light-sheet
microscopy
has
enabled
detailed
visualization
histological
changes
from
the
micrometer
to
millimeter
scale,
deepening
understanding
various
disease
processes.
However,
these
protocols
are
not
fully
optimized
for
animal
species
beyond
mice
or
organs
outside
brain.
Additionally,
lack
suitable
fluorescent
probes
target
molecules
limits
their
broader
application.
In
this
study,
we
present
a
protocol
whole-organ
clearing
rat
hearts
in
myocardial
infarction
model,
achieving
complete
transparency
and
enabling
label-free
imaging
collagen
fibers
wall
up
depth
∼5
mm
using
Second
Harmonic
Generation
(SHG)
microscopy.
For
first
time,
successfully
compared
fiber
orientations
between
infarcted
healthy
regions.
Our
approach
facilitates
high-resolution
tissue
remodeling
analysis
cardiovascular
research
without
need
antibody
staining,
demonstrating
that
feasible
even
limited
available
antibodies.
Summary
statement
We
novel
method
visualizing
deep
within
heart
advanced
microscopy,
providing
valuable
data
cardiac
research.
Language: Английский
Open-source antibodies as a path to enhanced research reproducibility and transparency
New Biotechnology,
Journal Year:
2025,
Volume and Issue:
unknown
Published: April 1, 2025
Antibodies
are
important
tools
with
diverse
uses
in
biomedical
research.
However,
open
access
to
reliable
sources
of
well-characterized
antibodies
unambiguous
molecular
identities
remains
an
obstacle
research
transparency
and
reproducibility.
We
propose
here
a
community
shift
towards
open-source
antibodies,
analogous
computer
software.
The
tenets
such
that
1)
they
available
researchers
ready
use
form,
2)
the
renewable
source
antibody
(e.g.,
hybridoma
cells
or
plasmid)
is
also
widely
ensuring
reproducible
cost-effective
same
antibody,
3)
sequence
publicly
available.
With
these
criteria
met,
can
be
used
transparent
assurance
associated
molecularly
defined
reagent,
code
edited
generate
variants
meet
researchers'
specific
needs.
(the
UC
Davis/NIH
NeuroMab
Facility,
Development
Studies
Hybridoma
Bank,
Addgene)
have
established
consortium
provide
large
collection
well
characterized
antibodies.
As
software
has
benefitted
both
users
developers,
we
suggest
will
similar
positive
impact
on
based
encourage
funding
agencies
support
initiatives
expand
resources,
utilize
contribute
them,
goal
enabling
more
pursuit
Language: Английский
Genetically encoded intrabody probes for labeling and manipulating AMPA-type glutamate receptors
Nature Communications,
Journal Year:
2024,
Volume and Issue:
15(1)
Published: Nov. 29, 2024
Tools
for
visualizing
and
manipulating
protein
dynamics
in
living
cells
are
critical
understanding
cellular
function.
Here
we
leverage
recently
available
monoclonal
antibody
sequences
to
generate
a
set
of
affinity
tags
labeling
AMPA-type
glutamate
receptors
(AMPARs),
which
mediate
nearly
all
excitatory
neurotransmission
the
central
nervous
system.
These
antibodies
can
be
produced
from
heterologous
exogenous
applications
or
directly
expressed
neurons
as
intrabodies,
where
they
bind
their
epitopes
endoplasmic
reticulum
co-traffic
cell
surface
visualization
with
impermeant
fluorescent
dyes.
We
show
these
reagents
do
not
perturb
AMPAR
trafficking,
function,
mobility,
synaptic
recruitment
during
plasticity
therefore
used
probes
monitoring
endogenous
neurons.
also
adapt
deplete
AMPARs
by
trapping
them
reticulum,
providing
simple
approach
loss
neurotransmission.
The
strategies
outlined
here
serve
template
generating
similar
targeting
diverse
proteins
more
become
available.
imaging
native
environments
understand
Here,
authors
develop
antibody-based
visualize
manipulate
Language: Английский
Semi-automated navigation for efficient targeting of electron tomography to regions of interest in volume correlative light and electron microscopy
bioRxiv (Cold Spring Harbor Laboratory),
Journal Year:
2024,
Volume and Issue:
unknown
Published: Nov. 30, 2024
Electron
microscopy
is
essential
for
the
quantitative
study
of
synaptic
ultrastructure.
At
present,
correlation
functional
and
structural
properties
same
synapse
extremely
challenging.
We
introduce
a
novel
integrated
workflow
designed
to
simplify
sample
navigation
across
spatial
scales,
allowing
identification
individual
synapses
from
optical
mouse
brain
image
stacks
that
can
be
targeted
analysis
using
electron
tomography
imaging.
developed
software
which
has
function
register
multimodal
images
segmentation-based
registration
algorithm
as
well
visualize
all
results.
Using
our
newly
we
streamline
mapping
high-resolution
imaging
onto
reference
maps
blood
vessels
endogenous
fiducial
marks.
Further
demonstrate
significant
improvements
on
ultramicrotomy
stage
volume
Correlative
Light
Microscopy
(vCLEM)
workflows,
providing
real
time
guidance
trimming
match
previously
acquired
Regions
Of
Interest
(ROIs),
reliable
estimates
cutting
depth
relative
ROI,
based
fluorescence
TEM
ready
ultrathin
sections.
this
workflow,
successfully
proximal
axonal
region
containing
Axon
Initial
Segment
identified
fluorescent
light
microscopy.
Language: Английский