Nature Communications,
Год журнала:
2023,
Номер
14(1)
Опубликована: Фев. 25, 2023
Abstract
Endosomal
escape
and
subsequent
cytosolic
delivery
of
small
interfering
RNA
(siRNA)
therapeutics
is
believed
to
be
highly
inefficient.
Since
it
has
not
been
possible
quantify
amounts
delivered
siRNA
at
therapeutic
doses,
determining
bottlenecks
total
efficiency
difficult.
Here,
we
present
a
confocal
microscopy-based
method
fluorescently
labeled
during
lipid-mediated
delivery.
This
enables
detection
quantification
sub-nanomolar
release
from
individual
events
with
measures
quantitation
confidence
for
each
event.
Single-cell
kinetics
siRNA-mediated
knockdown
in
cells
expressing
destabilized
eGFP
unveiled
dose-response
relationship
respect
induction,
depth
duration
the
range
several
hundred
thousands
molecules.
Accurate
siRNA,
establishment
intracellular
relationships,
will
aid
development
characterization
novel
strategies
nucleic
acid
therapeutics.
We
present
an
oblique
plane
microscope
(OPM)
that
uses
a
bespoke
glass-tipped
tertiary
objective
to
improve
the
resolution,
field
of
view,
and
usability
over
previous
variants.
Owing
its
high
numerical
aperture
optics,
this
achieves
lateral
axial
resolutions
are
comparable
square
illumination
mode
lattice
light-sheet
microscopy,
but
in
user
friendly
versatile
format.
Given
performance,
we
demonstrate
high-resolution
imaging
clathrin-mediated
endocytosis,
vimentin,
endoplasmic
reticulum,
membrane
dynamics,
Natural
Killer-mediated
cytotoxicity.
Furthermore,
image
biological
phenomena
would
be
otherwise
challenging
or
impossible
perform
traditional
geometry,
including
cell
migration
through
confined
spaces
within
microfluidic
device,
subcellular
photoactivation
Rac1,
diffusion
cytoplasmic
rheological
tracers
at
volumetric
rate
14
Hz,
large
view
neurons,
developing
embryos,
centimeter-scale
tissue
sections.
Nature Methods,
Год журнала:
2022,
Номер
19(4), С. 461 - 469
Опубликована: Март 21, 2022
The
promise
of
single-objective
light-sheet
microscopy
is
to
combine
the
convenience
standard
microscopes
with
speed,
coverage,
resolution
and
gentleness
microscopes.
We
present
DaXi,
a
microscope
design
based
on
oblique
plane
illumination
that
achieves:
(1)
wider
field
view
high-resolution
imaging
via
custom
remote
focusing
objective;
(2)
fast
volumetric
over
larger
volumes
without
compromising
image
quality
or
necessitating
tiled
acquisition;
(3)
fuller
coverage
for
large
samples
multi-view
(4)
higher
throughput
multi-well
coverslip
placement.
Our
instrument
achieves
450
nm
laterally
2
μm
axially
an
volume
3,000
×
800
300
μm.
demonstrate
view,
versatility
our
by
various
systems,
including
Drosophila
egg
chamber
development,
zebrafish
whole-brain
activity
embryonic
development
-
up
nine
embryos
at
time.
Neuron,
Год журнала:
2022,
Номер
110(17), С. 2815 - 2835.e13
Опубликована: Июль 8, 2022
Dynamin
mediates
fission
of
vesicles
from
the
plasma
membrane
during
endocytosis.
Typically,
dynamin
is
recruited
cytosol
to
endocytic
sites,
requiring
seconds
tens
seconds.
However,
ultrafast
endocytosis
in
neurons
internalizes
as
quickly
50
ms
synaptic
vesicle
recycling.
Here,
we
demonstrate
that
1
pre-recruited
sites
for
Specifically,
1xA,
a
splice
variant
1,
interacts
with
Syndapin
form
molecular
condensates
on
membrane.
Single-particle
tracking
1xA
molecules
confirms
liquid-like
property
vivo.
When
mutated
disrupt
its
interaction
do
not
form,
and
consequently,
slows
down
by
100-fold.
Mechanistically,
acts
an
adaptor
binding
stores
at
sites.
This
cache
bypasses
recruitment
step
accelerates
synapses.
Communications Biology,
Год журнала:
2023,
Номер
6(1)
Опубликована: Май 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.
