Resolving the Orientations of and Angular Separation Between a Pair of Dipole Emitters
Physical Review Letters,
Journal Year:
2025,
Volume and Issue:
134(9)
Published: March 7, 2025
We
prove
that
it
is
impossible
to
distinguish
two
spatially
coinciding
fluorescent
molecules
from
a
single
rotating
molecule
using
polarization-sensitive
imaging,
even
if
one
modulates
the
polarization
of
illumination
or
detection
dipole-spread
function
(DSF).
If
target
known
be
dipole
pair,
existing
imaging
methods
perform
poorly
for
measuring
their
angular
separation.
propose
simultaneously
modulating
excitation
and
DSF,
which
demonstrates
robust
discrimination
between
pairs
versus
molecules.
Our
method
improves
precision
centroid
orientation
by
50%
separation
two-
four-fold
over
techniques.
Language: Английский
Single-molecule orientation-localization microscopy: Applications and approaches
Quarterly Reviews of Biophysics,
Journal Year:
2024,
Volume and Issue:
57
Published: Jan. 1, 2024
Abstract
Single-molecule
orientation-localization
microscopy
(SMOLM)
builds
upon
super-resolved
localization
by
imaging
orientations
and
rotational
dynamics
of
individual
molecules
in
addition
to
their
positions.
This
added
dimensionality
provides
unparalleled
insights
into
nanoscale
biophysical
biochemical
processes,
including
the
organization
actin
networks,
movement
molecular
motors,
conformations
DNA
strands,
growth
remodeling
amyloid
aggregates,
composition
changes
within
lipid
membranes.
In
this
review,
we
discuss
recent
innovations
SMOLM
cover
three
key
aspects:
(1)
enabled
labeling
strategies
that
endow
fluorescent
probes
bind
targets
with
orientation
specificity;
(2)
advanced
techniques
leverage
physics
light-matter
interactions
estimation
theory
encode
information
high
fidelity
microscope
images;
(3)
computational
methods
ensure
accurate
precise
data
analysis
interpretation,
even
presence
severe
shot
noise.
Additionally,
compare
approaches,
hardware,
publicly
available
software
aid
community
choosing
best
implementation
for
specific
application.
Finally,
highlight
future
directions
SMOLM,
such
as
development
improved
photostability
specificity,
design
“smart”
adaptive
use
approaches
handle
large,
complex
datasets.
review
underscores
significant
current
potential
impact
deepening
our
understanding
dynamics,
paving
way
breakthroughs
fields
biophysics,
biochemistry,
materials
science.
Language: Английский
Stokes-based analysis for the estimation of 3D dipolar emission
Journal of the Optical Society of America A,
Journal Year:
2024,
Volume and Issue:
41(11), P. 2134 - 2134
Published: Sept. 25, 2024
We
provide
a
general
description
of
the
measurement
capabilities
systems
that
probe
3D
state
polarization
light
emitted
by
dipole
or
collection
dipoles.
This
analysis
is
based
on
generalization
Stokes
parameters
for
polarization,
and
its
goal
to
insight
into
what
constitutes
good
system
under
specific
circumstances,
through
definition
appropriate
merit
functions.
Three
cases
are
considered:
case
arbitrary
states
special
linear
full
partial
states,
even
more
dipoles
wobble
with
rotational
symmetry
around
central
direction.
Note
latter
two
interest
in
fluorescence
microscopy.
The
presented
here
illustrated
applying
it
different
approaches
used
commonly
orientation
microscopy:
PSF
engineering
ratiometric
measurements.
Language: Английский
Stokes-based analysis for the estimation of 3D dipolar emission
Published: Aug. 7, 2024
We
provide
a
general
description
of
the
measurement
capabilities
systems
that
probe
3D
state
polarization
light
emitted
by
dipole
or
collection
dipoles.
This
analysis
is
based
on
generalization
Stokes
parameters
for
polarization,
and
its
goal
to
insight
into
what
constitutes
good
system
under
specific
circumstances,
through
definition
appropriate
merit
functions.
Three
cases
are
considered:
case
arbitrary
states
special
linear
full
partial
states,
even
more
dipoles
wobble
with
rotational
symmetry
around
central
direction.
Note
latter
two
interest
in
fluorescence
microscopy.
The
presented
here
illustrated
applying
it
different
approaches
used
commonly
orientation
microscopy:
PSF
engineering
radiometric
measurements.
Language: Английский
Stokes-based analysis for the estimation of 3D dipolar emission
Published: Aug. 7, 2024
We
provide
a
general
description
of
the
measurement
capabilities
systems
that
probe
3D
state
polarization
light
emitted
by
dipole
or
collection
dipoles.
This
analysis
is
based
on
generalization
Stokes
parameters
for
polarization,
and
its
goal
to
insight
into
what
constitutes
good
system
under
specific
circumstances,
through
definition
appropriate
merit
functions.
Three
cases
are
considered:
case
arbitrary
states
special
linear
full
partial
states,
even
more
dipoles
wobble
with
rotational
symmetry
around
central
direction.
Note
latter
two
interest
in
fluorescence
microscopy.
The
presented
here
illustrated
applying
it
different
approaches
used
commonly
orientation
microscopy:
PSF
engineering
radiometric
measurements.
Language: Английский
Fluorescence Interference-Based Polarized Structured Illumination Microscopy for High Axial Accuracy Morphology Imaging of Dipole Orientations
ACS Photonics,
Journal Year:
2024,
Volume and Issue:
unknown
Published: Dec. 29, 2024
The
dipole
orientation
of
fluorescent
molecules
reveals
the
structural
organization
at
subcellular
level,
and
imaging
its
distribution
is
important
for
fundamental
cell
biology
studies.
Using
conventional
fluorescence
microscopy,
dipoles
can
be
obtained
by
demodulating
polarization
states
through
excitation
or
emission
processes.
recent
introduction
structured
illumination
microscopy
(SIM)
has
improved
axial
lateral
resolution
to
around
300
100
nm,
respectively.
However,
still
very
limited,
which
hinders
capabilities
offer
more
precise
measurements
information.
Here,
we
report
Fluorescence
Interference
based
polarized
(FI-pSIM),
provides
a
sub-30
nm
reconstruction
accuracy
maintains
same
comparable
SIM
only
9
raw
images.
By
synergizing
interference
via
4Pi
configuration
demodulation
illumination,
FI-pSIM
enables
three-dimensional
morphological
ensemble
orientations
in
structures
with
∼
reconstruction.
high
efficacy
four-dimensional
(4D)
modality
are
demonstrated
mapping
biological
microfilaments.
elucidating
orientations,
envision
may
open
up
new
avenues
demystifying
4D
organizations
structures.
Language: Английский