Fluorescence
Resonance
Energy
Transfer
(FRET)-based
approaches
are
unique
tools
for
sensing
the
immediate
surroundings
and
interactions
of
(bio)molecules.
FRET
imaging
Lifetime
Imaging
Microscopy
(FLIM)
enable
visualization
spatial
distribution
molecular
functional
states.
However,
conventional
FLIM
provide
average
information
over
an
ensemble
molecules
within
a
diffraction-limited
volume,
which
limits
information,
accuracy,
dynamic
range
observed
signals.
Here,
approach
to
obtain
super-resolved
based
on
single-molecule
localization
microscopy
using
early
prototype
commercial
time-resolved
confocal
microscope
is
demonstrated.
DNA
Points
Accumulation
in
Nanoscale
Topography
with
fluorogenic
probes
provides
suitable
combination
background
reduction
binding
kinetics
compatible
scanning
speed
usual
microscopes.
A
single
laser
used
excite
donor,
broad
detection
band
employed
retrieve
both
donor
acceptor
emission,
events
detected
from
lifetime
information.
Light Science & Applications,
Год журнала:
2024,
Номер
13(1)
Опубликована: Июнь 7, 2024
MINFLUX
has
achieved
extraordinary
resolution
in
superresolution
imaging
and
single
fluorophore
tracking.
It
is
based
on
localizing
fluorophores
by
rapid
probing
with
a
patterned
beam
that
features
local
intensity
minimum.
Current
implementations,
however,
are
complex
expensive
limited
speed
robustness.
Here,
we
show
combination
of
an
electro-optical
modulator
segmented
birefringent
element
such
as
spatial
light
produces
variable
phase
plate
for
which
the
can
be
scanned
MHz
timescale.
Bisected
or
top-hat
patterns
generate
high-contrast
compact
excitation
point-spread
functions
localization
x,
y,
z-direction,
respectively,
across
within
microsecond,
switched
60
microseconds
alternated
among
different
wavelengths.
We
discuss
how
to
compensate
non-optimal
performance
components
present
robust
3D
multi-color
module,
envision
integral
component
high-performance
cost-effective
open-source
MINFLUX.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2020,
Номер
unknown
Опубликована: Дек. 29, 2020
Abstract
With
rapid
developments
in
microscopy
methods,
highly
versatile,
robust
and
affordable
implementations
are
needed
to
enable
wide
adoption
by
the
biological
sciences
community.
Here
we
report
Squid,
a
quantitative
imaging
platform
with
full
suite
of
hardware
software
components
configurations
for
deploying
facility-grade
widefield
microscopes
advanced
features
like
flat
field
fluorescence
excitation,
patterned
illumination
tracking
microscopy,
at
fraction
cost
commercial
solutions.
The
open
modular
nature
(both
software)
lowers
barrier
deployment,
importantly,
simplifies
development,
making
system
configurable
experiments
that
can
run
on
easily
programmable.
Developed
goal
helping
translate
advances
microscopy-enabled
including
those
powered
deep
learning,
envision
Squid
will
simplify
roll-out
microscopy-based
applications
-
point
care
low
resource
settings,
make
new
or
otherwise
techniques
easier,
significantly
increase
available
microscope-hours
labs.
Nanoscale,
Год журнала:
2021,
Номер
13(44), С. 18421 - 18433
Опубликована: Янв. 1, 2021
Recent
advances
combining
FRET
and
super-resolution
techniques
are
presented.
This
merger
of
methods
is
opening
exciting
opportunities
to
study
the
spatial
organization
functional
state
(bio)molecules
materials
at
nanoscale.
Chemical Society Reviews,
Год журнала:
2022,
Номер
51(24), С. 9882 - 9916
Опубликована: Янв. 1, 2022
Following
an
overview
of
the
approaches
and
techniques
used
to
acheive
super-resolution
microscopy,
this
review
presents
advantages
supplied
by
nanoparticle
based
probes
for
these
applications.
Fluorescence
Resonance
Energy
Transfer
(FRET)-based
approaches
are
unique
tools
for
sensing
the
immediate
surroundings
and
interactions
of
(bio)molecules.
FRET
imaging
Lifetime
Imaging
Microscopy
(FLIM)
enable
visualization
spatial
distribution
molecular
functional
states.
However,
conventional
FLIM
provide
average
information
over
an
ensemble
molecules
within
a
diffraction-limited
volume,
which
limits
information,
accuracy,
dynamic
range
observed
signals.
Here,
approach
to
obtain
super-resolved
based
on
single-molecule
localization
microscopy
using
early
prototype
commercial
time-resolved
confocal
microscope
is
demonstrated.
DNA
Points
Accumulation
in
Nanoscale
Topography
with
fluorogenic
probes
provides
suitable
combination
background
reduction
binding
kinetics
compatible
scanning
speed
usual
microscopes.
A
single
laser
used
excite
donor,
broad
detection
band
employed
retrieve
both
donor
acceptor
emission,
events
detected
from
lifetime
information.
