Biology Methods and Protocols,
Journal Year:
2024,
Volume and Issue:
9(1)
Published: Jan. 1, 2024
Abstract
Microorganisms
are
widely
used
for
the
industrial
production
of
various
valuable
products,
such
as
pharmaceuticals,
food
and
beverages,
biofuels,
enzymes,
amino
acids,
vaccines,
etc.
Research
is
constantly
carried
out
to
improve
their
properties,
mainly
increase
productivity
efficiency
reduce
cost
processes.
The
selection
microorganisms
with
improved
qualities
takes
a
lot
time
resources
(both
human
material);
therefore,
this
process
itself
needs
optimization.
In
last
two
decades,
microfluidics
technology
appeared
in
bioengineering,
which
allows
manipulating
small
particles
(from
tens
microns
nanometre
scale)
flow
liquid
microchannels.
based
on
small-volume
objects
(microdroplets
from
nano
femtolitres),
manipulated
using
microchip.
chip
made
an
optically
transparent
inert
medium
material
contains
series
channels
size
(<1
mm)
certain
geometry.
Based
physical
chemical
properties
microparticles
(like
size,
weight,
optical
density,
dielectric
constant,
etc.),
they
separated
microsensors.
idea
accelerated
application
microfluidic
technologies
separate
mutants
after
mutagenesis.
This
article
discusses
possible
practical
implementation
separation
mutants,
including
yeasts
like
Yarrowia
lipolytica
Phaffia
rhodozyma
mutagenesis
will
be
discussed.
Small Science,
Journal Year:
2024,
Volume and Issue:
4(5)
Published: March 25, 2024
The
encapsulation
of
single
cells
has
emerged
as
a
promising
field
in
recent
years,
owing
to
its
potential
applications
cell‐based
therapeutics,
bioprinting,
vitro
cell
culture,
high‐throughput
screening,
and
diagnostics.
Single‐cell
units
offer
several
advantages,
including
compatibility
with
standard
imaging
techniques,
superior
diffusion
rates,
lower
material‐to‐cell
volume
ratios.
They
also
serve
effective
carriers
for
targeted
drug
delivery,
allowing
precise
administration
therapeutics
cell‐mediated
quantities.
Moreover,
single‐cell
exhibit
improved
circulation
throughout
the
vasculature,
reduced
likelihood
entrapment
compared
multicell
strategies.
However,
production
from
random
dispersion
follows
Poisson
distribution,
requiring
separation
empty
ones.
Various
methods
have
been
developed
address
this
challenge;
nevertheless,
majority
these
strategies
are
either
expensive
or
time‐consuming.
This
review
provides
an
in‐depth
analysis
advantages
limitations
their
applications,
well
comprehensive
overview
most
used
techniques
sorting
Microbial Cell Factories,
Journal Year:
2024,
Volume and Issue:
23(1)
Published: April 9, 2024
Abstract
Background
Single-cell
droplet
microfluidics
is
an
important
platform
for
high-throughput
analyses
and
screening
because
it
provides
independent
compartmentalized
microenvironment
reaction
or
cultivation
by
coencapsulating
individual
cells
with
various
molecules
in
monodisperse
microdroplets.
In
combination
microbial
biosensors,
this
technology
becomes
a
potent
tool
the
of
mutant
strains.
study,
we
demonstrated
that
genetically
engineered
yeast
strain
can
fluorescently
sense
agonist
ligands
via
heterologous
expression
human
G-protein-coupled
receptor
(GPCR)
concurrently
secrete
candidate
peptides
highly
compatible
single-cell
microfluidic
new
agonistically
active
peptides.
Results
The
water-in-oil
microdroplets
were
generated
using
flow-focusing
chip
to
encapsulate
coexpressing
GPCR
[i.e.,
angiotensin
II
type
1
(AGTR1)]
secretory
agonistic
peptide
(Ang
II)].
single
cultured
droplets
then
observed
under
microscope
analyzed
image
processing
incorporating
machine
learning
techniques.
AGTR1-mediated
signal
transduction
elicited
self-secreted
Ang
was
successfully
detected
fluorescent
reporter
cultures.
system
could
also
distinguish
analog
different
activities.
Notably,
further
culture
enabled
detection
rarely
existing
positive
II-secreting)
model
mixed
cell
library,
whereas
conventional
batch-culture
environment
shake
flask
failed
do
so.
Thus,
our
approach
provided
microculture
environments,
which
prevent
diffusion,
dilution,
cross-contamination
secreted
from
easy
identification
agonists.
Conclusions
We
established
droplet-based
integrated
biosensor
expressed
This
offers
individually
isolated
microenvironments
allow
secreting
these
gaging
their
signaling
activities,
Our
base
on
biosensors
will
be
widely
applicable
metabolic
engineering,
environmental
drug
discovery.
Journal of Pharmaceutical Analysis,
Journal Year:
2023,
Volume and Issue:
13(10), P. 1102 - 1116
Published: Aug. 23, 2023
Cellular
heterogeneity
is
crucial
for
understanding
tissue
biology
and
disease
pathophysiology.
Pharmacological
research
being
advanced
by
single-cell
metabolic
analysis,
which
offers
a
technique
to
identify
variations
in
RNA,
proteins,
metabolites,
drug
molecules
cells.
In
this
review,
the
recent
advancement
of
analysis
techniques
their
applications
metabolism
response
are
summarized.
High-precision
controlled
isolation
manipulation
provided
microfluidics-based
methods,
such
as
droplet
microfluidics,
microchamber,
open
microfluidic
probe,
digital
microfluidics.
They
used
tandem
with
variety
detection
techniques,
including
optical
imaging,
Raman
spectroscopy,
electrochemical
detection,
RNA
sequencing,
mass
spectrometry,
evaluate
changes
administration.
The
advantages
disadvantages
different
discussed
along
challenges
future
directions
analysis.
These
employed
pharmaceutical
studying
resistance
pathway,
therapeutic
targets
discovery,
vitro
model
evaluation.
Advanced Healthcare Materials,
Journal Year:
2025,
Volume and Issue:
unknown
Published: Feb. 24, 2025
Abstract
More
recently,
single‐cell
encapsulation
emerged
as
a
promising
field
in
biomedicine
due
to
its
potential
applications,
cell
analysis
and
therapy.
Traditional
techniques
involve
embedding
cells
crosslinked
polymers
create
continuous
microgels,
suitable
mainly
for
adherent
cells,
or
encapsulating
them
droplets
only
short‐term
analysis,
their
instability.
In
this
study,
we
developed
method
single
liquid‐core
microcapsules
address
these
limitations.
The
liquid
system
is
generated
an
all
aqueous
environment
through
polymeric
electrostatic
interactions.
Additionally,
design
innovative
low
cost
sorting
utilizing
magnetic
nanoparticles
(MNPs)
efficiently
select
encapsulated
units
further
applications.
This
tested
with
both
suspension
types,
demonstrating
cytocompatibility
no
abnormal
effects
on
behavior.
MNP‐based
achieved
nearly
80%
purity
of
the
population.
Overall,
technology
provides
highly
efficient
such
screening,
by
enabling
precise
short
medium‐term
real
time
monitoring,
high
resolution
imaging
cellular
Furthermore,
semipermeable
membrane
unlocks
new
advancing
therapy
offering
protection
while
ensuring
diffusion
therapeutic
factors,
paving
way
strategies.
Small,
Journal Year:
2025,
Volume and Issue:
unknown
Published: May 8, 2025
Abstract
Single‐cell
encapsulation
in
droplet
microfluidics
has
become
a
powerful
tool
precision
medicine,
single‐cell
analysis,
and
immunotherapy.
However,
generation
with
is
random
process,
which
also
results
large
number
of
empty
multi‐cell
droplets.
Current
sorting
technologies
suffer
from
drawbacks
such
as
fluorescent
labeling,
inability
to
remove
droplets,
or
low
throughput.
This
paper
presents
gray‐level
guided
image‐activated
sorter
(GL‐IADS),
enables
label‐free,
high‐accuracy
screening
droplets
by
rejecting
The
based
recognition
method
can
accurately
classify
images
(empty,
single‐cell,
droplets),
especially
differentiating
cell‐laden
(accuracy
100%).
Crucially,
this
reduces
the
image
processing
time
≈300
µs,
makes
GL‐IADS
possible
reach
an
ultra‐high
throughput
up
hundreds
even
KHz.
integrates
novel
detachable
acoustofluidic
system,
achieving
purity
97.9%,
97.4%,
>99%
for
multi‐cell,
respectively,
43
Hz.
holds
promise
numerous
biological
applications
that
are
previously
difficult
fluorescence‐based
technologies.
Communications Engineering,
Journal Year:
2025,
Volume and Issue:
4(1)
Published: May 13, 2025
Rare
particle
enrichment
plays
a
pivotal
role
in
advancing
numerous
scientific
research
areas
and
industrial
processes.
Traditional
methods
encounter
obstacles
such
as
low
efficiency,
high
cost,
complexity.
Acoustic
focusing,
optical
fiber
detection,
electrical
manipulation
have
shown
potential
microfluidics
for
analysis.
This
study
pioneers
the
integration
of
acoustic,
optical,
units
to
overcome
traditional
limitations.
The
cooperative
dynamics
acoustic
flow
focusing
are
explored.
fibers
with
an
enhanced
detection
algorithm
greatly
boost
sensitivity.
Furthermore,
droplet
charging
enhance
tip
phenomenon
is
complemented
validated.
sorting
accuracy
enriching
large-size
H22
cells
reached
99.8%
99.3%,
respectively,
target
cell
concentration
increased
by
nearly
86-fold.
Our
work
significantly
enhances
sensitivity
accuracy,
ultimately
offering
robust
reliable
solution
generating
droplets
enrich
rare
particles.