Download Neuronal Cell Health Assays

I N C U C Y T E ® L I V E - C E L L A N A LY S I S S Y S T E M
Neuronal Cell Health Assays
Real-time automated measurements of neuronal cell viability
and neurite dynamics inside your incubator.
I N C U C Y T E ® A S S AY S F O R N E U R O N A L C E L L H E A LT H
Know the whole story – Real-time neuronal cell viability
and neurite outgrowth analysis inside your incubator
IncuCyte® Neuronal Cell Health Assays enable automated, real-time multiplexed analysis of neurite
dynamics and cell viability. Combine label-free readouts with non-perturbing live-cell reagents to
simultaneously distinguish treatments that affect one or more aspects of neuronal cell health.
Get the answers you need
Save time and effort
Protect your cells
• Never miss a data point with
real-time continuous analysis.
• Set up, walk away and receive
unbiased, automated analysis.
• Robust, reproducible assay
formats suitable for screening
and profiling.
• Run experiments in six 96 or
384-well plates at one time.
Ask more questions
• Analyze stationary cultures
within the stable environment
of an incubator - ideal for
studies over days or weeks.
• Maintain optimal culture
conditions with label-free
analysis options and nonperturbing reagent formulations.
• Compatible with a wide
range of neurons (primary,
stem cell derived, cell lines) in
monoculture or in co-culture
with astrocytes.
• Multiplex neurite outgrowth
analysis with cell health
reagents for additional insight.
Confidently assess and measure neurite outgrowth and cell viability
• Rank compounds in drug discovery screens.
• Investigate the cellular changes that underlie neurodegeneration and neuronal injury.
• Assess factors affecting neural stem cell differentiation.
Measurements of neurite outgrowth and cell viability are
central to the study of drug candidates, culture conditions
and the impact of environmental factors on neuronal cell
health and function. Both neurite outgrowth and viability
assays are routinely used in drug discovery screens and may
be used to identify treatments that elicit neuroprotective
effects but do not induce neurotoxicity.
Neurite analysis
Cytotoxicity assays
Apoptosis assays
Measure neurite dynamics
automatically and in real-time in
neuronal monocultures and cocultures with astrocytes.
Real-time measurements of cell
viability using simple mix-and-read
protocols suitable for screening.
Detect apoptosis in living cells
and in real time using simple
mix-and-read protocols.
Learn more at www.essenbioscience.com/NeuroCellHealth
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I N C U C Y T E ® A S S AY S F O R N E U R O N A L C E L L H E A LT H
Real-time, long-term neurite analysis
• Visualize and analyze long-term changes
in neurite length and branch points with
the IncuCyte® live cell analysis system
and NeuroTrack™ software.
Day 0
Day 3
Day 5
• Study label-free monocultures, or use
IncuCyte® NeuroLight™ reagents or
NeuroPrime™ cell kits for selective
analysis of neurite dynamics in co-cultures.
Neurite length: 11.2 mm/mm²
Cell body cluster: 134/mm²
Branch points: 259.9/mm²
IncuCyte® NeuroTrack™ Software
Neurite length: 55.1 mm/mm²
Cell body cluster: 130/mm²
Branch points: 1368/mm²
Neurite length: 75.1 mm/mm²
Cell body cluster: 139/mm²
Branch points: 2048/mm²
Label-free analysis of neurons in mono-culture
Perform label-free analysis of neurons in mono-culture or use IncuCyte® Neurolight™ reagents
or NeuroPrime™ cell kits to study co-cultures. IncuCyte® NeuroTrack™ Software automatically
analyzes images to provide measurement of neurite length, branch points and cell body clusters.
Neuron specific live-cell labeling reagents and cell kits –
analyze neurite dynamics in co-cultures
• NeuroLight™ Red lentivirus reagent for efficient labeling of neurons without perturbing the underlying biology.
• NeuroPrime™ Cell Kit for monitoring neurite dynamics in co-cultures of primary rat neurons and astrocytes.
Fluorescence analysis of neurons in co-culture with astrocytes
Day 3
IncuCyte® NeuroPrime™ Cell Kit
Day 10
NeuroLight™ Red lentivirus reagent can be used to transduce your choice of
neuronal cell types including cell lines, primary and stem cell-derived cultures. The
NeuroLight™ Red lentivirus encodes a red fluorescent protein driven off a synapsin
promoter to strengthen neuronal expression and minimize non-neuronal crossover.
NeuroPrime Cell Kit: Cryopreserved vials of rat
forebrain neurons and astrocytes and one vial of
NeuroLight Red Lentivirus reagent. The kit includes
a validated protocol for optimal neuron labeling and
assay performance. Ideal for studies of neurotoxicity or
the biology of established neural networks.
Learn more at www.essenbioscience.com/NeuroCellHealth
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I N C U C Y T E ® A S S AY S F O R N E U R O N A L C E L L H E A LT H
Visualize and quantify cell viability in real time
–multiplex with neurite analysis
• IncuCyte® mix-and-read reagents for continuous monitoring of cytotoxicity and apoptosis in neuronal cultures.
• Simultaneous measurements of neurite outgrowth and neuronal cell health.
Cytotox Red reagent
Annexin V Green reagent
Rat forebrain neurons 333 µM
glutamate, 3 days after treatment.
Rat forebrain neurons in culture with astrocytes. 300 µM glutamate (left) vs. control (right),
5 hours after treatment.
Use IncuCyte® Cytotox and Annexin V reagents to positively mark and quantify apoptotic or cytotoxic cells.
IncuCyte® Cytotox Red reagent labels
dead rat forebrain neurons treated with
333 µM glutamate.
IncuCyte® Annexin V Green reagent identifies apoptotic cells in a rat forebrain neuron
and astrocyte co-culture in the presence and absence of 300 µM glutamate. Neurons are
selectively labeled with the IncuCyte® NeuroLight Red reagent. Multiplexed measurements
of cell viability and neurite dynamics can be made in real time.
Automatically generate time courses and reveal concentrationdependent responses
• Automatically generate metrics for 96- or 384-well plates.
• Correlate neurite dynamics with measures of neuronal cell health.
Quantify treatment effects automatically
and non-invasively.
Every well of a 96- or 384-well plate is imaged and analyzed
automatically to provide a microplate readout of cytotoxicity
over time (right). Time-courses reveal concentration-dependent
treatment effects on neurite dynamics and neuronal cell health.
Transform data into concentration-response curves to compare
pharmacology (below).
1 40
16
N eu ri te Lengt h
A nn ex in V
I C 5 0 = 55 µ M
E C 5 0 = 59 µ M
16
1 00 0 µ M
1 20
1 00 µ M
4 µM
1 µM
8
4
0
0
12
2
N eu ri te Le ng th (mm /m m )
10 µM
5
Time (h )
10
15
Learn more at www.essenbioscience.com/NeuroCellHealth
1 00
8
80
4
60
40
A nn ex in V Rea gent
40 µM
G re en O bj ect C onfl ue nc e
A nn ex in V Rea gent
G re en O bj ect C onfl ue nc e
3 00 µ M
12
-6
-5
-4
-3
0
Log [ Glut amate] ( M)
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I N C U C Y T E ® A S S AY S F O R N E U R O N A L C E L L H E A LT H
Quick Guides
Neuronal mono-cultures – label free protocol overview
1
PLATE NEURONS
2
Coat wells and
seed neuronal cells.
ADD TREATMENTS
3
Treat cells with neurite
modulating test agents.
LIVE CELL ANALYSIS
Monitor every well in
the IncuCyte® ZOOM
live-cell analysis system.
Neuronal co-cultures – fluorescence protocol overview
1
PLATE NEURONS
AND TRANSDUCE
Coat wells and seed
neurons. Transduce neurons
with NeuroLight™ Red
Lentivirus Reagent.
2
PLATE ASTROCYTES & MONITOR
NEUROLIGHT EXPRESSION
Remove the reagent from the
neurons and seed astrocytes.
Monitor NeuroLight™
expression in the IncuCyte®
live-cell analysis system.
3
ADD TREATMENTS
When NeuroLight™
expression is maximal,
treat cells with neurite
modulating test agents.
4
LIVE CELL
FLUORESCENT ANALYSIS
Monitor every well in
the IncuCyte® ZOOM
live-cell analysis system.
Assays compatible with your choice of neuronal cultures
• The IncuCyte® system provides the flexibility to work with your choice of cells and cell models.
Primary neurons
Stem-cell derived neurons
Neuronal-like cells
Learn more at www.essenbioscience.com/NeuroCellHealth
Neuronal monocultures
or co-cultures
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I N C U C Y T E ® A S S AY S F O R N E U R O N A L C E L L H E A LT H
Easy and complete cell health workflow with the IncuCyte® system
Simple sample preparation
Set up & walk away
Measure long term
Analyze in real-time
96/384-well protocols—
no washing,
no fixing, no lifting.
Setup automated acquisition
and analysis parameters with
confidence—no need to
pre-define the assay endpoint.
Flexibility to measure
from hours to weeks.
Make decisions about
your experiment as the
story unfolds.
See what else you can do with the IncuCyte® System
Learn more at www.essenbioscience.com/applications
Apoptosis
Immune Cell Killing
Detect apoptosis in living cells and in real time using simple mixand-read protocols.
Detect tumor cell death directly by counting NucLight™ nuclearlabeled cells or measuring apoptosis with IncuCyte® Caspase 3/7
reagent.
Cytotoxicity
Chemotaxis
Measure real-time cell viability with simple mix-and-read protocols
suitable for screening.
Visually confirm the chemotactic migration of immune cells
towards chemoattractants with ClearView 96-well plates.
Proliferation
Scratch Wound Migration and Invasion
Automatically measure label-free growth or count living cells with
NucLight™ nuclear labeling in real time.
Investigate treatment effects on neuronal cell migration across a
2D substrate or invasion through a 3D gel matrix.
Immune Cell Clustering
Phagocytosis
Visualize and quantify expansion and clustering without removing
your cultures from the incubator.
Continuously analyze and generate movies of immune cells
engulfing pHrodo® labeled target tumor cells.
Ordering information
Contact us at [email protected] to place an order or for more information.
APPLICATION
PRODUCTS
Neurite analysis
Measure neurite dynamics in monocultures
and co-cultures with astrocytes.
Apoptosis
Detect and quantify apoptotic cells in real time.
Reagents
Software
Reagents
IncuCyte® NeuroLight™ Red
IncuCyte® NeuroPrime™ Cell Kit
Detect and count non-viable cells in real time.
Cell lines
Cryopreserved neuronal cells and astrocytes.
Reagents
Neuronal
Cells
CAT. NO.
0.45 mL
4584
1 Kit
4585
IncuCyte NeuroTrack Software
9600-0010
IncuCyte® Annexin V Red for Apoptosis
1 vial, 100 tests
4641
IncuCyte® Annexin V Green for Apoptosis
1 vial, 100 tests
4642
20 μL
4440
IncuCyte® Cytotox Red
5 μL x 5
4632
IncuCyte® Cytotox Green
5 μL x 5
4633
Neuro-2a NucLight™ Green Cells
2 x 10 cells/vial
4511
Neuro-2a NucLight™ Red Cells
2 x 10 cells/vial
4512
NeuroPrime™ rAstrocytes
2 x 10 cells/vial
4586
NeuroPrime™ rForebrain Neurons
2 x 10 cells/vial
4587
Caspase-3/7
Cytotoxicity
QUANTITY
6
6
6
6
Learn more at www.essenbioscience.com/NeuroCellHealth
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