Download Let`s move cell health forward together

Let's move cell health forward together
Sensitive indicators of cell health
and stress
Mitochondrial stains
Mitochondria are prominent cellular markers,
and mitochondrial function is a highly sensitive
indicator of cell health and stress. Depolarization
of the inner mitochondrial membrane potential is a
reliable indicator of mitochondrial dysfunction, which
has been increasingly implicated in drug toxicity.
Life Technologies provides a range of Molecular
Probes® tools for tracking mitochondrial location and
investigating mitochondrial function using cell stains,
functional probes, and fluorescent proteins.
•Choice of technologies for optimized performance
in each application
•Wide range of colors for counterstaining with other
fluorescent reporters
•Functional probes for reactive oxygen species and
membrane potential
Fluorescent proteins:
Mitochondrial behavior can be
visualized dynamically in live
cells using CellLight® technology.
CellLight® reagents express
fusion proteins with red or green fluorescence for
imaging in live cells or after formalin fixation.
Antibody conjugates: With
Molecular Probes® antibody
conjugates you can label
mitochondria in fixed cells or
frozen tissue sections even after
paraffin embedding. (Unconjugated antibodies
require the use of secondary detection labels.)
MitoTracker® Probes: The
MitoTracker® family of probes
stains mitochondria in live cells
through a variety of mechanisms.
Some probes load according to
mitochondrial membrane potential and some are
retained after fixation.
Functional probes: MitoSOX™ Red
Mitochondrial Superoxide Indicator
is an indicator of superoxide
activity in mitochondria, and
MitoHealth stain can serve as a
marker for mitochondrial membrane potential in high
content screening applications.
For more information go to lifetechnologies.com/mitochondria
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0 13
Cellular scaffold and the ultimate
counterstain target
Cytoskeleton stains
The cytoskeleton has a role in organelle transport,
cell division, motility, and signaling, making it
central to normal growth processes and critical
in disease models. Cytoskeleton stains enable
research on the cellular scaffold, and they also
serve as counterstains or fiducial markers
for cellular orientation and co-localization in
both manual and automated imaging systems.
Molecular Probes® products are the most cited
cytoskeleton stains and support any application.
Alexa Fluor® conjugates, antibodies, and
fluorescent proteins provide the leading technology
solutions for labeling the major classes of
cytoskeletal proteins.
•Bright fluorescence for visualizing fibers and
tubules
•Choice of technologies for optimized performance
in any application
•Wide choice of colors for counterstaining with
other fluorescent reporters
Fluorescent proteins: Both
actin and tubulin behavior can
be visualized dynamically in live
cells using CellLight® technology.
CellLight® reagents express
fusion proteins with red or green fluorescence for
imaging in live cells or after formalin fixation.
Antibody conjugates: With
Molecular Probes® antibody
conjugates you can label actin
or tubulin in fixed cells or frozen
tissue sections even after paraffin
embedding. (Unconjugated antibodies require the use
of secondary detection labels.)
Phalloidin conjugates: The most
common actin label for fixed and
permeabilized cells is available
conjugated to a range of high
performance dyes. Brighter
staining and resistance to photobleaching in a wide
range of colors for multiplexing.
For more information go to lifetechnologies.com/cytoskeleton
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
TubulinTracker: This Taxol
derivative stains polymerized
tubulin for live cell imaging.
Classic counterstains and cell
death markers
Nuclear stains
The nucleus is routinely stained to serve as a fiducial
marker in automated imaging systems. In addition,
nucleic acid stains that show enhanced fluorescence
on binding to DNA are frequently used as markers of
membrane permeability and cell death.
•Choice of technologies for optimized performance
in each application
•Wide range of colors for counterstaining with other
fluorescent reporters
•Applications from dead-cell assays to
counterstaining
Counterstains: Nuclear
counterstains with blue
fluorescence like Hoechst
33342 and DAPI provide
spatial information about the
cell and leave popular spectral channels open for
multiplexing. SYTO® dyes stain DNA and RNA in live
or fixed cells. They serve as counterstains in a range
of colors from green to far-red.
Fluorescent proteins: Nuclear
behavior can be visualized
dynamically in live cells
using CellLight® nuclear and
histone constructs. CellLight®
reagents express fusion proteins with red or green
fluorescence for imaging in live cells or after formalin
fixation.
Antibody conjugates: With
Molecular Probes® antibody
conjugates you can label nuclear
components in fixed cells or frozen
tissue sections even after paraffin
embedding. (Unconjugated antibodies require the use
of secondary detection labels.)
SYTOX® dyes: SYTOX® dyes stain
nucleic acids in fixed cells with
high affinity and a 500x signal
enhancement on binding. They
can easily be multiplexed and serve as dead cell
stains.
For more information go to lifetechnologies.com/nucleus
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO0 013
Fluorescence for studying
morphology and outgrowth
Cell tracing tools
Cell morphology and connectivity are key
components in understanding neuronal function.
Connectivity and function are routinely studied
in cultured cells, fixed tissue sections, and live
tissues that are subsequently fixed for analysis.
The fluorescence technologies used to trace neural
networks can be divided into those for single cells or
cell populations. While the different classes of probes
listed here all label tissue sections and cultured
cells, there are unique performance features for
addressing each specific application and answering
each biological question.
•Bright fluorescence for visualizing fine neuronal
processes
•Choice of technologies for optimized performance
in any application
•Choice of colors for multiplexing with other
fluorescent reporters
Hydrazides and biocytins: Bright
polar tracers with multiple
colors are loaded into cells by
microinjection. They provide
fixable and photostable cellular
contrast and also track gap junctions.
Conjugated dextrans: Available
in a range of colors and sizes and
loaded by microinjection. Smaller
dextrans will track gap junctions,
while larger dextrans are slower
and more persistent for axonal tracing.
Lipophilic tracers: Formulated
as crystals or pastes for applying
directly to tissues, lipophilic
tracers will stain fresh or fixed
tissue sections as well as
live cells, and track neuronal outgrowths in both
directions.
Toxins and lectins: These
fixable conjugates all have
unique properties and a range
of fluorescent reporters. Wheat
germ agglutinin is a retrograde
tracer, while cholera toxin is anterograde. IB4
differentiates between different classes of neurons.
For more information go to lifetechnologies.com/neuronaltracers
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Studying migration, chemotaxis,
and invasion
Cell tracking tools
Cell migration studies are key to understanding
processes such as wound healing, immune
response, and tumor metastasis. Ideal tracking
tools should be bright enough for easy visualization,
should resist cell-to-cell transfer, and for longterm tracking, should remain in the cell for the
required number of divisions. Molecular Probes®
products offer solutions based on transgenic protein
expression, organic dyes, or Qdot® probes. Each
solution is available for a range of applications with
several wavelengths for multiplexed detection.
•Bright fluorescence for easy detection and
quantitation
•Choice of technologies for optimized performance
in any application
•Choice of colors for multiplexing with other
fluorescent reporters
CellLight® solutions
Transiently transduced cells
express fluorescent fusion
proteins at different locations
within the cell. Labeled cells are
easy to track for at least five days or up to two weeks
in slow-growing populations.
Learn more at lifetechnologies.com/celllight
CellTracker™ solutions
Organic dyes will label cell
populations easily without
the need for transduction.
CellTracker™ probes provide
bright, uniform labeling of cells in a range of colors
lasting for 3 to 6 generations.
Learn more at lifetechnologies.com/celltracker
Qtracker® solutions
Qtracker® probes are extremely
bright and resistant to
photobleaching. They provide
punctate labeling of cells in a wide
range of wavelengths that lasts for 6 to 10 generations
of tracking.
Learn more at lifetechnologies.com/qtracker
For more information go to lifetechnologies.com
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Tracking the endocytic pathway
with fluorescence
Phagocytosis and endocytosis are the processes by
which cells internalize particulate matter such as
microorganisms, and materials such as proteins and
ligands. They are important in immune responses
and the clearance of apoptotic cells in metabolism
and cell signaling. When cells internalize proteins
or particles, the endosome is acidic relative to the
extracellular environment, so pH indicators like
pHrodo™ dye are useful for monitoring stages in
the pathway from early endosome to lysosome
formation.
•Monitor phagocytosis with red or green
fluorescence
•Sensitive and robust assay with no washing or
quenching required
•Flexible workflow for imaging, flow cytometry, or
microplate assays
RAW cells were incubated
with zymosan BioParticles®
particles (containing pHrodo™
Green dye) and were imaged
in Live Cell Imaging Solution
(Cat. No. A14291DJ). Green
fluorescence indicates that
the particles have become
acidified.
Schematic of pHrodo™
dye–based detection
of phagocytosis and
endocytosis. Particles
(microorganisms or proteins)
labeled with pHrodo™ dye
are added to cells. Some
remain in solution or become
nonspecifically attached to
cells—they do not fluoresce
because of the neutral
pH of the extracellular
environment. Some are taken up by cells by phagocytosis or endocytosis
and become encapsulated in vesicles. As vesicles are processed, pH
decreases and the pHrodo™ dye–labeled particles fluoresce.
Phagocytosis
Endocytosis and pinocytosis
Selection guide
Application
Phagocytosis
Red
Green
Dextran (10K)
Zymosan
P35364
P35365
E.coli
P35361
P35366
S. aureus
A10010
Endocytosis
Red
Green
P10361
P35368
P35367
Reactive pHrodo dyes allow you to conjugate your own particles or proteins
Coupling
chemistry
Red
Avidin
P35362
Maleimide
P35371
SE
P35363
STP ester
For more information, go to lifetechnologies.com/phrodo
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Green
P35370
P35369
Monitoring oxidative stress and
reactive oxygen species
CellROX® reagents
Oxidative damage is a fact of life associated with
aging, disease, and cellular damage. To measure
oxidative stress in live cells, CellROX® reagents
generate a fluorescent green, orange, or deep red
signal. You can use flow cytometry, fluorescence
imaging, or microplate fluorometry to monitor the
signal and easily multiplex with immunostaining or
with other fluorescent reporters.
•Monitor reactive oxygen species with deep red,
orange, or green fluorescence
•Flexible workflow for imaging, flow cytometry, or
microplate assays
•Choice of colors for multiplexing with other
fluorescent reporters
Imaging oxidative stress with CellROX® Green Reagent. Human
osteosarcoma (U2-OS) cells expressing CellLight® Actin-RFP were
treated with 100 µM menadione. Cells were stained with CellROX®
Green Reagent (Cat. No. C10444) and NucBlue™ Live Cell Stain (Cat. No.
R37605), washed, and imaged with Live Cell Imaging Solution (Cat. No.
A14291DJ).
CellROX® reagent selection guide
Ex/Em max
Deep red
Orange
Green
640/665 nm
545/565 nm
485/520 nm
Live-cell compatible
Yes
Yes
Yes
Can be added to complete media
Yes
Yes
Yes
Formaldehyde fixable
Yes
No
Yes
Multiplex ready
No
No
Yes
Signal to noise
+++++
+++++
+++++
Photostability
+++
++++
++
GFP compatible
No
Yes
Yes
RFP compatible
Yes
No
Yes
Cat. No.
C10422
C10443
C10444
For more information go to lifetechnologies.com/cellrox
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Autophagy—cells under stress
Fluorescence-based tools for LC3B detection
The LC3B protein plays a critical role in autophagy.
Normally, LC3B resides in the cytosol, but
following cleavage and lipidation with phosphatidylethanolamine, the LC3B protein associates with the
phagophore and can be used as a general marker
for autophagic membranes. A series of Molecular
Probes® fluorescence-based tools are available to
help visualize and detect this critical protein.
•Premo Autophagy Sensor—detect LC3B
temporally and spacially in live cells
™
•LC3B Antibody Kit for Autophagy—complete kit to
visualize LC3B localization in fixed samples
•Premo™ Autophagy TR-FRET Assay—obtain
quantitative, plate reader measurement of
autophagy
Mitochondrion
Protein
Amino acids,
fatty acids
Peroxisome
Phagophore
Nucleus
Autolysosome
Autophagosome
LC3B
recruitment
LC3B
disassociation
Lysosome
Schematic depiction of the multistep autophagy pathway in a
eukaryotic cell. The first step involves the formation and elongation
of isolation membranes, or phagophores. In the second step, which
involves the LC3B protein, the cytoplasmic cargo is sequestered,
and the double-membrane autophagosome is formed. Fusion of a
lysosome with the autophagosome to generate the autolysosome is the
penultimate step. In the fourth and final phase, the cargo is degraded.
Premo™ Autophagy Sensor
The Premo™ Autophagy Sensor combines the
selectivity of an LC3B-fluorescent protein (LC3B-FP)
chimera, with the transduction efficiency of BacMam
technology, enabling unambiguous visualization of
this protein in live cells. Available in either GFP or
RFP configurations, the reagent also tolerates fixation
with formaldehyde and thus is compatible with
fixed-cell analysis, which is required for multiplex
experiments using antibodies, and is preferred for
HCS analysis workflows.
The LC3B Antibody Kit
Each LC3 Antibody Kit for autophagy includes a
rabbit polyclonal antibody against LC3B that has
been validated for use in fluorescence microscopy
and high-content imaging and analysis. The kit
also includes a control compound for inducing
autophagosomes—following treatment with
this compound, lysosomal pH increases and the
normal autophagic flux is disrupted, resulting in
autophagosome accumulation.
Premo™ Autophagy TR-FRET Assay
The Premo™ Autophagy TR-FRET Assay measures
changes in the autophagy activity of cells expressing
GFP-tagged LC3B using a Terbium (Tb)-based TRFRET plate reader immunoassay approach, providing
you with a quantitative, objective measurement of
autophagy. The expression kit provides BacMam
reagent for easily expressing GFP-LC3B in many
different cell types along with a Tb-labeled LC3B-II
specific detection antibody for performing the assay.
For more information go to lifetechnologies.com/autophagy
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Apoptosis—caspases bringing cell
death to light
CellEvent™ Caspase-3/7 detection reagent
Cysteine aspartic acid proteases, or caspases,
are enzymes that play a key role in initiating and
executing apoptosis. Activation of caspases is one
of the more definitive markers of programmed
cell death. The CellEvent™ Caspase-3/7 Green
detection reagent is a new fluorogenic substrate
for activated caspases 3 and 7 that is compatible
with microscopy, flow cytometry, or high-content
imaging, and is well suited for multiplex imaging in
live or fixed cells.
•Optimized caspase-3⁄7 substrate for apoptosis
analysis by microscopy, flow cytometry, or highthroughput screening
untreated
2-hr staurosporine
4-hr staurosporine
Traditional fluorescence imaging of oxidative stress and apoptosis.
HeLa cells were treated with 0.5 µM staurosporine for 0, 2, or 4 hr
in the presence of 7.5 µM CellEvent™ Caspase-3/7 Green detection
reagent. Cells were then stained with 5 µM CellROX ™ Deep Red reagent
and Hoechst 33342 nucleic acid stain for 30 min at 37°C, washed with
warm DPBS, and imaged immediately. Increased oxidative stress was
observed at 2 hr after staurosporine treatment (magenta), whereas
caspase-3/7 activation was not observed until 4 hr after treatment
(green).
•Simple, no-wash protocol helps preserve delicate
apoptotic cells
green nuclei, while cells without activated caspase 3/7
show minimal fluorescence.
•Compatible with both live cell fluorescence
imaging and formaldehyde-based fixation
methods
This robust assay is highly specific for caspase 3/7
activation and, as expected, nearly complete inhibition
of the CellEvent™ Caspase-3/7 Green detection
reagent signal was observed after pretreatment of
cells with Caspase 3/7 Inhibitor 1. The fluorescent
signal from the CellEvent™ Caspase-3/7 reagent
survives fixation and permeabilization, providing the
flexibility to perform end-point assays and probe for
other proteins of interest using immunocytochemistry.
The CellEvent™ detection reagent requires no wash
steps—simply add it to cells in complete culture
medium and incubate for 30 minutes. Samples are
ready for imaging using the standard FITC filter set.
Apoptotic cells with activated caspase 3/7 show bright
Product
CellEvent Caspase-3/7 Detection Reagent
™
Platform
Flow cytometry laser
Imaging filter set
Ex/Em (nm)
Cat No.
FC,I,HCS,HTS
Blue (488 nm)
FITC
502/530
C10423
To see CellEvent™ Caspase-3/7 reagent in action, go to
lifetechnologies.com/celleventvideo
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Apoptosis—DNA fragmentation
Click-iT® TUNEL Assays—the best and the brightest
Click-it® TUNEL is a breakthrough technology using
click chemistry for detection of DNA fragmentation.
For TUNEL assays to yield meaningful results,
it's necessary that the modified nucleotide is an
acceptable substrate for TdT, and that the detection
method is sensitive and avoids detrimental loss of
cells from the sample. Click-it® TUNEL Assays:
•Detect late-stage apoptosis before cells "pop"
•Make it easy to multiplex with additional apoptosis
markers
•Save you time with click chemistry
How the TUNEL assay works with click chemistry
Click-iT® TUNEL Imaging Assays
The Click-iT® TUNEL Imaging Assays employ dUTP
modified with an alkyne, a small, bio-orthogonal
functional group. The incorporated nucleotide is
detected through click chemistry—a copper-catalyzed
reaction between an azide and an alkyne.
Click chemistry
Click chemistry is an alternative way to label and
detect when methods such as direct labeling or
using antibodies are too destructive to the sample.
The small size of the Alexa Fluor® azide (MW ~1,000)
compared to that of an antibody (MW ~150,000)
enables effortless penetration of complex samples
with only mild fixation and permeabilization needed.
DNA strand breaks typical of late-stage apoptosis visualized using
the Click-iT® TUNEL Imaging Assay (red) on HeLa cells treated
with staurosporine. Activated caspase-3 was detected with a rabbit
polyclonal primary antibody for cleaved caspase-3 and labeled with
Alexa Fluor® 488 goat anti–rabbit IgG (green). Tubulin was detected
with a mouse monoclonal anti-tubulin antibody and labeled with Alexa
Fluor® 555 goat anti–mouse IgG (orange). Nuclei were stained with
Hoechst 33342 (blue). The cells on the right not only have a high level
of caspase-3 activity and DNA strand breaks, but also show a loss of
structural integrity consistent with cells undergoing apoptosis.
Click-iT® TUNEL click chemistry Alexa Fluor®
detection
Available in three different bright and photostable
Alexa Fluor® dyes
To unequivocally establish programmed cell death
in any given cell or tissue model, two or more
biomarkers of apoptosis must be observed. The
click chemistry Click-iT® TUNEL Imaging Assays are
available in three different bright and photostable
Alexa Fluor® dyes, ranging from green to far-red
fluorescence, providing flexibility when working with
other apoptosis detection reagents that are typically
available only in green- or red-fluorescent colors.
Detects DNA fragmentation in adherent cells
Each Click-iT® TUNEL kit contains all of the
components necessary to accurately and reliably
detect DNA fragmentation with simple click chemistry
reactions in adherent cells grown on coverslips or in
96-well microplates.
Product
Platform
Common imaging filter sets
Ex/Em (nM)
Cat No.
Click-iT® TUNEL Alexa Fluor® 488 Imaging Assay
I, HCS
FITC
495/519
C10245
Click-iT TUNEL Alexa Fluor 594 Imaging Assay
I, HCS
Texas Red
590/615
C10246
Click-iT® TUNEL Alexa Fluor® 647 Imaging Assay
I, HCS
Cy® 5
650/670
C10247
®
®
For more information go to lifetechnologies.com/tunel
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Simple, efficient, effective cell
viability determination
Cell viability—LIVE/DEAD® viability assays for mammalian cells
Molecular Probes® LIVE/DEAD® assays provide
complete solutions for easy, sensitive determination
of cell viability, cell vitality, and compound
cytotoxicity. These fluorescence-based assays can
be used to examine a variety of mammalian cell
types. Specific LIVE/DEAD® assays can be used for
flow cytometry, microscopy, or microplate formats.
Principle of assays
LIVE/DEAD® Viability/Cytotoxicity Kit (Cat. No.
L3224)— quick and easy two-color assay to
determine viability of cells in a population based on
plasma membrane integrity and esterase activity.
• Live stain: calcein AM (green)
Viability of kangaroo rat (PtK2) cells using the LIVE/DEAD® Viability/
Cytotoxicity Kit. Live and dead PtK2 cells were stained with ethidium
homodimer-1 and the esterase substrate calcein AM, both provided in the
LIVE/DEAD® Viability/Cytotoxicity Kit . Live cells fluoresce a bright green,
whereas dead cells with compromised membranes fluoresce red-orange.
• Dead stain: ethidium homodimer-1 (red)
LIVE/DEAD® Cell-Mediated Cytotoxicity Kit (Cat. No.
L7010)— measures cytotoxicity mediated by natural
killer cells, lymphokine-activated killer cells, and
T cells.
• Live stain: DiOC18(3) (green)
• Dead stain: propidium iodide (red)
LIVE/DEAD® Sperm Viability Kit (Cat. No. L7011)—
analyzes the viability and fertilizing potential of sperm
by fluorescence microscopy or flow cytometry.
• Live stain: SYBR® 14 dye (green)
• Dead stain: propidium iodide (red)
LIVE/DEAD® Cell Vitality Assay Kit (Cat. No.
L34951)—simple, two-color fluorescence assay that
distinguishes metabolically active cells from injured
cells and dead cells.
Distinguishing live from dead sperm with the LIVE/DEAD® Sperm
Viability Kit . Live sperm with intact membranes were labeled with
SYBR® 14 dye, a proprietary cell-permeant nucleic acid stain, and
fluoresce green. Dead sperm, which were killed by unprotected freezethawing, were labeled with propidium iodide and fluoresce red-orange.
Both dyes are components of the kit. The image was contributed by
Duane L. Garner, School of Veterinary Medicine, University of Nevada,
Reno, and Lawrence A. Johnson, USDA Agricultural Research Service.
• Live stain: C12-resazurin (red)
• Dead stain: SYTOX® Green (green)
For more information go to lifetechnologies.com/livedeadassays
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Live-cell imaging of cell cycle and division
Solutions for microscope imaging and HCS
Measuring a cell’s ability to proliferate and exploring
the mechanisms of cell growth are fundamental
methods used in developmental and stem cell
biology, cancer research, and drug discovery and
toxicology. Several Molecular Probes® assays are
now available to efficiently and accurately measure
the progression of a cell through the cell cycle,
whether you are exploring the mitotic pathway or
evaluating a novel compound.
Cell cycle measurement in live cells
Premo™ FUCCI Cell Cycle Sensor is a fluorescent
two-color sensor of cell cycle progression and
division in live cells. It allows accurate and sensitive
cell cycle analysis of individual cells or a population
of cells by fluorescence microscopy, flow cytometry,
or high-content imaging. The fluorescence
ubiquitination cell cycle indicator (FUCCI) is a
genetically encoded, two-color (red and green)
indicator that lets you follow cell division within a cell
population. We have incorporated the FUCCI genetic
constructs into the powerful BacMam gene delivery
system, creating a simple and efficient method for
labeling cells and following their division.
• Accurate—monitor changes in cell proliferation with
direct visual readout of cell cycle phase for every cell in
the population
• Highly efficient—>90% transduction of a wide range of
mammalian cell lines, including primary cells and stem
cells
Visualization of cell cycle phases with the Premo™ FUCCI Cell
Cycle Sensor over a 16-hour period. Initially, the cell in the center
of the image transitioned from green (G2/M) to red (G1) to yellow (S).
Approximately 6 hr after imaging was initiated, the cell at the top of
the visual field migrated down before undergoing mitosis (green) and
progressing into G1 (red).
Imaging cell cycle progression in live cells with Premo™ FUCCI
Cell Cycle Sensor. Schematic of cell cycle progression with nuclear
fluorescence changes.
• Fast and convenient—simply add Premo™ FUCCI Cell
Cycle Sensor to your cells in complete medium, incubate overnight
Ordering information
Product
Quantity
Premo™ FUCCI Cell Cycle Sensor (BacMam 2.0)
1 kit
For more information, go to lifetechnologies.com/fucci
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
Cat. No.
P36238
Cell proliferation and DNA replication
Multicolor imaging techniques for accurate cell proliferation assays
Cell proliferation assays are important tests to
determine whether a cell population is actively
dividing or has undergone division during an assay
period. Using fluorescent labels to detect new DNA
synthesis can provide rich data sets on imaging and
microplate platforms. With a choice of fluorescent
labels, assays can be multiplexed to explore other cell
health parameters and answer complex biological
questions.
•Superior performance—highly sensitive assays to
detect cell division
•Flexibility—multiple assays and colors available
for multiparametric studies
•Efficiency—quick, robust, easy-to-use labeling
techniques for multiple cell types
Stains for new DNA synthesis
The Click-iT® EdU Assay kits provide simple, robust
methods for analyzing DNA replication in proliferating
cells in as little as 90 minutes. Kits are available for
microscopic imaging or high-content screening (HCS)
analysis, with a choice of fluorescence wavelengths
for multiplexing or with Amplex® UltraRed signal
amplification to provide a fluorescence intensity
readout on microplates.
Multicolor imaging with Click-iT® EdU. Muntjac cells pulsed with EdU,
fixed and permeabilized. Labeled DNA was detected using the Click-iT®
EdU Alexa Fluor® 647 Imaging Kit (Cat. No. C10340). Tubulin (in blue)
is labeled with a mouse primary followed by Alexa Fluor® 350 goat
anti–mouse IgG antibody. Golgi (in green) is labeled with Alexa Fluor®
488 conjugate of lectin HPA, and peroxisomes (in orange) are labeled
with a rabbit primary followed by Alexa Fluor® 555 donkey anti–rabbit
IgG antibody.
Ordering information
Product
Size
Cat. No.
Click-iT® EdU Microplate Assay
1 kit/400 assays
C10214
Click-iT® EdU Alexa Fluor® 488 HCS
Assay
2 plates
C10350
Click-iT® EdU Alexa Fluor® 555 HCS
Assay
2 plates
C10352
Click-iT® EdU Alexa Fluor® 594 HCS
Assay
2 plates
C10354
Click-iT® EdU Alexa Fluor® 647 HCS
Assay
2 plates
C10356
Click-iT® EdU Imaging Kit with
Alexa Fluor® 488, 594, and 647
1 kit
C10086
Click-iT® EdU Alexa Fluor® 488
Imaging Kit
1 kit/50
coverslips
C10337
Click-iT® EdU Alexa Fluor® 555
Imaging Kit
1 kit/50
coverslips
C10338
Click-iT® EdU Alexa Fluor® 647
Imaging Kit
1 kit/50
coverslips
C10340
For more information, go to lifetechnologies.com/clickit
For research use only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0113
ReadyProbes™ ready-to-use
imaging reagents
No Calculations. No Dilutions. No Resuspensions.
Fluorescence imaging has never been easier
with ReadyProbes™ reagents.
• Dropper bottle formats
See the following example of a simple
ReadyProbes™ protocol.
Protocol for NucBlue® Live Cell Stain
• Stable at room temperature
• Simplified protocols
• No dilutions, resuspensions, calculations, or
aliquoting necessary
1. Add 2 drops per mL of medium
2. Incubate 5–20 min at 20–25ºC
3. Image cells
ActinGreen™ 488 ReadyProbes™ reagent. BPAE cells were fixed, permeabilized, and blocked using the Image-iT® Fixation/
Permeabilization Kit. Tubulin was labeled using an anti-tubulin antibody followed by detection with an Alexa Fluor ® 594 secondary.
Actin was stained using ActinGreen™ 488 ReadyProbes™ reagent and nuclei were stained with NucBlue® Fixed Cell Stain.
Available Reagents
Product
Nuclear stains
NucBlue® Live ReadyProbes™ Reagent
NucBlue® Fixed Cell ReadyProbes™ Reagent
NucGreen™ Dead 488 ReadyProbes™ Reagent
Propidium iodide ReadyProbes™ Reagent
NucRed™ Live 647 ReadyProbes™ Reagent
NucRed™ Dead 647 ReadyProbes™ Reagent
Actin stains
ActinGreen™ 488 ReadyProbes™ Reagent
ActinRed™ 555 ReadyProbes™ Reagent
Apoptosis reagent
CellEvent® Caspase 3/7 Green ReadyProbes™ Reagent
Imaging accessories
BackDrop® Background Suppressor ReadyProbes™ Reagent
Live Cell Imaging Solution*
Image-iT® Fixation/Permeabilization Kit*
Image-iT® FX Signal Enhancer ReadyProbes™ Reagent
*Not available in dropper bottle format
Filter
Ex/Em (nm)
Cat. No.
DAPI
DAPI
FITC
RFP
Cy®5
Cy®5
358/461
352/461
504/523
535/617
638/686
642/661
R37605
R37606
R37109
R37108
R37106
R37113
FITC
RFP
495/518
540/565
R37110
R37112
FITC
495/518
R37111
NA
NA
NA
NA
NA
NA
NA
NA
R37603
A14291DJ
R37602
R37107
Find out more at lifetechnologies.com/readyprobes
For Research Use Only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0313
Antifade reagents
Minimize photobleaching across the visible spectrum
Loss of fluorescent signal through irreversible
photobleaching processes can lead to a significant
reduction in sensitivity, particularly when target
molecules are of low abundance or when excitation light is of high intensity or long duration.
To minimize photobleaching of experimental
samples we have developed a series of antifade
reagents that increase the photostability of many
popular fluorophores and preserve signals for
higher sensitivity and extended analysis.
•Inhibit photobleaching across the spectrum
•Mounted samples stable for weeks to months
Both SlowFade® and ProLong® reagents are
available preformulated with DAPI as a nuclear
counterstain, eliminating the need for a separate
counterstaining step.
•Available with or without DAPI counterstain
Convenience
•Hard-setting or immediate-use formulations
•Ready-to-use benchtop formulation
•Optimized refractive index
Choosing the right antifade
Life Technologies antifade reagents are optimized
for a variety of sample requirements such as longterm storage or immediate imaging. SlowFade®
Antifade Reagent provides immediate imaging and
short-term (3–4 weeks) storage, while ProLong®
Antifade Reagent requires a curing period but
provides long-term stability.
Life Technologies antifade reagents are now formulated for room temperature storage and provided in dropper bottles for immediate benchtop
use.
Green chemistry
As well as providing improved convenience for
routine imaging, these reagents provide energysaving room temperature storage. Environmental and health risks have also been significantly
reduced by the elimination of sodium azide.
ProLong® Gold Antifade Reagent
SlowFade® Gold Antifade Reagent
Long-term storage
Yes
Yes
Yes
Yes
No
No
No
No
Immediate viewing
No
No
No
No
Yes
Yes
Yes
Yes
Counterstain
No
No
DAPI
DAPI
No
No
DAPI
DAPI
Quantity
5 x 2 mL
10 mL
5 x 2 mL
10 mL
5 x 2 mL
10 mL
5 x 2 mL
10 mL
Cat. No.
P36934
P36930
P36935
P36931
S36937
S36936
S36939
S36938
View all products online at lifetechnologies.com/antifades
For Research Use Only. Not for use in diagnostic procedures. ©2013 Life Technologies Corporation. All rights reserved. The trademarks
mentioned herein are the property of Life Technologies Corporation and/or its affiliate(s) or their respective owners. CO07205 0613