Oxidative Stress and Hypoxia Information

Measuring oxidative stress and hypoxia in live cells or tissue can be very challenging. Here you will find useful tips and other educational resources to help you overcome these issues.

Oxidative stress and hypoxia features

Application note

Application note

Guide to setting up hypoxic conditions on the EVOS FL Auto Imaging System with onstage incubator

Here we provide step-by-step instructions for setting up hypoxic conditions with the EVOS FL Auto Imaging System with Onstage Incubator.

On-demand webinar

Breathing new life into hypoxia research

In this webinar, we will discuss hypoxia in human diseases, classical methods for setting up hypoxic conditions, and novel instruments and reagents for imaging cells in hypoxic conditions.


Oxidative stress and hypoxia learning resources

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Type Title Categories
Application note (2014) Oxidative stress measurements made easy: CellROX reagents with simple workflow for high content imaging  ArrayScan, CellInsight, CellROX, high content analysis, oxidative stress
Application note (2015) Using time-lapse imaging with the EVOS FL Auto Imaging System EVOS FL Auto Microscope, fluorescence microscopy/fluorescence imaging, hypoxia, live-cell imaging, onstage incubator
Application note (2015) Guide to setting up hypoxic conditions on the EVOS FL Auto Imaging System with Onstage Incubator EVOS FL Auto Microscope, fluorescence microscopy/fluorescence imaging, live-cell imaging, onstage incubator, phagocytosis
BioProbes article (2007) Seeing superoxide in a radical new light cell structure-mitochondria, fluorescent dyes, fluorescence microscopy/fluorescence imaging, live-cell imaging, oxidative stress, sample stability
BioProbes article (2007) Mitochondria and cardiac disorders  antibodies, antibody labeling, cardiovascular system, cell structure-mitochondria, fluorescent dyes, fluorescence microscopy/fluorescence imaging, oxidative stress, western blotting
BioProbes article (2009) Multiparametric approaches for predictive hepatotoxicity  cell structure-mitochondria, cell structure-plasma membrane, flow cytometer/flow cytometry, fluorescent dyes, fluorescence microscopy/fluorescence imaging, high content analysis, oxidative stress, viability
BioProbes article (2009) Fluorescence technologies for high-content imaging and analysis  Alexa Fluor, cell health, Click-iT, drug discovery, flow cytometer/flow cytometry, fluorescent dyes, fluorescence microscopy/fluorescence imaging, high content analysis, oxidative stress, protein detection, viability
BioProbes article (2010) Putting autophagy into context autophagy, CellInsight fluorescence microscopy/fluorescence imaging, high content analysis, oxidative stress
BioProbes article (2010) A closer look at mitochondrial dysfunction with MitoSOX Red Mitochondrial Superoxide Indicator cell structure-mitochondria, oxidative stress
BioProbes article (2010) Alexa Fluor dyes shed light on a new actin assembly factor cell structure-actin, fluorescence microscopy/fluorescence imaging, high content analysis, oxidative stress
BioProbes article (2011) Fluorescence imaging of oxidative stress in live cells—CellROX Deep Red Reagent is a bright and photostable ROS sensor CellROX, fluorescence microscopy/fluorescence imaging, oxidative stress
BioProbes article (2011) Shedding light on oxidative stress—CellROX Deep Red Reagent for ROS detection ArrayScan, cell health, CellROX, fluorescence microscopy/fluorescence imaging, high content analysis, oxidative stress
BioProbes article (2012) Streamline your oxidative stress measurements—CellROX Green and CellROX Orange reagents with simple workflows CellROX, fluorescence microscopy/fluorescence imaging, microplate reader, oxidative stress
BioProbes article (2013) Oxidative stress from a cell's point of view—From CellROX ROS sensors to Image-iT and Click-iT lipid peroxidation detection kits CellROX, Click-iT, flow cytometer/flow cytometry, fluorescence microscopy/fluorescence imaging, live-cell imaging, oxidative stress
BioProbes article (2013) Checking vital signs: Don't let dead cells mislead you—Molecular Probes cell health assays for flow cytometry apoptosis, caspase substrate, cell cycle, cell health, cell proliferation, cell tracking, Click-iT, flow cytometer/flow cytometry,oxidative stress, viability
BioProbes article (2014) Image live cells through time and space—Optimizing conditions for time-lapse fluorescence microscopy cell structure, cell tracking, CellLight, EVOS, fluorescence microscopy/fluorescence imaging, high content analysis, live-cell imaging, oxidative stress
BioProbes article (2015) Tools to study mitochondrial morphology and function—Multiplex your mitochondrial data cell structure-mitochondria, fluorescence microscopy/fluorescence imaging, oxidative stress
BioProbes article (2015) Breathe new life into hypoxia research—Intracellular detection of low oxygen tension in live cells antibodies EVOS, fluorescence microscopy/fluorescence imaging, hypoxia, live-cell imaging
BioProbes article (2015) Formation of uniform and reproducible 3D cancer spheroids in high-throughput plates—Mimic life in three dimensions cancer, fluorescence microscopy/fluorescence imaging, hypoxia, oxidative stress, viability
Molecular Probes Handbook Probes for nitric oxide research—Section 18.3 NO, nitrate, nitric oxide (NO), oxidative stress, SNAP
Molecular Probes Handbook Introduction to reactive oxygen species—Section 18.1 oxidative stress
Molecular Probes Handbook Generating and detecting reactive oxygen species—Section 18.2 oxidative stress
Scientific poster (2010) Simultaneous analysis of cell death mechanisms and oxidative stress using Molecular Probes next generation reagents for imaging and flow cytometry  apoptosis, autophagy, caspase substrates, flow cytometry/flow cytometer, fluorescence microscopy/fluorescence imaging, fluorescent dyes, live-cell imaging, oxidative stress
Scientific poster (2010) Oxidative stress-induced protein modification: Application of clickable linoleic acid analogs  Alexa Fluor, ArrayScan, Click-iT, fluorescence microscopy/fluorescence imaging, fluorescent dyes, gel electrophoresis, high content analysis, microplate reader, oxidative stress, protein detection, protein enrichment
Scientific poster (2010) Simultaneous analysis of cell death mechanisms and oxidative stress using live cell fluorescence microscopy  apoptosis, ArrayScan, BacMam technology, caspase substrates, fluorescence microscopy/fluorescence imaging, fluorescent dyes, high content analysis, live-cell imaging, microplate reader, oxidative stress
Scientific poster (2011) Cell-based measurements of oxidative stress with a new near infrared emitting probe for reactive oxygen species: Applications for multiplexed evaluation of cell health  ArrayScan, cell health, flow cytometer/flow cytometry, fluorescence microscopy/fluorescence imaging, fluorescent dyes, high content analysis, microplate reader, oxidative stress, viability
Scientific poster (2011) Simultaneous analysis of cell death mechanisms and oxidative stress using high content imaging  Alexa Fluor, antibodies, apoptosis, ArrayScan, autophagy, caspase substrates, fluorescence microscopy/fluorescence imaging, fluorescent dyes, high content analysis, microplate reader, oxidative stress
Scientific poster (2011) The next generation of cell-based imaging assays for apoptosis, autophagy and oxidative stress from Molecular Probes  Alexa Fluor, antibodies, apoptosis, ArrayScan, autophagy, caspase substrates, fluorescence microscopy/fluorescence imaging, fluorescent dyes, high content analysis, microplate reader, oxidative stress
Scientific poster (2012) Cell-based analysis of oxidative stress, lipid peroxidation and lipid peroxidation-derived protein modifications using fluorescence microscopy  flow cytometer/flow cytometry, fluorescence microscopy/fluorescence imaging, high content analysis, lipid peroxidation, microplate reader, oxidative stress, viability
Scientific poster (2013) Cell-based measurements of oxidative stress with a new near infrared emitting probe for reactive oxygen species: Applications for multiplexed evaluation of cell health  apoptosis, ArrayScan, caspase substrates, cell health, flow cytometer/flow cytometry, fluorescence microscopy/fluorescence imaging, fluorescent dyes, fluorescent proteins, high content analysis, microplate reader, oxidative stress
Scientific poster (2013) Reactive oxygen probes—A broad range of colors with easier labeling: Novel CellROX reagents from Molecular Probes  cells, flow cytometer/flow cytometry, fluorescent dyes, multicolor flow cytometry, oxidative stress, viability
Scientific poster (2013) Reactive oxygen probes—A broad range of colors with easier labeling and compatibility with fixation: Novel CellROX reagents from Molecular Probes  cells, flow cytometer/flow cytometry, fluorescent dyes, multicolor flow cytometry, oxidative stress
Scientific poster (2015) Intracellular detection of hypoxia in live cells  EVOS, fluorescence microscopy/fluorescence imaging, hypoxia, EVOS, onstage incubator
Scientific poster (2017) Hypoxia measurements in live and fixed cells using fluorescence microscopy and high-content imaging  EVOS, fluorescence microscopy/fluorescence imaging, high content analysis, hypoxia, live-cell imaging, onstage incubator
Video Compilation of live cell imaging videos using Invitrogen fluorescent reagents  
This video demonstrates novel product brands from Invitrogen for live cell imaging include CellLight targeted fluorescent proteins, CellROX reagents for oxidative stress, CellEvent caspase 3/7 detection reagent, and many more fluorescent dyes and probes.
cell structure-mitochondria, CellEvent, CellLight, CellROX, fluorescence microscopy/fluorescence imaging, live-cell imaging, oxidative stress
Video Detection of oxidative stress with CellROX Green in U2-OS cells 
U2-OS cells were plated at 200,000 cells per dish on a 35 mm glass bottom dish (MatTek) and cultured overnight. The cells were rinsed once and loaded with 5 µM CellROX Green Reagent (Cat. No. C10444) and 100 nM TMRM in Live Cell Imaging Solution (LCIS, Cat. No. A14291DJ) for 15 minutes at 37 degrees C. After loading, cells were imaged live with no washing every 20 seconds for seventy minutes on the DeltaVision Core inverted microscope at 37 degrees C. Menadione was added to a final concentration of 100 uM after ten minutes of baseline acquisition. The time lapse data shows the loss of signal from TMRM in the red channel as mitochondrial function decreases, concomitant with the onset of a nuclear signal in the green channel as CellRox Green is oxidized and the reagent migrates to the nucleus to generate a fluorogenic response as the active form of the dye binds to DNA.
CellROX, fluorescence microscopy/fluorescence imaging, live-cell imaging, oxidative stress
Webinar Breathing new life into hypoxia research 
Although the significance of hypoxia in biological processes is well known, creating model systems with accurate control of hypoxic conditions is extremely difficult without access to elaborate systems that allow precise control and maintenance of temperature, humidity, and gases (CO2 and O2) during an experiment. In this webinar, we will discuss: Overview of hypoxia in human diseases Classical methods for setting up hypoxic conditions Novel instruments and reagents for imaging cells in hypoxic conditions
EVOS, fluorescence microscopy/fluorescence imaging, high content analysis, hypoxia, onstage incubator
Webinar From the hood to the microscope; Revolutionizing cell-based imaging 
Join us for an interactive, educational webinar that showcases some of the latest advancements in cell-based research and imaging. Learn as we explore how to obtain superior results through the careful selection of reagents and the optimization of your imaging workflow— from growing cells on suitable surfaces through to image capture. Particular attention will be given to growing and monitoring cells for imaging, fluorescent labeling of live cells, critical considerations for time-lapse imaging and optimizing live cell imaging. Topics include: Choose a suitable imaging culture vessel for studying cell growth and viability using bright-field microscopy Label cells to maximize signal-to-noise for fluorescence imaging Prepare culture conditions and capture images without losing temporal data for time-lapse imaging Achieve hypoxic culture conditions by modifying gas conditions or employing cell spheroid cultures
cell health, EVOS, FLoid, fluorescence microscopy/fluorescence imaging, hypoxia, live-cell imaging, onstage incubator