Hypoxia Incubator: “True hypoxia” in a bench-top incubator and workstation
Compact and efficient
Delivers “true hypoxia” using the partial pressure of oxygen
Imitates the oxygen state in vivo
Chamber rapidly equilibrates while consuming less gas
HEPA filtration scrubs the air in seconds
Ergonomically engineered with built-in touch-screen controls
Optional – integrated in situ dissolved oxygen measurements with the OxyLite add-on
Optional – digital microscope add-on
Within tissue culture research, there has recently been increasing awareness and need for approaches capable of producing accurate, reproducible, and physiologically relevant cell environments vital for reliable analysis of relevant metabolism and function in various cellular processes closer to in vivo situations.
To reproduce more relevant physiological environments in cell-based research, a workstation, free of contamination, offering accurate and continuous control of O2, CO2, humidity and temperature is essential. To this end, the HypoxyLab has been developed as an exceptional, fully-featured, ergonomically designed and practical combined hypoxia incubator and workstation that delivers a highly stabilized HEPA-filtered environment in which levels of O2, CO2, relative humidity and temperature are all accurately controlled.
The HypoxyLab is an easy to use compact, bench-top hypoxia workstation and incubator perfect for day to day use.
Representative research topics of in vitro cellular applications for the HypoxyLab include:
- Cancer and radiation research
- Stem cell research
- Virology research
- Drug development and pharmacology
- Oxygen deprivation effects on specific tissues
Hypoxia and Cell Culture Rationale
Cells can respond in vastly different ways both morphologically and metabolically depending on exposure to different environmental influences.
In tissue culture research, reliably replicating cellular environments in vivo is essential for an accurate evaluation of cell function, structure and metabolism.
Most cells in vivo are exposed to tissue type dependent O2 concentrations ranging from 5 – 80 mmHg (0.5 – 10% oxygen). By contrast, the majority of in vitro cell culture investigations are still carried out in incubators where cells are only subjected to atmospheric oxygen levels (≈21%), a hyperoxic state for most native type of cells. Conversely, the oxygen concentration “seen” by those cells in conventional incubators could be twice that found in their respective ‘natural’ tissues.
Subjecting cells to higher O2 concentrations outside of their ‘normal’ physiological range (even for a brief time) can have profound repercussions on cell metabolism and signalling pathways. For example, the hypoxia inducible factor (HIF) pathways exceptionally demonstrate the potential for precise oxygen pressures to drastically change intra-cellular metabolism.
Furthermore, it’s important to realize that cells “react to” the partial pressure of oxygen and not just the percentage of oxygen concentration. The HypoxyLab uses this essential scientific principle and regulates the chamber ‘atmospheric composition’ using the precise partial pressure of oxygen (pO2) expressed in mmHg or kPa; because the partial pressure of oxygen varies not only with oxygen concentration but also with altitude and prevailing atmospheric pressure. This valid scientific approach significantly improves the HypoxyLab’s performance accuracy when compared to other hypoxia chambers and workstations that rely on percentage oxygen alone.
Combined with the HEPA-filtrated, uncontaminated environment, HypoxyLab supplies a precise and continuous control of O2, as well as of CO2, humidity and temperature. As such, a new and improved solution is available to academic and industry professionals looking to more accurately and controllably replicate the characteristic physiological environments in cell-based investigations.
HypoxyLab, unlike other systems that simply express oxygen concentration in a percentage value, uses the absolute partial pressure of O2 (in kPa or mmHg) as a determinant to adjust its internal atmosphere. This scientifically rigorous approach has been used to eliminate substantial errors that would otherwise arise from changes in either barometric pressure and altitude. This provides academic researchers and industry professionals with high reliability that they can emulate ‘real-life’ hypoxia in vitro with a high level of precision.
Rapid changes and highly efficient
HypoxyLab can generate fully humidified, temperature stable hypoxic conditions (pO2 of 10 mmHg) in under 20 minutes from turning it on and can supply a stable, near-anoxia environment (pO2 of 1-2 mmHg) in under an hour, all while using surprisingly small quantities of N2 gas.
Dissolved oxygen measurements in situ
HypoxyLab provides a connection with the OxyLite oxygen sensor option allowing the user to determine oxygen availability in the current microenvironment where cells are growing, thereby uncovering the oxygen gradient between the HypoxyLab’s atmosphere and the cell layer. This is achieved by providing a connection within the chamber to attach the OxyLite’s oxygen sensor. The OxyLite sensor is a glass fiber-based, highly precise, non-oxygen consuming sensor that when positioned anywhere within the cell culture or media can provide continuous dissolved O2 measurements. This data is conveniently displayed the HypoxyLab’s touch-screen along with chamber O2, CO2, temperature and relative humidity.
An internal connection for an OxyLite sensor facilitates in situ dissolved oxygen measurements. Optional OxyLite system is required
Live cell imaging
HypoxyLab provides the user the option to image their cells without having to remove them from the chamber with the CytoSMART 2™ microscope. The affordable and compact CytoSMART 2 microscope fits perfectly into the HypoxyLab and is designed to withstand long-term exposure to the cell culture environmental conditions. The system supports time-lapse recording with cloud-based access to all data from any browser-capable computer or smart device in addition to the provided tablet. This tablet, which is attached outside the HypoxyLab can display live cell culture imaging.
CytoSMART 2 digital microscope for imaging cells within the HypoxyLab
The HypoxyLab uses a replaceable HEPA filtration unit that continuously cleans the chambers atmosphere. This HEPA filter can achieve an atmosphere equivalent to, or better than ISO 14644-1 Class 2 standard within 30 seconds from start-up. This ensures that cell culture, media and components within the chamber are protected from becoming contaminated by air-circulation factors. Furthermore, rather than a water pan the internal humidification is achieved by ultrasound nebulization, removing another common source of contamination.
Replaceable HEPA filtration unit
HypoxyLab’s intuitive touch-screen allows for set point values of the partial pressure of oxygen (pO2), CO2, temperature and humidity to be adjusted and tightly controlled by auto-calibrating sensors, and by advanced digital gas flow regulators. By incorporating these state-of-the-art technologies, HypoxyLab is the most reliable and accurate system on the market today, fully capable of regulating the partial pressure of oxygen with precision of up to +/- 1 mmHg (approximately 0.13%)
HypoxyLab’s modern color touch-screen supports convenient control over all instrument settings with concurrent real-time data presentation in a digital and graphical format. The screen displays essential information such as the actual and set-point values for O2, CO2, humidity and temperature. Appropriate warnings alert the user with advanced notice of a tank running low on gas or to top up the water reservoir.
The HypoxyLab has an internal flash memory where recorded parameters are stored and saved continuously. This data can be exported to a USB through a port near the rear of the HypoxyLab any time. These exported files are compatible with the free LabChart Reader by ADInstruments.
All data from the HypoxyLab can be saved to USB flash memory
An easy-entry hatch provides the user a quick and simple way to pass cell cultures, media and other items in or out of the HypoxyLab chamber. This letterbox sized hatch combined with slight atmospheric over-pressure removes any need for overly-complicated and expensive air locks or isolation hatches. Once the hatch is opened, the chamber immediately compensates by applying the precise gas composition injection required to maintain a low oxygen environment.
A direct easy-entry hatch without disturbing the controlled environment of the HypoxyLab. A time-consuming and expensive transfer hatch is not needed!
Lightweight removable cover
HypoxyLab has been engineered with a sturdy lightweight cover allowing a single person to effortlessly detached it from the base. This means the HypoxyLab not only allows for easy loading of larger volumes of media and consumables, but also makes regular cleaning and disinfecting of the chamber an uncomplicated task.
The light and sturdy cover easily lifts off the base
The HypoxyLab has been ergonomically engineered to ensure the user a natural and relaxed operational position. Internally there are height-adjustable shelves, LED lighting and a USB power socket to power various accessories like the CytoSMART 2 microscope. Externally the angled display panel provides excellent visibility while working within the chamber. The easy to use cuff and sleeve system allows the user to choose between bare hand, gloved or gauntlet (plugged) operation.
Programmable oxygen profiles
HypoxyLab’s intuitive user-interface provides researchers the ability to develop customized pO2 profiles that automatically cycle across eight user defined oxygen settings. Knowing the HypoxyLab adjusts the hypoxia equilibrium within minutes, as opposed to hours, adds real value to the notion of ‘oxygen profiling’ all while preserving gas expenditure.
Program 8 oxygen conditions via the touch-screen to run consecutively
Low operational and maintenance cost
The HypoxyLab’s small footprint minimizes dead-space and assures rapid internal atmospheric equilibration to programmed set-points with minimal gas expenditure. Its long-life, automatic calibrating sensors and user exchangeable HEPA filters simplify scheduled maintenance minimizing operational and maintenance costs.
Touch Screen Views
The HypoxyLab’s fully functional color touch-screen displays real-time data presented in both digital and graphical formats. Through the screen the user has full control over all gas, humidity and temperature set-points, including programmable O2 profiles and a wider array of system settings via a control panel.
All programable parameters (O2, CO2, humidity and temperature) are continuously presented in separate graphic trace format
Icons at the bottom of the display allow values for O2, CO2, relative humidity, and temperature to be set appropriately through a simple dialogue box pop-up
The HypoxyLab can be programmed, with an 8-step continuous cycling program, O2 adjustable profile. These steps may be looped as many times as the user requires
The control panel further offers a wide range of system functions and settings
“The experience with the HypoxyLab has been great. It’s footprint is very small, a distinct advantage. The quality of technical support/service available is unmatched. Issues have been addressed by Oxford Optronix very swiftly and accurately along with a very personalized service.”
– Dr. Anurag Kulkarni, Department of Medicine, Imperial College, London, UK
“Working with the HypoxyLab is great. It is reliable, easy to use and requires quite little gas (as compared to larger chambers). Especially the possibility of using pre-defined oxygenation profiles is an important additional benefit. During these profiles the composition of the gas is changed rapidly and the actual gas fractions are well documented continuously. This feature allows interesting hypoxia/re-oxygenation experiments.”
– Prof. Oliver Thews, Faculty of Medicine, Martin-Luther-University, Halle-Wittenberg, Germany
(Last updated July 2018)
Riemann A, Reime S and Thews O. Tumor Acidosis and Hypoxia Differently Modulate the Inflammatory Program: Measurements In Vitro and In Vivo. Neoplasia (2017); 19(12), 1033-1042 [LINK]
Riemann A, Güttler A, Haupt V, Wichmann H, Reime S, Bache M, Vordermark D and Thews O. Inhibition of Carbonic Anhydrase IX by Ureidosulfonamide Inhibitor U104 Reduces Prostate Cancer Cell Growth, But Does Not Modulate Daunorubicin or Cisplatin Cytotoxicity. Oncology Research (2018); 26, 191-200 [LINK]
Kulkarni A, Mateus M, Thinnes C, McCullagh J, Schofield C, Taylor G and Bangham C. Glucose Metabolism and Oxygen Availability Govern Reactivation of the Latent Human Retrovirus HTLV-1. Cell Chemical Biology (2017); 24(11), 1377-1387 [LINK]
Gagner J, Lechpammer M and Zagzag D. Induction and Assessment of Hypoxia in Glioblastoma Cells In Vitro. Methods Molecular Biology (2018); 1741, 111-123 [LINK]
Specifications for HypoxyLab
|Gas control||Microprocessor controlled, 3 channel Digital Electronic Flow Controller (DEFC)|
|Gases||Air, Nitrogen, CO2 (4 bar inlet pressure)|
|Contamination control||Integrated HEPA filtration (generating equivalent of ISO 14644-1 Class 2 atmosphere within 30 s)|
|Chamber volume||Approx. 90L|
|Chamber working area||500 mm (W) x 280 mm (D) (approx. 1,100 cm² / 170 inch² usable surface area)|
|Chamber storage capacity||Up to 40 multi-well plates or 10 cm Petri dishes (assumes 2 shelf units; excludes storage on working area)|
|Maximum gas flow rate||15L/minute/gas|
|Arm ports||Cuff and sleeve or bare-hand operation|
|Transfer hatch||‘Easy-Entry’ letterbox with internal flap (dimension 200 mm (W) x 88 mm (H))|
|Power||Auto-switching 110 – 240V AC 50/60 Hz, 500W max|
|Enclosure||PET-G. Lightweight and removable|
|External USB port||Integrated. Supports data recording and firmware upgrades|
|Internal, powered USB port||Integrated|
|Physical dimensions||800 mm (H) x 590 mm (W) x 690 mm (D)|
|Weight||25 Kg / 55 lbs|
|Operating temperature range|
|15 – 30 °C|
|Oxygen control method||Feedback algorithm with auto PID – using DEFC and optical O2 sensor|
|Oxygen control range||1 – 140 mmHg; user programmable via touch-screen|
|Oxygen calibration||None required|
|Oxygen accuracy||± 0.5 mmHg (1-20 mmHg)|
± 1 mmHg (20-40 mmHg)
± 2 mmHg (40-80 mmHg)
± 3 mmHg (80-140 mmHg)
|Oxygen resolution||1 mmHg|
|CO2 control method||Feedback algorithm with auto PID – using DEFC and IR CO2sensor|
|CO2 control range||0.0 – 20.0%; user programmable via touch-screen|
|CO2 accuracy||± 0.25%|
|Temp control method||Feedback auto PID – using Class A Pt sensor|
|Temp control range||Ambient +5°C – 42 °C; user programmable via touch-screen|
|Temp accuracy||± 0.5 °C|
|Temp resolution||0.1 °C|
|Temp gradient across chamber||± 0.5 °C|
|Humidity control method||Nebuliser with auto PID using pre-calibrated capacitive sensor|
|Humidity control range||Ambient – 90% RH; user programmable via touch-screen|
|Humidity accuracy||± 2% RH|
|Humidity resolution||1% RH|
Frequently Asked Questions
What is the HypoxyLab?
The HypoxyLab is a bench-top cell culture incubator and workstation that can precisely mimic and maintain “true” physiologically relevant oxygen exposure conditions in addition to regulating CO2, humidity and temperature conditions comparable, if not better than, other standard cell culture incubators on the market.
How does HypoxyLab differ from other hypoxia workstations?
Unlike other hypoxia workstations the HypoxyLab is compact and can be placed on any ordinary lab bench. Being compact, allows it to quickly modulate and compensate for O2 changes within, thereby excluding the need for an expensive transfer hatch. The HypoxyLab features a built-in HEPA filtration system, active ultrasound nebulizer for humidification and sensors which accurately regulate the O2 environment using the partial pressure of oxygen.
Why use mmHg O2 as opposed to percent oxygen?
Since barometric (atmospheric) pressure changes daily, or even hourly, with weather patterns, so does the oxygen concentration if percentage oxygen is used to determine oxygen concentration regulation. The cells within these chambers can therefore unknowingly be exposed to less or more molecular oxygen than the user is aware of. HypoxyLab, by using the partial pressure of oxygen offsets barometric pressure fluctuations (or even variances in laboratory atmospheric pressure, if working in an over- or under-pressure environment due to safety concerns), ensuring that cultured cells ‘experience’ consistent and more determinable oxygen concentrations. Therefore, employing the partial pressure principle for regulating oxygen concentration in the HypoxyLab is the most scientifically precise thing to do!
How do I get items in and out of the system?
During operation of the HypoxyLab objects like cell culture plates, medium, or pipettes can be moved in and out of the HypoxyLab through the easy-entry hatch at the front of the chamber. Alternatively, simply removing the lightweight enclosure cover items can be placed onto the HypoxyLab base prior to the start of a study.
When I use the easy-entry hatch does external air enter the HypoxyLab?
The HypoxyLab stringently minimizes external air entry by instantly detecting the opening of the easy-entry hatch and pushes nitrogen into the system which generates a slight over-pressure within the chamber. The minimal amount of air that does enter is quickly corrected for by the system and should have no bearing on the cells or media within.
Are gloves necessary to operate the HypoxyLab?
Wearing gloves represents good laboratory practice with all cell culture work, but there is no requirement for the user to wear them.
How can I clean the HypoxyLab?
By removing the enclosure cover to expose all internal parts, the HypoxyLab can be disinfected and cleaned using 70% ethanol or hydrogen peroxide-based cleaners. The lightweight cover can be quickly and easily taken off by one person.
What gases are required to operate HypoxyLab?
Laboratory grade quality 20% oxygen (balanced with nitrogen; a.k.a. ‘synthetic air’), 100% CO2, and 100% nitrogenstocked at approximately 4 bar pressure (60 PSI).
Is the user protected from substances inside the HypoxyLab?
The HypoxyLab is not a suitable substitute for a biological safety cabinet or fume hood because the HEPA filtration that scrubs the internal atmosphere vents externally, meaning the user cannot be fully protected from the substances within.
How internal environment clean?
Yes, when activated the HEPA filter accomplishes an atmosphere equivalent to, or better than, ISO 14644-1 Class 2 within 30 seconds of start-up. This ensures that cell cultures and media are at very low risk of contamination by air.
Can I measure dissolved oxygen levels directly within my cell culture media?
Yes, with the addition of our OxyLite™ oxygen monitor. The HypoxyLab features a dedicated internal connector that supports a fibre-optic sensor capable of detecting absolute dissolved oxygen in units of mmHg directly by placing the sensor fibre in media or cell culture dishes.
What is the typical N2 consumption rate of the HypoxyLab?
Nitrogen is used to create and sustain the internal O2 set-point environment, and to generate an over-pressure when opening the easy-entry hatch. Therefore, of the three required gases it is used up the quickest. While usage rates will generally be dependent on the set-point usage ‘profile’ conditions being maintained, we estimate with daily use that a large (50L water volume) cylinder of compressed nitrogen will last approximately 1 week.
Can I access the trace data displayed on the touch-screen?
Yes, this data is automatically and continuously stored to internal memory and can be copied to an external USB media at any time. The internal memory can support approximately 2 weeks of continuous recordings, at which point old data files will be overwritten. The data format can be read by free LabChart Reader software,from ADInstruments.
The compact CytoSMART 2 microscope is perfectly created for live cell imaging and monitoring. Cell cultures can be recorded and watched via the cloud, so they can be viewed anytime, anywhere on a smart device or computer.
Dissolved oxygen sensor – OxyLite system
The OxyLite is the ‘gold standard’ for physiological and biological applications. This oxygen monitor functions by means of a small, fibre-optic sensor to determine very precise measurements of dissolved oxygen from within cell cultures and medium through a dedicated connector positioned within the HypoxyLab.
The OxyLite technical specifications can be found on the system page.
A stretchy thermal jacket is now included with the HypoxyLab. This thermal jacket permits the application of higher humidity thresholds reducing evaporation from cell cultures and lowering power consumption.
Accelerates the oxygen conditioning of tissue culture media.
The HypoxyCOOL is a, scientifically authenticated, protocol-driven way to condition oxygen content of tissue culture media to be used within the HypoxyLab chamber. It can rapidly and accurately reduce media oxygen concentrations to target values while maintaining media sterility, pH and temperature.
Thereby HypoxyCOOL improves research results by eliminating media conditioning errors.
|Refrigeration temperature||2 – 6 ºC, adjust at 0.1 ºC increments|
|O2 control||0.5 – 15.0%, adjust at 0.1% increments|
|CO2 control||0 – 10.0%, adjust at 0.1% increments|
|Conditioning cycle range||0.5 – 16 hours|
|Shaker speed range||70 – 120 rpm|
|Gas requirements||N2, CO2 (2.5 – 5 bar inlet pressure)|
|Media conditioning capacity||Up to nine 500 ml bottles|
|Power supply||AC 120 or 240V, 15A, 500W, 60Hz|
|Dimensions||87 cm x 51 mm x 63 cm (H x W x D)|
Customized Multi-stage regulator
Multi-stage, 5.0 purity regulator. Outlet: 15 µm in-line filter with ¼” NPT F.
Compressed Synthetic Air with inlet connection standard CGA590 (US);
Compressed Nitrogen with inlet connection standard CGA580 (US);
Compressed CO2 with inlet connection standard CGA320 (US).