Authentic physoxia in a bench-top hypoxia incubator/workstation.
In the field of tissue culture, there is a rapidly growing appreciation that a physiological cellular oxygen environment is essential for the analysis of cellular processes with respect to function and metabolism.
Rising to this challenge, Oxford Optronix has developed the HypoxyLab™ - a fully-featured, ergonomically engineered and easy to use normoxia/hypoxia workstation that provides a HEPA-filtered environment in which oxygen, carbon dioxide, temperature and humidity can be precisely controlled.
In short, HypoxyLab™ is a compact hypoxia workstation and incubator for everyday use, suitable for applications including,
As every scientist knows, it’s the partial pressure of oxygen that reflects the oxygen that cells actually ‘see’. The partial pressure of oxygen varies not only with oxygen concentration in the atmosphere, but also with altitude and prevailing weather conditions. For this reason we believe it is insufficient to express hypoxia in terms of a barometrically uncompensated percent oxygen concentration alone. HypoxyLab is therefore the world’s first and only hypoxia workstation that directly regulates its environment using the partial pressure of oxygen, expressed in absolute units of mmHg or kPa, making it insensitive to meteorological conditions or the altitude of the laboratory. This scientifically rigorous approach provides researchers the ability to replicate physiological oxygen in vitro, at the highest levels of accuracy and reproducibility.
See our related article, "The importance of oxygen partial pressure in cell culture"
HypoxyLab uses purpose-designed gas control hardware and an advanced Model Predictive Control (MPC) gas control algorithm, developed in-house, to create and maintain a fully humidified, highly stable oxygen-controlled environment for tissue culture. It responds rapidly to single or multiple setpoint changes, with minimal or zero overshoot characteristics, all while consuming surprisingly little nitrogen gas.
Dissolved oxygen can optionally be sampled directly and continuously from cell cultures (e.g. from the cell monolayer itself) and/or from media being conditioned within the HypoxyLab, via a miniature optical sensor that can be fed into the chamber through a dedicated, sealable side-gland. Oxygen readings (in mmHg) can be read from the OxyLite™ monitor or can be displayed and recorded by the HypoxyLab itself. OxyLite™ monitor required.
HypoxyLab’s 'letterbox' hatch entry system affords quick and easy transfer of cell plates and accessories into or out of the chamber without the need for an air lock or isolation hatch. HypoxyLab senses when items are passed through the hatch, temporarily generating a slight over-pressure to minimize ingress of external atmosphere. This immediate response ensures that oxygen concentrations within the chamber remain stable even when transferring items directly into or out of the chamber.
See our related article and video demonstration, "The HypoxyLab easy-entry system"
HypoxyLab includes a built-in UV source within the water reservoir and a user replaceable HEPA filter that continuously scrubs the chamber atmosphere, ensuring that cultures and media are protected from contamination.
Chamber oxygen, CO2, humidity and temperature are all set and controlled from the intuitively designed user interface and integrated touchscreen, which simultaneously displays the current real-time levels of these parameters in both digital and trace formats.
HypoxyLab’s lightweight and durable cover can be easily removed. This not only allows for easy loading of media and consumables but makes routine cleaning and disinfecting of the workstation a practical reality. Its ergonomic design ensures natural, relaxed operation, with an angled vision panel combined with adjustable LED illumination that provide excellent visibility.
In support of powered devices deployed within the HypoxyLab (digital microscopes, shakers, micro-centrifuges, etc.) the HypoxyLab features two dedicated thru-ports for the transfer of power and/or data cables up to 7 mm in diameter through the chamber wall without compromising the internal environment. The HypoxyLab ships with a pair of blanking glands.
The optional thru-port gland kit is described here.
A 6 mm stainless steel pass-thru waste port is also built into the chamber wall as standard, intended for the aspiration of media, supernatants etc. from within the chamber via an externally connected vacuum pump.
Internal oxygen, CO2, temperature and humidity parameters are continuously recorded onto internal flash memory and can be exported to a USB flash drive. Data files can subsequently be analysed and played back using the free LabChart® Reader by ADInstruments.
HypoxyLab’s compact design and advanced gas control algorithms ensure rapid and ultra-smooth transitions to setpoints, with industry-leading gas consumption performance. Together with service-free sensors and a user changeable HEPA filter, these characteristics minimize the long term cost of ownership.
Our confidence in the reliability of the HypoxyLab is reflected in our comprehensive manufacturer's warranty, covering defects in materials and workmanship for a period of 2 years from the date of delivery. Extended warranty packages, tied into preventative maintenance servicing are also available.
Technical and application support is both comprehensive and free throughout the lifetime of our products. It is provided by highly experienced support staff who pride themselves in providing personal and responsive assistance via on-site visits, video calls, or email and will 'go the extra mile' to ensure that our users can extract the very best value and opportunity from our products.
Cells react in different ways both metabolically and morphologically depending on the environmental factors maintaining and interacting with them.
In tissue culture, faithfully reproducing the in vivo cellular environment is vital for meaningful analysis of both cell metabolism and cell function.
Cells in vivo experience oxygen concentrations in the range of 5 – 80 mmHg (approx. 0.5 – 10% oxygen), depending on the tissue type. Yet in vitro cell biology is typically performed in incubators in which cells are exposed to atmospheric levels of oxygen (21%), a 'hyperoxic' state for most cell types. In other words, the oxygen concentration typically encountered by cells in traditional incubators is 2 - 20 times that experienced in living tissues!
Maintaining cells in such an oxygen-rich environment can have profound implications on cell metabolism, as illustrated by the large body of scientific evidence surrounding the hypoxia-inducible factor (HIF) pathways.
The HypoxyLab™ reproduces physiological conditions in cell-based research by delivering a contamination-free tissue culture environment that offers precise control of oxygen, CO2, temperature, and humidity. And it does so in a remarkably compact and ergonomic form-factor that offers unmatched ease-of-use and economical gas consumption.
Meanwhile, it is the partial pressure of oxygen, not merely the proportion of oxygen in the environment (% oxygen) that cells in culture actually ‘see’. HypoxyLab adopts this fundamental scientific principle and controls the chamber environment using the partial pressure of oxygen (pO2) expressed in units of mmHg or kPa.
Since the partial pressure of oxygen varies not only with oxygen concentration but also with altitude and prevailing atmospheric pressure, this scientifically rigorous approach substantially enhances HypoxyLab’s accuracy relative to other hypoxia chambers and workstations that rely on % oxygen concentration control alone.
Furthermore, this approach addresses a secondary shortcoming affecting traditional hypoxic/physoxic workstations in use today: reproducibility. By intrinsically correcting for both altitude and atmospheric pressure, data generated by one HypoxyLab user can ALWAYS be directly compared to data generated by another HypoxyLab user, wherever they may be situated.
(Selection only; click here for a full list)
by Justin Croft, January 2025
by Justin Croft, December 2024
by Justin Croft, April 2024
by Justin Croft, November 2023
Presented by Michael Rau, 23 September 2022
Effect of hypoxia on Cystic Fibrosis Transmembrane conductance Regulator channel corrected by Elexacaftor/Tezacaftor/Ivacaftor (2024). Pascarel K, Colas J, et al., and Vandebrouck C. Medical Research Archives
Poison cassette exon splicing of SRSF6 regulates nuclear speckle dispersal and the response to hypoxia (2023). de Oliveira Freitas Machado C, Schafranek M, Brüggemann M et al. PNAS
Discovery of endogenous nitroxyl as a new redox player in Arabidopsis thaliana (2023). Arasimowicz-Jelonek, M., Floryszak-Wieczorek, J., Suarez, S., et al. Nature Plants
A combined experimental and computational framework to evaluate the behavior of therapeutic cells for peripheral nerve regeneration (2022). Eleftheriadou D, Berg M, Phillips JB and Shipley RJ. Biotechnol Bioeng
Impact of the acidic environment on gene expression and functional parameters of tumors in vitro and in vivo (2021). Rauschner M, Lange L, Hüsing T, Reime S, Nolze A, Maschek M, Thews O & Riemann A. J Exp Clin Cancer Res
“The HypoxyLab has been a great addition to our laboratory. It’s very easy to use, making it straightforward to achieve accurate and consistent levels of hypoxia, has a small footprint and uses less gas than most other workstations.”
Dr Dan Peet, Associate Professor at the School of Biological Sciences, The University of Adelaide, Australia
“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
“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
Gas control | 3 channel gas mixer controlled by a Model Predictive Control (MPC) algorithm |
Gases | Air, Nitrogen, CO2 (1-4 bar inlet pressure) |
Contamination control | Integrated HEPA filtration |
Chamber working volume | approx. 130L |
Chamber working area | approx. 500 mm (W) x 390 mm (D) |
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 |
Enclosure (removable) | Polyethylene terephthalate G (PET-G) |
Transfer hatch | ‘Easy-Entry’ letterbox with internal flap (usable dimensions 204 mm (W) x 88 mm (H)) |
OxyLite thru-gland | 1 x integrated thru-gland for OxyLite oxygen sensor |
Waste port | 1 x integrated stainless steel 6 mm waste port pass-thru (for vacuum line) |
Display | High contrast 800 x 480 pixel LCD touchscreen |
External USB ports | 1 x Type A (data recording and firmware upgrades) ; 1 x Type mini-B (pass-thru for internal USB port) |
Internal, powered USB port | Integrated |
Power | 110 – 240V AC 50/60 Hz, 500W max |
External dimensions | Width 590 mm, Depth 690 mm, Height 800 mm |
Weight | 28 Kg / 62 lbs |
Operating temperature range (external) | 15 – 30 °C |
Oxygen control method | MPC using an optical oxygen sensor |
Oxygen control range | 1 – 175 mmHg (0.1 - 23%); user programmable via touchscreen |
Oxygen calibration | Manual, recommended 6-monthly |
Oxygen accuracy | ± 0.5 mmHg (<20 mmHg) ; ± 1 mmHg (<40 mmHg) ; ± 2 mmHg (<80 mmHg) ; ± 3 mmHg (<130 mmHg) ; ± 5 mmHg (<175 mmHg) |
Oxygen resolution | 1 mmHg |
CO2 control method | MPC using an IR CO2 sensor |
CO2 control range | 1.0 – 18.0%; user programmable via touchscreen |
CO2 calibration | Manual, recommended monthly |
CO2 accuracy | ± 0.25% (<10%); ± 0.5% (<18%) |
CO2 resolution | 0.10% |
Temp control method | PI control algorithm using a band-gap temperature sensor |
Temp control range | Ambient +5°C (min 20°C) – 42 °C; user programmable via touchscreen |
Temp calibration | Pre-calibrated, none required |
Temp accuracy | ± 0.5 °C |
Temp resolution | 0.1 °C |
Temp gradient across chamber | ± 0.5 °C |
Humidity control method | Modified PI control algorithm using a capacitive RH sensor |
Humidity calibration | Pre-calibrated, none required |
Humidity control range | Ambient – 85% RH; user programmable via touchscreen |
Humidity accuracy | ± 2.5% RH |
Humidity resolution | 1% RH |
*Specifications subject to change without notice
What is HypoxyLab™?
HypoxyLab is a cell culture incubator and hands-on workstation that accurately mimics and maintains true physiological oxygen conditions, as well as maintaining temperature, humidity and CO2 conditions like any standard cell culture incubator.
How does HypoxyLab™ differ from other hypoxia workstations?
The HypoxyLab is highly compact, can fit on an ordinary laboratory bench, does not require the use of a dedicated transfer hatch, features built-in HEPA filtration, active humidification and regulates its oxygen environment using the absolute partial pressure of oxygen for maximum precision.
Why mmHg instead of % oxygen?
Using the partial pressure of oxygen to control the oxygen environment within HypoxyLab is simply the more scientifically rigorous thing to do! Since atmospheric (barometric) pressure changes with weather patterns, as well as with laboratory altitude, so does the oxygen concentration if it is merely maintained as a percentage thereof. Instead, HypoxyLab uses the partial pressure of oxygen to compensate for barometric pressure, ensuring that cells cultured within it 'experience' a consistent oxygen concentration whatever the weather and laboratory location. It ensures that data generated in HypoxyLab can be compared with and/or accurately reproduced by any other HypoxyLab user around the world.
How do I get items in and out of the system?
Cell culture plates, medium, pipettors and other objects can be moved in an out of the HypoxyLab via the simple to operate easy-entry hatch at the front, or can be placed in the chamber prior to the start of the experiment by simply lifting off the whole enclosure cover.
Does external air enter the HypoxyLab™ when I open the easy-entry hatch?
The HypoxyLab strictly minimizes the ingress of external air by detecting the opening of the hatch and transiently generating a slight over-pressure. This completely prevents ingress of external air in the vast majority of scenarios. Only at very low chamber oxygen levels and where the hatch is opened for an unusually long time will a temporary, limited oxygen rise be noticeable. This is then corrected so quickly that no adverse effect on cells being cultured can be observed.
Do I need to wear gloves to operate HypoxyLab?
There is no requirement per se for the operator to wear gloves, however, as with all cell culture work, wearing gloves represents good laboratory practice.
How can I clean the HypoxyLab™?
The HypoxyLab can be easily cleaned/disinfected using 70% ethanol or hydrogen peroxide based surface cleaners by removing the enclosure cover to expose all internal surfaces.
What gases are required to operate HypoxyLab™?
The HypoxyLab requires a source of synthetic air (20% oxygen, balanced in nitrogen), 100% CO2, and 100% nitrogen. All gases should be laboratory grade (99.98% ('N3.8') or greater) and be supplied at between 1 - 5 bar (15 - 70 PSI) pressure.
Is the user protected from substances inside the HypoxyLab™?
No. While the HypoxyLab features a built-in HEPA filter that continuously ‘scrubs’ the internal atmosphere, the system does vent externally. The HypoxyLab is NOT therefore a substitute for a fume cupboard or a biological safety cabinet.
How clean is the internal environment?
The built-in HEPA filtration system achieves an atmosphere equivalent to ISO 14644-1 Class 2 within a few minutes of power-up, helping to protect cultures and media from the risk of contamination.
Can I measure dissolved oxygen levels directly from within my cell cultures?
Yes. The HypoxyLab features a dedicated side-gland that supports our fibre-optic dissolved oxygen sensors. These are capable of reading absolute dissolved oxygen in units of mmHg directly from media or cell culture dishes within the chamber. OxyLite™ oxygen monitor required.
Does the user need to be standing to operate the HypoxyLab™?
The system can be operated standing up or from a seated position. A height-adjustable stool is recommended for seated operation.
What is the typical nitrogen consumption rate of the HypoxyLab™?
Nitrogen is used both to generate and to maintain the internal hypoxic environment, as well as to generate the temporary over-pressure when the easy-entry hatch is operated. Of the 3 required gases, nitrogen is therefore consumed the quickest. While usage rates are extremely dependent on the conditions being maintained and the usage ‘profile’ being employed, a standard (50L) laboratory gas cylinder of compressed nitrogen can be expected to last 2-4 weeks in most cases.
Do I have access to the trace data displayed on the touch-screen?
Yes. Trace data are automatically and continuously stored to internal memory and can be copied to USB media at any time. Data files are automatically overwritten after 6 months. Data are written in a format that can be read by LabChart Reader software, available for free from ADInstruments.
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