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. We believe it is simply implausible to express hypoxia in terms of a (barometrically uncompensated) percent oxygen concentration alone. HypoxyLab is the world’s first and only hypoxia chamber that directly regulates its environment using the partial pressure of oxygen, expressed in units of mmHg or kPa, and is thereby insensitive to changing climatic conditions or the altitude of the laboratory. This scientifically rigorous approach enhances HypoxyLab's accuracy and provides researchers with scientific certainty and the ability to replicate ‘real-life’ hypoxia in vitro at the highest level of accuracy achievable.
HypoxyLab creates a fully humidified, stable hypoxia environment less than 20 minutes from switch-on and can deliver stable, near-anoxia conditions in under an hour, all while consuming surprisingly little nitrogen gas. Environmental conditions are delivered using digital gas flow controllers and state-of-the-art sensors for unrivalled performance with an oxygen accuracy of up to +/- 1 mmHg.
HypoxyLab supports the measurement of dissolved oxygen directly from media or cell cultures using any of our OxyLite™ oxygen monitors and a dedicated, sealable side-gland via which an OxyLite oxygen sensor can be fed into the chamber. Considered the ‘gold standard’ for biological and physiological applications, the OxyLite sensor is a tiny, highly accurate, non oxygen consuming sensor that can be accurately positioned anywhere within media or cell cultures to obtain continuous dissolved oxygen measurements. Oxygen readings (in absolute units of mmHg) can be read from the OxyLite display, or can optionally be displayed and recorded by the HypoxyLab itself.
We have teamed with CytoSMART™ to offer the Lux2 digital microscope system, an extremely compact digital microscope solution that can be deployed within the HypoxyLab and is built to withstand long-term exposure to tissue culture environments. The system is supplied with software for installation on a PC kept outside the HypoxyLab and on which live cell culture images are displayed. The CytoSMART™ Lux2 supports time-lapse recording and cloud-based data access.
HypoxyLab includes a built-in, 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 set and controlled from the integrated touch-screen, which simultaneously displays the current real-time levels of these parameters in both digital and trace formats.
An fully customizable oxygen profile program is available from the touch screen, with which the HypoxyLab can be set to automatically subject cells to up to 8 sequentially defined oxygen concentrations, including an option to keep looping the program.
The 4 key environmental parameters are continuously recorded onto internal flash memory and can be exported to a USB flash drive via a port at the rear of the HypoxyLab at any time. Data files can be analyzed and played back using the free LabChart® Reader by ADInstruments.
HypoxyLab’s Easy-Entry hatch system affords quick and convenient transfer of cells, culture-ware and media into the chamber without the need for complicated air locks and isolation hatches. While items are passed through the hatch, HypoxyLab applies the precise gas composition necessary to maintain steady-state conditions. This immediate response eliminates fluctuations in the pO2 of cell media during operation of the Easy-Entry system. An internal ‘letterbox’ style flap acts as an additional barrier against gas displacement when the hatch is opened.
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.
HypoxyLab’s compact and considered design ensures rapid equilibration to set points combined with frugal gas consumption. Service-free oxygen sensors and a user changeable HEPA filter simplify routine maintenance and minimize the long term cost of owning a HypoxyLab.
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 cell environment is vital for accurate analysis of both cell metabolism and cell function.
It is commonly accepted that cells in vivo experience oxygen concentrations in the range of 5 – 80 mmHg (approx. 0.5 – 10% oxygen), depending on the tissue type. Yet the vast majority of cell biology research is still performed in incubators in which cells are exposed to atmospheric oxygen levels (circa 21%), officially a 'hyperoxic' state for most cell types. In other words, the oxygen concentration typically encountered by cells in traditional incubators is at least twice that experienced in normal tissues.
Culturing cells in such rich oxygen environments can have profound implications on cell metabolism and signalling pathways. Famously the hypoxia-inducible factor (HIF) pathways clearly illustrate the importance of environment oxygen in cellular metabolism.
By delivering a contamination-free environment that offers precise and continuous control of oxygen, as well as of CO2, temperature and humidity, HypoxyLab delivers a powerful solution to research and industry looking to reproduce representative physiological conditions in cell-based research.
Meanwhile, it is the partial pressure of oxygen, not merely the % oxygen concentration 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 directly in SI 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 performance accuracy relative to other hypoxia chambers and workstations that rely on % oxygen concentration control alone.
by Justin Croft, 6 September 2021
by Justin Croft, 17 August 2021
“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||Microprocessor controlled, 3 channel Digital Electronic Flow Controller (DEFC)|
|Gases||Air, Nitrogen, CO2 (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|
|Arm ports||Cuff and sleeve or bare-hand operation|
|Transfer hatch||‘Easy-Entry’ letterbox with internal flap (usable dimensions 204 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 ports||1 x Type A (data recording and firmware upgrades) ; 1 x Type mini-B (pass-thru port for internal digital devices)|
|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 (21-40 mmHg) ; ± 2 mmHg (41-80 mmHg) ; ± 3 mmHg (81-140 mmHg)|
|Oxygen resolution||1 mmHg|
|CO2 control method||Feedback algorithm with auto PID – using DEFC and IR CO2sensor|
|CO2 control range||0.1 – 10.0%; user programmable via touch-screen|
|CO2 calibration||Manual, user calibration via touch-screen|
|CO2 accuracy||± 0.25%|
|Temp control method||Feedback auto PID – using Class A Pt sensor|
|Temp control range||Ambient +5°C (min 20°C) – 45 °C; user programmable via touch-screen|
|Temp calibration||None required|
|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 calibration||None required|
|Humidity control range||Ambient – 85% RH; user programmable via touch-screen|
|Humidity accuracy||± 2.5% RH|
|Humidity resolution||1% RH|
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.
The HypoxyLab is highly compact, can fit on an ordinary lab bench, does not require the use of a transfer hatch, features built-in HEPA filtration, active humidification and regulates its oxygen environment using the absolute partial pressure of oxygen for maximum precision.
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 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 fluctuations (or differences in laboratory altitude), ensuring that cells in culture 'experience' a consistent oxygen concentration.
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 into the chamber prior to the start of the experiment by simply lifting off the whole enclosure cover.
The HypoxyLab strictly minimizes the ingress of external air by detecting the opening of the hatch and transiently generating a slight over-pressure. Some external air will enter the chamber but the system is so quick to correct for any atmospheric oxygen ingress that there can be no adverse effect on cell cultures being maintained.
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.
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 parts. The same applies to the enclosure cover, which can be easily and quickly removed.
3 gases are required, namely: synthetic air (20% oxygen balanced in nitrogen), 100% CO2 and 100% nitrogen. All gases should be of typical laboratory grade purity (we suggest 99.995% ('N4.5') or better) and be supplied at between 1 - 4 bar (15 - 60 PSI) pressure for synthetic air and CO2, and at between 3 - 5 bar (45 - 75 PSI) pressure for nitrogen.
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.
The built-in HEPA filtration system achieves an atmosphere equivalent to, or better than, ISO 14644-1 Class 2 within 60 seconds of power-up, ensuring that cultures and media are protected from the risk of contamination.
Yes. The HypoxyLab features a dedicated side-gland that supports our fibre-optic dissolved oxygen sensors, which are capable of reading absolute dissolved oxygen in units of mmHg directly from media or cell culture dishes. This requires any one of our OxyLite™ oxygen monitors.
The system can be operated standing up or from a seated position. A height-adjustable stool is recommended for seated operation.
Nitrogen is used both to generate and maintain an internal hypoxic environment and 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 will be extremely dependent on the conditions being maintained and the usage ‘profile’, we estimate that a large (50L water volume) bottle of compressed nitrogen will last 1-2 weeks with typical daily use of the system.
Yes. Data are automatically and continuously stored to internal memory and can be copied to an external USB media at any time. The internal memory is sufficient for approx. 2 weeks of continuous recordings, at which point old data files are progressively overwritten. Data are written in a format that can be read by LabChart Reader software, available for free from ADInstruments.
(Updated: June 2021)
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1, 2 and 4‑channel dissolved oxygen and temperature monitorsOxyLite™ oxygen monitors