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Frequently Asked Questions

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Product Background and Usage

• How do you set up an experiment in the Modular Incubator Chamber (MIC-101)?

Simply place your tissue culture cells in the chamber using one or more of the trays, add a container with filtered water for humidity, attach a flow meter to the unit, flush for several minutes with the desired gas mixture, then seal the chamber and place in your 37 ̊C incubator. See the instruction sheet for detailed protocols.

• How do I know when I have completely flushed the Modular Incubator Chamber?

Typically, at a flow rate of 50 liters/minute (LPM), 2 minutes is sufficient to fully exchange the chamber gas with your desired mixture, such that a total of 100 L of gas flush through the MIC-101 will obtain a 100% exchange of gases.

• Why is there no Edge Effect when using plates in the Modular Incubator Chamber (MIC-101)?

In conventional continuous airflow CO2 incubators, 96-well plates and 24-well plates typically experience increased evaporation in the outside wells, known as the Edge Effect. This is due to increased movement of air experienced mostly by the outside wells, resulting in lower liquid volumes, changes in pH, and altered nutrient, salt and reagent concentrations. The longer the incubation times, the more pronounced the Edge Effect becomes. Because there is no airflow inside the MIC-101, the Edge Effect is eliminated.

• What is the difference between the terms Physioxic, Normoxic, Hypoxic, and Hyperoxic?

Physioxic refers to the level of oxygen found under normal physiologic conditions within tissues. Depending on the organ or tissue, physioxic levels are oxygen typically between 1 and 8%, while blood oxygen is normally 6-16%. Normoxic is most often used to describe the level of atmospheric oxygen, 21%, but depending on the context, is also used to describe physioxic oxygen levels. Since normoxic can have multiple meanings for the level of oxygen implied, it is important to understand the context of how the term is used. We suggest that this term never be used due to the potential for confusion. Hypoxic is used to describe a condition when oxygen is insufficient at the tissue level to maintain adequate homeostasis, typically less than 1%. Although, “hypoxic” is also commonly used to describe any oxygenation level less than the 21% atmospheric oxygen. Again, depending on the context, hypoxic could be referring to any oxygen level less than 21%. Hyperoxic refers to any oxygen percentage greater than 21% atmospheric oxygen.

• How do I create low oxygen media?

Low oxygen media can be created using the MIC-101 chamber. Place your tissue culture flasks with media into the chamber using one or more of the trays, add a container with sterile water for humidity, attach a flow meter to the unit, flush for 2 minutes at 50 L/min with a low oxygen gas mixture, and then seal the chamber. Allow the oxygen from the media to diffuse into the air in the chamber for 1 hour. After 1 hour, you can flush the chamber again with the low oxygen gas mixture to ensure low oxygen in the media or use the media with cells for experimentation at this point. Refer to the detailed instructions for the MIC-101 chamber and flow meters as needed.

• How do I humidify the Modular Incubator Chamber (MIC-101)?

To humidify the chamber, place an open petri dish or similar container containing 10-20 ml of water into the chamber. The petri dish fits below the trays on the base of the chamber or just place the petri dish on one of the trays.

• How do I adjust the retaining clamp on the Modular Incubator Chamber if it doesn’t seem to be sufficiently tightening?

If the clamp needs to be tightened, use a 3/8” open end box wrench to turn the adjustment bolt clockwise. The bolt is only accessible when the clamp is in the fully open position.

• How does the flow meter work?

Our flow meters are designed to accurately measure the rate of airflow movement passing from the gas source into the modular incubator chamber, which is measured in liters per minute (LPM). A moving float rises as the air pressure from the gas moves through the flow meter. The vertical position of the float aligns with the labeled scale, measuring 5-50 LPM. Do not exceed the maximum 50 LPM of the flow meter.

• What type of regulator should I use for my gas canister?

You can use either a one-stage or two-stage regulator. The recommended inline pressure necessary to create a flow rate of approximately 50 LPM is between 2 and 4 PSI. You can obtain them through Fisher Scientific or wherever you purchase your laboratory supplies. Usually, suppliers of bottle gases will also carry the appropriate regulators.

• How is the MIC-101 better than a standard CO2 incubator with an opening door?

Because of the fully closed environment, the Modular Incubator Chamber provides distinct advantages over standard CO2 incubators. Firstly, the lack of air currents prevents the edge effect, where the outside wells of multi-well plates experience extensive evaporation. Secondly, the closed environment helps to protect against contamination, which is a common pitfall for standard incubators. Lastly, the frequent opening of standard incubator doors creates fluctuations in oxygenation, temperature, and humidity, which can create inconsistencies in experimental results. The Modular Incubator Chamber maintains a consistently stable environment throughout the incubation period; something that cannot be achieved with standard incubators.

• How do I get rid of toxic gases when I clean the incubator?

If toxic gases are used as part of the experiment, the user needs to turn off the gas supply, then purge the chamber for 2 minutes (see purging suggestions above).

• Can I autoclave the Modular Incubator Chamber (MIC-101)?

We do not recommend autoclaving the incubator chamber or its components. Autoclaving may cause cracking or warping of the polycarbonate material, resulting in the loss of an air-tight chamber.

• I already have a low oxygen incubator, so why would I need the MIC-101?

Great! You are on your way to achieving optimal cell culture but there are some additional benefits of having an air-tight environment for cell culture using the MIC-101. First, the MIC-101 has no air flow, so it eliminates the Edge Effect observed when the outer wells of multi-well plates experience extensive evaporation. Second, the MIC-101 minimizes contamination risk, a common pitfall of open door incubators. Third, it maintains a consistent environment in terms of humidity, oxygenation, and temperature regardless of door openings. Lastly, its small size minimizes the total consumption of expensive tri-gas mixes compared to a continuous flow incubator.

• How does the Embrient incubator chamber compare to a "glove box incubator”

Each type of incubator has its advantages. A glove box incubator allows the user to perform some manipulation of the plates without disturbing the environment, serving as a workbench and hood, while directly accessing a 37°C controlled environment. Because of the complex setup, initial equipment costs, as well as routine gas costs, will be high. Additionally, these units require dedicated floor space in your tissue culture room. The Embrient MIC-101 incubator is a compact, sealed unit, and after desired conditions are set, the unit can be placed in a standard incubator. A separate workbench, tissue culture hood, and 37°C incubator are required for the workflow. The cost of the MIC-101 is significantly less than that of standard glove box incubators and it requires no additional space, only inside an existing incubator. Additional benefits of the MIC-101 over standard continuous flow incubators is covered in another FAQ above and in this Tech Note.

• How do I gas (flush or purge) the Modular Incubator Chamber?

The Modular Incubator Chamber can be gassed with pre-mixed gases from a single source or separate gases can be combined using the Dual Flow Meter. Follow our instructions for the Single Flow Meter or the Dual Flow Meter, depending on your setup. Typically, at a flow rate of 50 liters/minute (LPM), 2 minutes is sufficient to fully exchange the chamber gas with your desired mixture, such that a total of 100 L of gas flush through the MIC-101 will obtain a 100% exchange of gases.

• How long can I maintain the environment inside the Modular Incubator Chamber (MIC-101)?

Once sealed, the MIC-101 maintains consistent gas levels for at least 72 hours, confidently offering extended experiment durations without environmental disturbance. Longer incubations with sustained conditions may be possible, but each unique set of experimental conditions would need to be verified for performance.

• Are there references citing the use of the Modular Incubator Chamber (MIC-101)?

There are over 4500 citations that reference the Embrient/Billups-Rothenberg Modular Incubator Chamber. View our citations page to learn more and view citations by research area or cell type.

• What percentage of O2 or pO2 should I use?

Optimal oxygen percentage or oxygen pressure for cell culture depends on your cell type and the conditions of the experimentation. Read our Tech Note and view the chart to compare published oxygen levels for different cell types and the observed effects on cells.

• How do I heat the Modular Incubator Chamber (MIC-101)?

There are multiple options for getting your MIC-101 to the desired temperature. You can place the chamber into a standard continuous flow CO2 incubator, a heat only incubator, or a warm room at the desired temperature. It will take a few minutes for the chamber and its contents to equilibrate to the external temperature.

• What accessories should I consider when purchasing the MIC-101?

Depending on the experimentation you plan to do and your current setup, some accessories may be required. Flow meters are essential if you are planning to use custom air mixtures, as these allow you to measure the flow of air for flushing the chamber. Consider additional tygon tubing to ensure sufficient connections from the gas canister(s) to the flow meter and then to the chamber. Additional air filters are required when air flow is reduced in any current air filters or as part of a regular maintenance routine. Lastly, additional backup tubing clamps may be required as these may crack, break, or become faulty with extended use. To make purchasing products simpler, Embrient provides a variety of bundles of the MIC-101 chamber with combinations of accessory products.

• How can I tell if the inlet and outlet ports of the chamber are fully sealed?

When the white clamps are fully tightened, the tubing should be fully compressed preventing any air from entering or exiting the chamber. View the tubing from a side angle to ensure that the tubing is fully compressed. If a clamp is cracked or worn from wear and tear, replace it with a new one.

• Where do I obtain gas mixtures?

In most cases, the purchasing department at your institution has agreements with local mixed-gas suppliers. Custom gas mixtures can be ordered through most suppliers.

• Why should I perform my tissue culture experiments under physiological conditions?

Almost all mammalian cells exist in environments with much lower oxygen levels than in the atmosphere. For example, oxygen levels in the brain are usually between 0.5-7%; in the eyes, it’s between 1-5%, and in major organs like the liver, heart, and kidneys, it ranges from 4-12%. Even blood has oxygen levels between 6-16%. Research shows that using atmospheric oxygen levels in cell cultures can mess with critical processes like metabolism, development, and activation. For instance, normoxic conditions at 21% oxygen can suppress the function of Hypoxia Inducible Factor (HIF-1), which affects numerous cellular processes, such as metabolism and differentiation, notably reducing the viability and differentiation of stem cells. Thus, culturing cells at non-physiological atmospheric oxygen levels creates artificial and inaccurate results that do not reflect true biology. Read more on this important topic in our Tech Note.

• How do I get rid of Volatile Organic Compounds (VOC) and other contaminants when I clean the incubator?

Fortunately, the Embrient incubator is designed so that very few contaminants are allowed to enter, which reduces the effort required for cleaning. That said, diluted alcohol (70% ethanol, 30% distilled water) can be used to remove any remaining contaminants, after use, and prior to next use.

• Can I use titanium oxide to clean the MIC-101 incubator?

The MIC-101 is made from polycarbonate and as such, titanium oxide cannot be used for cleaning and removing contaminants.

• Which chemicals are compatible and which are not compatible with polycarbonate?

The MIC-101 polycarbonate is resistant to many acids, bases, salts, greases, oils, detergents, hydrocarbons, and alcohols. It can also withstand aluminum chloride solution, aluminum sulfate solution, citric acid solution, Freon 12, formic acid solution, glycerine, hydrochloric acid solution, hydrofluoric acid solution, and hydrogen peroxide. Polycarbonate is not compatible with acetone, benzene, chlorobenzene, chloroform, ethers, ethyl acetate, ketones, m-cresol, pyridine, tetrachloroethane, and ammonia gas. These substances can cause polycarbonate to swell, change color, or develop stress cracks that can lead to failure. For larger lists of chemical compatibility, search Google for "polycarbonate chemical compatibility".

• Is the use of 2-ME (2-mercaptoethanol) in cell culture the same as low oxygen?

No, 2-mercaptoethanol (also known as beta-mercaptoethanol or BME) is not the same as low oxygen in cell culture; while both can affect cellular redox balance, 2-mercaptoethanol acts as a reducing agent by directly donating electrons to counteract oxidative stress, while low oxygen simply reduces the amount of oxygen available to cells, more accurately reflecting physiologic oxygen.

Ordering

• What payments do we accept?

Visa, Mastercard, and most Credit Cards, Wire, ACH, and Checks.

• How do I place an order if I can't use a credit card?

For domestic orders (US only) we do accept some orders with a Purchase Order number. Please email us at info@embrient.com to inquire about payment options. All international orders must be prepaid either by Wire, Check, ACH, or Credit Card before an order can be processed for shipment.

• What information do I need to get a proforma invoice/ quote?

In order to provide you with an accurate quote, we kindly request that you provide us with the following details: Shipping Information: - Person Receiving Order - Address (Street, City, State/Province, Country, Postal Code) - Phone Number - Email Billing Information (if different from Shipping Information): - Billing Contact - Address (Street, City, State/Province, Country, Postal Code) - Phone Number - Email Please also provide us with the details of the product(s) you are interested in, the catalog number, and the desired quantity for each item.

• How do I place an order with a credit card?

You can place an order via our website in the shop. You will need to create an account to do so. All international orders must be prepaid either by Wire, Check, ACH, or Credit Card before an order can be processed for shipment.

• What are our Net Terms?

We have a 15 day Net Term and also a 30 day Net Term for approved universities/company domestic orders. All international orders must be prepaid either by Wire, Check, ACH, or Credit Card before an order can be processed for shipment.

• What are my options for ordering internationally, outside of the US?

There are several available options for ordering internationally. You can order directly from us and we will ship your order from our facility in San Diego, CA. This process requires that you pay any import and customs fees. Alternatively, you can purchase our products through our distributors. While list pricing may be higher with distributors, they typically handle import and customs fees, resulting in potentially lower overall costs. View our current distributors.