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Glove Box Design

The Ossila Glove Box.

You can use Laboratory glove boxes (or gloveboxes) to either protect sensitive materials from an outside environment (isolation), or to protect yourself from infectious samples or radioactive materials (containment). The glove box is made up from a clear panel which allows you to see the contents of the chamber, while the gloves themselves allow you to manipulate it. Smaller, secondary chambers called antechambers allow you to add or remove items without exposing the inside of the glove box to the atmospheric conditions.

Inert atmosphere isolation glove boxes are the most commonly used type of glove box and are one of the most critical pieces of equipment found in any laboratory. This is because they provide you with the means to work in an environment that is completely inert, eliminating the presence of reactive species such as oxygen and water. This is crucial for applications where the presence of small quantities of these elements can result in significant chemical or physical changes in the properties of the materials that you are using. Common uses for inert atmosphere glove boxes include chemical synthesis, organic electronics, additive manufacturing, materials handling and storage, development of battery technology, and perovskite electronics.

To maintain an inert atmosphere inside the glove box, it is important that you make sure that the inner atmosphere is exposed to as little moisture and oxygen as possible. There are two main ways that the inner atmosphere can be compromised; via physical leaks in the box, or by the ingression of moisture and oxygen through the materials that make up the chamber.

You can remove contaminants from a glove box by purging the system with a continuous flow of inert gas, or by circulating the atmosphere through a filtration system that removes air and water5. Over time, however, they can re-enter the glove box via a few different pathways.

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Leak Detection and Testing

Unfortunately, as it is impossible to create a perfectly closed system, glove box leaks are inevitable. However, by understanding how and where leaks occur, you can often identify and repair them at their source. Some leaks can be prevented entirely through careful monitoring and maintenance, while ingression rates can often be reduced by ensuring that all glove box materials have low a oxygen and moisture permeability. With the right combination of barrier materials, permanent and non-permanent seals, and effective leak management and mitigation, the internal environment in a glove box can remain inert for extended periods of time.

Any leak in a glove box, however small, has the potential to result in contaminants entering the chamber. This will compromise the inert environment inside. The frequency and severity of leaks can be managed by choosing a well-designed glove box and following good maintenance and operational procedures. These are discussed in this guide.

Managing glove box leaks comes down to a few simple rules:

  1. Use appropriate seals for joints between surfaces.
  2. Introduce standard glove box operating procedures to reduce occurrences of human error.
  3. Monitor gas usage so that leaks can be detected early.
  4. Schedule tests and inspections of seals.
  5. Have procedures in place to reduce the impact of containment failure.

Glove boxes are typically held at a positive pressure of a few millibar (mb) relative to atmosphere. This means that if there is a leak anywhere, the pressure difference will cause gas to flow outwards and a low O2/H2O atmosphere can be maintained within. A glove box with a small leak will be able to maintain overpressure by replenishing the gas within the leaking chamber, so initially the contamination levels will not increase substantially.

Leaks can have other impacts in addition to the risk of contamination. For example, increased gas flow into a glove box can result in turbulent airflow within the chamber. For systems with HEPA filtration, this can result in stagnation points which can leave areas unfiltered.

The increased gas usage for maintaining an over pressure can also often result in a depletion of the nitrogen source, the internal and external pressures equalising and water and O2/H2O entering the system. Therefore, while initially a small leak is no cause for alarm, identifying a leak, locating its origin, and fixing it are critical in maintaining the atmosphere inside the glove box.

Small leaks can also develop into larger leaks which can be dangerous, not only for the glove box functionality, but for the users of the system. Glove boxes are often used to isolate materials that can either be dangerous if exposed to air, or otherwise be harmful to the user. Large leaks have the potential to result in users being exposed to harmful materials unknowingly.

Additionally, if the glove box is continually purging to maintain an overpressure, this constant outflow can affect the oxygen levels in the room. Inert gasses, such as nitrogen, cannot be detected in the air by sight or smell, and the effects of oxygen depletion mean that it is difficult for those exposed to know they are in imminent danger. For this reason, it is important that glove boxes are used in rooms that are well ventilated and fitted with oxygen-level monitors.

Causes of Leaks

The most common time for the internal atmosphere of a glove box to be compromised is when you are transfer samples in and out via the antechamber. This happens in the event of incorrect cycling of a failure to purge the antechamber.

It is important that correct training is given to users of the glove box on how to work with and maintain it. It is also important to have standard glove box operating procedures available for commonly performed tasks as well as a plan of action incase issues occur. Even with thorough procedures and careful users, leaks can still happen. Because of this, it is also important to ensure that in the event of atmospheric exposure, samples are safe and a plan of action is in place to quickly get the system back to inert conditions.

Glove box Antechamber
The antechamber on the Ossila Glove Box is equipped with highly accurate sensors. The in-built display indicates when it is safe to open.

Damage to the glove box, such as a punctured glove or damage to the seals, can also cause leaks.

Dirt or debris can cause seals to leak. Even if they are well maintained and kept clean, the seals will lose elasticity over time, especially through repeated use. Other mechanisms where damage to the seals can occur, thus resulting in formation of micro cracks. An example of this is ozone cracking of rubbers, which can occur at surfaces exposed to air. However, this can be mostly eliminated through correct material choice in the glove box design.

Some seals, like knife-edge gaskets, will need to be replaced after use as they are not built for repeat usage. Other gaskets, such as rubber O-rings, are more resilient to multiple uses. However, these are usually made from flexible materials so are less resistant to chemical stress, extreme temperatures, or pressure. It is therefore important to continually assess the quality of your seals, this will reduce the likelihood of serious leaks. Simply looking at the seals each time you open a door can prevent a serious compromise in your glove box environment.

Glove Box Leak Tests

There are many versions of leak test that can be performed, depending on your system, as outlined in the ISO 10648-2 standard. However, the most used leak rate test is the positive pressure test, which involves increasing the internal pressure of the glove box and monitoring how it changes over time. A general overview of the process is shown below.

Each manufacturer will specify a way of testing the glove box; the Ossila Glove Box internal software has an automated test which completes the process.

In some glove box systems, a negative pressure leak test can be performed. It is possible, but unusual, for leaks to be seen under negative pressures but not positive pressures. For this, you should follow a procedure like the one described above but use a pressure of between -5 mbar and -10 mbar. If there is a leak in the system, the pressure will increase towards atmospheric pressure and the rate of this increase can be used to determine the leak rate of the system.

Leak testing a glove box
General process for positive pressure leak test. A set over pressure is created in the glove box (1) and this pressure is monitored over a given time period. If the box is sufficiently sealed, the pressure will maintain (2); if there is a significant leak, the pressure will drop quickly as the gas is pushed out (3).

These leak tests can identify if there a significant leak within the system but are not thorough enough to provide exact leak rate or demonstrate where the leak is coming from.

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Gaskets and Seals

Glove box leaks, seals, and sealant
Two hard surfaces with and without sealant materials. Even if there is no visible damage at the interface, there may be small gaps between the surfaces. At these joins sealant material is used to fill in these gaps.

Stationary and Permanent Seals

At the interface between two hard surfaces, there will always be a pathway for gas to pass through. On a microscopic level, the two mating surfaces have a high degree of roughness relative to the size of gas molecules.

It is not economically feasible to get the two surfaces to a level of flatness that would allow for a gas tight seal. For this reason, a softer material is compressed between the two surfaces to fill these gaps and reduce leaks. For stationary and permanent joins, silicone sealants can be used along with mechanical fixtures to ensure a tight fit. Alternatively, metals can be welded together to create a seamless join.

However, there are a several parts of a glove box that require non-permanent seals, such as doors for pass-throughs and feed-throughs for services. For this, gaskets are extremely useful.

Types of Gasket for Non-Permanent Seals

A gasket is a piece of material that can be placed between two surfaces to form a seal. In the context of a glove box, their main purpose is to create an air-tight seal against gas/liquids entering and leaving the system. There are many types of gasket each suited for different situations. The three most commonly used in glove boxes are O-rings, flat gaskets, and knife edge gaskets. These are shown in the figure below.

Flat gaskets

Flat gaskets are used to form a seal between two surfaces as shown in the figure above. Flat gaskets can be cut to a wide variety of shapes, and they can be made from many types of materials. They are most commonly made with rubber and expanded rubber foams. Flat gaskets are often used for washers to help make seals, or rectangular joins such as windows and doors. They are especially beneficially when low clamping forces are used.


O-rings are doughnut-shaped mechanical gaskets that sit within a groove that is milled into one of the surfaces. These are usually made out of harder elastomeric materials such as nitrile or Viton. Compression of an O-ring creates an air-tight seal that can withstand high pressures and temperatures. These are often used in regions where high-pressure differentials occur such as inlet piping, or for the doors of vacuum chambers integrated into a glove box system.

Glove box gaskets used to prevent leaks
Diagrams of Flat and O-Ring Gaskets (above). Knife-Edge Gaskets (below) before and after sealing. Two hard metal "knife-edges" (blue) indent the soft copper gasket inbetween (red) forming seal.

Knife-edge metal gaskets

Knife-edge metal gaskets are often used in situations where a seal needs to withstand very low vacuum pressures1. Knife-edge seals are nearly perfect seals as the materials used have extremely low ingress rates and can withstand extremely high clamping forces. To achieve this, there are "knife-edge" indentations that are on the surfaces of the two joining parts. The gasket itself is made of a much softer metal such as copper. When these two surfaces are pressed together, the knife-edges of the hard metal indent the softer metal gasket in between.

Knife-edge gaskets create a flawless metal-metal seal. However, this does result in a permanent denting of the gasket, thus limiting the amount of times the seal can be broken and resealed. In addition, the production of the knife-edge is expensive and so is limited to smaller surface couplings. In glove box systems, these are typically reserved for connecting components in integrated vacuum chambers.

Permeability and Impact of Barrier Materials

The physical properties of the materials used for the solid barriers in a glove box are the biggest determination of the amount of oxygen and moisture permeation that will occur. As we must keep these levels as low as possible, the choice of material for each aspect of the glove box is equally important.

How Does Ingress Through Barrier Materials Occur?

Ingress, in the case of glove boxes, is the diffusion of unwanted oxygen and water into a sealed chamber. This typically occurs as moisture and oxygen pass directly through the materials used in the construction of the glove box. When discussing ingress, we need to examine the permeability each material4. This is a measure of the total transport of matter through a system.4 Or in this case, the O2 or moisture through the glove box walls.

The ingress process can be split into several different steps. The figure below shows the stages of a permeant travelling through a solid barrier. First (Step 1), the vapour molecules must reach the first barrier interface, B1. Then, the permeant is absorbed at B1 (Step 2), before diffusing through the solid barrier (Step 3). The permeant then undergoes desorption at the B2 (Step 4) and diffuses away from the barrier (Step 5).

The processes in Step 2/4 are mainly determined by solubility, S, while step 3, is determined by the Diffusion coefficient, D0.

Glove box ingress: stags of permeability
The different stages that a permeant must go through to ingress through a barrier material.

Steps 2 and 4 can be described by the following adaptation of Henry’s Law:

Henry's Law

Where the Solubility, S, is determined by the partial pressure (pPM) of the permeant and Ksol is Henry’s law constant4–6. This essentially describes how quickly a permeant can be adsorbed or desorbed by the solid barrier.

The permeants movement through the solid barrier is governed by diffusion (step 3). This is often the slowest process; therefore it acts as the rate limiting step for permeability. This diffusion process can be described by Fick’s laws. Fick’s first law denotes the relationship between concentration gradient and movement of permeants across a space dx is:

Fick’s first law

Where J is the flux of permeants moving through the barrier, D is the diffusion constant, and dC/dx is the concentration difference across the barrier (from x à x+dx).

However, as permeant diffuses through a material, the concentration of the permeant at both point X and point (X+dx) will change. It is therefore necessary to consider how this process changes with time. This leads to Fick’s second law of diffusion:

Fick's second law diffusion

Where EA is the activation energy, T is temperature and R is the gas constant. This EA essentially represents the energy required for a permeant to squeeze through a solid.7 This also means ingress depends exponentially on temperature.

The permeability coefficient, P0, is a product of a systems S and D0 values. This is the property used to describe a barriers permeability to a specific permeant.

How to measure the permeability of barrier materials

Transmission rates of O2 and water vapor can be measured using a gas transmission cell.

For O2 transmission rates, a thin layer of barrier material is mounted in the centre of the transmission cell to divide the two chambers11. An oxygen sensor is placed in one chamber, and this chamber is then purged with nitrogen gas to remove all O2. Meanwhile, high oxygen levels are maintained with a constant N2/O2 supply. The increase in O levels detected by the sensor as oxygen diffuses through the semi-barrier can be used to determine oxygen permeation per unit time, or the oxygen (gas) transmission rate (OTR or O2GTR).

Glove box ingress MVTR measurement
Different methods of measuring Moisture Vapour Transmission Rate. A gas transmission cell (left) can be use to measure extremely low MVTR (1. N2/O2 or N2/H2O flow; 2. N2 flow; 3. Vent to atmosphere; 4. N2 flow + H2O or O2 to Sensor). Simpler methods such as the Desiccant and Water methods (right) can be used where moisture ingress is significant.

You can also use this method to track moisture ingress. Instead of using high oxygen levels in the first chamber, use wet nitrogen. This maintains high moisture levels and you can then use an infrared sensor to detect the increase in humidity.

Another common method to measure moisture vapor transmission rates is by using the desiccant or water methods (ASTM E96). For these methods, fill a dish with either desiccant material or water and cover with a barrier film layer. Measure this dish before and after a significant period of time. In the desiccant method, you use mass gain to determine MVTR through the barrier material. In the water method, you use mass to calculate MVTR. The desiccant/water methods are much simpler to construct, but using a gas transmission cell will determine MVTR and OTR more accurately.

The units used for these measurements will vary depending on how they have been measured. Some metrics are outlined below:

  1. Moisture vapour transmission rate, MVTR, (or water vapour transmission rate, WVTR) is a measure of the rate of water passing through a film or barrier (usually per day). This is measured in g/m2/day. One way to calculate this is to measure the uptake of water via the change in mass of the material.11 However, different methods can give different values for MVTR11, so to state the conditions measured in is very important. MVTR is generally described using this equation12:
    Moisture vapour transmission rate formula
    Where Pis the permeation constant for the barrier, p1-p2 is the partial pressure difference of water across the material, R is the ideal gas constant, EA is the activation energy and T is temperature.
  2. Moisture vapour permeability is a measure of how much moisture will penetrate a barrier per given thickness. Therefore, for this article, we will assume that MVTR ~ P0/(barrier thickness).
  3. Additionally, oxygen transmission permeability is a measure of the amount of oxygen passage through a film or barrier per day and is measured in

Permeability of various possible glove box barrier materials

The table below lists some coefficients of Oxygen and Moisture Permeability, and some MVTR for various materials. The footnotes detail the conditions under which these values are accurate.

Material O2. Permeability** ( 2O Permeability** ( MVTR (g/m2.d.atm)
PMMA 5.8-7.2 ^12,13 5.25 ^12 55.20 ***14 -
PET 1.2-2.4 ^11 0.39-0.51 ***11 -
PTFE 222 ^11 0.006 ^15 -
Silicone rubber 3940 -4330 ^11,12 1.73-3.31 ***11
Butyl rubber 7.8-85.4 0.006 ^15 -
Aluminum foil <0.006 16 - <0.01 16
Glass Coverslip(0.1-0.2mm) - - 4 x 10-6 ^^^17
Steel N/A* N/A* N/A*

*These materials behave as near “perfect barriers” for oxygen and moisture 18,19 – most of the ingression with these materials is at the joints, depends on the bonding method used or grain boundaries within the material. **Permeability = (volume of permeant)*(film thickness) / (surface area)*(time)*(pressure drop across barrier) ***At 37°C at 90% RH ^ At 25°C at 1 atm^^ at 20 and 40°C respectively^^^ at 30°C at 90% RH.

Oxygen and water ingress through polymers

An early prototype of the Ossila Glove Box built using polymer plastic; the final version uses high quality stainless steel and glass to achieve low ingress rates that are not possible with plastics.

Polymers are a popular choice for barrier materials as they are cheap, strong, and can have a wide range of material properties.

The most important thing to consider when thinking about diffusivity is the fractional free volume of the barrier material. This depends on its packing factor. Essentially, the more space there is between molecules in a barrier, the more room the gas has to diffuse through it. For this reason, permeability also depends on the size and structure of the permeant8.

Polymers consist of long chains and therefore have a low packing factor. However, whether a polymer is classed as glassy, crystalline or rubbery will affect this packing factor. Furthermore, the quality of a polymer will depend on its on its structure at room temperature. A glassy polymer has glass transition temperature, Tg>room temperature, RT, meaning that the polymer chains are locked into one position and the material will be characteristically brittle.9 A rubbery polymer will have Tg<RT, so is more malleable.

In glassy polymers, solubility is increased as they have an excess non-equilibrium free volume10. In other words, the polymers chains may be locked in awkward positions, leaving holes for moisture uptake. However, rubbery polymers tend to have a higher free volume, as they have a more disorganized structure. Also, their polymer chains have more room to move around, facilitating diffusion through the material. For this reason, rubbery polymers tend to have higher permeability.

Crystalline materials in general have a much better packing density than rubbery/glassy polymers therefore permeants have a harder time diffusing through them. When crystalline materials form, there is a point where two crystals meet. This is called a grain boundary. There can often be atomic mismatch at this grain boundary leading to slightly larger free volume, thus, there may be a small ingression of moisture or oxygen at grain boundaries7.

In materials such as metal or glass, as used in the production version of the Ossila Glove Box, this amount is almost negligible.

Impact of Moisture Ingress on Glove box Atmosphere

To show how the permeability of a material will result in a rise in the water content of a glove box, we have simulated how the water content inside of a glove box increases over time for different materials. We have used a wall thickness of 5mm, the values from the table above, as well as the moisture vapour transmission rate equation to model the transfer of moisture into a box with external dimensions of 60 x 60 x 50 cm in diameter.

For simplicity, it is assumed that this is a closed system at a constant temperature, with 0.01ppm of H2O in initially the system. We have also assumed that the change in concentration gradient across the walls (barriers) with time is negligible. Thus, by the time a non-negligible change in concentration gradient has occurred, the atmosphere in the glove box is already classed as compromised. Therefore, we have excluded the effect from this example. We have also considered separately the effect of ingress through rubber gloves using two common materials.

Glove box ingress moisture variation with material
Log-plot of moisture levels within a sealed box, shown in parts per million by weight (ppmW) over a period of 14 days. Inset (shown on left) show linear plot of moisture levels in ppm over 12 hours.

The resulting plots show how easily the levels of moisture can build in a closed glove box environment. In a real glove box, there would be additional purging of N2/Ar gas or atmosphere filtration, which would reduce the impact of this ingression. However, constantly purging an environment can quickly become expensive, and filtration media can become saturated - requiring frequent regeneration of the material. Therefore, when using a glove box, it is crucial to consider which materials are being used when working in this inert atmosphere.

Inert Atmosphere Glove Box

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  19. Flexible Organic Electronic Devices On Metal Foil Substrates For Lighting, Photovoltaic, And Other Applications, B. W. D’Andrade et al., Handb. Flex. Org. Electron., 315–341 (2015); DOI: 10.1016/B978-1-78242-035-4.00013-0.

Further Reading

An Aid to the Design of Ventilation of Radioactive areas., G. Hall et al. (2009) Nuclear Industry Guidance. Accessed at:

Contributing Authors

  • Jon Griffin
  • Mary O'Kane
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