Helpful Hints for Those Who Will Be Working with anaerobic chamber

Having the ability to work in anaerobic chamber is a very specialized skill set. Only if you are dealing with oxygen-sensitive compounds is it necessary for you to do so. For instance, some metalloproteins can only remain in their reduced state in the absence of oxygen, whereas other metalloproteins are sensitive to oxygen and can even become reactive in its presence. When conducting experiments in anaerobic chambers, one needs to devote a longer amount of time to the task at hand, as the process is typically more laborious than when working in oxygenated environments. Working through a pair of gloves slows everything down, and it takes a long time to prepare and make sure that your sample and environment are deoxygenated in the first place. If you ever find yourself working in an anaerobic environment, the following are some tips to keep in mind. We can only hope that this makes them a little less intimidating than they initially seemed.

 


Here are some pointers that I've picked up over the years, but don't forget to also get some training from someone who has experience working in an anaerobic chamber. Each one is a little bit unique in its own way.

The Anaerobic Chambers Are Comparable To Large Glove Boxes

There are two main types of anaerobic chambers: rigid boxes and flexible polyvinyl bags.

Chambers come equipped with either one or two sets of gloves, making them suitable for use singly or in tandem.

 

They are equipped with an airlock transfer chamber so that they can bring things in without disrupting the atmosphere inside.

 

  • The anaerobic chamber is filled with a gas mixture that is composed of 95% nitrogen and 5% hydrogen



A palladium catalyst can remove all traces of oxygen from the atmosphere, bringing the level down to 0–5 ppm.

RecommendationsA Moment Ahead of Entering the Transfer Chamber

Create a list of the things you need to bring with you so that you can reduce the number of times you have to enter and exit the chamber. An excessive number of times results in the loss of time and gas as well as the contamination of oxygen.

Bring any samples or pieces of equipment you intend to use in the anaerobic chambers through the transfer chamber first so that the oxygen can be removed from the environment. The inner chamber shouldn't be opened for at least three full cycles before you begin cycling the transfer chamber between a vacuum and a gas mix consisting of 95% nitrogen and 5% hydrogen. Before you open the door, check to see that the chamber is not currently going through the vacuum cycle. Don't forget to keep the inner door closed and the seal in place at all times!

At the very least, you should bring any necessary apparatus into the chamber the night before an experiment is scheduled to start.

Before bringing buffers or solutions into the intake chamber, they must first be purged or degassed for a period of thirty minutes to one hour using an inert gas such as argon or nitrogen.

Sometimes the catalyst will'shut down' and the oxygen detector will begin to pick up on the presence of oxygen. Don't be concerned; simply bake your product and reactivate your catalyst in accordance with the instructions.

In the event that you are working with a protein that is stable enough to not require disulfide reduction, you should allow the protein to incubate anaerobically with the reductant. After that, remove the reductant from the chamber by dialyzing it overnight. Use a desalting spin column as an alternative to dialysis in order to quickly remove any residual reductant.

If the protein is not stable enough to undergo dialysis in the chamber overnight, first purge the chamber for a short period of time with an inert gas, and then allow it to equilibrate for 30 minutes to an hour before proceeding with the disulfide reduction.

During This Time That You Are Working in the Chamber

Please don't forget to take out all of your trash.

Ice releases oxygen, so you shouldn't bring it into the  with you. Instead, you should keep the ice packs and use them in a container made of Styrofoam.

For the purpose of archiving, you can flash-freeze samples inside the chamber using liquid nitrogen; however, you should be aware of the following special considerations:

Because liquid nitrogen emits nitrogen gas while it is contained within the chamber, the chamber must be evacuated of a significant portion of the gas (approximately half) prior to the introduction of the liquid nitrogen. Otherwise, the chamber will quickly become overcrowded. Because of the increased pressure, doing so can be extremely risky.

No matter where you are working, you should never handle liquid nitrogen without first taking all of the necessary safety precautions.

Fill your sleeping quarters, or at the very least get it ready for the next occupant. Please bring some of the following items into the building:

Pipette tips for all different sized stock pipettes

Beakers for the waste (one each for the dump and the stock)

Micro centrifuge tubes, 15 mL conical tubes, and 50 mL conical tubes (stock) are the types of tubes that are included here.

Pipettes (if at all possible, store them in the interior at all times)

A centrifuge designed for use on a tabletop, which should be stored indoors whenever possible.

There are some chambers that have oxygen present.

You might come across other chambers while exploring the laboratory, but not all of them serve anaerobic functions. It is important to differentiate the following from anaerobic chambers:

The dry boxes

Complete regulation of the humidity

Oxygen tissue culture glove boxes

Oxygen research on animals, glove boxes

If you have never been inside of an anaerobic chamber or seen one in action, it can be quite frightening. When working with gasses, regulators, seals, vacuums, redox states, and proteins that precipitate or even sometimes evaporate, they can be difficult to work with, and there is a significant learning curve associated with this type of work. However, if the anaerobic chambers are maintained properly, they can last through a number of PhDs, which can result in some fascinating scientific findings.

Posted in Default Category on August 29, 2022 at 11:55 PM

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