Introduction: DIY Anaerobic Chamber (aka Glove Box)

This DIY Anaerobic chamber is considered a fully functional piece of Bio Art, As a statement from the Maker DIY community to the large Institutions that have a tendency to withhold knowledge. It was designed and built to grow cultures of Geobacter Sulfurreducens bacteria, to do research and experimentation on my other Bio Art project: a Microbial Fuel Cell project called "BactElecTric".

It is important to state the fact that this is piece of equipment intended to be used in a Bio Safety Level 1 lab environment.

Also, it was conceived to keep a low oxygen level atmosphere inside of the chamber, it is constantly pumped with inert gas such as Nitrogen or Argon.

It is NOT(repeat...not) intended to keep organisms from going outside of the chamber, it is intended to keep a desired atmosphere inside, with positive pressure.

It is NOT air tight, the positive pressure and small leaks keeps outside air from going in.

This is a functional piece of biology lab equipment, it was welcomed inside GENSPACE, the first community Biolab, based in Brooklyn, New York.

Step 1: Chamber Parts Breakdown

Here is the breakdown of the parts of the my DIY Anaerobic Chamber:

1- Big plastic container (36 gal), as clear as possible, as big and square as you can find.
2- Gas intake
3- Relief valve
4- Double hatch system (two plastic food containers)
5- Double gasket with metal reinforcements
6- Pressure gauge
7- Rubber arms (flex tube) and gloves
8- C clamps
9- Insulation foam + silicon + duct tape

More detailed material list and building procedures ahead.

Step 2: Materials Used

This are most of the materials used to build the Chamber. Some other materials (like liquid rubber spray) have been used to do some adjustments along the way.

Step 3: Footers

This pieces are made to lift the chamber a little from any surface or table it is on, this is mainly because of the C-clamps used to close the lid tightly.

I suggest using half inch screws.

I used 1/16 inch rubber to make small gaskets, reinforced with Lexel.
(!!!note on the Lexel: It takes very....very long to dry (curate), bu is is 1000x better than silicon!!!)

Step 4: Closing the Lid

Use the 1" C-clamps (x10) to hold the lid very tight. This combined with insulating foam (1" and heavy duty) works pretty well.

The corners are a little tricky but with patience and a good x-acto knife, the insulating foam works.
You can use as many C-clamps as needed.

The idea  is to NOT close the lid permanently, so that the chamber can be opened for cleaning, maintenance, modifications or any other need, without loosing integrity.

(You can also get a big sheet of 1/8" rubber and tailor a nice big gasket...)

Step 5: In Goes the Gas

It took me some time and help from a employee at the hardware store to get all this pieces to fit together properly, but it was worht it.

I used quick release valves so that you could change hose, or gas type without compromising the integrity of the chamber and to be able to be stand alone in case the hose (or gas tank) needs to be removed or exchanged.

On the inside of the chamber, the valve has a brass connector to screw a barb to connect a hose (you can find size adapters), this is if you need to sparge (blow bubbles) a liquid medium (as it was my case) with a certain gas.

I used a power drill, starting with a small diameter bit, incrementing the size bit by bit. It is important that you insert the drill while its moving at high speed (do not start the drill while touching the plastic). I also used fabric tape to help support the plastic.

Remember to clamp a piece of wood on the inside and the whole chamber to a corner of a table, so that you can drill properly.

!!!!!RECOMMENDED:
Latter on I used the Dremel, using a pointy polishing stone to make the first hole, and then, using the Dremel and a sanding paper tip, to make the whole to the desired size.

To help seal the hole properly and make it air tight, I used a gromit.

finally, I caulked some Lexel around the brass valves, inside and outside. (remember to let it dry for a very long time)


!!!!!    IMPORTANT   !!!!!
To prepare your oxygen free (almost) atmospherey
To sequestrate the oxygen of the normal atmosphere, I used "food preserving tablets" they are used to preserve dry food for extended periods of time.

I got them on line and you can easily calculate how many you need for your chamber size. I just put them in a corner of the chamber in a way that they did not interrupt my work... and later on i got them out of the chamber through the double hatch mechanism.

Step 6: Just in Case, Pressure Relief Valve

Just in case, I decided to include a pressure relief valve, so that the chamber could not blow up or have its structure compromised in the event of gas being pumped excessively....you never know.

As mentioned before, this time I used a Dremel to make the hole. I recommend this technique over the power drill, to work with plastic containers.

The same finishing touches of the gas intake valve where made to this pressure relief valve.

I decided to put both valves on the same corner of the chamber, so that I would not have another corner used up and could not place elements in it.

Step 7: Accordion Arms

This is the most funny element of the DIY anaerobic chamber.

Funny thing is... they worked!!!

Make sure you make a prototype of your arm size and distance between arm holes, before you get to work on the chamber.
Also, be sure to use enough accordion tubing, to reach the inside of the double hatch while you are working.

The best way to make the big holes for the arms is with the Dremel, as explained before. (not a plastic cutter as I first did)

Since the accordion tube, had a spiral structure, it did not seal very nicely against the edges of the holes on the plastic wall of the container. To deal with this, i used expanding sealing foam... and a lot of Lexel.

It worked ;)

Step 8: Rubber Gloves

I used regular dish washing gloves, with latex (be sure to know if your are allergic to latex) .

Because of the spiral structure of the accordion tube, A LOT of patience and time was needed to seal the union of the gloves and the accordion hose. I cut and tailored pieces of gorilla tape independently as I worked, so that they made a "scale mail" pattern, like scales on a fish of armor.

Additional support was provided by zip ties...and the gloves own rubber.

This elements have been optimized with liquid rubber spray, to strengthen their structure.

During lab work, it is strongly advised that you use your lab gloves while using the chamber, your hands sweat a lot, and getting them out of the chamber's gloves can be tricky.

Step 9: Double Hatch Ensemble

I used two identical food containers (with rubber tight seals and latches on four edges), attached back to back, big enough to let a tall lab bottle fit inside.

This dual hatch system was designed so that minimize loss of atmosphere while working, and provide a way to get elements needed into and out of the chamber at any given time. (without having to open the whole chamber and loosing all your atmosphere)

Be very careful making your holes. For this I made templates made of masonite, so that every hole drilled was on the same spot.

You'll see on the next step, this is very important to make the dual gasket system for the dual hatches.

Step 10: Hatch Gaskets

The rubber gaskets were laser cut.

The wholes on the metal frames where drilled with a Drill Press. The metal frames where made so that  screws could be used to really tight the sandwich of layers that this step dealt with, without compromising the integrity of the plastic containers. In other words, this was done so that the plastic didn't crack when you tight the screws as it is needed for the air tight sealing of this large square hole on the chamber.

Everything was done using the same templates that where made at the beginning of the double hatch process.

For everything to come together correctly, you have to use clamps, to make all the layers be tight and pressed against each other.

layers (in order of appearance):
metal, plastic container 1, rubber, plastic wall of main chamber, rubber, plastic container 2, metal

The finishing touch was to caulk Lexel on the outside edge, and Silicon on the inside. I only used silicon because the caulking gun was too big to be used on the inside of the chamber

If you can TIG weld your metal frame so that it is one solid piece, it is best, I want to do it for my next chamber :)

Step 11: Horiozontal Surface for the Hatch

A flat horizontal surface had to be made for the inside of the double hatch.

Since the walls of the main chamber are tapered, the double hatch is inclined downward towards the inside of the chamber. There is also prominent edges because of the gaskets.

I did the pieces with plexi, on a laser cutter, but they can be made by hand. They three pieces were glued using "weld-on" and the notches on the middle were made,by hand, very carefully, with the Dremel.


Step 12: Pressure Gauge

By now this procedure is self explanatory.

The finishing touch was made with silicon...inside and outside

The only comment is that I want to replace this gauge with another one with a smaller scale, up to a 100 is enough.
The pressure relief valve works at 70 psi.

Comments

author
LizZ5 made it! (author)2017-02-18

Thanks for the hot tip about the rubber gloves. Really saved the day when our anaerobic chamber started leaking through it's degraded cuffs.
I noticed you don't have an oxygen sensor per say, did you know you can make a solution of reduced methylene blue that will turn blue in the presence of oxygen. I think you can also purchase strips to act as indicators.

Dishwasher gloves.jpg
author
rolonrene made it! (author)2016-09-27

Hi,

I am not very sure about how to manage nitrogen or argon, so my question is: are all the valves you used especially made for the use of this types of gases? Is it dangerous to use solenoids valves?

author
frix00 made it! (author)2014-10-13

Hi,

Just to confirm, you are not using any vacuum at all here ? The only way to get the oxygen level down is to pump in inert gas right ?

Did you use an oxygen detector at all ? Do you know what levels can be reached in terms of % ?

Thanks

author
ponckk made it! (author)ponckk2014-11-04

Hello
Thank you for your message, interest and support :)
Sorry for the delay.

So...
No vacuum at all.
only pumping inert gas ( the chamber is not air tight, it has some leakage, so it is actually good because you create a positive pressure situation and no outside air comes in.
using O2 sequestrating tablets (used to preserve food), and eve a small candle...to burn O2, but his is kind of risky.

Sadly I have no O2 detection... this is the big weakness of my effort.

Thank yo for your interest.

May I have more info on your project? and if you go an build one of these, could you please send me some documentation :D

I would love to se it replicated.

Best wishes

Nelson Ramon

author
frix00 made it! (author)frix002014-11-05

Hey Nelson,

Thanks for the reply :-)
We've done a very very similar project, without the box on the side to act as a intermediate chamber, but using an O2 detector :
http://www.amazon.co.uk/SPD201-Oxygen-Detector-Tester-Deterctor/dp/B00I6AOV8I/

We can get down to less tha 1% oxygen, and with a good seal everywhere, we manage to maintain that level for several days without using much of a positive pressure inside.

I've attached a couple of photos too :-)

We're currently refining it further, building it with perspex (plexiglass) which we weld together using a a solvent, and then replication professional lab glove ports, but cutting the cost using 3D printing. Will share some photos of that once we're done.

Thanks for the inspiration !

photo 1.JPGphoto 2.JPG
author
CowGuy made it! (author)CowGuy2015-05-08

Hello, any progress with the perspex? I'd like to fabricate my own chamber

author
frix00 made it! (author)2014-10-13

Hi,

We're building this at the moment, to do some re-bottling of food products, that need extremely low levels of oxygen in order to avoid oxydation.

I just wanted to clarify something: how do you reach low levels of oxygen ? Is it purely the use of oxygen scavengers and pumping inert gas ? Do you use a vacuum pump at all at any point ?

Do you have an idea of the levels of oxygen you can reach (in %) ?

Many thanks- impressive effort!

author
frix00 made it! (author)2014-10-13

Hi,

We're building this at the moment, to do some re-bottling of food products, that need extremely low levels of oxygen in order to avoid oxydation.

I just wanted to clarify something: how do you reach low levels of oxygen ? Is it purely the use of oxygen scavengers and pumping inert gas ? Do you use a vacuum pump at all at any point ?

Do you have an idea of the levels of oxygen you can reach (in %) ?

Many thanks- impressive effort!

author
adamvan2000 made it! (author)2013-09-17

I'm seriously considering adapting this to use as a portable indoor spray-painting bsh a filtered air intake through the walls of the container to allow air to flow in, and a vacuum attached to another filter to remove fumes and particulates from spraying.

Do you have any suggestions for keeping the viewing panel I'd have to add clear from the overspray? I was thinking some sort of clear panel sandwich with small holes in the inside facing layer that I could hook up to an air pump to blow into the chamber. I can either make something like that, or see if there's something available that would do the trick.

Also, now that I think of it, I could also make one for sandblasting too, or make a single multi-purpose box.

Thanks!

author
ponckk made it! (author)ponckk2014-02-25

:D

author
mikeyc.2009 made it! (author)mikeyc.20092013-09-19

When I built my airbrush paint booth I used a fan with an A/C air filter on the exhaust side and another filter on the intake side. You would rather have airflow rather than a large pressure difference. For the viewing window I silicone caulked a piece of glass to a piece of plexi that I could bolt to the side of the box. Made for a GREAT dust-free painting environment.

author
andrae cameron made it! (author)2014-02-21

So sweet!!! I'm a medicinal and gourmet mushroom grower that was in need of a clean room that was portible by far this one that u guys put together blew everything out of the water! I'm in the process of constructing mine as we speak but having a Huge problem finding the double sided gaskets that where used? Did u custom build those? Been to Home Depot along with auto parts stores and no one knows where to get these? Also for the glove sleeves I used 4" female PVC pipe to add for a self seal affect! I will post when I'm done. But yes please help with the gasket issue as I'm inoculating live culture in the next couple days!! Nuhealthsolutionsllc@yahoo.com. Thanks again
Andrae

author
danbemp made it! (author)2013-04-05

Very creative solution -- really impressive. I keep hearing great things about GENSPACE, and this level of ingenuity proves the rumors are true!

author
ponckk made it! (author)ponckk2013-04-11

Thank you for your comment. GENSPACE is a great citizen scientist community, you will find very interesting smart people open to share and learn all the time. It's a privilege to have maker spaces like that.

author
danbemp made it! (author)danbemp2013-04-11

Definitely. Is it mostly biology-focused? Cell-culture focused? Any genetics work going on?

author
ponckk made it! (author)ponckk2013-04-12

all of the above :), as long as it fits in a BSL1 lab

author
rangefinder made it! (author)2013-02-03

Now remind me "why" I need to make one of these ?

Do I "really" need an anaerobic chamber ?

author
mattbeowulf made it! (author)mattbeowulf2013-02-05

The same *reason* that you *really* need to make... [selected 'ibles from rangefinder's "favorites" list] an Electric Umbrella! or an Instructables Plush Robot! or some delicious asian tomato noodle soup! Do you see what I'm getting at? ;-)
If you don't know what you'd do with an anaerobic chamber, well... maybe you ought to start doing some cooler stuff ^_^

author
mwuchevich made it! (author)mwuchevich2013-02-14

like your comment.

author
ponckk made it! (author)ponckk2013-02-06

:8)

author
ponckk made it! (author)ponckk2013-02-06

I needed it for a larger experiment which required cultures of a strain of bacteria that doesn't tolerate normal levels of oxygen at early stages of biofilm.
New chambers are way too expensive for me.
Used ones are still expensive and you have to still get some parts to complete them (at least the ones I was able to find)
... so, what does a man do... he make his own anaerobic chamber !!!, and than shares it. :)

author
DeusXMachina made it! (author)2013-02-04

This is awesome! Wish I had this idea when I needed to package some chemical compounds in a moisture-free environment, but my bottles were too big for our regular glovebox.

author
ponckk made it! (author)ponckk2013-02-06

thanks :D

author
cagstorm made it! (author)2013-02-04

Would this be a suitable "lab" for harvesting and growing yeast in clean conditions? I'm a home brewer and would like to harvest yeast, but need a clean environment to do so in a way that is safe for the yeast not to get contaminated.

author
ponckk made it! (author)ponckk2013-02-06

I would say.. yes, it can work for your cultures.
Good brewing! send me a sample jeje

author
CodfishCatfish made it! (author)2013-02-03

My use would be so simple, applying screen protectors to screens of phones. Every time I go to apply a screen protector it always seems to have accumulated some kind of lint of a stray speck of dirt now I can get some nice clean applications....awesome idea as every and gets my vote.
PS: I had thought about this so many times in the past and now I have the push to build it rather than think about it. Excellent!!

author
ponckk made it! (author)ponckk2013-02-06

it could work .. I think.. with a small modification to remove particles...
see my reply on "The Mighty El Rondo" post

Go and make one! just show me how it goes :)

author
Spokehedz made it! (author)Spokehedz2013-02-03

Your problem is static electricity. When you peel off the plastic, you generate static and it pulls the dust to the newly sticky surface. When I put screen protectors on, I make sure to do it in the bathroom after I let the shower run on hot for a few minutes. The steam/moisture in the air makes it very static unfriendly and I have done several perfect applications in there.

author
ponckk made it! (author)ponckk2013-02-06

nice :)

author
james.m.k made it! (author)james.m.k2013-02-03

You'd bring in the dust when you opened it up to put in the phone and protector.

This isn't a clean room (box) anyway, just a place to grow stuff that doesn't like oxygen, or grows too fast in oxygen.

author
kaytracy made it! (author)2013-02-03

Well done!This looks good! I can see that you might have trouble at the sleeve junction on the box, and I would suggest a flange, for the sleeve to fasten too, this could be as simple a cutting down some large yogurt tubs, though they do make some larger flanges for black PVC pipes. This would help reduce sleeve breakaway during use. Also at the glove end, to allow for easy replacement of the gloves, I suggest a solid non corrugated junction, again the yogurt tub, or aluminum dryer vent line section, fastened to the sleeve and well taped, then the gloves can go on and off with more ease, and a good seal as well. I used to hate changing the gloves on the boxes at work!

author
ponckk made it! (author)ponckk2013-02-06

thank you for your thorough description, it helps a lot, for upgrades or later versions :).
I think the main difficulty is that the flexible tubbing has a spiral backbone (like a slinky)... it is not like an accordion design (I could not find an affordable one with my time frame), which could make your DIY coupling (for sleeves and gloves) much more easier to implement.
Thank you.

author
The Mighty El Rondo made it! (author)2013-02-03

Any chance you might have some method for a DIY cleanroom box that I can make and use to work on hard drives?

author
ponckk made it! (author)ponckk2013-02-06

This could work...maybe making another valve, an OUT valve in which you can vacuum atmosphere from the chamber (at a smaller rate, and carefully... so the whole chamber doesn't implode), at one point you could have very clean air inside...and stop the vacuum.
just a thought... :)

author
codongolev made it! (author)2013-02-03

I was really hoping this post would start with "so I was planning on stealing the Declaration of Independence because there's a map on the back, and I needed to outfit the back of my van with a lab..."

great instructable. very thorough.

author
ponckk made it! (author)ponckk2013-02-06

jejejeje, thanks.

Still this is a part of a larger project that could potentially change the world ;)

author
robertmurdoch made it! (author)2013-02-03

I love this... I don't need it yet but I wave wanted to set up a lab for my daughter and I to do experiments. Very cool.....

author
ponckk made it! (author)ponckk2013-02-06

awesome, let me know how you do!

author
harristotle made it! (author)2013-02-03

Impressive effort! I was going to suggest a tray with steel wool and vinegar as an oxygen scrubber, then I saw your oxygen removal tablets. Watch that you don't use silicone tubing to connect your N2 cylinder - it is quite O2 permeable. Ditto silicone gloves.

What a brilliant idea, and excellently executed! I would have liked to use this for some long term sampling of a Winogradski column. Ahhh, in another life!

author
ponckk made it! (author)ponckk2013-02-06

OOOO this is great!!! thank you for that O2 sequestering tip!
Good info on the tubbing and gloves material, thanks!, Im using vinyl tubing.
Thank you for the encouraging comments.
:D

author
toad made it! (author)2013-02-03

Great idea, I need to replace the platters on my harddrive and was looking for a way to keep the dust out. This will also work for lens repair. Thanks for the tip.

author
ponckk made it! (author)ponckk2013-02-06

great!
the only remark I have to make is that since the large container is not completely clear, detailed work might be a little bit hard.
So this is another upgrade that I'm pending:
Cutting a large rectangular space above the gloves, and use plexi, and for the borders, rubber gaskets (or insulation foam could be used), and the metal frame to be able to apply pressure to the screws that hold everything together, without cracking the plexi or the container.

author
Tachyon made it! (author)2013-02-02

Just an idea, but imagine putting this inside another, larger, clear box with one shared wall. The wall containing the gloves. Now make the shared wall air tight. Next make the outside box a flow through negative pressure box. Vent that safely outside through appropriate filters.
You now have a protected, positive pressure interior box that protects the environment of the experiment and a negative pressure outer box that protects the experimenter.

author
ponckk made it! (author)ponckk2013-02-06

Sounds great! Just be safe, remember this was built for a Bio Safety Level 1 lab, where you don't interact with anything harmful for humans :)

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