Bioluminescent Bacterial Lightbulb / Water Pollution Tester

 by drdan152
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Bioluminescent Bacterial Lightbulb

This instructable will show you how to grow and culture your own glowing bacteria and use it to reinvent the light bulb!

I"ll show you how to grow your very own Vibrio phosphoreum or Vibrio fischeri from fresh sea fish (squid work well) and culture it onto special agar gel that you can make at home or simply buy ready-to-pour. The gel can be formed inside all kinds of things that you want to make glow a cool greenish/blue color !!!!

The glowing vibrio bacteria are marine life forms that live alone or in a symbiotic relationships with fish and squid inside their light organs or as parastites. Alone the bacteria don't really glow but in groups of large enough number something amazing happens called quorum sensing ..... The bacteria upon realizing they have enough of each other around turn on genes that allow them to GLOW !!!! The light that is emitted is super efficient around 98 % meaning that only 2 % is lost through heat, thus the light is cool and that's very COOL. 

Here is some good info on v. fischeri
http://www.bionity.com/en/encyclopedia/Vibrio_fischeri.html

It's these amazing little guys (1/1,000,000 of a meter long) that will power our lightbulb !!!! If properly cultured your lightbulb will glow for about a month using zero electricity and zero pollution!!!!  Granted its not incredibly bright but you can save electricity  costs if you replaced a night light , or you could use it to read maps or books while camping and not worry about changing batteries in your flashlight and disposing of the batteries causing more pollution !!!! 

Water Pollution Tester 

This instructable will also demonstrate how glowing  bacteria can test  how toxic water is. The methods used are much more Eco friendly than chemicals and processing ..... So not only does bioluminescent bacteria reduce pollution , they can also DETECT IT !!!!  The glowing process is a direct result of the bacteria's metabolism. Fresh clean water does not affect its cellular metabolism or how bright it glows. However even slight toxins that we can't smell , see or taste can alter the bacteria's metabolism and will reduce the amount of light they can produce. By comparing the amount of light emmited from bacteria treated with clean water to that of the sample at hand ..... Say from a stream near a power plant we can assess the level of toxicity in the water !!!! 

Here is a video of what the bacteria look like ALIVE under high magnification:
 
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Step 1: What you'll need

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  • Fresh (but dead) Sea fish, squid or shrimp (if you don't have access to fresh fish you can simply buy the bacteria from Carolina biological supply here: http://www.carolina.com/product/vibrio+anguillarum%2C+living%2C+tube.do?keyword=Vibrio&sortby=bestMatches)
  • Aquarium salt (from pet store) or fresh sea water 
  • Agar growth medium ( dehyrated tryptic soy agar, or homemade, or ready to pour photobacterium agar...you get to choose if you want to make it or buy it.)
  • Sterile Petri Dishes (or glass bowls that can withstand being boiled)
  • Sterile Q tip or loop of solid wire
  • Pressure Cooker (optional but highly recomended)
  • Large Beaker or saucepan
  • Med size Erlenmeyer flask (optional)
  • Distilled Water
If making your own agar gel see step 4 for supplies
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drdan152 (author) says: Aug 5, 2012. 12:20 PM
I've been doing some experimenting on adding things to the agar to make it glow more...it seems so far that by adding a crushed calcium vitamin (as calcium carbonate, CaCO3) AND some glycerol it glows brighter!!! I'll add the new recipe as soon as testing is done.
ksmith89 says: Mar 29, 2013. 11:08 PM
earlier you said "Just FYI I'm experimenting with a new agar to increase bioluminescence ..... It has more nutrients that should support more glowing ..... Calcium carbonate , yeast extract , and glycerol !!!!" so did you ever figure out your new agar? what amounts of the Calcium carbonate , yeast extract , and glycerol do you add to the ager recipe for the homeade ager method #2
qharding says: Feb 23, 2013. 7:16 PM
Hi. I know its been a long time since anyone has commented but I really need your help I'm currently growing Vibrio Ficheri and I have so many question. Mainly how did you get them to show up in pictures? Please help
drdan152 (author) in reply to qhardingMar 14, 2013. 8:49 PM
HEY! Glad to see people growing bacteria ....haha. Well the best way to take a picture is in a completely dark room and get an SLR camera or any camera that you can adjust the shutter speed. Mount the camera on a stable surface or tripod and set the shutter to stay open for as long as possible...maybe even 30 seconds. I set the timmer so it counts down 5...4...3...2....1 then take the picture so i dont bump the camera and get a blurry shot. BIggest thing is the shutter speed though .....think LONG or SLOW shutter speed and keep the camera steady!
ozzyfast says: Jan 4, 2013. 5:43 PM
Hi friends,
where can i buy v. fischeri's culture from Europe?

regards
ryan8816 says: Nov 17, 2012. 12:33 AM
For anyone who would like to see the Tryptic Soy Agar label on the bottle
IMAG0770.jpg
nano nite says: Sep 24, 2012. 9:07 PM
Do u have any idea about biological shops in India where i can find theese bioluminescent bacterias ? Be soon buddy its for my science exibition.
nano nite says: Sep 24, 2012. 11:19 AM
hey can u give a clear comparison between bioluminescent light bulb and an electrical light bulb ? including all aspects.
dragonito says: Sep 2, 2012. 8:34 PM
This might be a bit of a stretch... but could a concentrated, sterilized dirt/compost extract combined with digested skim milk (and perhaps some yeast extract) be used to make a suitable photobacterium broth? The digested milk and yeast would amino acids and vitamins, while the dirt extract would provide (trace) minerals. And keeping the bacteria in a continuously agitated medium would also increase the oxygen content, possibly maximizing glow.
drdan152 (author) in reply to dragonitoSep 5, 2012. 8:10 AM
Dude that sounds really cool
..... Not sure it of will work but I think it would be an awesome experiment to see if it does ..... As Bill Nye the Science guy says "TRY IT" !!!!
pchubb says: Aug 16, 2012. 5:37 PM
Any reason to use tryptic soy agar? I just mixed up agar into lamb broth made by boiling an old bone with a bit of meat on it. I added salt to make it 3% w/w, and I have the bacteria growing well.
drdan152 (author) in reply to pchubbAug 19, 2012. 6:23 AM
Well to be honest I'm not sure if boiled bone has all the nutrients that photo bacteria need..they are kinda picky eaters...I've noticed that the bacteria will readily GROW in different agar mediums (even polluted ones) but they will only GLOW if they have juuuuust the right conditions and nutrients to support their glowing reactions....thats why it makes such a good "water tester" cause they amount they glow is directly related to how good thier environment is. TSA has tryptones (short chains of amino acids) and other good stuff that fastidious (picky) bacteria like.
pchubb in reply to drdan152Aug 19, 2012. 3:39 PM
The colonies are growing and glowing well. It's my experience that most bacteria are not fussy eaters (unlike my ten y-o).
drdan152 (author) in reply to pchubbAug 19, 2012. 4:47 PM
Oh wow !!!! I'm sorry I thought you meant they were growing but not GLOWING !!!! Excellent !!!! Isn't amazing how glowing bacteria can come from a dead squid !!!! Good job culturing from different areas I only used the left over ink from the bag the squid was in. Can u post a picture of your glowin microbes ???? Would make a cool post for the instructable :-)
pchubb in reply to pchubbAug 16, 2012. 5:40 PM
Oh, and I tried culturing from different parts of the squid. I got nothing from the ink sac, but plenty from the body cavity (inside the `tube' of the squid. Our local squid don't glow.
drdan152 (author) in reply to pchubbAug 19, 2012. 6:28 AM
Try adding a pinch of powdered calcium carbonate (from a vitamin crushed into a powder...it shoul say on the back "calcium as calcium carbonate") to the medium you made...or maybe a few ml of glycerol for a carbon source?? also try letting your eyes adjust in PURE darkness for 5 min...if you find a glowing colonie , fish it out and reculture it in fresh agar......BUT it may be that the squid was previously frozeon OR you just got a bad egg...or uh squid. DO you have acess to a microscope? If so you can see if the bacteria you grew is indeed the a vibrio species.
pchubb in reply to drdan152Aug 19, 2012. 3:40 PM
As I said before, the bacteria cultured from the head and body cavities (under the mantle) are glowing well. It's just that I found no colonies growing from the ink sac.
yellowcatt says: Aug 17, 2012. 9:00 AM
Great instructable.
It got a mention on the This Week in Microbiology podcast:

http://www.microbeworld.org/index.php?option=com_content&view=category&layout=blog&id=107&Itemid=275
drdan152 (author) in reply to yellowcattAug 19, 2012. 6:18 AM
REALLY !! thats so awesome i love microbe world hahaha thanks for the info!
yellowcatt in reply to drdan152Aug 19, 2012. 7:18 AM
Your instructable was mentioned right at the end of podcast 39, when they were reading out the listeners emails, (at 1hour 11minutes and forty seconds)

Vincent Racaniello described it as "really quite amazing".

Mushroom man says: Aug 12, 2012. 7:30 AM
Aw dude this is awesome i've been trying to grow this for so long and this just simplified it sorry i'm new so I don't know how images work
drdan152 (author) in reply to Mushroom manAug 19, 2012. 6:30 AM
Sweeeet! Do you have acess to fresh non frozen fish or squid???!
techno guy says: Aug 15, 2012. 8:30 PM
did the idea for testing water pollution with this come from the idea that the 15 year old who made the cancer test strips had
drdan152 (author) in reply to techno guyAug 19, 2012. 6:30 AM
No, it's a pretty common experiment with v. fisheri.....i wish i came up with the idea but its been around :-)
drdan152 (author) says: Aug 3, 2012. 6:38 AM
Just FYI I'm experimenting with a new agar to increase bioluminescence ..... It has more nutrients that should support more glowing ..... Calcium carbonate , yeast extract , and glycerol !!!!
jcbeaver7 says: Jul 29, 2012. 5:43 PM
Why do the bacteria only last a month? if you kept them fed would they last longer? That would be cool if instead of electricity you used fish :D
drdan152 (author) in reply to jcbeaver7Jul 29, 2012. 6:10 PM
Well you CAN keep them growing longer if you continue re culture them from the brightest glowing spots into fresh agar :-) You could tell all your friends " yeah my house is powered by dead fish ..... How bout yours huh ? "
jcbeaver7 in reply to drdan152Aug 2, 2012. 2:42 PM
haha! If only i had the work ethic and supplies to finish any of the projects on instructables.....
Super Cronk says: Jul 30, 2012. 5:53 AM
yes, i like it a lot. I think I might try this with my kids. One question, is it possible to suspend or slow their growth once the colony has been established? For instance, can it be refrigerated when it's not needed or would that kill the bacteria?
drdan152 (author) in reply to Super CronkJul 30, 2012. 7:12 AM
I'm actually experimenting right now with refrigeration and storage ..... Note the last step on making "glycerol frozen stocks" ..... Also In all my research I found that v.fischeri can live and slowly grow at 4 deg Celsius which is household refrigerator temp. I'm experimenting with how long they live they way. How ever I think the BEST way to keep your bacteria alive is to re culture it every 48 to 36 hours ..... Which is kinda a pain ..... That's why I included instructions on how to make homemade gel cause store bought stuff can get expensive.
drdan152 (author) in reply to drdan152Jul 31, 2012. 4:52 PM
Ok so far the bacteria have survived and even noted some growth in my refrigerator for 6 days, wrapped in newspaper (so its dark). They still glow! My fridge is about 4 Celsius.
fuddle says: Jul 31, 2012. 4:21 AM
This is a superlative Instructable.
You have managed to demystify cutting edge biotech (OLEDS etc) an distill it in manner easily accessibly to many.

I have one question- is it possible to increase the brightness?
drdan152 (author) in reply to fuddleJul 31, 2012. 4:48 PM
So far I have found brightness to be a factor of density, oxygen, and strain of microbe...not sure yet if we can do anything to increase the brightness on a biological level....but I've been thinking of a highly reflective enclosure with perhaps a Fresnel lens built in to intensify the light, kinda like a light house has lenses or how flashlights have a reflective dome surrounding the bulb???
jbaker22 says: Jul 31, 2012. 7:35 AM
Can you breed it on solar panels?
drdan152 (author) in reply to jbaker22Jul 31, 2012. 4:45 PM
I don't think the wave length of light is the right amount to be of any use to solar panels ....but don't let that stop you....experiment!!
o2bjake says: Jul 31, 2012. 3:15 PM
Cool instructable. I used some of these bacteria a couple years ago trying to find a cheap/fast/effective way to detect quorum sensing molecules in the lung fluid of critically ill patients. It didn't work reliably, but I'm not sure whether it was because the bacteria (detected by culture and molecular genetics) weren't producing the molecules or simply that the fluid was too dilute to activate the bioluminescence.

I'd love to see something like a neon sign using recirculated V. fischeri culture in tygon tubing, though the bacterial density (and therefore light output per area or volume) in culture is far lower than you get on solid medium, so it may or may not produce decent results (except maybe in total darkness, which would still be cool).
drdan152 (author) in reply to o2bjakeJul 31, 2012. 4:44 PM
wow.... again these bacteria impress me! Yeah things that glow are just cool no matter what haha.
twhaley says: Jul 30, 2012. 12:55 PM
I just drank all my bacteria water by accident, will i be ok?
drdan152 (author) in reply to twhaleyJul 30, 2012. 6:08 PM
If you develop severe diarrhea ..... ESP if it's glowing bright blue in the toilet ..... Im sorry ..... You have 6 months to live.
sciencetor2 in reply to drdan152Jul 31, 2012. 12:58 PM
i believe the photobacterium you are working with is a food poisoning agent, but not a pathogen
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