GMU:DIY Biolab “Driver’s License”/Mana Mazidy

Which medium ??

bioluminescence

Bioluminescence is the ability of a living organism to produce and emit light, so to send it out. This is possible through certain chemical reactions that take place within the organism. A luminescent substance (luciferin) is oxidized to oxyluciferin with the help of an enzyme (luciferase) (consumption of oxygen and energy). The decomposition of oxiluziferin produces energy that is converted into light of a specific wavelength (λmax 530 nm) almost without loss. Due to the wavelength, we perceive the glow as green.

distribution

Animals

mushrooms

protozoa

bacteria

In this project I use mushrooms and bacteria to illuminate the drawings

bioluminescent chemicals

So we have a connection between humans, media and living organism. You can see the normal photos on banknote, but when the light is off, the bacteria will glow and you'll see glowing photos telling the right story.

Bacteria

Ability

they only glow when they are alive . so you can know are they dead or alive ! in this project we can use this Ability to draw . The positive area should be filled with live bacteria and negative area with dead bacteria.

Make Bacteria glowing :

BIOLUMINESCENT BACTERIAL LIGHTBULB

http://www.instructables.com/id/Bioluminescent-Bacterial-Lightbulb-Water-Polluti/

Step 1: What You'll Need

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 [7]

Step 2: Get Your Hands on Some Dead Sea Fish

If you live by the ocean, GREAT. Just go down to the local fish market and get some fresh ( never frozen) squid, shirmp or fish from the sea. I tend to think squid is the best, then shrimp then sea fish since vibrio fischeri etc live inside the light organ of the bobtail squid, but there are tons of these guys all over the ocean. With a little luck the seafood you pick up will have the strains on it, if not, just try again or buy online like the rest of the land locked people. If you are land locked but visiting the beach you may be able to put the fish in a jar, then pack the jar inside a cooler with ice untill you get home but its not a guarntee. Or you can take some sterile agar culture plates with you and grow some bacteria while your on vacation and take it back home to your lab....your family may think your a bit strange but it's worth it ( I did it on a recent trip to Cali muwahahaha).

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  Fresh fish [1]

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Step 3: Make Some Bacteria Food Method # 1

Like you and me bacteria need food to grow so let's make some food that our bacteria will LOVE. Its full of vitamins, amino acids, carbon, minerals and of course salt, without salt they won't live (they live in the ocean after all).

TSA agar gel recipe:

Per Liter of deionized water: (you can always reduce it by 1/2 or 1/4 if you don't need the whole liter)

40g TSA agar (or directions per liter depending on which company makes it) 30g Salt

Mix together in a large beaker or saucepan on a heating plate or stove.

If you are going to use a pressure cooker you just have to heat it up until it mixes because using the pressure cooker to autoclave it will sterilize it...if you don't have a pressure cooker bring the mix to a steady controlled rolling boil to sterilize it.

Pressure cooker instructions (wannabe autoclave)

Pour dissolved mixture of TSA, and Salt in a flask with a rag SLIGHTLY in the mouth and place in cooker. Place glass petri dishes (not needed if you bought some sterile plastic sealed ones) in as well. Fill cooker with about 1-2 inches of water. Seal pressure cooker and set heat to high When the top valve starts to dance and hiss start a timer for 15 min When time is up turn off heat and let cool down until pressure valve retracts

Your agar gel and petri dishes are now completely sterilized !!! ( the steam molecules created under pressure bounces around and kills any organisms that were present)

Step 4: Bacteria Food Method # 2

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A second method if you want to make bacteria food is to make your own nurtient agar from scratch (funner and cheaper). We will use PANCREATIN to digest the casein in milk into nutrients the bacteria can use and add some salt for minerals and agar-agar flakes to thicken it into a gel.

Pancreatin can be found at your local vitamin store...it's used by people that have trouble digesting food due to problems with thier pancreas. The pancreas releases digestive enzymes. The pancreatin pills contain porcine (PIG!!!) amylase, lipase and protease that disolve protien, fat and carbs. OINK OINK.

You will need:

• Pancreatin (I used 325mg capsules)
• Skim/Lowfat milk 7g
• 100ml Distilled water
• Aquarium salt 3g
• Agar-Agar seaweed flakes 2g

Pour 7g of skim milk into a beaker. Add 100ml of distilled water and mix. Then add the contents of 1 pancreatin capsule and mix. Allow to sit for 8 hours with occasional stirring. Next add 3g of salt and 2g agar-agar flakes. Heat up mixture until salt and agar-agar disolves (maybe a SLOW boil, you don't want to burn your agar). If you have an autoclave er uh.... I mean pressure cooker, sterilize it for 15 min. as per previous instructions. Pour still hot agar into sterile plates. Alow to cool into a gel and flip the plates over so gel is on the top.

Step 5: Bacteria Food Method # 3

you can buy ready-to-pour "photobacterium" agar from Carolina Biological for about 7 bucks for 125ml....it's pricey but is the best for luminescent bacteria

Step 6: Let the Fish Glow

Remove the squid from the bag and allow the ink to remain. Heat up your loop until it glows, allow it to cool for min, or use your sterile q-tip. Dip the loop/q-tip into the ink. Next streak the petri dish of bacteria food that you made with the loop or q-tip. (if you did good sterile technique, your dishes should not have any bacterial colonies growing on them when you streak it.) Streak the plates into 3-4 zones using an interrupted pattern as shown in the pic, this will help isolated colonies grow. Separate each streak with heating up your loop or using a new sterile q-tip. (The bacteria don't like to glow unless they are in isolated colonies, they are kinda shellfish uh I mean selfish) Place the plate in a cool dark (18-25 C) place and watch it grow and glow! I wrapped mine in newspaper to make sure they are in the dark. Note: Depending on temperature of the room and other factors, a newly streaked colony will take about 18-48 hours to grow large enought to turn on their glowing genes (lux genes).

bacteria need food to grow It should to be full of vitamins, amino acids, carbon, minerals and of course salt, without salt they won't live !

Agar is made from the cell walls of some algae species , mainly from East Asia. In microbiology, microbial growth media are almost invariably solidified with agar instead of the gelatin used until the end of the 19th century. Agar is more resistant to gelatin than the high temperature used in sterilization, and gelatin gels become liquid at higher incubation temperatures. Agar gel becomes liquid at 95 ° C and is more stable at higher temperatures than gelatin gels; it solidifies at 45 ° C. Thus, the addition of thermolabile substances before solidification is possible. Because some microorganisms can digest gelatin, this is another reason.

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6.Desember

I tryed to Make Some Bacteria Food :

TSA agar medium

• NaCI 2g .
• Yeast Extract 1g .
• Peptone 2g .
• Agar 3g .
• Fill with H2O up till 200 ml

Photobacterium

They are a genus of Gram-negative bacteria and belong to the family of Vibrionaceae. Some members of this genus have the ability to produce light and are therefore bioluminescent

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  7.December. it is not glowing :(

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  14.Desember. they are growing but not glowing

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  20.Desember. I use this bacteria to draw. I want to see how many details are visible when they are growing

The illuminated mushroom

This mushroom has developed an amazing feature, it shines in the dark. In our native forests sometimes infected by the Hallimasch root pieces. This phenomenon is referred to as bioluminescence and is also known by the fireflies. For most observers of yesteryear, the glowing wood was mysterious and eerie, and associated with wizards, fairies, and elves. In this mushroom species shine both the mycelium and the fruiting bodies. Interestingly, however, only the American style of Panellus stipticus shines. He has a creamy white color and 1-2 cm large, shell-like fruiting bodies that spread throughout the trunk. Every year new fruiting bodies appear.

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Nothing happens without reason in nature. So the extraordinary appearance of the mushrooms has a biological function. The emitted green light is visible to insects and attracts them. Similar to the "bees and flowers principle", the insects land on the mushrooms and transport on their onward flight spores that have stuck to them. Thus, the mushrooms receive by their striking luminous advantages in the propagation and distribution.

Panellus stipticus

The light mushroom (Panellus stipticus), often called fairy-fire or ghost-mushroom, belongs to a small group of mushrooms, which glow in the dark. Through natural bioluminescence, similar to fireflies, the tormenting twining (Panellus stipticus) enlightens its surroundings in a unique light. The mycelium is already recognizable, but the fruiting bodies are even more intense.

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Grow mushrooms yourself

Ordered at : https://dikarbion.eu

The mushroom is also ideal for terrariums with reptiles, amphibians and invertebrates, if they are regularly moistened

• temperature in the house should be all year round up to 25 ° C

I ordered the shining mushrooms from https://dikarbion.eu . I got it after a week. I first had to lay log in the water mixed with vinegar. It had to stay in there for 18 hours. Then I made the holes and put the mushrooms in.

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Light fungus Finished culture in the WECK © Glas

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Modification of green fluorescent

I noticed that most artists used either bioluminecenses or fluresences bactera.

But what's new: modified with fluorescent protein! I tried to know, what is it? and how can I do that?

is a protein composed of 238 amino acid residues and shows a bright green fluorescence when exposed to blue to ultraviolet light. The artists have used it for art projects and scientists have used it to highlighting specific structures.

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Different proteins produce different fluorescent colors when exposed to ultraviolet light. (Picture taken by Erik A. Rodriguez.) https://en.wikipedia.org/wiki/Green_fluorescent_protein#/media/File:Fluorescence_from_Fluorescent_Proteins.jpg

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Results

The world is huge with so many hidden organism that are alive.This project let me to know this new world. helped me to learn how to control these little animals and learn the differences from them. Nothing happens without reason in nature. These little animals have helped me with their skills to explain my story. A co-operation to explain a collaboration between humans and nature to history.

Related projects

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  [https://www.uncommongoods.com/product/bioluminescent-aquarium

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  This beautiful dress has a glowing secret: bacteria

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  Glowing trees could light up city streets

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  Glowing trees, no more colonoscopies and other wonders of synthetic biology

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  This painting is a living thing–literally. Artists at the World Science Festival in Brisbane, Australia, are creating art from marine bacteria known as Aliivibrio fischeri.

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  Student project of chicago university. fluoresced bacteria - meta-bacterial

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  With Bioluminescent Bacteria - gallery at Montana State University

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  from artist Miya Ando and photographer L. Young. With a different kind of bioluminescence

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  composed of multiple petri dishes of bacteria modified with fluorescent protein under UV light

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  http://www.ohiotrm.org/glow-in-the-dark-table/glow-in-the-dark-table-with-photo-luminescent-resinyoutube/

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  http://www.ohiotrm.org/glow-in-the-dark-table/glow-in-the-dark-table-with-photo-luminescent-resinyoutube/

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  https://www.huntercole.org/news/2017/10/12/donate-to-make-hunter-coles-art

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  http://www.annebrodie.com/#/exploringtheinvisible/

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  https://www.mnn.com/lifestyle/arts-culture/blogs/scientist-hunter-cole-bioluminescent-bacteria-creates-works-art

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  https://www.catersnews.com/stories/quirky/watch-where-youre-glowing-artist-creates-worlds-first-solar-powered-cycle-path/

Art / Design / Products using luminous bacteria

• Simon Park - Exploring the invisible
• Bioglyphs
• Hunter Cole
• Rebecca Klee
• Kathy M. Takayama
• Anne Brodie
• Tim Otto Roth
• Teresa Van Dongen
• Marianne Engel
• Thomas Feuerstein
• Anna Dumitriu

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References :

http://www.katrinrodegast.de/money-art/ https://www.designboom.com/design/money-design-and-history/ https://makezine.com/2013/05/16/diy-synthetic-biology-making-your-own-glowing-plants https://www.huffingtonpost.com/2012/02/13/glowing-sushi_n_1273131.html https://dikarbion.eu/epages/96a7356b-10eb-4075-b76d-a2feb5b9cba5.sf/de_DE/?ObjectPath=/Shops/96a7356b-10eb-4075-b76d-a2feb5b9cba5/Categories http://www.instructables.com/id/Grow-Your-Own-Bioluminescent-Algae/ https://www.wikihow.com/Grow-Bioluminescent-Algae-at-Home http://www.instructables.com/id/Bioluminescent-Bacterial-Lightbulb-Water-Polluti/ https://en.wikipedia.org/wiki/Panellus_stipticus https://gl.wikipedia.org/wiki/Photobacterium https://en.wikipedia.org/wiki/Bioluminescence https://oceanservice.noaa.gov/facts/biolum.html https://www.britannica.com/science/bioluminescence