Biolumin: Blue Light Green World How I made the World’s First Bioluminescence Illuminated Christmas Tree Edward A. Quinto
The World’s First Bioluminescence Illuminated Christmas Tree
If the year 1992 was for her Excellency Queen Elizabeth II an “ Annus Horribilis” or horrible year, the following year 1993 was for me my “ Annus Mirabilis” or wonderful year. In that year, I discovered up close and personal one of nature’s most aweinspiring living wonders. As I entered a dark laboratory in Germany, I vividly saw for the first time nutrient broths contained in Erlenmeyer flasks shining brightly with a bluegreen hue of captivating spectacle. Swimming in the broths were the teeming trillions of shining cells of the marine luminous bacterium: Aliivibrio fischeri . The combined radiance of the astronomical number of cells each shining like tiny light bulbs generates the homogeneously luminous blue liquid or plate colonies we humans see as the biological phenomenon of bioluminescence. While electrical light bulbs generate heat
over time, bioluminescence does not, thereby aptly earning for it the title of “cold light”. Since, Aliivibrio fischeri and many other luminous microbes are the bearers of light in the world of the smallest; I fondly called them the “Microbes of Light”. Bioluminescence can reveal itself in rare occasions and on a grand scale as the mysterious light illuminating a vast region of the powerful nighttime sea. This eerie shining sea, not to be confused with St. Elmo’s Fire, was mentioned by Jules Verne in his book “20,000 Leagues Under the Sea”. Sailors through the ages chronicled this mystifying light of the sea in their logbooks now known in science as the phenomenon of the “Milky Sea”. If one were cruising in the midst of a “Milky Sea”, the sea will appear shiny white like fresh milk stretching out far into the horizons. That is how this uncommon marine occurrence got its name. The satellite of the U.S. Naval Research Laboratory captured the image of a large “Milky Sea”, the size of Palawan Island, for the first time on 25, 26, and 27 January 1995. Seamen in a merchant ship at that time located off the coast of the horn of Africa also witnessed this unusually bright “Milky Sea”. For a bioluminescent phenomenon close to my heart to fall on a 26 January, my natal day, is indeed an honor! Bioluminescence is the “Light of Life” resulting from chemical reactions taking place inside the cells of living organisms. It renders the lantern fishes and the firefly squids luminous in the darkness of the depths. Furthermore, in the perpetually dark abyss, which covers a large part of the oceans’ basin, bioluminescence holds a significant role in the interactions of living organisms. Bioluminescent organisms have played out beautiful pyrotechnic flashes to deep-sea research submersibles. The “Light of Life” abounds in nature but is far more common in the hydrosphere than on the lithosphere. On land, bioluminescence unfurls its beauty by the twinkling of Fireflies, glimmering of mushrooms and the glowing of some worms. However, in the dark void of the continental ocean basin it is the only means of communication for marine denizens grand and small. Fishes flash, squids shine, Jellyfishes flicker, and a host of invertebrates shimmer in this dark largely unexplored world. Indeed, as an amateur astronomer I realized that the “The Microbes of Light” are like heavenly bodies called stars for they produce their own radiant energy in the darkness of their own deep-sea universe. These marine organisms like fishes and squids can produce their own lights by utilizing special “Light Organs” harboring dense loads of symbiotic luminous microbes. The Flashlight fish carries light organs under its eyes shining like headlights in the dark. My favorite luminous animal, the “Hawaiian Bobtail Squid” used light organs located in its belly to shine thereby canceling its shadow during full moons and so disappear from the lurking eyes of hungry predators swimming beneath it. These luminous microbes were the very same microbes that I saw in the shining broths in Germany in 1993. I was also fortunate to have isolated these luminous organisms from local sources here in the Philippines. Having successfully cultivated these luminous
microbes, I stored and preserved them in the microbiology laboratory in UST. These microbes are identified by their scientific names: Aliivibrio fischeri, Aliivibrio harveyi, Photobacterium phosphoreum, Photobacterium leiognathi and many others. When they are isolated and cultivated in special microbiological broths and agar media, they can be made to shine with a strong and enduring luminosity of alluring wonder. Scientists have estimated that each cell acting like miniature light bulbs generate 10,000 to 100,000 photons per second. A photon is the particle of light carrying discrete packets of energy. In the dark, our own scotopic vision (Dark Vision) is so sensitive to the bluish-green color of bioluminescence. Photoreceptor cells called rods in human eye can visualize as little as one photon! One culture agar plate of these marine luminous microbes can produce enough light to enable one to read books in the dark! When stained and observed through a microscope, the one-celled luminous microbe Aliivibrio fischeri appears as a plump rod occurring singly or in pairs. This “heavenly microbe” can also be found non-luminously in the intestinal contents and on the bodily surfaces of squids, fishes, crustaceans, and seaweeds. The bioluminescent animals use light to attract their mates and prey, as well as to distract and evade their predators. When the luminous microbes are isolated and grown in special nutrient broths, the population increases until a critical size is reached when every cell of about five billions per drop of broth shine all at the same time. What an awesome sight it must be to behold this phenomenon at the cellular level! This ability of microbes to communicate to one another to determine how big their population size is, comparable to conducting their own roll call, is termed “Quorum Sensing”. Imagine the action of five billion cells switching on their lights simultaneously! So stand aside, the lighting up of the light bulbs of the New York Rockefeller Christmas Tree. Evolution chose the emitted light of bioluminescence to be blue because this color is more energetic than green, yellow, and red thereby enabling it to travel farthest in the sea. To travel far in the sea, makes blue an efficient means of signaling among living organisms in a world where astronomical open spaces is the norm. Red and yellow “living lights” also occur in nature though minimally. It is also the reason why the marine world appears with a cool bluish tinge under white light or sunlight.
Glowing colonies of a plate culture of of Vibrio Vibrio fischeri
Brightly shining broth of Vibrio of Vibrio fischeri in fischeri in a flask
Upon my return to the Philippines in 1993, after successfully finishing my “Aufbaustudium” in Germany, I thought then that it would be the end of my work with microbial bioluminescence. I never thought of bringing the luminous microbes home with me though it is possible to do so. However, my desire to work again with microbial bioluminescence was simply too strong to be side stepped. So armed with my knowledge of microbiology, I embarked on an exciting quest to seek out these tiny luminous creatures in our locality. What a fulfilling quest it surely was! To my delight, I realized that the wet markets of the Philippines are virtual goldmines of these marine luminous microbes. Squids, saltwater fishes, crustaceans, mollusks, and even seawater itself are excellent sources sources of these bioluminescent microbes. In 1995, not long after I started scouting for them, I venerably held and beheld once more the radiance of the Philippine strains of the marine luminous microbes. Alone in a dark room, I just found myself staring at the luminous plate cultures shining as bright as the ones I saw in Germany. Sentiments of euphoria overwhelmed me for it felt so much like having found a long lost cherished friend. It was an unforgettable exhilarating experience as I felt their life’s radiant energy piercing the darkness and illuminating my improvised dark room. When it comes to catching people’s attention, my “Microbes of Light” did not let me down. The cool blue light of bioluminescence always generates gasps of amazement whenever I show them to friends and colleagues. My research started and centered on how I can simplify the utilization of these luminous microbes to uplift the quality of life. The rest as what they say is definitely history! Results of my research showed that these luminous microbes can be used inexpensively as a biological sensing device for the detection of toxic chemicals in water, foods, produce, and other samples. If these shining microbes are added to a water sample containing minute amounts of toxicants like heavy metals, detergents, pesticides, etc., their generated light fades significantly or may even black out completely. The strongly toxic a substance is and the higher its concentration in the
water sample tested, the faster is the complete inhibition of light production. Thus, checking drinking water, foods, and produce for safeness and the biomonitoring of environmental water for pollution are the main applications that I developed from my research work with microbial bioluminescence. Numerous blessings poured in from all aspects of my work with the “Microbes of Light” in the form of international and national awards, publications, thesis completion, and recognition from the Philippine Scientific Community. PIBiT (Paper-disc Immobilized Bioluminescence Technology) and TuBET (Tube Bioluminescence Extinction Method) were the simple and green innovations that came out of my research work. Through the years, high school students from UST, Assumption (Makati), Quezon City Science, Manila Science, Immaculate Concepcion (Davao), Parañaque Science, Philippine Science, Imus Science, and La Consolacion worked with microbial bioluminescence and many won in various local, regional, national, and international science fair contests. Undergraduate and graduate students in UST also utilized the luminous microbes for the successful completion of their theses. Many others and I surely did have the time of our lives with these unique and wonderful creatures, which have become a part of my life ever since. Playing around with them, I can proudly say that I was able to light up the world with the good blue light of bioluminescence. The benevolent sea has granted me the privilege to borrow this beautiful “Light of the Sea” that I used on land for many practical health-promoting benefits not only for people but for our environment as well. Indeed, I was able to light up so many things with bioluminescence. The first materials I made bioluminous were of course, water samples, then cotton plugs, small paper-discs, raw foods, metal plates, and to various produce like meat and beverages and would you believe even a small Christmas Tree!
Rich sources of the luminous microbes found in wet markets
So one cool memorable night on 28 November (Wed), as Christmas 2007 was fast approaching, I attempted the creation of the first Bioluminescent Christmas Tree in history or the World’s First Bioluminescence Illuminated Christmas Tree. Several days before, the idea of a bioluminescent Christmas Tree got me so excited as a Scuttle Fly. In the past, people used candles to illuminate Christmas trees. Then electricity was discovered, and light bulbs of various designs and colors completely replaced the use of candles. Now, a sophisticated electrical novelty for Christmas Trees like the use of fiber optic has dazzled the public. Two years ago, the Rockefeller Christmas Tree introduced the use of LED lights to promote a greener Christmas because of its small electrical consumption. An idea struck me one day; why not use bioluminescence to illuminate a Christmas Tree. Surely, it will be a first, it will be splendid and it will epitomize something really green because there is no consumption of electricity nor will greenhouse gases be emitted. It can symbolize all the “Green Movement” against Global Warming. So one afternoon, I excitedly visited the malls in and around Manila looking for the right tree. At the Gotesco Grand Central Department Store, I found a two feet tall dark green pine tree standing quite robust and thick. The smaller the tree the easier it will be for me to light it up entirley with microbial bioluminescence. I started the cultivation of my microbial friend, Aliivibrio fischeri in nutrient agar plates a day before I was to use them to light up my newly bought tree. The big night soon came; in the laboratory, I made a brightly shining liquid by suspending the luminous colonies from the plate culture in saline. Then, I rapidly filled twenty-five small glass vials the size of thumbs with the brightly shining suspensions. Next, I immediately capped the vials and tied short black strings around their necks. One by one, I hung the shining vials like light bulbs all over the branches of the tree. I hurriedly did it because I knew that once the dissolved oxygen in the liquid inside was consumed by the luminous cells the light would go out instantly. I just don’t know how fast that will occur with the vial size and physicochemical conditions present at that time. If I can just be given 20 minutes to pour, cap, tie, hang and photograph them up on the tree then I reckon that I can pull it off. Like fire, bioluminescence is powered by oxygen and without it, bioluminescence is extinguished immediately. Half way through the process of putting the vials up on the tree, the expected happened abruptly. One by one, the shining vials were “blacking out” indicating the depletion of the dissolved oxygen. I shook the vials in a desperate attempt to aerate the liquid inside. Aerating the liquid inside the vials should light them all up again though fleetingly. Since the vials are small, the dissolved oxygen is low, the quicker it is consumed by the luminous microbes and the faster the light will fade out. It was a futile exercise as the vials were “blacking out” again much faster than I can keep them alight. The entire exercise that night was so utterly disappointing. Using my digital camera, I frantically took long-exposure photographs of a dimly lit tree with half of the
twenty-five vials’ light already gone. The mossy green color of the robust tree must have also exacerbated the results, as the tree appeared so dark and gloomy. The dark green color seemed to be absorbing the light of bioluminescence and not reflecting it.
The robust dark-green Christmas Tree
In darkness with the luminous glass vials
But hope indeed springs eternal. From the lessons of this failure, I tried my best to come up with a better tree in my next attempt. Another opportunity has to be forthcoming I said to myself. The next time around, I thought of using long cotton strands wetted with luminous suspensions instead of putting them inside closed vials. Previous experiences with cotton plugged flasks of luminous broth cultures that accidentally got wet in the shaking produced shining cottons plugs for hours. In closed vials, the oxygen supply is small and limited it comes mainly in dissolved form in the liquid suspension of the luminous cells. However, I realized that in cotton impregnated luminous suspensions an abundant supply of oxygen should come from the surrounding air. This infinite supply of oxygen in the air should be able to sustain bioluminescence strongly and brightly over a long period. The mere thought of brightly shining suspensions in cotton reinvigorated me once more in my quest to make a bioluminescence illuminated Christmas tree. I cultivated once again my Aliivibrio fischeri in nutrient agar plates to produce the luminous colonies and then the luminous suspensions. I also replaced my dark bulky green tree with a two feet tall white plastic pine tree that I bought from Robinsons Place Manila. White should give a more brilliant and reflective appearance. I extended open the main branches of the tree from the central stem and started loading the entire length with thick strands of absorbent cotton. The tree was also decorated with small shiny orange Christmas balls. Then I freely sprinkled a brightly shining suspension of the luminous microbe on the
cotton strands using a glass dropper. At around 6 PM, I was finished and that time close to Christmas time in Manila, it’s already quite dark due to the gradually lengthening night. Luckily, it was also a holiday since its Andres Bonifacio Day, November 30 and so I was all alone in the research complex. I moved back to the door, took a deep breath and said to myself that this is it! I turned off the lights and what I saw from afar was truly an awe-inspiring scene. On a working bench, fronting the wall stood a Christmas tree brightly illuminated by the light of microbial bioluminescence. Feelings of relief and accomplishment seethed inside me. To capture the historical moment that could be over in hours, I swiftly prepared my digital camera and took several long exposure shots. The camera screen showed shining strands of cotton with a dramatic bluish color and cool snowy effect resting tenderly on the branches of the tree. One can even see luminous drops of the shining suspensions at the base of the tree. After taking the photos, I decided to transfer them immediately to my laptop for better viewing and storage. When I saw the photos on the screen of my laptop, I was so amazed and thrilled. All the efforts and stress that went into days and nights of preparation were definitely worth what I accomplished that night. Once again, in my thoughts as I beheld the beautiful bioluminescent photos, I expressed my deep gratitude to my “Microbes of Light”. Indeed, once again they did not fail me in my efforts to promote bioluminescence to the public for a greener world. The night of 30 November 2007 was truly memorable because aside from being the night after the foiled coup d’ etat in Makati, I can proudly say that I succeeded in creating the world’s first bioluminescence illuminated Christmas tree. Surely, it was not a “Franken Christmas Tree”, a living pine tree incorporating a bioluminescence gene from another organism introduced by DNA Recombinant Technology. A genetically modified brightly shining Christmas tree will still probably take years to be developed and the financial resources to make it must be quite big. Not to mention the intricate laws and regulations that one need to secure from the appropriate national authorities to be able to perform genetic manipulation experiments in the Philippines. Besides, what I wanted to make was a natural and more acceptable Christmas tree to the general public. And the best way to do it was to use microbial bioluminescence in its simplest and purest form. When I left late that night, I placed the tree in front of a large and long glass window of our laboratory exposed to the outside for others from the neighboring high-rise condo to perhaps see it as well. Three hours after I created it, the bioluminescent Christmas tree was still shining brightly and I was sure for many more hours after I left the laboratory. The university was practically empty of students and personnel that night as I walked out of the campus alone through the almost dark streets. I made a Christmas card of the photo bearing a brief “green” message highlighting a greater awareness in our fight against global warming. I immediately sent the card to the prestigious American Society for Microbiology (ASM), the largest US life science
organization, of which I am a full member. Since the light is from microbial bioluminescence, I was sure that the ASM would take notice of it. It was also sent to the venerable “International Society for Bioluminescence and Chemiluminescence” (ISBC) because my work dealt with bioluminescence. Suffice it to say, both the ASM and the ISBC, acknowledged what I did and featured the photo of the first bioluminescent Christmas tree in history in their famous websites. ASM featured it in their popular microbeworld.org website for 3 weeks in the run up to the holidays of 2007. For that, I am eternally grateful to both of them; it was the best Christmas gift that I received in a long while. Searching the net, I found out that Edward H. Johnson in his home in New York introduced the first electrically illuminated Christmas tree in 1882. Well another Edward made the world’s first bioluminescence illuminated Christmas tree in Manila in 2007. I disseminated the unique card by email to all my friends, colleagues, and people who I believed would be interested in it and I must confess that the replies were highly encouraging.
The Bioluminescence illuminated Christmas tree in darkness and under fluorescent white light
These were some of the encouraging and generous messages: “Hello Edwar d, Ver y many thanks f or your email and lovely BL (BioLuminescent) Chr istmas Tr ee! I have today sent the image to my colleague at the Univer sity of Bologna. He is the per son who maintains the ISBC web site. Best r egar ds,” Philip 03 DEC 07 (Dr . Philip Stanley of Cambr idge Resear ch & Technology Tr ansf er Ltd, Cambr idge, England). “Hi Edwar d! Ang ganda naman (It’s so beautif ul)! Ver y uniquely micr o. Thanks f or the Chr istmas gr eetings,” Rose (Dr . Rose Monsalud, Pr esident of the Philippine Society f or Micr obiology). “Hi Edwar d, ver y nice and ver y intr iguing. Congr atulations!” Toby (Pr of . Fabian Dayr it, Ph.D., Dean of Ateneo de Manila Univer sity’s School of Science & Engineer ing and a highly r espected tower ing f igur e of f P Philippine Chemistr y).
Hi Sir EQ, I am gr eatly honor ed to r eceived such amazing photo f r ro m one and only gr eener pastur e micr obes ..... ang ganda ganda!!!!! Regar ds to ever ybody.....miss you all, Tess (Ter esa Calwit, a f r ri end and teaching colleague in UST, now r esiding in Canada) Hi Sir Quinto, Salamat po Sir Quinto. Congr ats po sa bioluminescent chr istmas tr ee! Kr istine (Kr istine Hutalle, a f or mer r s student pur suing Ph.D. in Ger many) Hi Ed, Gr eat Chr istmas tr ee! Mer r r a all of ry Xmas and mor e bioluminiscence to you and f or us! Vivian T. (Pr of . Vivian Tolentino of the Depar tment of Biology, Ateneo de Manila Univer sity, a colleague and f r ri end). Hi Edwar d, Your Chr istmas tr ee is beautif ul. Can I f or war d it to some member s of apsap (Asia Pacif ic society f or applied phycology) outside the countr y. Thanks Tito Evangelista (A colleague and an of f fi cer of the Association of Systematic Biologists of the Philippines). The message in the Christmas card that I sent reads “No electricity to illuminate, just the cold and clean blue-green radiance of the “Microbes of Light” shining in the dark for hours in water and at room temperature”. Wishing one and all a Merry Blue-green Christmas. The photo can surely make a good screen saver for the coming yuletide season. You can email me at
[email protected] or
[email protected] and I will send you the photo for your desktop and laptop computers.
A wonderful screen saver for your computer this Christmas time
Bioluminescence holds so much promise for providing green solutions and practical applications to safeguard public health and to protect environment from pollution. So let us explore and try to uncover the full potentials of this promising blue light. In a world beset with so many problems on energy and pollution, it is perhaps wise to the see the world in a different light, the light of Bioluminescence. Last year for 2008, I sent this card of a digital-camera enhanced photo of a Bioluminescence illuminated Gift-bearing Santa Claus.
A heartf elt wish of a Merry Bioluminescent Christmas to One and All.
Let us all wage war on Global Warming to ensure the continued healthy existence of our beautif ul Biosphere long into the distant f uture!!!
