{"id":22,"date":"2017-12-18T20:11:55","date_gmt":"2017-12-18T20:11:55","guid":{"rendered":"https:\/\/www.sciencesg.com\/amrit\/?p=22"},"modified":"2019-06-20T07:18:35","modified_gmt":"2019-06-20T07:18:35","slug":"fluorescence-light-beneath-your-eyes","status":"publish","type":"post","link":"https:\/\/www.sciencesg.com\/scientificity\/fluorescence-light-beneath-your-eyes\/","title":{"rendered":"I Am the Light beneath Your Eyes ~ I am Fluorescence"},"content":{"rendered":"

I am Fluorescence<\/h2>\n

You have seen me in the sparkles of rubies, emeralds and the Hope diamond when under short wave UV light. You have marvelled at me when you saw a greeneye swimming in the water. You have seen me in the iridescent light emitting “black box” entertainments and environments of theatres and shrines. Have you ever wondered who I can be? Hello, I am Fluorescence<\/strong>!<\/p>\n

My earliest occurrence was found in a Mexican tree in 1565. Nicol\u00e1s Monardes, a Spanish physician and botanist, discovered a peculiar blue tinge that I added to water when filled in cups made from that Mexican tree. Contemporarily, Bernardino de Sahag\u00fan, a Franciscan missionary also reported the presence of a blue colour and named it \u2018coatli\u2019 and proclaimed my powers to be beneficial for the urine. I was also called \u2018kidney wood\u2019 (Lignum nephriticum) by Charles de L\u2019\u00c9cluse because I was considered effective in treating kidney ailments. The chemical compound responsible for my existence is matlanine, which is the oxidation product of one of the flavonoids found in this wood.
\nIn 1819, Edward D. Clarke and in 1822, Ren\u00e9 Juste Ha\u00fcy described fluorescence in fluorites, Sir David Brewster described the phenomenon for chlorophyll in 1833 and Sir John Herschel did the same for quinine in 1845.<\/p>\n

In my presence, the visible light component -sometimes known as “white light”- tends to be reflected and perceived normally, as colour; while the UV component of light is modified, ‘stepped down’ energetically into longer wave-lengths, producing additional visible light frequencies, which are then emitted alongside the reflected white light. Human eyes perceive these changes as the unusual ‘glow’ of fluorescence.<\/p>\n

I am different<\/h3>\n

My type of luminance is significantly different from the natural bio-luminescence of bacteria, insects and fish such as the case of the fire-fly, etc. This involves no reflection at all but true, living generation of light (via the chemistry of Luciferin).<\/p>\n

<\/p>\n

I am versatile<\/h3>\n

My wonders did not cease in treating ailments alone. If I had the colour blue associated with me, I could also make light appear yellow when passing through the aqueous infusion of this wood and yet I seemed to emit blue light which reflected from the very same infusion.<\/p>\n

You might wonder that I was restricted to botanical pursuits only, but you may be pleased to know that more recently, several highly fluorescent glucosyl-hydroxychalcones were isolated from this plant.
\nHave you heard of lapis solaris? It is a widely known stone called Barium sulphate. Galileo Galilei, after having received inspiration from the work of a Bolognian shoemaker, Vincenzo Casciarolo, confirmed my presence in the famous stone. Galilei said, “It must be explained how it happens that the light is conceived into the stone, and is given back after some time, as in childbirth.”<\/p>\n

\"Fluorescence<\/a>
Fluorescence in Lapis Solaris<\/figcaption><\/figure>\n

You must be remembering that I talked about kidney wood a little while ago. My existence was further investigated into by Robert Boyle in 1664 when he was inspired by Monardes\u2019 report and investigated this system more fully. He discovered that after many infusions the wood lost its power to give colour to the water and concluded that there was some \u201cessential salt\u201d in the wood responsible for the effect. He also discovered that addition of acid abolished the colour and that addition of alkali brought it back.<\/p>\n

I do not limit myself<\/h3>\n

And if you thought that I could limit myself to yellow and blue light alone, you are mistaken, my friend, for how can the colours not multiply when something as \u2018moving\u2019 as alcohol is around. And who could brew alcohol better than Mr. Brewster himself? David Brewster, in 1833, described that when a beam of white light passed through an alcohol solution of leaves a red beam could be observed from the side (which was of course chlorophyll fluorescence). He considered the effect due to \u201cdispersion\u201d.
\nJohn Herschel went a step further by calling me \u201cepipolic dispersion\u201d when he observed me for the first time in quinine sulphate. George Gabriel Stokes (1852) published his massive treatise \u201cOn the Change of Refrangibility of Light\u201d \u2013 more than 100 pages. He initially used the term \u201cdispersive reflection\u201d to describe the phenomenon presented by quinine sulphate. Stokes used a prism to disperse the solar spectrum and illuminate a solution of quinine. He noted that there was no effect until the solution was placed in the ultraviolet region of the spectrum.
\nThese observations led Stokes to proclaim that I am of longer wavelength than the exciting light, which led to this displacement being called the Stokes Shift. He also seems to have been the first to propose, in 1864, of my use as an analytical tool, in a lecture “On the application of the optical properties to detection and discrimination of organic substances\u201d.<\/p>\n

I did not create this illumination from quinine sulphate alone. I went on to emit light from calcium sulphate also upon being excited in the UV. This was noted by Becquerel where he says that the emission occurs at a wavelength longer than that of the incident light.
\nAdolph Von Baeyer, a German chemist, synthesized Spiro[isobenzofuran-1(3H),9′-[9H]xanthen]-3-one, 3′,6′-dihydroxy dervatives in 1871. He apparently coined the name \u201cfluorescein\u201d, from \u201cfluo\u201d and resorcin (resorcinol), which he reacted with phthalic anhydride.<\/p>\n

All these famous scientists should give me credit for the fame that I have brought them, you know. For it is because of me that, in 1905, Adolph Von Baeyer was awarded the Nobel Prize in Chemistry<\/a> “in recognition of his services in the advancement of organic chemistry and the chemical industry, through his work on organic dyes and hydroaromatic compounds”.
\nFollowing this were several incidents which resulted in the discovery and production of various compounds where I happened to be the principal characteristic factor.<\/p>\n

<\/p>\n

Haven’t you seen me in Paints?<\/h3>\n

In 1856, at the age of 18, William Henry Perkin set out with idea of making quinine by oxidizing allytoluidine \u2013instead he accidentally produced the synthetic dye, mauve, a derivative of coal tar with an aniline base.
\nI can be present in the form of paints or dyes emitting the \u2018glow\u2019 that you so marvel at. When I am present in such paints, they are called fluorescent paints.
\nFluorescent paints offer a wide range of pigments and chroma which also ‘glow’ when exposed to the long-wave “ultraviolet” frequencies (UV). These UV frequencies are found in sunlight and some artificial lights. But they and their glowing-paint applications are popularly known as Black Light and ‘black-light effect’, respectively.
\nThere are both visible and invisible fluorescent paints. The visible appear under white light to be any bright colour, turning peculiarly brilliant under black lights. Invisible fluorescent paints appear transparent or pale under daytime lighting, but will glow only under UV light and in a limited range of colours. Since these can seem to ‘disappear’, they can be used to create a variety of clever effects.
\nBoth types of fluorescent painting benefit when used within a contrasting ambience of clean, matte-black backgrounds and borders. Such a “black out” effect will minimize other awareness, and cultivating the peculiar luminance of UV fluorescence. Both types of paints have extensive application where artistic lighting effects are desired, particularly in “black box” entertainments and environments such as theatres, bars, shrines, etc. Out-of-doors however UV wavelengths are rapidly scattered in space or absorbed by complex natural surfaces, dulling the effect. Furthermore, the complex pigments will degrade quickly in sunlight.<\/p>\n

Fluorophores<\/h3>\n

Then followed a series of compounds which added to the list where my prevalence was noteworthy.
\nH. Caro (1874): eosin
\nC. Liebermann (1880): polycyclic aromatic hydrocarbons
\nPaul Erlich (1882) used uranin (the sodium salt of fluorescein) to track secretion of the aqueous humour in the eye. First in vivo use of fluorescence.
\nK. Noack (1887) published a book listing 660 compounds arranged according to the colour of their fluorescence.
\nNow, there had to be the classification of compounds and chemical groups having me as a strong property in them. So, R. Meyer (1897) used the term \u201cfluorophore\u201d to describe chemical groups which tended to be associated with fluorescence; this word was analogous to \u201cchromophore\u201d which was first used in 1876 by O.N. Witt to describe groups associated with colour.
\nIn 1867, F. Goppelsr\u00f6der introduced the term form \u201cFluoreszenzanalyse\u201d and performed the first fluorimetric analysis in history: the determination of Al(III) by the fluorescence of its morin chelate.<\/p>\n

\"Fluorophore<\/a>
A panel of 30 common fluorescent dyes spanning the ultraviolet, visible, and near-infrared spectra regions<\/figcaption><\/figure>\n

Could one have imagined that one of the first uses of fluorescein could be found from ground-water? In 1877, in a major ground-water tracing experiment in southern Germany, it was found that River Danube and Rhine are connected by underground streams. I was placed in the form of Fluorescein in the Danube and about 60 hours later I appeared as an affluent in the Rhine. 10 Kilograms of fluorescein were used!<\/p>\n

<\/p>\n

Instruments and me<\/h3>\n

Now, naturally, with all these discoveries, one needed to experiment directly with me and thus needed to observe me. Otto Heimstaedt and Heinrich Lehmann (1911-1913) developed the first microscopes for studying my properties as an outgrowth of the UV microscope<\/strong> (1901-1904). The instrument was used to investigate the autofluorescence of bacteria, protozoa, plant and animal tissues, and bioorganic substances such as albumin, elastin, and keratin.
\nStanislav Von Prowazek (1914) employed the fluorescence microscope<\/strong> to study dye binding to living cells.<\/p>\n

In the inorganic field, Nichols and Slattery (1929) report the first observation of the intense fluorescence of uranium in a NaF matrix. Hieger (1930) found that the fluorescence spectrum (measured photographically) of 1,2-benzanthracene resembled the spectra of some cancer-producing coal tars. Cook and his associates (1933) isolated a few grams of 3,4-benzpyrene from two tons of coal tar and demonstrated that its fluorescence spectrum was identical to that of the active tars. Much later, Gregorio Weber (1952) synthesized dansyl chloride for attachment to proteins and used polarization to study protein hydrodynamics – these studies initiated the field of quantitative biological fluorescence.<\/p>\n

If we now look at the field of drugs, it will be interesting to note that Albert Coons (1941) labelled antibodies with FITC, thus giving birth to the field of immunofluorescence. Brodie and Udenfriend (1943) introduce a simple method for the determination of quinine, and its dextro-rotatory stereoisomer quinidine, in plasma. Saltzman (1948) introduces fluorimetric methods for salicylates in blood.
\nShimomura, Johnson and Saiga (1962) discovered Green Fluorescent Protein in the Aequorea victoria jellyfish.<\/p>\n

You must be knowing that during World War II, there was a huge breakout of malaria. The United States government issued a desperate call to scientists and doctors to find a treatment for malaria. Since Japan had taken over most of the world’s supply of quinine — the best known treatment— Allied forces in the Pacific Theatre needed a new drug, and fast. With an instrument called a fluorometer<\/strong>, Brodie and Udenfriend could measure how much of the drug was in a patient’s plasma sample. Much later, the spectrophotofluorometer was devised by Aminco-Bowman (Silver Spring, MD) and Farrand Optical Co. (Walhalla, NY) which costs about $100,000 in 2009 !<\/p>\n

\"Instrument
I used Fluoromax 3 by HORIBA Scientific during Ph.D.<\/figcaption><\/figure>\n

Several scientists have contributed their lives in discovering my properties and occurrences. Would you like to know more?<\/p>\n

Suggested Reading:<\/h4>\n

[1] Joseph R. Lakowicz, Fluorescence Spectroscopy, 3rd edition, Springer, 2006.
\n[2] Bernard Valeur, Mario N. Berberan-Santos, Molecular Fluorescence; Principles and Applications, 2nd edition, Wiley-VCH Verlag & Co.,2012.
\n[3] M. Sameiro, T. Goncalves, Fluorescent Labeling of Biomolecules with Organic Probes, Chemical Reviews, Vol.109, pp.190\u2013212, 2009.
\n[4] F.W.D. Rost, Fluorescence Microscopy, Volume 1, Cambridge University Press., 1992.
\n[5] Amrit Krishna Mitra, I Am the Light Beneath Your Eyes… A Short History of the Discovery and Properties of Fluorescence, Resonance \u2013 Journal of Science Education, Vol.24, pp.623-632, 2019<\/p>\n","protected":false},"excerpt":{"rendered":"

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