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| {{Chembox new | | {{Details0|Fluorescein (injection)}} |
| | ImageFile2 = Fluorescein-sample.jpg
| | {{Details0|Fluorescein (Ophthalmic)}} |
| | IUPACName =
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| | OtherNames = Fluorescein, resorcinolphthalein, C.I. 45350, solvent yellow 94, D & C yellow no. 7, angiofluor, Japan yellow 201, soap yellow, Pyoverdin
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| | Section1 = {{Chembox Identifiers
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| | CASNo = 2321-07-5
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| | EINECS = 219-031-8
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| | PubChem = 16850
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| | SMILES = OC(C1=C(C(C(C=CC(O)=C3)<br>=C3O2)=C4C2=CC(C=C4)=O)C=CC=C1)=O
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| | MeSHName = Fluorescein
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| }}
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| | Section2 = {{Chembox Properties
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| | Formula = C<sub>20</sub>H<sub>12</sub>O<sub>5</sub>
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| | MolarMass = 332.306 g/mol
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| | Appearance =
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| | Density =
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| | MeltingPt = 314 - 316 °C
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| | BoilingPt =
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| | Solubility = Slightly
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| }}
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| | Section3 = {{Chembox Hazards
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| | MainHazards =
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| | FlashPt =
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| | RPhrases =
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| | SPhrases = {{S26}} {{S36}}
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| | Autoignition =
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| }}
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| }}
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| {{SI}}
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| '''Fluorescein''' is a [[fluorophore]] commonly used in [[microscopy]], in a type of [[dye laser]] as the [[gain medium]], in [[forensics]] and [[serology]] to detect latent blood stains, and in dye tracing. Fluorescein has an [[Absorption (optics)|absorption]] maximum at 494 nm and [[Fluorescence|emission]] maximum of 521 nm (in water). Also, fluorescein has an isosbestic point (equal absorption for all [[pH|pH values]]) at 460 nm. Fluorescein is also known as a color additive ([[Federal Food, Drug, and Cosmetic Act|D&C]] Yellow no. 7). The disodium salt form of fluorescein is known as [[Federal Food, Drug, and Cosmetic Act|D&C]] Yellow no. 8.
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| ==Chemical and physical properties==
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| [[Image:Florescein.jpg|655px|Left|Fluorescein under [[Ultraviolet|UV]] illumination]]
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| The fluorescence of this molecule is very high, and excitation occurs at 494 [[Nanometre|nm]] and emission at 521.
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| Fluorescein has a [[Acid dissociation constant|pK<sub>a</sub>]] of 6.4 and its ionization equilibrium leads to pH-dependent [[Absorption (optics)|absorption]] and [[Fluorescence|emission]] over the range of 5 to 9. Also, the fluorescence lifetimes of the protonated and deprotonated forms of fluorescein are approximately 3 and 4 ns, which allows for pH determination from non-intensity based measurements. The lifetimes can be recovered using time-correlated single photon counting or phase-modulation fluorimetry.
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| ==Derivatives==
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| There are many fluorescein derivatives, for example [[fluorescein isothiocyanate]], often abbreviated as '''FITC'''. FITC is the original fluorescein molecule functionalized with an [[isothiocyanate]] group ('''-N=C=S'''), replacing a [[hydrogen]] atom on the bottom ring of the structure. This derivative is reactive towards [[amine]] groups on proteins inside cells. A [[succinimidyl-ester]] functional group attached to the fluorescein core, creating [[NHS-fluorescein]], forms another common amine reactive derivative.
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| :[[Image:FITC-2D-skeletal.png|200px|Fluorescein isothiocyanate]]
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| Other derivatives of fluorescein include Oregon Green, Tokyo Green, SNAFL, and carboxynaphthofluorescein. These derivatives, along with newer fluors such as [[Alexa (fluor)|Alexa 488]] and [[DyLight Fluor|DyLight 488]], have been tailored for various chemical and biological applications where higher [[Photobleaching|photostability]], different spectral characteristics, or different attachment groups are needed.
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| ==Synthesis==
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| Fluorescein was first synthesized by [[Adolf von Baeyer]] in 1871. It can be prepared from [[phthalic anhydride]] and [[resorcinol]] in the presence of [[zinc chloride]] via the [[Friedel-Crafts reaction]].
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| [[Image:ZnCl2 fluorescein.png|left|800px]]
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| A second method to prepare fluorescein uses [[methanesulfonic acid]] as a Lewis acid catalyst. This route has a high yield under milder conditions. <ref>Sun, W. C.; Gee, K. R.; Klaubert, D. H.; Haugland, R. P., Synthesis of Fluorinated Fluoresceins. Journal of Organic Chemistry 1997, 62, (19), 6469-6475.</ref> <ref>Yuichiro Ueno; Jiao, G.-S.; Burgess, K., Preparation of 5- and 6-Carboxyfluorescein. Practical Synthetic Procedures 2004, 31, (15), 2591-2593.</ref>
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| ==Applications==
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| ===Uses in river systems===
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| One of its more recognizable uses was in the Chicago River, where fluorescein was the first substance used to dye the river green on St. Patrick's Day in 1962. In 1966 environmentalists forced a change to a vegetable based dye to protect the thousands of goldfish that populate the river.
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| Other uses of fluorescein include using it as a water-soluble dye added to rainwater in environmental testing simulations to aid in locating and analyzing any water leaks, and in Australia and New Zealand as a [[methylated spirit]] dye.
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| ===Biochemical research===
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| In cellular biology, the [[isothiocyanate]] derivative of fluorescein is often used to label and track [[cell (biology)|cells]] in [[fluorescence microscopy]] applications (for example, [[flow cytometry]]). Additional biologically active molecules (such as [[antibody|antibodies]]) may also be attached to fluorescein, allowing biologists to target the fluorophore to specific proteins or structures within cells. This application is common in [[yeast display]].
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| Fluorescein can also be conjugated to [[nucleoside triphosphate]]s and incorporated into a [[hybridization probe|probe]] for [[in situ hybridisation]]. Fluorescein-labelled probes can be imaged using [[fluorescent in situ hybridization|FISH]], or targeted by [[antibodies]] using [[immunohistochemistry]]. The latter is a common alternative to [[digoxigenin]], and the two are used together for labelling two genes in one sample <ref>{{cite journal | author = Noga E. J., Udomkusonsri, P. | date = 2002 | title = Fluorescein: A Rapid, Sensitive, Nonlethal Method for Detecting Skin Ulceration in Fish | journal = Vet Pathol | volume = 39 | issue = | pages = 726–731(6) | pmid = 12450204 | doi = 10.1354/vp.39-6-726| url = http://www.vetpathology.org/cgi/reprint/39/6/726.pdf | accessdate = 2007-07-16}}</ref>.
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| ===Health care applications===
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| Fluorescein sodium is used extensively as a diagnostic tool in the field of [[ophthalmology]], where topical fluorescein is used in the diagnosis of [[corneal abrasion]]s, [[corneal ulcer]]s, [[herpetic corneal infection]]s.
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| [[Image:Fluorescin in dropper.jpg|thumb|Fluorescein in dropper used for eye examination.]]
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| Intravenous or oral fluorescein is used in [[fluorescein angiography]] in research and to diagnose and categorize vascular disorders in e.g. legs, including retinal disease [[macular degeneration]], [[diabetic retinopathy]], inflammatory intraocular conditions, and intraocular [[tumor]]s , and increasingly during surgery for [[brain tumor]]s.
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| ==Safety==
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| Topical, oral, and intravenous use of fluorescein can cause [[Adverse effect (medicine)|adverse reaction]]s including [[nausea]], [[vomiting]], [[urticaria|hives]], [[acute (medicine)|acute]] [[hypotension]], [[anaphylaxis]] and related [[anaphylaxis|anaphylactoid reaction]],<ref>The Diagnosis and Management of Anaphylaxis - XXI. Anaphylactoid reactions to fluorescein.
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| J Allergy Clin Immunol 1998;101:S465-528. [http://www.jcaai.org/PP/anaph_21_ana_reacs.asp Free full text]; see also [http://www.guideline.gov/summary/summary.aspx?ss=15&doc_id=6887&nbr=4211 The diagnosis and management of anaphylaxis: an updated practice parameter.] from the National Guideline Clearinghouse.</ref> [[cardiac arrest]],<ref name="pmid8952662">{{cite journal
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| | author = el Harrar N, Idali B, Moutaouakkil S, el Belhadji M, Zaghloul K, Amraoui A, Benaguida M
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| | title = [Anaphylactic shock caused by application of fluorescein on the ocular conjunctiva]
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| | language = French
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| | journal = Presse Med
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| | volume = 25
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| | issue = 32
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| | pages = 1546–7
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| | year = 1996
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| | pmid = 8952662
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| | doi =
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| | issn =
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| }}</ref> and sudden death.<ref name="pmid10356782">{{cite journal
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| | author = Fineschi V, Monasterolo G, Rosi R, Turillazzi E
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| | title = Fatal anaphylactic shock during a fluorescein angiography
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| | journal = Forensic Sci. Int.
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| | volume = 100
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| | issue = 1-2
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| | pages = 137–42
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| | year = 1999
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| | pmid = 10356782
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| | doi = 10.1016/S0379-0738(98)00205-9
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| }}</ref><ref name="pmid15296251">{{cite journal
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| | author = Hitosugi M, Omura K, Yokoyama T, Kawato H, Motozawa Y, Nagai T, Tokudome S
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| | title = An autopsy case of fatal anaphylactic shock following fluorescein angiography: a case report
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| | journal = Med Sci Law
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| | volume = 44
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| | issue = 3
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| | pages = 264–5
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| | year = 2004
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| | pmid = 15296251
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| | doi =
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| | issn = | |
| }}</ref>
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| Intravenous use has the most reported adverse reactions, including sudden death, but this may reflect greater use rather than greater risk. Both oral and topical uses have been reported to cause anaphylaxis,<ref name="pmid3195657">{{cite journal
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| | author = Kinsella FP, Mooney DJ
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| | title = Anaphylaxis following oral fluorescein angiography
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| | journal = Am. J. Ophthalmol.
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| | volume = 106
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| | issue = 6
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| | pages = 745–6
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| | year = 1988
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| | pmid = 3195657
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| | doi =
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| | issn =
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| }}</ref><ref name="pmid1882930">{{cite journal
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| | author = Gómez-Ulla F, Gutiérrez C, Seoane I
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| | title = Severe anaphylactic reaction to orally administered fluorescein
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| | journal = Am. J. Ophthalmol.
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| | volume = 112
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| | issue = 1
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| | pages = 94
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| | year = 1991
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| | pmid = 1882930
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| | doi =
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| | issn =
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| }}</ref> including one case of anaphylaxis with cardiac arrest ([[Cardiopulmonary resuscitation|resuscitated]]) following topical use in an eye drop.<ref name="pmid8952662"/> Reported rates of adverse reactions vary from 1% to 6%<ref name="pmid16451256">{{cite journal
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| | author = Kwan AS, Barry C, McAllister IL, Constable I
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| | title = Fluorescein angiography and adverse drug reactions revisited: the Lions Eye experience
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| | journal = Clin. Experiment. Ophthalmol.
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| | volume = 34
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| | issue = 1
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| | pages = 33–8
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| | year = 2006
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| | pmid = 16451256
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| | doi = 10.1111/j.1442-9071.2006.01136.x
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| }}</ref><ref name="pmid7930354">{{cite journal
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| | author = Jennings BJ, Mathews DE
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| | title = Adverse reactions during retinal fluorescein angiography
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| | journal = J Am Optom Assoc
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| | volume = 65
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| | issue = 7
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| | pages = 465–71
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| | year = 1994
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| | pmid = 7930354
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| | doi =
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| | issn =
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| }}</ref><ref name="pmid1891225">{{cite journal
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| | author = Kwiterovich KA, Maguire MG, Murphy RP, Schachat AP, Bressler NM, Bressler SB, Fine SL
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| | title = Frequency of adverse systemic reactions after fluorescein angiography. Results of a prospective study
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| | journal = Ophthalmology
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| | volume = 98
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| | issue = 7
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| | pages = 1139–42
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| | year = 1991
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| | pmid = 1891225
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| | doi =
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| | issn =
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| }}</ref><ref name="pmid8644545">{{cite journal
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| | author = Matsuura M, Ando F, Fukumoto K, Kyogane I, Torii Y, Matsuura M
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| | title = [Usefulness of the prick test for anaphylactoid reaction in intravenous fluorescein administration]
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| | language = Japanese
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| | journal = Nippon Ganka Gakkai Zasshi
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| | volume = 100
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| | issue = 4
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| | pages = 313–7
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| | year = 1996
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| | pmid = 8644545
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| | doi =
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| | issn =
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| }}</ref> The higher rates may reflect study populations that include a higher percentage of persons with prior adverse reactions. The risk of an adverse reaction is 25 times higher if the person has had a prior adverse reaction.<ref name="pmid1891225"/> The risk can be reduced with prior ([[prophylaxis|prophylactic]]) use of [[antihistamine]]s<ref name="pmid7257056">{{cite journal
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| | author = Ellis PP, Schoenberger M, Rendi MA
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| | title = Antihistamines as prophylaxis against side reactions to intravenous fluorescein
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| | journal = Trans Am Ophthalmol Soc
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| | volume = 78
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| | issue =
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| | pages = 190–205
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| | year = 1980
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| | pmid = 7257056
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| | doi =
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| | issn =
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| }}</ref> | |
| and prompt emergency management of any ensuing anaphylaxis.<ref name="pmid17698436">{{cite journal
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| | author = Yang CS, Sung CS, Lee FL, Hsu WM
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| | title = Management of anaphylactic shock during intravenous fluorescein angiography at an outpatient clinic
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| | journal = J Chin Med Assoc
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| | volume = 70
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| | issue = 8
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| | pages = 348–9
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| | year = 2007
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| | pmid = 17698436
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| | doi = 10.1016/S1726-4901(08)70017-0
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| }}</ref> A simple prick test may help to identify persons at greatest risk of adverse reaction.<ref name="pmid8644545">{{cite journal
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| | author = Matsuura M, Ando F, Fukumoto K, Kyogane I, Torii Y, Matsuura M
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| | title = [Usefulness of the prick test for anaphylactoid reaction in intravenous fluorescein administration]
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| | language = Japanese
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| | journal = Nippon Ganka Gakkai Zasshi
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| | volume = 100
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| | issue = 4
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| | pages = 313–7
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| | year = 1996
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| | pmid = 8644545 | |
| | doi =
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| | issn =
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| }}</ref>
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| The most common adverse reaction is nausea, due to a difference in the pH from the body and the pH of the sodium fluorescein dye. The nausea usually is transient and subsides quickly. Hives can range from a minor annoyance to severe. A single dose of antihistamine may give complete relief. Anaphylactic shock and subsequent cardiac arrest and sudden death are very rare but because they occur within minutes, a health care provider who uses fluorescein should be prepared to perform emergency resuscitation.
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| ==See also==
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| *Chemical derivatives of fluorescein
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| **[[Eosin]]
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| **[[Fluorescein isothiocyanate]]
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| **[[Erythrosine]]
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| **[[DyLight Fluor]], a product line of fluorescent dyes
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| *''[[Pseudomonas aeruginosa]]'', a bacterium that secretes fluorescein
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| *[[Fluorescein diacetate hydrolysis]], a biochemistry laboratory test
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| ==References==
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| {{Reflist|2}}
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| ==External links==
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| *[http://omlc.ogi.edu/spectra/PhotochemCAD/html/fluorescein(EtOH).html Absorption and Emission Spectra of Fluorescein in Ethanol] and [http://omlc.ogi.edu/spectra/PhotochemCAD/html/fluorescein-dibase.html Basic Ethanol] at [[OGI School of Science and Engineering]]
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| *[http://probes.invitrogen.com/handbook/figures/0571.html Fluorescein Ionization Equilibria] at [[Invitrogen]]
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| *[http://physchem.ox.ac.uk/MSDS/FL/fluorescein.html MSDS at Oxford University]
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| *[http://omlc.ogi.edu/spectra/PhotochemCAD/html/fluorescein-dibase.html Absorption spectra and fluorescence emission spectra]
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| [[Category:Staining dyes]]
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| [[Category:Laser gain media]]
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| [[ca:Fluoresceïna]]
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| [[de:Fluorescein]]
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| [[eo:Fluoreskeino]]
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| [[fr:Fluorescéine]]
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| [[ja:フルオレセイン]]
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| [[pl:Fluoresceina]]
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| [[pt:Fluoresceína]]
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| [[ru:Флуоресцеин]]
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| [[sv:Fluorescein]]
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| {{WH}}
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| {{WS}}
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