Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “FLUORESCEINS”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 253 records · Page 14Linked to original sources
PubMed1982

Measurement of fluorescein binding in human plasma using fluorescence polarization.

The binding of fluorescein was measured in human plasma using four methods: steady-state dialysis, pressure dialysis, equilibrium dialysis, and fluorescence polarization. All methods indicated that fluorescein is approximately 85% bound throughout a wide range of concentrations. Technical difficulties were encountered with the fluorescence polarization method above fluorescein concentrations of 10(-5) g/mL, but below this concentration, the method gave reproducible and reliable results compared with the three other methods. The number of binding sites available in plasma was estimated to be 4 x 10(-3) M, and the dissociation constant was estimated to be 6 x 10(-4) M. These estimates imply that the bound-free ratio of fluorescein in plasma is almost constant at concentrations below 10(-4) g/mL. Fluorescein binding can affect the fluorescent intensity or the rate at which fluorescein penetrates the blood-ocular barriers.

Blood Chemical Analysis↗
PubMed1982

Carboxyfluorescein. A probe of the blood-ocular barriers with lower membrane permeability than fluorescein.

We have examined the permeability of the blood-ocular barriers to carboxyfluorescein, a dye similar in spectral properties but more polar than fluorescein. Octanol-buffer partition ratios of carboxyfluorescein, measured as an indication of lipid solubility, were approximately 1,000 times lower than those of fluorescein at pH values between 6.40 and 8.03. The partition ratios of both dyes show pronounced pH dependence. We also evaluated intraocular dye distribution by fluorescence microscopy after intravenous (IV) injection in rats. Carboxyfluorescein does not penetrate ciliary or iris epithelial cells, whereas fluorescein prominently stains these cells. Quantitative measurement of fluorescence intensity demonstrates that carboxyfluorescein does not enter the retina even when high doses are administered. Fluorescein, in contrast, can be detected throughout the retina with fluorescence intensity levels proportional to the IV dose administered. The relative inability of carboxyfluorescein to penetrate the blood-ocular barriers is not caused by greater binding to plasma proteins, since the plasma concentration of free carboxyfluorescein is greater than that of fluorescein. We conclude that carboxyfluorescein has potential experimental and clinical use as a probe of the blood-ocular barriers. Because of its low membrane permeability, it may yield a better definition of the nature of barrier abnormalities than is now possible with fluorescein.

Animals↗
PubMed1998

Characterization of fluorescein-oligonucleotide conjugates and measurement of local electrostatic potential.

The properties of fluorescein are substantially altered upon conjugation to nucleic acids, affecting not only the molar absorptivities and fluorescence quantum yields but also the protolytic equilibrium constant and fluorescence lifetimes. Around neutral pH, the fluorescein moiety is present as both mono- and dianion, and the pKa relating them is increased from 6.43 for free fluorescein to about 6.90 for fluorescein attached to both single- and double-stranded oligonucleotides of at least 12 bases/base pairs. This difference reflects the local electrostatic potential around the nucleic acid, which is calculated to -28 mV. The molar absorptivities and spectral responses of the conjugated fluorescein protolytic species are also determined, from which the concentrations of fluorescein-oligonucleotide conjugates can be calculated by assuming: epsilon 494 = 62000/[1 + 10-(pH-6.90)] + 12000/[1 + 10(pH-6.90)] (M-1 cm-1). The fluorescence quantum yield of the conjugates depends, in a complex way, on temperature, environment and oligonucleotide length, sequence and conformation, and must be determined for each experimental situation.

Fluoresceins↗
PubMed2004

Adsorption of fluorescein by protein crystals.

Adsorption characteristics of native and cross-linked lysozyme crystals were examined using fluorescein as model adsorbate. The adsorption isotherms exhibited Langmuir or linear behavior. The affinity constant (b1) and the adsorption capacity (Qsat) for fluorescein were found to depend on the type and concentration of co-solute present in the solution. The dynamics of adsorption isotherm transition from Langmuir to linear showed that affinity of lysozyme for solutes increases in the order 2-(cyclohexylamino)ethanesulphonic acid (CHES), 4-morpholinepropanesulphonic acid (MOPS), acetate, fluorescein. Furthermore, the crystal morphology, the degree of cross-linking of the crystals, and, in particular, solution pH were identified as factors determining fluorescein adsorption by the lysozyme crystals. These factors seem to affect crystal capacity for the solute more than affinity for the solute. Adsorption of fluorescein by cross-linked tetragonal lysozyme crystals was exponentially dependent on the lysozyme net charge calculated from the final solution pH. The 3-5-fold increase in the fluorescein adsorption as a result of cross-linking is presumably due to the increasing hydrophobicity of the lysozyme crystal.

Adsorption↗
PubMed1980

Visualization of thrombin receptors on mouse embryo fibroblasts using fluorescein-amine conjugated human alpha-thrombin.

The localization of thrombin receptors on mouse embryo (ME) cells has been examined by direct fluorescence microscopy using a fluorescein amine-labeled thrombin. Two fluorescein amines, 4-(N-2-aminoethyl thioureal)-fluorescein and 4-(N-5-aminohexyl thioureal)-fluorescein, were synthesized and attached to the carbohydrate moiety of highly purified human alpha-thrombin by periodate oxidation of the carbohydrate and selective reduction of the Schiff's base using sodium cyanoborohydride. Preparations of fluorescent thrombin with from 1 to 4 fluoresceins per molecule of thrombin retained their ability to proteolytically cleave fibrinogen to form fibrin clots, to bind to thrombin receptors on ME cells, and to initiate cell division. After incubating mitogenic concentrations of the fluorescein amine labeled thrombin with ME cells at 4 degrees C, a diffuse fluorescent pattern was observed over the surface of the ME cells. This diffuse pattern was specific: it was not observed on cells from parallel cultures incubated with fluorescent thrombin plus a 20-fold excess of unlabeled thrombin. Thus, thrombin receptors appear to be distributed randomly over the surface of ME cells prior to interaction with thrombin. Increasing the temperature to 37 degrees C following binding at 4 degrees C resulted in a rapid dissociation of the fluorescent pattern from the cells leaving only the autofluorescent vesicles. This result may reflect the unique ability of thrombin to proteolytically cleave its own receptor.

Animals↗
PubMed1982

Fluorescein kinetics in perilymph and blood: a fluorophotometric study.

A modified slit lamp fluorophotometer was used to determine fluorescein concentration changes in the perilymph, cerebrospinal fluid and blood of chinchillas after intravenous injection of 0.2 ml of fluorescein sodium. This new technique provides a means of determining quantitative changes of fluorescein concentration in the perilymph without the need to withdraw fluid samples through the round window membrane of cochlear wall. Fluorescein was observed to enter the perilymph between 1 and 2 minutes after injection, and it reached its peak concentration in the mean time of 23 minutes. The mean peak concentration was 4.20 X 10-6 g/ml. Both increasing and decreasing fluorescein concentration changes in the perilymph followed an exponential time course. Although the observations of cerebrospinal fluid fluorescence were thought to represent a composite of the fluorescence of the cerebrospinal fluid itself and the underlying brain stem blood vessels, the peak fluorescence did not exceed that observed in the perilymph. These observations support the view that most of the perilymph is produced in the cochlea by ultrafiltration from the cochlear blood vessels. The slit lamp fluorophotometer appears to be a satisfactory means of recording fluorescein concentration changes in the perilymph without disturbing the cochlear physiology by penetrating the labyrinth to obtain fluid samples.

Animals↗
PubMed2006

Time-resolved detection of hot electron-induced electrochemiluminescence of fluorescein in aqueous solution.

Strong electrogenerated chemiluminescence (ECL) of fluorescein is generated during cathodic pulse polarization of oxide-covered aluminum electrodes and the resulting decay of emission is so sluggish that time-resolved detection of fluorescein is feasible. The present ECL in aqueous solution is based on the tunnel emission of hot electrons into the aqueous electrolyte solution, which probably results in the generation of hydrated electrons and hydroxyl radicals acting as redox mediators. The successive one-electron redox steps with the primary radicals result in fluorescein in its lowest excited singlet state. The method allows the detection of fluorescein (or its derivatives containing usable linking groups to biomolecules) over several orders of magnitude of concentration with detection limits well below nanomolar concentration level. The detection limits can still be lowered, e.g., by addition of azide or bromide ions as coreactants. The results suggest that the derivatives of fluorescein, such as fluorescein isothiocyanate (FITC), can be detected by time-resolved measurements and thus be efficiently used as electrochemiluminescent labels in bioaffinity assays.

Aluminum↗
PubMed2005

Penetration of topically applied fluorescein into eyes with avascular filtering bleb after trabeculectomy.

PURPOSE: To evaluate intraocular penetration of topical fluorescein in eyes with avascular blebs after trabeculectomy. DESIGN: Case control study. METHODS: The study included 11 eyes with open-angle glaucoma and functioning avascular blebs, six of which were treated with topical anti-glaucomatous medications and had no history of surgery, 15 with open-angle glaucoma, and untreated eyes suspected of having open-angle glaucoma. The fluorescein concentration in the superior peripheral and central corneal stroma and anterior chamber was determined 30 and 60 minutes after fluorescein instillation. RESULTS: The fluorescein concentration in the superior cornea was significantly higher in eyes with blebs or those topically treated than in untreated eyes (P <.01); there was no significant difference in the central cornea. The fluorescein concentration in the anterior chamber was much higher in eyes with blebs than in those that were untreated or topically treated (P <.001). CONCLUSION: The presence of avascular filtering blebs greatly enhances intraocular penetration of topically instilled fluorescein.

Administration, Topical↗
PubMed2005

Intestinal uptake of nateglinide by an intestinal fluorescein transporter.

Nateglinide, a novel oral hypoglycemic agent, rapidly reaches its maximum serum concentration after oral administration, suggesting that it is rapidly absorbed in the intestine. However, nateglinide itself is not transported by MCT1 or PEPT1. The aim of this study was to characterize the transporters on the apical side of the small intestine that are responsible for the rapid absorption of nateglinide. It has been reported that the uptake of fluorescein by Caco-2 cells occurs via an H+-driven transporter and that the intestinal fluorescein transporter is probably not MCT1. We examined the contribution of the fluorescein transporter to the uptake of nateglinide by Caco-2 cells. Fluorescein competitively inhibited H+-dependent nateglinide uptake. All of fluorescein transporter inhibitors examined reduced the uptake of nateglinide. Furthermore, nateglinide inhibited fluorescein uptake. We conclude that the intestinal nateglinide/H+ cotransport system is identical to the intestinal fluorescein/H+ cotransport system.

Absorption↗
PubMed2004

Three-dimensional structures of idiotypically related Fabs with intermediate and high affinity for fluorescein.

Multi-disciplinary studies of fluorescein-protein conjugates have led to the generation of a family of antibodies with common idiotypes and affinities for fluorescein ranging over five orders of magnitude. The high affinity 4-4-20 prototype traps the ligand in a highly complementary binding slot, which is lined by multiple aromatic side-chains. An antibody (9-40) of intermediate affinity belongs to the same idiotypic family as 4-4-20 and shares substantial amino acid identities within the VL and VH domains. To establish the structural basis for the affinity differences, we solved the crystal structure of the 9-40 Fab-fluorescein complex at a resolution of 2.3A. Similar to 4-4-20, 9-40 binds fluorescein in a tight aromatic slot with its xanthenonyl ring system accommodated by end-on insertion. However, the combined effects of the amino acid substitutions have resulted in reorganization of the binding site, with the HCDR3 loops showing the greatest differences in conformations. Access to the binding site of 9-40 is substantially more open, leaving the fluorescein's phenylcarboxylate moiety partially exposed to solvent. In addition to the usage of a different D (diversity) mini-gene encoding the HCDR3 loop, the decrease in fluorescein affinity in the 9-40 antibody family appears to be correlated with the substitution of histidine (9-40) for arginine (4-4-20) in position 34 of the antibody light chains.

Amino Acid Sequence↗
PubMed2006

Comparison of 2D reconstructions of surgically excised subfoveal choroidal neovascularization with fluorescein angiographic features: SST report No. 15.

OBJECTIVES: To compare topographic features of surgically excised subfoveal choroidal neovascularization with preoperative and postoperative fluorescein angiographic features from Submacular Surgery Trials (SST) patients, and to compare histological and angiographic features with preoperative and postoperative visual acuities (VAs). METHODS: Patients enrolled in the SST Groups N, B, and H trials between October 1999 and September 2001 and assigned to the surgery arm had surgically removed choroidal neovascularization sent to the SST Pathology Center. Grossly intact specimens were sectioned serially for 2-dimensional reconstruction and were assigned to growth pattern groups based on topographic mapping of the location of cellular components relative to the retinal pigment epithelium (RPE): sub-RPE, subretinal, combined, or indeterminate. These features were compared with preoperative fluorescein angiographic features. The histological choroidal neovascularization growth pattern was compared with preoperative VAs. RESULTS: Two-dimensional reconstructions of surgically excised choroidal neovascularization could not be matched point for point to fluorescein angiographic features. Among the 52 specimens selected, the growth pattern could be determined by 2-dimensional reconstruction in 34 instances (65%), including 28 (80%) of 35 Group N specimens, 2 (40%) of 5 Group B specimens, and 4 (33%) of 12 Group H specimens. Among the choroidal neovascularization growth patterns that could be determined from specimens submitted, the majority of Group N specimens were combined, and the majority of Group H specimens were subretinal. In most instances for Group B specimens, the growth pattern was indeterminate. The postoperative abnormalities on fluorescein angiography were generally larger than measurements of excised specimens. The subretinal growth pattern was associated with the smallest decrease in 3-month postoperative average VA. CONCLUSIONS: Among the 52 specimens from the SST with adequate tissue to try to evaluate the growth pattern of the neovascular lesion, 34 had patterns that could be determined. The subretinal growth pattern tended to correspond with lesions that were classic choroidal neovascularization without occult choroidal neovascularization on fluorescein angiography. The most favorable visual outcome occurred if the choroidal neovascularization had a subretinal growth pattern. The combined growth pattern tended to correspond with lesions on fluorescein angiography that had both classic choroidal neovascularization and occult choroidal neovascularization. The conclusions must be viewed within the context that 18 (35%) of the 52 participants could not be characterized for histologic growth pattern.

Adult↗
PubMed1998

Choroidopathy after blunt trauma to the eye: a fluorescein and indocyanine green angiographic study.

PURPOSE: To describe the uses of fluorescein angiography and indocyanine green angiography in highlighting traumatic choroidopathy. METHODS: In a prospective study, 21 patients (21 eyes) who presented with traumatic retinal opacity ophthalmoscopically underwent fluorescein angiography and indocyanine green angiography. With indocyanine green angiography, the subtraction method was used for detailed examination. RESULTS: In 11 of 21 eyes, fluorescein angiography showed no abnormalities. On indocyanine green angiography, delayed filling in the choroid was found locally in nine of these 11 eyes. Delayed filling of the choroidal veins was clearly visualized by the subtraction method. In six eyes, intrachoroidal indocyanine green leakage around the choroidal vessels, including vortex veins, was found. In the remaining 10 of the 21 eyes, fluorescein angiography showed fluorescein leakage or a salt-and-pepper appearance in the region of retinal opacity. On indocyanine green angiography, a triangular-shaped area of delayed filling of the choroidal arteries was observed in four eyes. Delayed filling of the choroidal veins was visualized in all 10 eyes, intrachoroidal indocyanine green leakage was found in eight, and dilation and/or narrowing of the choroidal veins and changes in choroidal vasculature were visualized in five eyes. Furthermore, in regions with fluorescein leakage, indocyanine green leakage or hypofluorescence was observed, suggesting damage to the choriocapillaris. CONCLUSION: Indocyanine green angiography allows the analysis of various degrees of choroidal vascular damage. Because the choroidal circulation nourishes the outer retinal layer, traumatic choroidopathy may play a role in the prognosis for visual recovery in eyes affected by contusion retinal opacities.

Adolescent↗
PubMed1991

Fluorescein diacetate assessment of embryo viability after ultrarapid freezing of human multipronucleate embryos.

OBJECTIVE: To assess embryo viability after ultrarapid freezing-thawing. DESIGN: We studied the fluorescence pattern of 35 ultrarapidly frozen-thawed multipronucleate human embryos exposed to fluorescein diacetate. SETTING: All the embryos were obtained from the Medical Center for Fertility Diagnostics and In Vitro Fertilization and Embryo Transfer at Leuven (Belgium), a private care center. PATIENTS, PARTICIPANTS: None. INTERVENTIONS: None. MAIN OUTCOME MEASURE: The fluorescence pattern was evaluated at room temperature after a 1-minute incubation in fluorescein diacetate solution, 4 hours and 24 hours after thawing. RESULTS: Healthy human multipronucleate embryos, when exposed to fluorescein diacetate, accumulated intracellular fluorescein and fluoresced brightly under ultraviolet light. On the other hand, embryos presenting submicroscopic cell membranes damage caused by different processes (e.g., high or low temperatures) lost the ability to accumulate intracellular fluorescein. All the ultrarapidly frozen embryos with normal looking blastomeres fluoresced brightly after a short exposure to fluorescein diacetate. CONCLUSIONS: Our experiments indicate an intact cell membrane permeability and an integrity of the intracytoplasmatic esterase enzyme activity of human embryos ultrarapidly frozen.

Blastocyst↗
PubMed1997

Fluorescein angiographic evaluation of the effect of latanoprost treatment on blood-retinal barrier integrity: a review of studies conducted on pseudophakic glaucoma patients and on phakic and aphakic monkeys.

Endogenous prostaglandins (PGs) have been claimed to play a role in the development of cystoid macular edema (CME). Two fluorescein angiographic studies evaluating the effect of latanoprost, a new ocular hypotensive PG analogue, on blood-retinal barrier integrity are, therefore, reviewed here. In the first study, six of eight unilaterally aphakic cynomolgus monkeys were treated bilaterally once daily for six months with 0.035% latanoprost (seven times the clinically used oculohypotensive concentration). Two of the animals served as controls. Fluorescein angiography of the fundus after one, three and six months of treatment revealed no leakage of fluorescein in any of the 16 eyes. In another study, pseudophakic eyes of 16 glaucoma patients who received twice-daily treatment with 0.006% latanoprost for four weeks were compared to eight patients treated with placebo. Biomicroscopic examination did not reveal any signs of CME and only one placebo-treated eye revealed a slight perifoveal leakage of fluorescein. These studies suggest that topically-applied latanoprost does not have a fluorescein angiographically detectable direct effect on the integrity of the blood-retinal barrier system in phakic or aphakic monkey eyes or in pseudophakic human eyes. This does not rule out the occurrence of CME in eyes more susceptible to CME, due to vitreous loss, posterior capsulotomy, or other postoperative situations. Especially in those eyes a study with latanoprost is proposed. Since, fluorescein angiography is a rather crude method of detecting abnormalities of the blood-retinal barriers, vitreous fluorometry in addition is suggested.

Animals↗
PubMed1998

Development of a fluorescein operative microscope for use during malignant glioma surgery: a technical note and preliminary report.

BACKGROUND: Fluorescein has been used in the field of neurosurgery; however, fluorescein enhancement or contrast proved to be inadequate because of a lack of appropriate light sources or filters. A new operative microscope system, in which the microscope itself is equipped with excitation and barrier filters, and the application of this system to surgery for malignant glioma are reported. METHODS: BP 450-490, a glass interference filter used as the excitation filter for the light source optical system, and a Kodak Wratten No. 12 filter used as the barrier filter for the microscope optical system, were incorporated in the operative microscope. A switching apparatus was devised so that filters could be inserted instantly when fluorescence was to be observed. Ten cases in which the location of malignant glioma was enhanced by computed tomography (CT) or magnetic resonance imaging (MRI) were selected for this study. After incision of the dura mater, 8 mg/Kg body weight of fluorescein Na was injected intravenously. Tumor removal was begun some 20 min after the injection with the aid of this newly developed fluorescein operative microscope system. RESULTS: Fluorescence enhancement and contrast were remarkable when this system was used. It enabled surgical maneuvering while viewing the fluorescent image of objects. The boundaries between the tumor areas enhanced by CT or MRI and the surrounding brain could be clearly distinguished in the fluorescent image; the tumor was totally removed, except for deep lesions, without any neurological deterioration. When the tumor was relatively hard, the area surrounding the tumor was aspirated with a cavitron ultrasonic surgical aspirator; as a result, the tumor could be removed en bloc; otherwise, the fluorescent tumor was removed piece by piece. CONCLUSIONS: This system provides adequate fluorescent enhancement and contrast and is useful for observing intravenously injected fluorescein during an operation. Though long-term follow-up of such cases is needed, the conditions of our patients immediately after surgery for malignant glioma were satisfactory. These results suggest that our fluorescein operative microscope system is highly effective in surgery for malignant glioma.

Adult↗
PubMed2003

The value of combining anterior segment fluorescein angiography with indocyanine green angiography in scleral inflammation.

PURPOSE: To determine the value of anterior segment indocyanine green (ICG) angiography combined with anterior segment fluorescein angiography in scleral inflammation. DESIGN: Comparative observational case series. PARTICIPANTS: The study included 18 patients with various forms of scleral and episcleral disease and a single normal subject. METHODS: Anterior segment angiography using both ICG and fluorescein was performed to identify any vascular abnormalities and pathologic changes in the anterior segment. MAIN OUTCOME MEASURE: The pathologic criteria for anterior segment fluorescein angiography described by Watson and Bovey (1995) were used to compare and contrast the results of the angiograms. RESULTS: Fluorescein angiography and ICG angiography provide different and complementary information. Both dyes have different leakage patterns caused by their difference in optical and chemical properties. Areas of slow flow are more readily determined with fluorescein angiography. ICG angiography determines damage to and patency of individual vessels. CONCLUSION: Fluorescein angiography and ICG angiography detect areas of damage not clinically visible and can be useful in the differential diagnosis, the selection of appropriate medication, and monitoring and regulation of treatment in scleritis. To obtain the most information both investigations should be performed sequentially.

Adult↗
PubMed2002

Digital image analysis of trypan blue and fluorescein staining of anterior lens capsules and intraocular lenses.

PURPOSE: To study the intensity of trypan blue and fluorescein staining over time in anterior lens capsules and various intraocular lens (IOL) materials. SETTING: Fritz Eye Associates, Geislingen, Germany. METHODS: Excised anterior lens capsules and IOLs were exposed to trypan blue 0.1% and fluorescein 2% solution to assess the time correlation of color saturation. A technique of color differentiation using red-green-blue analysis was used. RESULTS: In the anterior lens capsules, trypan blue saturation was 17% after 60 seconds and 71% after 24 hours; with fluorescein, there was no visible staining after 5 minutes and 17% saturation after 24 hours. Red and green light transmission was reduced with trypan blue and high with fluorescein. Blue light transmission was reduced with fluorescein. In poly(methyl methacrylate) and silicone IOLs, there was no to minimal staining and in acrylic IOLs, there was intense uptake of both dyes. CONCLUSIONS: The implications of this surgical technique are to use a short exposure time with trypan blue and avoid acrylic IOLs in cases of questionabale dye loss to the vitreous. Fluorescein should be used as a dye only when injected subcapsularly.

Acrylic Resins↗
PubMedDate not supplied

Thiolate and phosphorothioate functionalized fluoresceins and their use as fluorescent labels.

We report the syntheses of two new fluorescein derivatives, 3',6'-dihydroxy-3-oxo-2-[(phosphonothio)-acetyl]spiro[isobenzof uran- 1(3H),9'-9H-xanthene]-6-carboxylic acid hydrazide, disodium salt, a phosphorothioate fluorescein, and 3',6'-dihydroxy-3-oxo-2-(mercaptoacetyl)spiro[isobenzofuran-1(3H), 9'-9H- xanthene]-6-carboxylic acid hydrazide, a mercaptoacetyl fluorescein. The latter is derived from the first compound by hydrolysis of the phosphate. Direct nonenzymatic labeling of the maleimide-derivatized IgG molecule by the novel mercaptoacetyl fluorescein is discussed. We also present a new method of bioconjugating phosphorothioate-functionalized fluorophores to a maleimide-derivatized protein, based on the alkaline phosphatase-catalyzed hydrolysis of the S-P bond of the phosphorothioate and the concomitant liberation of the fluorophore thiolate. This last species reacts in situ with the maleimide on the protein. A high degree of conjugation control is achieved in that modulation of the stoichiometry of the label and enzyme results in incorporation from seven to eight fluorophores per protein, depending on the ratio of the phosphorothioate fluorescein to alkaline phosphatase. The quantum yield of the mercaptoacetyl fluorescein relative to 6-carboxyfluorescein is 0.22 and lambda exc = 494 nm and lambda em = 517 nm.

Alkaline Phosphatase↗