Analysis of mitochondria isolated from single cells.
AuthorsJohnson RD,Navratil M,Poe BG,Xiong G,Olson KJ,Ahmadzadeh H,Andreyev D,Duffy CF,Arriaga EA
JournalAnalytical and bioanalytical chemistry
PubMed ID16937092
Fluorescent indicators of ion concentrations.
AuthorsTsien RY
JournalMethods Cell Biol
PubMed ID2538708
Relative binding properties of fluorescein and 9-hydroxyphenylfluoron (HPF) with murine monoclonal anti-fluorescein antibodies.
AuthorsBedzyk WD, Swindlehurst CA, Voss EW
JournalBiochim Biophys Acta
PubMed ID1540631
'Contribution of the fluorescein (F1) carboxyl group to hapten binding by idiotypically related murine monoclonal anti-F1 antibodies 4-4-20, 9-40 and 12-40 was studied by comparing relative liganded active site properties with bound Fl or 9-hydroxyphenylfluoron (HPF). Kinetic studies revealed similar association rate constants between Fl and HPF to 4-4-20 (approximately ... More
Selective cell targeting with light-absorbing microparticles and nanoparticles.
AuthorsPitsillides CM, Joe EK, Wei X, Anderson RR, Lin CP
JournalBiophys J
PubMed ID12770906
'We describe a new method for selective cell targeting based on the use of light-absorbing microparticles and nanoparticles that are heated by short laser pulses to create highly localized cell damage. The method is closely related to chromophore-assisted laser inactivation and photodynamic therapy, but is driven solely by light absorption, ... More
Metabolite anion carriers mediate the uptake of the anionic drug fluorescein in renal cortical mitochondria.
AuthorsTerlouw SA, Tanriseven O, Russel FG, Masereeuw R
JournalJ Pharmacol Exp Ther
PubMed ID10688611
'The fluorescent organic anion fluorescein (FL) accumulates in proximal tubular cells of the kidney during renal secretion. In freshly isolated and permeabilized proximal tubular cells, the uptake was reduced but still sensitive to probenecid, suggesting a concentrative mechanism that is associated with intracellular compartments. Previous studies have shown that one ... More
Inhibition of nitric oxide synthase does not alter basal permeability of the blood-brain barrier.
AuthorsMayhan WG
JournalBrain Res
PubMed ID10650141
'The goal of the present study was to determine the role of basal synthesis/release of nitric oxide on the basal permeability characteristics of the blood-brain barrier to various sized molecules in vivo. We examined the pial microcirculation in rats using intravital fluorescence microscopy. Permeability of the blood-brain barrier was quantitated ... More
Ca2+ fluorescence imaging with pico- and femtosecond two-photon excitation: signal and photodamage.
AuthorsKoester HJ, Baur D, Uhl R, Hell SW
JournalBiophys J
PubMed ID10512842
'The signal and limitations of calcium florescence imaging using nonresonant multiphoton absorption of near-infrared femto- and picosecond laser pulses were examined. The fluorescence changes of various Ca(2+)-indicators induced by transient increases of the intradendritic calcium concentration were evaluated by evoking physiological activity in neocortical neurons in rat brain slices. Photodamage ... More
A study of the fluorescence measurement using a 96-well microplate by a remodelled parallel luminescent measuring system.
AuthorsKumae T
JournalLuminescence
PubMed ID10602312
'To develop an index to measure alveolar macrophage activity, the fluorescent technique for detection of calcium flux was paid special attention. In this study, a parallel luminescence measuring system was remodelled for fluorescence measurement using a 96-well microplate. The fluorescence indicators widely used to measure cytosolic free calcium ion concentration ... More
Cell-to-cell diffusion of fluorescent dyes in paired ventricular cells.
AuthorsImanaga I, Kameyama M, Irisawa H
JournalAm J Physiol
PubMed ID3812712
'The intracellular and cell-to-cell diffusion of fluorescent dyes of various molecular sizes were studied in enzymatically isolated paired ventricular cells of the guinea pig heart. Fluorescein sodium (mol wt 332), 6-carboxyfluorescein (mol wt 376), Lucifer yellow CH (mol wt 457), lissamine rhodamine B-200 (mol wt 559), and tetraglycine-conjugated lissamine rhodamine ... More
In vitro model of the outer blood-retina barrier.
AuthorsSteuer H, Jaworski A, Stoll D, Schlosshauer B
JournalBrain Res Brain Res Protoc
PubMed ID15063838
'The outer blood-retina barrier (BRB) is formed by the retinal pigment epithelium (rpe) and functions similarly to the blood-brain barrier (BBB). In contrast to the BBB, which is composed of a myriad of capillaries, the rpe can in principle be prepared as an intact planar tissue sheet without disruption of ... More
Continuous measurement of mitochondrial pH gradients in isolated hepatocytes by difference ratio spectroscopy.
AuthorsThomas PJ, Gaspers LD, Pharr C, Thomas JA
JournalArch Biochem Biophys
PubMed ID1898020
'The pH gradient, delta pH, present across the inner mitochondrial membrane in isolated rat hepatocytes was continuously monitored with a novel spectroscopic technique that utilizes the weak acid fluorescein. Unlike most cytosolic pH indicators, such as 2'',7''-bis(carboxyethyl)-5,(6)-carboxyfluorescein (BCECF), fluorescein freely distributes between the cytosolic and mitochondrial compartments. As is typical ... More
Confocal imaging of organic anion transport in intact rat choroid plexus.
AuthorsBreen CM, Sykes DB, Fricker G, Miller DS
JournalAm J Physiol Renal Physiol
PubMed ID11934698
'We used confocal microscopy and quantitative image analysis to follow the movement of the fluorescent organic anion fluorescein (FL) from bath to cell and cell to blood vessel in intact rat lateral choroid plexus. FL accumulation in epithelial cells and underlying vessels was rapid, concentrative, and reduced by other organic ... More
Use of fluorescent Ca2+ dyes with green fluorescent protein and its variants: problems and solutions.
AuthorsBolsover S, Ibrahim O, O'luanaigh N, Williams H, Cockcroft S
JournalBiochem J
PubMed ID11368760
'We have studied the degree to which fluorescent Ca(2+) indicator dyes, and green fluorescent protein and its variants, can be used together. We find that the most commonly used fluorescent protein, enhanced green fluorescent protein (EGFP), seriously contaminates fura 2 signals. We suggest two alternative combinations for which there is ... More
Fluorogenic label for biomolecular imaging.
AuthorsLavis LD, Chao TY, Raines RT
JournalACS Chem Biol
PubMed ID17163679
'Traditional small-molecule fluorophores are always fluorescent. This attribute can obscure valuable information in biological experiments. Here, we report on a versatile "latent" fluorophore that overcomes this limitation. At the core of the latent fluorophore is a derivative of rhodamine in which one nitrogen is modified as a urea. That modification ... More
Localized sampling of cytoplasm from Xenopus oocytes for capillary electrophoresis.
AuthorsLuzzi V, Lee CL, Allbritton NL
JournalAnal Chem
PubMed ID9406526
'Continued progress in cellular physiology requires new measurement strategies which can be applied to solitary cells. Since many cellular signaling pathways act on time scales of a few seconds, there is a critical need for single-cell techniques with subsecond time resolution. Capillary electrophoresis shows great promise as a tool for ... More
Fluorescent modification of the cysteine 202 residue of Escherichia coli transcription termination factor rho.
AuthorsSeifried SE, Wang Y, von Hippel PH
JournalJ Biol Chem
PubMed ID2458348
'The lone cysteine residue (Cys-202) of transcription termination factor rho has been modified with the sulfhydryl-specific dyes 5-iodoacetamidofluorescein and 5-(2-[iodoacetyl)amino)ethyl)aminonaphthalene-1-sulfonic acid. Labeling with both dyes is specific for the Cys-202 residue and is at least 90% complete. Rho protein is an RNA-dependent ATPase and exists as a hexamer of identical ... More
Inter- and intramolecular fluorescence quenching of organic dyes by tryptophan.
AuthorsMarmé N, Knemeyer JP, Sauer M, Wolfrum J
JournalBioconjug Chem
PubMed ID14624626
'Steady-state and time-resolved fluorescence measurements were performed to elucidate the fluorescence quenching of oxazine, rhodamine, carbocyanine, and bora-diaza-indacene dyes by amino acids. Among the natural amino acids, tryptophan exhibits the most pronounced quenching efficiency. Especially, the red-absorbing dyes ATTO 655, ATTO 680, and the oxazine derivative MR 121 are strongly ... More
In vivo mammalian brain imaging using one- and two-photon fluorescence microendoscopy.
AuthorsJung JC, Mehta AD, Aksay E, Stepnoski R, Schnitzer MJ
JournalJ Neurophysiol
PubMed ID15128753
'One of the major limitations in the current set of techniques available to neuroscientists is a dearth of methods for imaging individual cells deep within the brains of live animals. To overcome this limitation, we developed two forms of minimally invasive fluorescence microendoscopy and tested their abilities to image cells ... More
Gateable nanofluidic interconnects for multilayered microfluidic separation systems.
'The extension of microfluidic devices to include three-dimensional fluidic networks allows complex fluidic and chemical manipulations but requires innovative methods to interface fluidic layers. Externally controllable interconnects, employing nuclear track-etched polycarbonate membranes containing nanometer-diameter capillaries, are described that produce hybrid three-dimensional fluidic architectures. Controllable nanofluidic transfer is achieved by controlling ... More
Temperature dependence of membrane ion conductance analyzed by using the amphiphilic anion 5/6-carboxyfluorescein.
AuthorsBramhall J, Hofmann J, DeGuzman R, Montestruque S, Schell R
JournalBiochemistry
PubMed ID3427008
'The rate of efflux of trapped 5/6-carboxyfluorescein from sealed lipid vesicles showed a marked dependence on (a) temperature, (b) phospholipid acyl chain composition, and (c) the nature of co-trapped counterions. When the dye was salted with sodium, at pH greater than 7, the rate of dye permeation showed a discrete ... More
Photobleaching kinetics of fluorescein in quantitative fluorescence microscopy.
AuthorsSong L, Hennink EJ, Young IT, Tanke HJ
JournalBiophys J
PubMed ID7647262
'An investigation on the photobleaching behavior of fluorescein in microscopy was carried out through a systematic analysis of photobleaching mechanisms. The individual photochemical reactions of fluorescein were incorporated into a theoretical analysis and mathematical simulation to study the photochemical processes leading to photobleaching of fluorescein in microscopy. The photobleaching behavior ... More
Saturation effects in polarized fluorescence photobleaching recovery and steady state fluorescence polarization.
AuthorsHellen EH, Burghardt TP
JournalBiophys J
PubMed ID8011921
'The time-resolved anisotropy produced in polarized fluorescence photobleaching recovery experiments has been successfully used to measure rotational correlation times in a variety of biological systems, however the magnitudes of the reported initial anisotropies have been much lower than the theoretically predicted maximum values. This small time-zero anisotropy has been attributed ... More
Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements.
AuthorsPrior RL, Wu X, Schaich K,
JournalJ Agric Food Chem
PubMed ID15884874
'Methods available for the measurement of antioxidant capacity are reviewed, presenting the general chemistry underlying the assays, the types of molecules detected, and the most important advantages and shortcomings of each method. This overview provides a basis and rationale for developing standardized antioxidant capacity methods for the food, nutraceutical, and ... More
Platelet GPIIb/IIIa receptor occupancy studies using a novel fluoresceinated cyclic Arg-Gly-Asp peptide.
AuthorsTsao PW, Bozarth JM, Jackson SA, Forsythe MS, Flint SK, Mousa SA
JournalThromb Res
PubMed ID7624840
'DMP 728 is a potent and specific platelet GPIIb/IIIa antagonist. Like all GPIIb/IIIa antagonists, DMP 728 has a steep dose-response relationship in inhibiting platelet aggregation. In this study the relationships between receptor occupancy, platelet aggregation and bleeding time was determined in anesthetized dogs after intravenous infusion of DMP 728 (0.01 ... More
Wavelength-resolved fluorescence detection in capillary electrophoresis.
AuthorsTimperman AT, Khatib K, Sweedler JV
JournalAnal Chem
PubMed ID7864386
'A multichannel laser-induced fluorescence detector for capillary electrophoresis is described. The detection system combines yoctomole limits of detection with the simultaneous acquisition of entire fluorescence emission spectra. An Ar/Kr mixed-gas ion laser provides great flexibility in excitation wavelengths, and a holographic grating and charge-coupled device detector combination allows a 500-nm ... More
Monolithic integration of optical waveguides for absorbance detection in microfabricated electrophoresis devices.
AuthorsMogensen KB, Petersen NJ, Hübner J, Kutter JR
JournalElectrophoresis
PubMed ID11700723
'The fabrication and performance of an electrophoretic separation chip with integrated optical waveguides for absorption detection is presented. The device was fabricated on a silicon substrate by standard microfabrication techniques with the use of two photolithographic mask steps. The waveguides on the device were connected to optical fibers, which enabled ... More
Stopped-flow enzyme assays on a chip using a microfabricated mixer.
AuthorsBurke BJ, Regnier FE
JournalAnal Chem
PubMed ID12713034
'This paper describes a microfabricated enzyme assay system including a micromixer that can be used to perform stopped-flow reactions. Samples and reagents were transported into the system by electroosmotic flow (EOF). Streams of reagents were merged and passed through the 100-pL micromixer in < 1 s. The objective of the ... More
Multiplexed capillary zone electrophoresis and micellar electrokinetic chromatography with internal standardization.
AuthorsXue G, Pang HM, Yeung ES
JournalAnal Chem
PubMed ID10424161
'Although liquid chromatography and gas chromatography are the main workhorses in the analytical laboratory, samples can only be analyzed consecutively in an instrument. In this study, capillary zone electrophoresis and micellar electrokinetic chromatography separations are performed in a 96-capillary array system with laser-induced fluorescence detection. Migration times of four kinds ... More
Serial quantitative image analysis and confocal microscopy of hepatic uptake, intracellular distribution and biliary secretion of a fluorescent bile acid analog in rat hepatocyte doublets.
AuthorsKitamura T, Gatmaitan Z, Arias IM
JournalHepatology
PubMed ID2124205
'To characterize the poorly understood mechanisms of intracellular transport of bile acids, fluorescein isothiocyanate-glycocholate was synthesized and its ring-OH-linked structure established by fast atom bombardment, mass spectroscopy and 13C nuclear magnetic resonance. Biliary secretion of fluorescein isothiocyanate-glycocholate and [14C]-labeled glycocholate in rats was similar, in contrast to the biliary secretion ... More
Thin-layer chromatography and electrospray mass spectrometry coupled using a surface sampling probe.
AuthorsVan Berkel GJ, Sanchez AD, Quirke JM
JournalAnal Chem
PubMed ID12510741
'A combined surface sampling probe/electrospray emitter was used for the direct readout of thin-layer chromatography plates by electrospray mass spectrometry. The technique was demonstrated with reversed-phase C18 plates using a three-dye mixture composed of methylene blue, crystal violet, and rhodamine 6G for positive ion mode detection and a separate dye ... More
Novel fluorescence sensing methods for high throughput screening.
AuthorsLakowicz JR, Gryczynski I, Gryczynski Z
JournalJ Biomol Screen
PubMed ID10894755
'We describe two new methods of fluorescence sensing for use in high throughput screening (HTS). Modulation sensing transforms analyte-dependent intensity changes into a change in the low-frequency modulation signal. Polarization sensing transforms an intensity change into a change in polarization. Both methods are internally calibrated by using a reference film ... More
A microplate reader-based nonisotopic histone deacetylase activity assay.
AuthorsHeltweg B, Jung M
JournalAnal Biochem
PubMed ID11878795
'Recent years have brought an enormous increase in knowledge concerning the involvement of histone deacetylase (HDAC) in gene regulation and the potential use of its inhibitors in transcription therapy. This also stimulates research toward new methods for the determination of HDAC activity and thus the potency of potential inhibitors. We ... More
Size-dependent electrophoretic migration and separation of liposomes by capillary zone electrophoresis in electrolyte solutions of various ionic strengths.
AuthorsRadko SP, Stastna M, Chrambach A
JournalAnal Chem
PubMed ID11140762
'The size-dependent electrophoretic migration and separation of liposomes was demonstrated and studied in capillary zone electrophoresis (CZE). The liposomes were extruded and nonextruded preparations consisting of phosphatidylcholine/phosphatidylglycerol/cholesterol in various ratios and ranging from 125 to 488 nm in mean diameter. When liposomes of identical surface charge density were subjected to ... More
Construction and evaluation of a frequency-domain epifluorescence microscope for lifetime and anisotropy decay measurements in subcellular domains.
AuthorsVerkman AS, Armijo M, Fushimi K
JournalBiophys Chem
PubMed ID1873470
'The measurement of time-resolved fluorescence parameters in living cells provides a powerful approach to study cell structure and dynamics. An epifluorescence microscope was constructed to resolve multi-component fluorescence lifetimes and complex anisotropy decay rapidly in labile biological samples. The excitation source consisted of focused, polarized laser light modulated by an ... More
The fluorescent indicator pyranine is suitable for measuring stromal and cameral pH in vivo.
'Three pH-sensitive fluorescent indicators (fluorescein, bis-carboxyethyl-carboxyfluorescein and pyranine) were studied to determine their usefulness as probes for the living cornea and anterior chamber. Of the three dyes, pyranine had properties which render it almost ideally suited for this purpose. Adequate concentrations are reached in the cornea and anterior chamber after ... More
Novel fluorometric assay for hydroxyl radical prevention capacity using fluorescein as the probe.
'A novel fluorometric method has been developed to evaluate hydroxyl radical prevention capacity using fluorescein (FL) as the probe. The hydroxyl radical is generated by a Co(II)-mediated Fenton-like reaction, and the hydroxyl radical formation under the experimental condition is indirectly confirmed by the hydroxylation of p-hydroxybenzoic acid. The fluorescence decay ... More
Inhibition of in vitro growth of Plasmodium falciparum by a brief exposure to the cationic rhodamine dyes.
AuthorsIzumo A, Tanabe K
JournalAnn Trop Med Parasitol
PubMed ID3541806
'The effects of eight permeant fluorescent dyes on the in vitro growth of Plasmodium falciparum was investigated. First, P. falciparum-infected human erythrocytes were synchronized with D-sorbitol and treated with the cationic fluorescent dye rhodamine 123 at 37 degrees C for 30 minutes, and the growth of the treated parasites monitored ... More
Continuous photobleaching in vesicles and living cells: a measure of diffusion and compartmentation.
AuthorsDelon A, Usson Y, Derouard J, Biben T, Souchier C
JournalBiophys J
PubMed ID16428281
'We present a comprehensive and analytical treatment of continuous photobleaching in a compartment, under single photon excitation. In the very short time regime (t<0.1 ms), the diffusion does not play any role. After a transition (or short time regime), one enters in the long time regime (t>0.1-5 s), for which ... More
Probing ligand protein binding equilibria with fluorescence fluctuation spectroscopy.
AuthorsChen Y, Müller JD, Tetin SY, Tyner JD, Gratton E
JournalBiophys J
PubMed ID10920037
'We examine the binding of fluorescent ligands to proteins by analyzing the fluctuation amplitude g(0) of fluorescence fluctuation experiments. The normalized variance g(0) depends on the molecular brightness and the concentration of each species in the sample. Thus a single g(0) measurement is not sufficient to resolve individual species. Titration ... More
Interaction of fluorescein derivatives with glibenclamide binding sites in rat brain.
AuthorsHolemans S, Feron O, Octave JN, Maloteaux JM
JournalNeurosci Lett
PubMed ID7739789
'In rat brain, [3H]glibenclamide binds with high affinity to sulfonylurea receptors associated with ATP-sensitive potassium (KATP) channels. KATP channels may play a modulatory role in neurotransmitter release and are involved in acute pathological events occurring in the brain. Fluorescein derivatives, which are suitable tools for the labelling of nucleotide binding ... More
Velocity measurement of particles flowing in a microfluidic chip using Shah convolution Fourier transform detection.
AuthorsKwok YC, Jeffery NT, Manz A
JournalAnal Chem
PubMed ID11338588
'A noninvasive radiative technique, based on Shah convolution Fourier transform detection, for velocity measurement of particles in fluid flows in a microfluidic chip, is presented. It boasts a simpler instrumental setup and optical alignment than existing measurement methods and a wide dynamic range of velocities measurable. A glass-PDMS microchip with ... More
A diffusion wake model for tracer ultrastructure-permeability studies in microvessels.
AuthorsFu BM, Curry FE, Weinbaum S
JournalAm J Physiol
PubMed ID8594924
'We developed a time-dependent diffusion model for analyzing the concentration profiles of low-molecular-weight tracers in the interendothelial clefts of the capillary wall that takes into account the three-dimensional time-dependent filling of the surrounding tissue space. The model provides a connecting link between two methods to investigate transvascular exchange: electron-microscopic experiments ... More
Using an AraC-based three-hybrid system to detect biocatalysts in vivo.
'Recent methods to create large libraries of proteins have greatly advanced the discovery of proteins with novel functions. However, one limitation in the discovery of new biocatalysts is the screening or selection methods employed to find enzymes from these libraries. We have developed a potentially general method termed QUEST (QUerying ... More
The rate of diffusion of fluorophores through the corneal epithelium and stroma.
AuthorsAraie M, Maurice D
JournalExp Eye Res
PubMed ID3556451
'The time taken to cross the rabbit corneal epithelium and stroma was estimated for fluorescein (F), carboxyfluorescein (CF), rhodamine B (RB), and sulforhodamine B (SRB). Paired corneas were mounted in vitro; one was intact and the dye solution was kept in continuous contact with its epithelial surface; the epithelium was ... More
Angiography with fluorescein-labeled dextrans in a primate model of uveitis.
AuthorsLightman SL, Caspers-Velu LE, Hirose S, Nussenblatt RB, Palestine AG
JournalArch Ophthalmol
PubMed ID2437896
'Sequential fluorescein angiography, using fluorescein-labeled dextran molecules of several different sizes, was carried out in monkeys with intraocular inflammation induced with interphotoreceptor retinoid-binding protein. The vascular leakage seen with the dextrans was compared with that seen with standard fluorescein sodium angiography. The angiograms demonstrated that different-sized leaks appear in the ... More
Influence of the triplet excited state on the photobleaching kinetics of fluorescein in microscopy.
AuthorsSong L, Varma CA, Verhoeven JW, Tanke HJ
JournalBiophys J
PubMed ID8744334
'The investigation in this report aimed at providing photophysical evidence that the long-lived triplet excited state plays an important role in the non-single-exponential photobleaching kinetics of fluorescein in microscopy. Experiments demonstrated that a thiol-containing reducing agent, mercaptoethylamine (MEA or cysteamine), was the most effective, among other commonly known radical quenchers ... More
One-step concentration of analytes based on dynamic change in pH in capillary zone electrophoresis.
AuthorsWei W, Xue G, Yeung ES
JournalAnal Chem
PubMed ID11924995
'A novel strategy for one-step concentration of analytes during capillary electrophoresis (CE) is presented. A short platinum wire was inserted into the 75-microm-i.d. separation capillary. When a high voltage was applied for CE separation, a sharp pH gradient along the capillary was created dynamically by the electrolysis of water in ... More
Determination of the molar absorption coefficient of fluorescein sodium.
AuthorsBoets PM, van Vreeswijk H, Verhaegen A, den Hartigh J, van Best JA
JournalExp Eye Res
PubMed ID1541332
Efficient mixing and reactions within microfluidic channels using microbead-supported catalysts.
AuthorsSeong GH, Crooks RM
JournalJ Am Chem Soc
PubMed ID12418869
Corrected emission spectra and quantum yields for a series of fluorescent compounds in the visible spectral region.
AuthorsVelapoldi RA, Tønnesen HH,
JournalJ Fluoresc
PubMed ID15617389
This communication describes the determination of corrected fluorescence emission spectra and fluorescence quantum yields for fluorescein, sulforhodamine 101 and other standard fluorescent dyes.
Interaction of organic cations with organic anion transporters.
AuthorsAhn SY, Eraly SA, Tsigelny I, Nigam SK,
JournalJ Biol Chem
PubMed ID19737926
Studies of the organic anion transporters (Oats) have focused mainly on their interactions with organic anionic substrates. However, as suggested when Oat1 was originally identified as NKT (Lopez-Nieto, C. E., You, G., Bush, K. T., Barros, E. J., Beier, D. R., and Nigam, S. K. (1997) J. Biol. Chem. 272, ... More
A chemiluminescent assay for quantitation of beta-galactosidase in the femtogram range: application to quantitation of beta-galactosidase in lacZ-transfected cells.
AuthorsJain VK, Magrath IT
JournalAnal Biochem
PubMed ID1807155
An optimized chemiluminescent assay for beta-galactosidase using a chemiluminescent substrate AMPGD (3-(4-methoxyspiro[1,2-dioxetane-3,2'-tricyclo-[3.3.1. 1(3,7)]decan]-4- yl)phenyl-beta-D-galactopyranoside) is described. This assay is rapid and sensitive and can detect as little as 2 fg of beta-galactosidase. Its use for the quantitation of beta-galactosidase in cells transfected with lacZ-expressing vectors is described. It is possible ... More
Immobilized culture of nonadherent cells on an oleyl poly(ethylene glycol) ether-modified surface.
AuthorsKato K, Umezawa K, Funeriu DP, Miyake M, Miyake J, Nagamune T
JournalBiotechniques
PubMed ID14628675
Microarrays of living cells are an emerging tool in systems such as reverse transfection. These studies are limited to adherent cells partly because of the difficulty of cell immobilization. Using a newly developed reagent, the biocompatible anchor for membrane (BAM), we show herein the rapid and strong attachment of living ... More
Fluorescence-based enzymatic assay by capillary electrophoresis laser-induced fluorescence detection for the determination of a few beta-galactosidase molecules.
AuthorsCraig D, Arriaga EA, Banks P, Zhang Y, Renborg A, Palcic MM, Dovichi NJ
JournalAnal Biochem
PubMed ID7785765
beta-Galactosidase can be assayed by monitoring the generation of the fluorescent products, fluorescein-mono-beta-D-galactopyranoside and fluorescein, when the fluorogenic substrate fluorescein-di-beta-D-galactopyranoside is used. We have used capillary electrophoresis with ultrasensitive laser-induced fluorescence detection to monitor the formation of the fluorescent products of off-column enzymatic reactions. By analyzing as little as 40 ... More
Diacylglycerol inhibits gap junctional communication in cultured epidermal cells: evidence for a role of protein kinase C.
AuthorsGainer HS, Murray AW
JournalBiochem Biophys Res Commun
PubMed ID3156592
Incubation of mouse epidermal HEL-37 cells with 1-oleoyl-2-acetylglycerol (OAG) caused a dose-dependent and transient inhibition of the transfer of microinjected fluorescein between contacting cells. Soluble extracts of HEL-37 cells contained protein kinase C activity after fractionation on DEAE-cellulose and translocation of this activity to the particulate fraction occurred after 10 ... More
Injection of ultrasmall samples and single molecules into tapered capillaries.
AuthorsChiu DT, Hsiao A, Gaggar A, Garza-López RA, Orwar O, Zare RN
JournalAnal Chem
PubMed ID9164157
A novel injection scheme is described in which ultrasmall samples in the attoliter (10(-18) L) and low femtoliter (10(-15) L) range, or even single molecules, are controllably introduced into a tapered capillary so that electrophoretic separation can be carried out. To match the dimensions of the capillary inlet with that ... More
Binding of fluorescein monoglucuronide to human serum albumin.
AuthorsNagataki S, Matsunaga I
JournalInvest Ophthalmol Vis Sci
PubMed ID4019111
The binding to human albumin of fluorescein monoglucuronide, a fluorescent metabolite of fluorescein, was studied using two methods: pressure dialysis and fluorescence polarization. Both methods indicated that fluorescein monoglucuronide binds to human albumin more loosely than fluorescein. The free fraction in human plasma estimated from the dissociation constant and the ... More
Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe.
AuthorsOu B, Hampsch-Woodill M, Prior RL,
JournalJ Agric Food Chem
PubMed ID11599998
An improved method of oxygen radical absorbance capacity (ORAC) assay has been developed and validated using fluorescein (3',6'-dihydroxyspiro[isobenzofuran-1[3H],9'[9H]-xanthen]-3-one) as the fluorescent probe. Our results demonstrate that fluorescein (FL) is superior to B-phycoerythrin. The oxidized FL products induced by peroxyl radical were identified by LC/MS, and the reaction mechanism was determined ... More
Permeability of the isolated dog retinal pigment epithelium to carboxyfluorescein.
AuthorsTsuboi S, Pederson JE
JournalInvest Ophthalmol Vis Sci
PubMed ID3793409
Outward (retina to choroid) and inward (choroid to retina) permeabilities of carboxyfluorescein and fluorescein in the isolated dog retinal pigment epithelium (RPE)-choroid were determined. Outward permeability was 9 and 47 times larger than inward permeability for carboxyfluorescein and fluorescein, respectively. The outward permeability of carboxyfluorescein was seven times lower than ... More
Flow patterns of blood cells in the retinal capillaries. Retinal capillary flow patterns.
AuthorsBen-nun J, Alder V, Thompson D, Constable IJ
JournalInt Ophthalmol
PubMed ID1587699
We present a new technique for retinal blood cell angiography which was used to investigate flow patterns of blood cells in the retinal capillary net of normal cats. Capillary paths were defined to be the path taken by the labelled blood cells from the terminal arteriole to the entry point ... More
Comparative study of three fluorescent dyes for angiography: sodium fluorescein, carboxyfluorescein, and calcein.
AuthorsFang T, Naguib KS, Peyman GA, Khoobehi B
JournalOphthalmic Surg
PubMed ID2362751
To compare the characteristics of carboxyfluorescein (CF) and calcein (Calc) with those of sodium fluorescein (Naf), the only fluorescent dye currently in clinical use, we performed angiography in rabbits and primates using these three dyes. The circulation decay time of all dyes was longer in primates than in rabbits. In ... More
Chemical transformations in individual ultrasmall biomimetic containers.
AuthorsChiu DT, Wilson CF, Ryttsén F, Strömberg A, Farre C, Karlsson A, Nordholm S, Gaggar A, Modi BP, Moscho A, Garza-López RA, Orwar O, Zare RN
JournalScience
PubMed ID10082457
Individual phospholipid vesicles, 1 to 5 micrometers in diameter, containing a single reagent or a complete reaction system, were immobilized with an infrared laser optical trap or by adhesion to modified borosilicate glass surfaces. Chemical transformations were initiated either by electroporation or by electrofusion, in each case through application of ... More
Construction of a two-photon microscope for video-rate Ca(2+) imaging.
AuthorsNguyen QT, Callamaras N, Hsieh C, Parker I
JournalCell Calcium
PubMed ID11728133
We describe the construction of a video-rate two-photon laser scanning microscope, compare its performance to a similar confocal microscope, and illustrate its use for imaging local Ca(2+) transients from cortical neurons in brain slices. Key features include the use of a Ti-sapphire femtosecond laser allowing continuous tuning over a wide ... More
Fluorescein as a model molecular calculator with reset capability.
AuthorsMargulies D, Melman G, Shanzer A,
JournalNat Mater
PubMed ID16155575
The evolution of molecules capable of performing boolean operations has gone a long way since the inception of the first molecular AND logic gate, followed by other logic functions, such as XOR and INHIBIT, and has reached the stage where these tiny processors execute arithmetic calculations. Molecular logic gates that ... More
Multi-level analysis of organic anion transporters 1, 3, and 6 reveals major differences in structural determinants of antiviral discrimination.
AuthorsTruong DM, Kaler G, Khandelwal A, Swaan PW, Nigam SK,
JournalJ Biol Chem
PubMed ID18174163
Long-term exposure to antivirals is associated with serious cellular toxicity to the kidney and other tissues. Organic anion transporters (OATs) are believed to mediate the cellular uptake, and hence cytotoxicity, of many antivirals. However, a systematic in vitro and ex vivo analysis of interactions between these compounds with various OAT ... More
Fluorescein transport in isolated proximal tubules in vitro: epifluorometric analysis.
An epifluorometric method was used to quantify the bidirectional fluxes of fluorescein across the basolateral surfaces of nonperfused rabbit tubule segments in vitro. Proximal S2 segments, but not cortical collecting tubules or cortical thick ascending limbs, accumulated fluorescein to levels in cytoplasm over 100-fold greater than in the external medium. ... More
Indirect fluorescence determination of lactate and pyruvate in single erythrocytes by capillary electrophoresis.
AuthorsXue Q, Yeung ES
JournalJ Chromatogr A
PubMed ID8136909
A scheme of using fluorescein as the fluorophore for indirect detection of anions was demonstrated. This system is quite stable at a fluorescein concentration of 100 microM even without any other buffer components. Different injection modes affect the limit of detection (LOD). A LOD of about 20 amol was obtained ... More
Altered states: involvement of phosphorylated CagA in the induction of host cellular growth changes by Helicobacter pylori.
AuthorsSegal ED, Cha J, Lo J, Falkow S, Tompkins LS
JournalProc Natl Acad Sci U S A
PubMed ID10588744
Helicobacter pylori, present in half of the world's population, is a very successful pathogen. It can survive for decades in the human stomach with few obvious consequences to the host. However, it is also the cause of gastric diseases ranging from gastritis to ulcers to gastric cancer and has been ... More
Fibre optic confocal imaging (FOCI) for subsurface microscopy of the colon in vivo.
AuthorsDelaney PM, King RG, Lambert JR, Harris MR
JournalJ Anat
PubMed ID8157487
Fibre optic confocal imaging (FOCI) is a new type of microscopy which has been recently developed (Delaney et al. 1993). In contrast to conventional light microscopy, FOCI and other confocal techniques allow clear imaging of subsurface structures within translucent objects. However, unlike conventional confocal microscopes which are bulky (because of ... More
Carboxyfluorescein. A dye for evaluating the corneal endothelial barrier function in vivo.
AuthorsAraie M
JournalExp Eye Res
PubMed ID3699104
The relationship of the cornea-aqueous distribution ratio (r ca) and concentration in vitro was established for fluorescein and carboxyfluorescein. The value of rca for fluorescein was found to fall from 3.20 +/- 0.25 (mean +/- S.D., n V 6) to 1.78 as the concentration of the free fluorescein in the ... More
Barley aleurone cells contain two types of vacuoles. Characterization Of lytic organelles by use of fluorescent probes
AuthorsSwanson SJ, Bethke PC, Jones RL
JournalPlant Cell
PubMed ID9596630
Light microscopy was used to study the structure and function of vacuoles in living protoplasts of barley (Hordeum vulgare cv Himalaya) aleurone. Light microscopy showed that aleurone protoplasts contain two distinct types of vacuole: the protein storage vacuole and a lysosome-like organelle, which we have called the secondary vacuole. Fluorescence ... More
Binding of fluorescein and carboxyfluorescein by normal and glycosylated human serum proteins.
AuthorsRockey JH, Li W
JournalOphthalmic Res
PubMed ID7162770
The binding of fluorescein and 6-carboxyfluorescein by normal, diabetic and in vitro glycosylated human serum proteins was analyzed by absorption and fluorescence spectroscopy, gel filtration and equilibrium dialysis. The absorption spectra of bound fluorochromes showed red shifts and hypochromic changes, and fluorochrome fluorescence was reduced by serum proteins. Bio-gel P-6 ... More
Human corneal endothelial permeability to fluorescein and fluorescein glucuronide.
AuthorsSeto C, Araie M, Sawa M, Takase M
JournalInvest Ophthalmol Vis Sci
PubMed ID3623832
The corneal endothelial permeability coefficient (Pac) for fluorescein and fluorescein glucuronide was determined in ten normal young volunteers. After oral administration of fluorescein, the apparent concentrations of both dyes in the corneal stroma and the anterior chamber were measured by differential fluorometry. The apparent dye levels calculated directly from the ... More
Lateral phase separation of phospholipids as a basis for increased permeability of membranes towards fluorescein and other chemical species.
AuthorsHumphries GM, Lovejoy JP
JournalJ Membr Biol
PubMed ID6438340
Using mouse spleen cells, before and after treatment with glutaraldehyde or mild hyperthermia, we observe a strong correlation between permeability to fluorescein and susceptibility to staining with N epsilon-dansyl-L-lysine (irrespective of the cells' ability to exclude trypan blue). We observe the same correlation using liposomes prepared from phosphatidylcholine and varying ... More
Pressure pulse injection: a powerful alternative to electrokinetic sample loading in electrophoresis microchips.
AuthorsSolignac D, Gijs MA
JournalAnal Chem
PubMed ID12705598
We present a new and simple capillary electrophoresis microsystem in which the sample is injected hydrodynamically using a pressure pulse. This technique maintains the sample composition, in contrast to a classical electrokinetic injection, in which the magnitude of the electric field in the sample reservoir in combination with variations in ... More
Electrophoretic behavior of individual nuclear species as determined by capillary electrophoresis with laser-induced fluorescence detection.
AuthorsGunasekera N, Musier-Forsyth K, Arriaga E
JournalElectrophoresis
PubMed ID12210266
We determined the feasibility of using capillary electrophoresis with postcolumn laser-induced fluorescence (CE-LIF) detection to characterize electrophoretic properties of isolated cell nuclei and impurities present in nuclear fractions. These fractions were isolated from NS-1 mouse hybridoma cells, stained with hexidium iodide, a DNA intercalating dye, and analyzed by CE-LIF detection. ... More
Motility of bile canaliculi in the living animal: implications for bile flow.
AuthorsWatanabe N, Tsukada N, Smith CR, Phillips MJ
JournalJ Cell Biol
PubMed ID2040644
Modern fluorescence microscopic techniques were used to image the bile canalicular system in the intact rat liver, in vivo. By combining the use of sodium fluorescein secretion into bile, with digitally enhanced fluorescence microscopy and time-lapse video, it was possible to capture and record the canalicular motility events that accompany ... More
Electroinjection of colloid particles and biopolymers into single unilamellar liposomes and cells for bioanalytical applications.
AuthorsKarlsson M, Nolkrantz K, Davidson MJ, Strömberg A, Ryttsén F, Akerman B, Orwar O
JournalAnal Chem
PubMed ID11128948
A combined electroporation and pressure-driven microinjection method for efficient loading of biopolymers and colloidal particles into single-cell-sized unilamellar liposomes was developed. Single liposomes were positioned between a approximately 2-microm tip diameter solute-filled glass micropipet, equipped with a Pt electrode, and a 5-microm-diameter carbon fiber electrode. A transient, 1-10 ms, rectangular ... More
Time-resolved fluorescence properties of fluorescein and fluorescein glucuronide.
AuthorsLarsen M, Johansson LB
JournalExp Eye Res
PubMed ID2714409
The use of fluorescein as a tracer in the study of the blood-ocular barriers is complicated by the metabolic production of fluorescein monoglucuronide, the excitation and fluorescence spectra of which overlap with those of fluorescein. Time-resolved fluorescence measurements provide a means of detecting the two substances in a mixture. Fluorescein ... More
On-the-fly frequency-domain fluorescence lifetime detection in capillary electrophoresis.
AuthorsLi LC, McGown LB
JournalAnal Chem
PubMed ID8794914
On-the-fly fluorescence lifetime detection in capillary electrophoresis (CE) is demonstrated. Virtually continuous detection is achieved by interfacing a commercial CE instrument with a commercial, multiharmonic Fourier transform lifetime fluorometer (MHF). The CE capillary cartridge was modified to allow the capillary to pass through a specially constructed capillary column mount capable ... More
Two-photon thermal bleaching of single fluorescent molecules.
AuthorsChirico G, Cannone F, Baldini G, Diaspro A
JournalBiophys J
PubMed ID12524312
We have studied the fluorescence emission by two-photon excitation of four dyes widely used for bioimaging studies, rhodamine 6G, fluorescein, pyrene and indo-1 at the single molecule level. The single dye molecules, spread on a glass substrate by spin coating, show a constant fluorescence output until a sudden transition to ... More
Altering the biochemical state of individual cultured cells and organelles with ultramicroelectrodes.
AuthorsLundqvist JA, Sahlin F, Aberg MA, Strömberg A, Eriksson PS, Orwar O
JournalProc Natl Acad Sci U S A
PubMed ID9724707
We describe an efficient technique for the selective chemical and biological manipulation of the contents of individual cells. This technique is based on the electric-field-induced permeabilization (electroporation) in biological membranes using a low-voltage pulse generator and microelectrodes. A spatially highly focused electric field allows introduction of polar cell-impermeant solutes such ... More
Transepithelial transport of fluorescein in Caco-2 cell monolayers and use of such transport in in vitro evaluation of phenolic acid availability.
AuthorsKonishi Y, Hagiwara K, Shimizu M
JournalBiosci Biotechnol Biochem
PubMed ID12506986
Fluorescein is a marker-dye customary applied to the evaluation of tight-junctional permeability of epithelial cell monolayers. However, the true mechanism for the permeation has not been elucidated. Transepithelial transport of fluorescein in Caco-2 cell monolayers was therefore examined. Fluorescein transport was dependent on pH, and in a vectorical way in ... More
A fluorogenic substrate for detection of organophosphatase activity.
AuthorsSoukharev S, Hammond DJ
JournalAnal Biochem
PubMed ID15033522
A new fluorogenic substrate for the specific detection of organophosphatase (OPase) activity has been designed and evaluated. Our results indicate that 7-diethylphospho-6,8-difluor-4-methylumbelliferyl (DEPFMU) is hydrolyzed specifically by the OPases, mammalian serum paraoxonase and bacterial organophosphorus hydrolase (OPH). The apparent K(m) of DEPFMU is 29 microM for OPH and 91 and ... More
We demonstrate that a novel high-pressure cell is suitable for fluorescence correlation spectroscopy (FCS). The pressure cell consists of a single fused silica microcapillary. The cylindrical shape of the capillary leads to refraction of the excitation light, which affects the point spread function of the system. We characterize the influence ... More
Fluorescence polarization of six membrane probes in embryonal carcinoma cells after differentiation as measured on a FACS II cell sorter.
AuthorsSchaap GH, de Josselin de Jong JE, Jongkind JF
JournalCytometry
PubMed ID6201328
Fluorescence polarization measurements on a FACS II cell sorter were compared with static measurements on a spectrofluorimeter using calibration solutions and Hoechst 33258-labeled cells. For the flow cytometric measurements on the FACS we used a pseudodepolarizer for normalization of the output of the two photomultipliers. The results showed that fluorescein ... More
Caffeine interaction with fluorescent calcium indicator dyes.
AuthorsMuschol M, Dasgupta BR, Salzberg BM
JournalBiophys J
PubMed ID10388782
We report that caffeine, in millimolar concentrations, interacts strongly with four common calcium indicator dyes: mag-fura-2, magnesium green, fura-2, and fluo-3. Fluorescence intensities are either noticeably enhanced (mag-fura-2, fura-2) or diminished (magnesium green, fluo-3). The caffeine-induced changes in the fluorescence spectra are clearly distinct from those of metal ion binding ... More
The photon counting histogram in fluorescence fluctuation spectroscopy with non-ideal photodetectors.
AuthorsHillesheim LN, Müller JD
JournalBiophys J
PubMed ID12944307
Fluorescence fluctuation spectroscopy utilizes the signal fluctuations of single molecules for studying biological processes. Information about the biological system is extracted from the raw data by statistical methods such as used in fluctuation correlation spectroscopy or photon counting histogram (PCH) analysis. Since detectors are never ideal, it is crucial to ... More
Sample stacking in laboratory-on-a-chip devices.
AuthorsYang H, Chien RL
JournalJ Chromatogr A
PubMed ID11521862
Sample stacking is a very important sample concentration technique. It has been used widely in capillary electrophoresis (CE). There are many different stacking techniques. One of the most popular techniques is called "field-amplified sample stacking" where an electric field discontinuity is set up across a concentration boundary. Charged analytes will ... More
RNA quantitation by fluorescence-based solution assay: RiboGreen reagent characterization.
AuthorsJones LJ, Yue ST, Cheung CY, Singer VL
JournalAnal Biochem
PubMed ID9882416
We describe the development of a sensitive fluorescence-based solution assay for RNA using a new dye, RiboGreen RNA quantitation reagent. RiboGreen reagent exhibits >1000-fold fluorescence enhancement and high quantum yield (0.65) upon binding nucleic acids, with excitation and emission maxima near those of fluorescein. Unbound dye is essentially nonfluorescent and ... More
The loss of fluorescein, fluorescein glucuronide and fluorescein isothiocyanate dextran from the vitreous by the anterior and retinal pathways.
AuthorsAraie M, Maurice DM
JournalExp Eye Res
PubMed ID1714398
The pathways by which fluorescein (F), fluorescein glucuronide (FG) and fluorescein dextran (FD) leave the vitreous body of the rabbit were examined by measuring the concentration distribution of the injected fluorophores in sections of the frozen eyes. The contours of F, as already known, show that it leaves the vitreous ... More
Fluorescence quantum yields and their relation to lifetimes of rhodamine 6G and fluorescein in nine solvents: improved absolute standards for quantum yields.
AuthorsMagde D, Wong R, Seybold PG
JournalPhotochem Photobiol
PubMed ID12003120
Absolute fluorescence quantum yields are reported for the rhodamine 6G cation and the fluorescein dianion dyes in nine solvents. This information is combined with previously reported fluorescence lifetimes to deduce radiative and nonradiative decay rates. Along the alcohol series from methanol to octanol, rhodamine 6G displays an increasing radiative rate, ... More
Reconstitution of holin activity with a synthetic peptide containing the 1-32 sequence region of EJh, the EJ-1 phage holin.
AuthorsHaro A, Vélez M, Goormaghtigh E, Lago S, Vázquez J, Andreu D, Gasset M
JournalJ Biol Chem
PubMed ID12464609
Pneumococcal EJ-1 phage holin (EJh) is a hydrophobic polypeptide of 85 amino acid residues displaying lethal inner membrane disruption activity. To get an insight into holin structure and function, several peptides representing the different topological regions predicted by sequence analysis have been synthesized. Peptides were structurally characterized in both aqueous ... More
Fluorescein uptake by a monocarboxylic acid transporter in human intestinal Caco-2 cells.
AuthorsKuwayama K, Miyauchi S, Tateoka R, Abe H, Kamo N
JournalBiochem Pharmacol
PubMed ID11754877
The fluorescein transport characteristics of the human intestinal epithelial Caco-2 cell line were examined in monolayer cultures. The initial uptake rate was concentration-dependent and saturable; the Michealis constant and the maximum velocity were 0.40 mM and 1.32 nmol/min/mg protein, respectively. A protonophore, carbonyl cyanide m-chlorophenyl-hydrazone, reduced uptake significantly. The replacement ... More
Predicting the behaviour and selectivity of fluorescent probes for lysosomes and related structures by means of structure-activity models.
AuthorsRashid F, Horobin RW, Williams MA
JournalHistochem J
PubMed ID1743993
Cultured rat fibroblasts were exposed to 50 fluorescent probes of varied physicochemical characteristics. Probe concentrations, fluorochrome excitation wavelength and period of illumination, and cell-probe contact time were varied. Structure-activity relationships defining a number of classes of fluorescent probes for lysosomes and related processes and compartments were demonstrated. Numerical specifications are ... More
Underlying mechanisms of symmetric calcium wave propagation in rat ventricular myocytes.
Calcium waves in heart cells are mediated by diffusion-coupled calcium-induced calcium release. The waves propagate in circular fashion. This is counterintuitive in view of the accepted ultrastructure of the cardiac myocyte. The density of calcium release sites in the transverse direction is four times higher than in the longitudinal direction. ... More
Fluorescence imaging study of organic anion transport from renal proximal tubule cell to lumen.
AuthorsMiller DS, Letcher S, Barnes DM
JournalAm J Physiol
PubMed ID8853412
The mechanisms driving organic anion transport from cell to lumen were studied in intact killifish proximal tubules using fluorescence microscopy. Three fluorescent substrates were used as follows: 1) fluorescein (FL); 2) carboxyfluorescein (CF), generated intracellularly from carboxyfluorescein diacetate (CFDA); and 3) bimane-S conjugates, generated intracellularly by conjugation of monochlorobimane (MCB) ... More
A compartment model for fluorometric indication of tissue perfusion.
AuthorsOstrander LE, Lee BY, Silverman DG, Groskopf RW
JournalIEEE Trans Biomed Eng
PubMed ID2249861
Sodium fluorescein is an in vivo blood perfusion indicator for soft tissue. When fluorescein dye is introduced into the blood, it distributes throughout the vasculature and the extravascular space. Incident light from an external source causes the dye to fluoresce, the level of which is monitored by a photodetector placed ... More
Activation and inhibition of ATP-sensitive K+ channels by fluorescein derivatives.
Authorsde Weille JR, Müller M, Lazdunski M
JournalJ Biol Chem
PubMed ID1311312
Fluorescein derivatives are known to bind to nucleotide-binding sites on transport ATPases. In this study, they have been used as ligands to nucleotide-binding sites on ATP-sensitive K+ channels in insulinoma cells. Their effect on channel activity has been studied using 86Rb+ efflux and patch-clamp techniques. Fluorescein derivatives have two opposite ... More
Measurement of transmission of ultraviolet and visible light in the living rabbit cornea.
AuthorsMcLaren JW, Brubaker RF
JournalCurr Eye Res
PubMed ID8670741
Corneal transmittance in pigmented rabbits was measured at ultraviolet and visible wavelengths from the ratio of fluorescence of dyes in the anterior chamber to fluorescence of the same dyes in a quartz cuvette. Aqueous humor was drained through a limbal incision and the anterior chamber was reformed with a mixture ... More