Observation of solution-induced corneal staining with fluorescein, rose bengal and lissamine green

Ever since sodium fluorescein (‘fluorescein’ [FL]) was first used to investigate the ocular surface over a century ago, the term ‘staining’ has been taken to mean the presence of ocular surface fluorescence [1]. This term has not been necessarily taken to infer any particular mechanism of causation, and indeed, can be attributed to a variety of possible aetiologies [2]. In recent times, there has been considerable interest in a form of ocular surface fluorescence seen in association with the use of certain combinations of soft contact lenses and multipurpose solutions. The first clinical account of this phenomenon was reported by Jones et al. [3], which was followed by a more formal investigation by the same author in 2002 [4]. Jones et al described this appearance as a ‘classic solution-based toxicity reaction’. Subsequently, this appearance has come to be known as ‘solution-induced corneal staining’ or more recently by the acronym ‘SICS’ [5]. The term SICS is potentially problematic in that from a cell biology point of view, there is an inference that ‘staining’ means the entry of a dye into corneal epithelial cells. Morgan and Maldonado-Codina [2] noted there was no foundation of solid scientific literature underpinning our understanding of the true basic causative mechanisms of this phenomenon; since that time, further work has been published in this field [6] and [7] but questions still remain about the precise aetiology of this phenomenon...

Encapsulation of fluorescein into MCM-41 mesoporous molecular sieve by a sol-gel method

MCM-41-hosted fluorescein mesophase was prepared by addition of the dye into the sol-gel mixture for the synthesis of MCM-41 mesoporous molecular sieve under microwave radiation. The as-synthesized organo-silica-surfactant material possessed hexagonal mesostructure with short-range symmetry and a uniform nanosize of about 30 nm. Furthermore, fluorescence spectrum, increase in lifetime and lack of aggregation at high concentration were discussed in terms of the effect of the host-guest interaction on these properties. (C) 2001 Published by Elsevier Science B.V.

Preparation and spectra study of TiO2 gel doped with fluorescein

TiO2 gel doped with fluorescein was prepared by a sol-gel method, and the fluorescence of fluorescein and its effect on formation of the: gel were investigated with absorption, fluorescence, IR spectroscopies and TG-DTA analysis. The results indicated that FL incorporated into TiO2 gel had exhibited big changes compared to that in ethanol solution both in the absorption and the emission spectra, and the formation of TiO2 gel was influenced greatly by FL.

Morphologic fluorescein angiographic, and light microscopic features of experimental choroidal neovascularization

We induced choroidal neovascularization in the rhesus monkey by impoverishing the blood supply to the inner retina and producing defects in Bruch's membrane by photocoagulation. Fourteen of 46 eyes undergoing photocoagulation developed neovascular fronds which were identified and categorized by histopathologic examination and fluorescein angiography. All new vessels gained access to the retina through defects in Bruch's membrane at the site of photocoagulation marks. In eight eyes the new vessels remained localized to the immediate vicinity of photocoagulation marks. In four eyes neovascular fronds infiltrated the subretinal space for distances up to 6 disk diameters from the point of entry into the retina. In the two eyes choroidovitreal neovascular complexes developed but rapidly regressed shortly after gaining the vitreous cavity. Fluorescein angiography demonstrated that all neovascular fronds were grossly incompetent to dye but that formed feeding channels had some degree of integrity. Light microscopic studies showed the proliferating networks to be composed of capillaries with well-formed basement membranes and more mature vessels with the basic structure of choroidal arteries and veins.

On the barrier properties of the cornea: a microscopy study of the penetration of fluorescently labeled nanoparticles, polymers, and sodium fluorescein

Overcoming the natural defensive barrier functions of the eye remains one of the greatest challenges of ocular drug delivery. Cornea is a chemical and mechanical barrier preventing the passage of any foreign bodies including drugs into the eye, but the factors limiting penetration of permeants and nanoparticulate drug delivery systems through the cornea are still not fully understood. In this study, we investigate these barrier properties of the cornea using thiolated and PEGylated (750 and 5000 Da) nanoparticles, sodium fluorescein, and two linear polymers (dextran and polyethylene glycol). Experiments used intact bovine cornea in addition to bovine cornea de-epithelialized or tissues pretreated with cyclodextrin. It was shown that corneal epithelium is the major barrier for permeation; pretreatment of the cornea with β-cyclodextrin provides higher permeation of low molecular weight compounds, such as sodium fluorescein, but does not enhance penetration of nanoparticles and larger molecules. Studying penetration of thiolated and PEGylated (750 and 5000 Da) nanoparticles into the de-epithelialized ocular tissue revealed that interactions between corneal surface and thiol groups of nanoparticles were more significant determinants of penetration than particle size (for the sizes used here). PEGylation with polyethylene glycol of a higher molecular weight (5000 Da) allows penetration of nanoparticles into the stroma, which proceeds gradually, after an initial 1 h lag phase.

The use of fluorescein for labeling genomic probes in the checkerboard DNA-DNA hybridization method

Molecular methods that permit the simultaneous detection and quantification of a large number of microbial species are currently employed in the evaluation of complex ecosystems. The checkerboard DNA-DNA hybridization technique enables the simultaneous identification of distinct bacterial. species in a large number of dental samples. The original technique employed digoxigenin-labeled whole genomic DNA probes which were detected by chemiluminescence. In this study, we present an alternative protocol for labeling and detecting whole genomic DNA probes in the Checkerboard DNA-DNA hybridization method. Whole genomic DNA was extracted from five bacterial species and labeled with fluorescein. The fluorescein labeled whole genomic DNA probes were hybridized against whole genomic DNA or subgingival plaque samples in a checkerboard hybridization format, followed by chemiluminescent detection. Our results reveal that fluorescein is a viable and adequate alternative labeling reagent to be employed in the checkerboard DNA-DNA hybridization technique. (c) 2007 Elsevier GmbH. All rights reserved.

Electronic spectroscopy of biomolecules in solution: fluorescein dianion in water

A combined and sequential Monte Carlo-quantum mechanics methodology is used to describe the electronic absorption spectrum of the fluorescein dianion in water. Different sets of 100 statistically relevant configurations composed of the solute and several solvent molecules are sampled from the Monte Carlo simulation for a posteriori quantum mechanical calculations of the spectra. In the largest case the configurations are composed of fluorescein and 90 explicit water molecules embedded in the electrostatic field of all remaining water molecules within a distance of 11.3 angstrom. These configurations include 305 atoms and 842 valence electrons, justifying the use of a semi-empirical approach. The electronic spectrum is then calculated using the INDO/CIS method. The solvatochromic shift of fluorescein in water, compared with in isolation, is calculated using the discrete and explicit solvent models. The use of electrostatically embedded explicit water molecules, in INDO/CIS calculations, gives a good description of the spectral shift of the fluorescein dianion in aqueous environment. The results are verified to converge both statistically and with respect to the number of explicit solvent molecules used.

Sodium fluorescein for early detection of skin ulcers in Aeromonas hydrophila infected Piaractus mesopotamicus

The increase in aquaculture global fish production has been associated with an increase in infectious diseases affecting aquacultured species.. The aim of this study was to identify ulcers in pacu (Piaractus mesopotamicus) with sodium fluorescein before it was possible to observe them visually. Twenty one fish with no apparent skin lesions were divided into three groups. Group 1 were injected with saline 0.65% (Group 1), Group 2 with 6 x 10(8) CFU of Aeromonas hydrophila and group 3 with 6 x 10(8) CFU of Streptococcus agalactiae into the peritoneal cavity. All animals showed a positive reaction to fluorescein. A. hydrophila infected fish presented with large round marks with irregular borders, the characteristic ulcers caused by this agent. However, the other groups only displayed markings associated with handling caused by the netting procedures. We concluded that sodium fluorescein may be useful for an early and fast evaluation of cutaneous infection of A. hydrophila in pacu.

Use of fluorescein and indocyanine green angiography in polypoidal choroidal vasculopathy patients following photodynamic therapy

BACKGROUND: Exudative age-related macular degeneration (AMD) is a sight-threatening event in many elderly people. Some patients have a much better outcome in visual acuity (VA) than others after treatment with photodynamic therapy (PDT) with verteporfin. The combination of fluorescein angiography (FA) and indocyanine green (ICG) angiography using the Heidelberg Retina Angiograph II (HRA 2) should make a delineation of distinct pattern(s) possible in order to better select and assess therapy. METHODS: This is a retrospective, case-control, single-centre study. We identified a total of 168 eyes of 168 patients from July 2003 to June 2006, including 30 eyes of 30 patients with better visual outcome, defined in this study as VA < or = 0.48 logMAR (> or =20/60 Snellen chart) at the end of the study. Best-corrected VA, maximal central retinal thickness as measured by optical coherence tomography, and results of the FA/ICG angiography using the HRA 2 were analyzed. In this article, we discuss patients with polypoidal choroidal vasculopathy (PCV) and their characteristics. RESULTS: The average follow-up time was 15.3 months (range 4-28 months). Seventeen (57%) of the 30 patients with better visual outcome had PCV. All patients in the group with better visual outcome needed fewer PDT treatments compared with our control group of patients with an exudative AMD. INTERPRETATION: Simultaneous FA/ICG angiography using the HRA 2 allowed delineation of a subgroup of patients with PCV who showed a better visual outcome compared with those with other types of exudative AMD, after treatment with PDT.

Optimization of anterior eye fluorescein viewing

Purpose: To optimize anterior eye fluorescein viewing and image capture. Design: Prospective experimental investigation. Methods: The spectral radiance of ten different models of slit-lamp blue luminance and the spectral transmission of three barrier filters were measured. Optimal clinical instillation of fluorescein was evaluated by a comparison of four different instillation methods of fluorescein into 10 subjects. Two methods used a floret, and two used minims of different concentration. The resulting fluorescence was evaluated for quenching effects and efficiency over time. Results: Spectral radiance of the blue illumination typically had an average peak at 460 nm. Comparison between three slit-lamps of the same model showed a similar spectral radiance distribution. Of the slit-lamps examined, 8.3% to 50.6% of the illumination output was optimized for >80% fluorescein excitation, and 1.2% to 23.5% of the illumination overlapped with that emitted by the fluorophore. The barrier filters had an average cut-off at 510 to 520 nm. Quenching was observed for all methods of fluorescein instillation. The moistened floret and the 1% minim reached a useful level of fluorescence in on average ∼20s (∼2.5× faster than the saturated floret and 2% minim) and this lasted for ∼160 seconds. Conclusions: Most slit-lamps' blue light and yellow barrier filters are not optimal for fluorescein viewing and capture. Instillation of fluorescein using a moistened floret or 1% minim seems most clinically appropriate as lower quantities and concentrations of fluorescein improve the efficiency of clinical examination. © 2006 Elsevier Inc. All rights reserved.

Optimal time following fluorescein instillation to evaluate rigid gas permeable contact lens fit

PURPOSE: To examine the optimum time at which fluorescein patterns of gas permeable lenses (GPs) should be evaluated. METHODS: Aligned, 0.2mm steep and 0.2mm flat GPs were fitted to 17 patients (aged 20.6±1.1 years, 10 male). Fluorescein was applied to their upper temporal bulbar conjunctiva with a moistened fluorescein strip. Digital slit lamp images (CSO, Italy) at 10× magnification of the fluorescein pattern viewed with blue light through a yellow filter were captured every 15s. Fluorescein intensity in central, mid peripheral and edge regions of the superior, inferior, temporal and nasal quadrants of the lens were graded subjectively using a +2 to -2 scale and using ImageJ software on the simultaneously captured images. RESULTS: Subjectively graded and objectively image analysed fluorescein intensity changed with time (p<0.001), lens region (centre, mid-periphery and edge: p<0.05) and there was interaction between lens region with lens fit (p<0.001). For edge band width, there was a significant effect of time (F=118.503, p<0.001) and lens fit (F=5.1249, p=0.012). The expected alignment, flat and steep fitting patterns could be seen from approximately after 30 to 180s subjectively and 15 to 105s in captured images. CONCLUSION: Although the stability of fluorescein intensity can start to decline in as little as 45s post fluorescein instillation, the diagnostic pattern of alignment, steep or flat fit is seen in each meridian by subjective observation from about 30s to 3min indicating this is the most appropriate time window to evaluate GP lenses in clinical practice.