The Effect of High-Intensity Focused Ultrasound on Aqueous Humor Dynamics in Patients with Glaucoma.

PURPOSE: To investigate the effects of high-intensity focused ultrasound (HiFU) on aqueous humor dynamics in patients with glaucoma. DESIGN: Comparative, nonrandomized, interventional study. PARTICIPANTS: Adult patients with a diagnosis of open-angle glaucoma or ocular hypertension with suboptimal intraocular pressure (IOP) control despite maximum medical treatment who required further IOP optimization. METHODS: All patients underwent comprehensive ophthalmic examination before aqueous humor dynamics study measurements, including fluorophotometry and digital Schiøtz tonography. All patients received 6 seconds of HiFU therapy. Aqueous humor dynamics studies were repeated 3 months after the treatment (patients had 4-week washout from their glaucoma medication before their aqueous humor dynamics study measurements at baseline and the 3-month visit). MAIN OUTCOME MEASURES: Intraocular pressure, facility of topographic outflow, aqueous flow rate, and uveoscleral outflow. RESULTS: Thirty eyes of 30 patients were included in the study. At the 3-month postoperative visit, the mean postwashout IOP was reduced by 16% (31.7±5.3 vs. 26.6±4.8 mmHg, P = 0.004), and aqueous flow rate was decreased by 15% (2.07±0.73 vs. 1.77±0.55 µl/min, P = 0.05) from baseline. Neither the tonographic outflow facility nor the uveoscleral outflow was significantly different from baseline. There is a 20% risk of treatment failure (those who needed further glaucoma surgical intervention) within 1 month after a single HiFU treatment (n = 6). Only 25 patients (80%) were able to undergo post-treatment washout measurements, and in these eyes, only 26.6% of eyes achieved >20% IOP reduction at 3 months compared with baseline. CONCLUSIONS: We investigated the aqueous humor dynamics effects of a cyclodestructive procedure and specifically HiFU in patients with uncontrolled open-angle glaucoma on maximum tolerated medical therapy. High-intensity focused ultrasound reduced IOP 3 months postoperatively by 16% and aqueous flow decreased by 15% without any significant effect on tonographic outflow facility and uveoscleral outflow.

Noninvasive Monitoring of Choroid-Retina Autofluorescence and Intravitreal Nanoparticle Disposition in Royal College of Surgeon Rats of Different Ages and Retinal Thinning.

Purpose: To determine the baseline choroid-retina fluorescence signal in Royal College of Surgeon (RCS) rats of various ages with different degrees of retinal degeneration and assess the persistence of intravitreal nanoparticles. Methods: In RCS rats of age 6, 12, and 20 weeks and Sprague Dawley (SD) rats of age 6 and 20 weeks, baseline eye tissue fluorescence and retinal thickness were recorded noninvasively using fluorophotometry and optical coherence tomography (OCT), respectively. Further, 20-nm carboxylate-modified fluorescent particles were injected intravitreally in the above groups of rats, and the depth-wise fluorescence signal was monitored over 7 days using fluorophotometry and confocal laser scanning ophthalmoscopy (cSLO). Additionally, 200 nm particles of the same material were injected intravitreally into about 7-week-old RCS rats and the fluorescence signal was monitored up to 35 days using fluorophotometry. Results: Reduction in retinal thickness and an increase in choroid-retina and lens baseline fluorescence was observed with increasing age of RCS and SD rats. The 20 nm particles persisted in the vitreous of animals from all age groups for at least 7 days postadministration, irrespective of the differences in retinal thickness. cSLO confirmed nanoparticle persistence in the eye. The fluorescence signal from 200 nm particles persisted for 35 days in the vitreous humor. Conclusions: Choroid-retina and lens autofluorescence monitored using fluorophotometry increase with age. Intravitreally injected nanoparticles can be monitored noninvasively in rats using fluorophotometry and cSLO imaging. Both 20 and 200 nm particles persist in the back of the eye tissues, for several days following intravitreal injection.

Parameter Estimation for Evaporation-Driven Tear Film Thinning.

Many parameters affect tear film thickness and fluorescent intensity distributions over time; exact values or ranges for some are not well known. We conduct parameter estimation by fitting to fluorescent intensity data recorded from normal subjects' tear films. The fitting is done with thin film fluid dynamics models that are nonlinear partial differential equation models for the thickness, osmolarity and fluorescein concentration of the tear film for circular (spot) or linear (streak) tear film breakup. The corresponding fluorescent intensity is computed from the tear film thickness and fluorescein concentration. The least squares error between computed and experimental fluorescent intensity determines the parameters. The results vary across subjects and trials. The optimal values for variables that cannot be measured in vivo within tear film breakup often fall within accepted experimental ranges for related tear film dynamics; however, some instances suggest that a wider range of parameter values may be acceptable.

Strip meniscometry tube: a rapid method for assessing aqueous deficient dry eye.

BACKGROUND: The aim of this study was to investigate the utility of strip meniscometry tube (SMTube) for the quantitative assessment of the tear film, by comparing it to measurements of tear turnover rate using the gold standard method, fluorophotometry. Also, to determine the viability of this test as a diagnostic tool for aqueous deficient dry eye (ADDE), to inform appropriate clinical management. METHODS: Thirty-two participants (15 ADDE; 17 non-ADDE) were recruited. Tear turnover rate of the right eye of each subject was conducted with an automated scanning fluorophotometer and SMTube test was conducted. Tear meniscus height was assessed using a slitlamp biomicroscope and eyepiece graticule. RESULTS: Significant differences between the ADDE and the non-ADDE groups were found for all measures: tear turnover rate 7.9 ± 1.8 versus 19.6 ± 5.9 per cent/minute (p < 0.001), SMTube 3.2 ± 1.1 versus 5.7 ± 2.3 mm (p = 0.001) and tear meniscus height 0.18 ± 0.05 versus 0.23 ± 0.04 mm (p = 0.004). Moreover, significant correlations were found between tear turnover rate and SMTube (rho = 0.78, p < 0.001), tear turnover rate and tear meniscus height (rho = 0.54, p < 0.001) and SMTube and tear meniscus height (rho = 0.47, p < 0.01). Using a receiver operating characteristic curve, SMTube showed a sensitivity of 67 per cent and a specificity of 88 per cent for the diagnosis of ADDE. CONCLUSION: Given its performance, availability, speed and the fact it is relatively cheap, the study shows that the SMTube can be used as an alternative to fluorophotometry to assess tear production. It appears from the results that SMTube is a viable minimally invasive test for the diagnosis of ADDE.

Rapid and visual detection of berberine hydrochloride based on a water-soluble pyrene derivative.

In this study, a rapid method for the detection of berberine hydrochloride (BRH) was developed based on a water-soluble pyrenyl probe, 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS). This method features low cost, good selectivity, high sensitivity and a fast response. The sensing mechanism of this probe is attributed to the formation of a complex between HPTS and BRH induced by electrostatic interaction and π-π stacking. To the best of our knowledge, this is the first fluorescent sensor for BRH based on organic materials that has low cost and a visual response. The detection limit of this method was as low as 1.24 µM and the linear response range is 2-50 µM. This method also allowed rapid detection of BRH real samples.

A measure of tear inflow in habitual scleral lens wearers with and without midday fogging.

PURPOSE: The purpose of this pilot study was to evaluate tear inflow in a scleral lens system using fluorophotometry, and indirectly assess the exchange of the tear reservoir in habitual scleral lens wearers with the presence or absence of midday fogging (MDF). METHODS: Habitual scleral lens wearers (n=23) and normal scleral lens neophytes (n=10) were recruited. Of the 23 habitual wearers, 11 of them experienced MDF and 12 did not have a diagnosis of MDF. Contact lens-fitting characteristics were evaluated using ocular coherence tomograpy (OCT) and biomicroscopy. High molecular weight fluorescein (FITC) Dextran was instilled into the tear reservoir beneath the scleral lens, and the tear fluid fluorescein concentration was measured using the Fluorotron fluorophotometer. Calculated fluorescein concentrations were plotted over time to measure the fluorescein decay rate of the tear fluid beneath the scleral lens, which was used to calculate the tear exchange rate. RESULTS: There was no significant difference in tear inflow between the MDF group (mean: 0.111%) and the non-MDF group (mean: 0.417%), and there was a high amount of variability seen in the rates (p = 0.26). In addition, there was no significance between the tear reservoir thickness in the MDF (283um) and non-MDF (326um) groups (p = 0.53). CONCLUSIONS: The relationship between the amount of tear exchange during scleral lens wear and the incidence of MDF was not significant. Additional studies are needed to further examine the role of tear exchange in MDF and address the causes of variability to improve measurement techniques with fluorophotometry in the scleral lens system.

An Automated Grading and Diagnosis System for Evaluation of Dry Eye Syndrome.

This article describes methods used to determine the severity of Dry Eye Syndrome (DES) based on Oxford Grading Schema (OGS) automatically by developing and applying a decider model. The number of dry punctate dots occurred on corneal surface after corneal fluorescein staining can be used as a diagnostic indicator of DES severity according to OGS; however, grading of DES severity exactly by carefully assessing these dots is a rather difficult task for humans. Taking into account that current methods are also subjectively dependent on the perception of the ophtalmologists coupled with the time and resource intensive requirements, enhanced diagnosis techniques would greatly contribute to clinical assessment of DES. Automated grading system proposed in this study utilizes image processing methods in order to provide more objective and reliable diagnostic results for DES. A total of 70 fluorescein-stained cornea images from 20 patients with mild, moderate, or severe DES (labeled by an ophthalmologist in the Keratoconus Center of Yildirim Beyazit University Ataturk Training and Research Hospital) used as the participants for the study. Correlations between the number of dry punctate dots and DES severity levels were determined. When automatically created scores and clinical scores were compared, the following measures were observed: Pearson's correlation value between the two was 0.981; Lin's Concordance Correlation Coefficients (CCC) was 0.980; and 95% confidence interval limites were 0.963 and 0.989. The automated DES grade was estimated from the regression fit and accordingly the unknown grade is calculated with the following formula: Gpred = 1.3244 log(Ndots) - 0.0612. The study has shown the viability and the utility of a highly successful automated DES diagnostic system based on OGS, which can be developed by working on the fluorescein-stained cornea images. Proper implemention of a computationally savvy and highly accurate classification system, can assist investigators to perform more objective and faster DES diagnoses in real-world scenerios.

A paper-based competitive lateral flow immunoassay for multi ß-agonist residues by using a single monoclonal antibody labelled with red fluorescent nanoparticles.

An ultrasensitive paper based lateral flow assay is described for rapid and simultaneous fluorometric detection of several ß-agonists including clenbuterol and its chemical analogues (mabuterol, brombuterol, cimaterol, cimbuterol, bromchlorbuterol and banbuterol). A nonspecific monoclonal antibody (mAb) against clenbuterol and its analogues was prepared and employed in a competitive immunoassay where mAb conjugated to fluorescent nanoparticles and free ß-agonists compete for the binding sites. This enables rapid screening for the 7 ß-agonists in a single run that takes about 8 min. Detection limits for the seven ß-agonists are <50 pg g-1 of pork. Recoveries ranged from 69.5% to 102.4%, and relative standard deviations were ±15%. The assay was applied to the analysis of both using spiked and unspiked pork for ß-agonists, and the results compare well to those obtained by HPLC-MS. Graphical abstractSchematic presentation of an ultra sensitive fluorescent nanoparticle based paper based assay for rapid detection of multi ß-agonists in pork tissue.

Effect of oral administration of robenacoxib on inhibition of paracentesis-induced blood-aqueous barrier breakdown in healthy cats.

OBJECTIVE To determine the effect of oral administration of robenacoxib on inhibition of anterior chamber paracentesis (ACP)-induced breakdown of the blood-aqueous barrier (BAB) and assess whether robenacoxib can cross an intact BAB in healthy cats. ANIMALS 12 healthy adult domestic shorthair cats. PROCEDURES Cats received robenacoxib (6-mg tablet in a treat, PO; n = 6) or a control treatment (treat without any drug, PO; 6) once daily for 3 days, beginning 1 day before ACP. One eye of each cat served as an untreated control, whereas the other underwent ACP, during which a 30-gauge needle was used to aspirate 100 µL of aqueous humor for determination of robenacoxib concentration. Both eyes of each cat underwent anterior chamber fluorophotometry at 0 (immediately before), 6, 24, and 48 hours after ACP. Fluorescein concentration and percentage fluorescein increase were used to assess extent of ACP-induced BAB breakdown and compared between cats that did and did not receive robenacoxib. RESULTS Extent of BAB breakdown induced by ACP did not differ significantly between cats that did and did not receive robenacoxib. Low concentrations of robenacoxib were detected in the aqueous humor (mean, 5.32 ng/mL; range, 0.9 to 16 ng/mL) for 5 of the 6 cats that received the drug. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that oral administration of robenacoxib did not significantly decrease extent of BAB breakdown in healthy cats. Detection of low robenacoxib concentrations in the aqueous humor for most treated cats indicated that the drug can cross an intact BAB.

Advances in dry eye imaging: the present and beyond.

New advances in imaging allow objective measurements for dry eye as well as define new parameters that cannot be measured by clinical assessment alone. A combination of these modalities provides unprecedented information on the static and dynamic properties of the structural and functional parameters in this multifactorial disease. A literature search was conducted to include studies investigating the use of imaging techniques in dry eye disease. This review describes the application of non-invasive tear breakup time, optical coherence tomography, meibomian gland imaging, interferometry, in vivo confocal microscopy, thermography and optical quality assessment for this condition.

Classification of Fluorescein Breakup Patterns: A Novel Method of Differential Diagnosis for Dry Eye.

PURPOSE: To investigate the relationship between fluorescein breakup patterns (FBUPs) and clinical manifestations in dry eye cases. DESIGN: Cross-sectional study. METHODS: In 106 eyes of 106 subjects (19 male, 87 female; mean age: 64.2 years), FBUPs were categorized into 1 of the following 5 break (B) types: area (AB, n = 19); spot (SB, n = 22); line (LB, n = 24); dimple (DB, n = 19); random (RB, n = 22 eyes); and dry eye-related symptoms using the visual analog scale (VAS, 100 mm = maximum), tear meniscus radius (TMR, mm), tear film lipid layer interference grade (IG) (grades 1-5; 1 = best) and spread grade (SG) (grades 1-4; 1 = best), tear film noninvasive breakup time (NIBUT, seconds), fluorescein breakup time (FBUT, seconds), corneal-epithelial damage (CED) score (15 points = maximum), ocular surface epithelial damage (OSED) score (9 points = maximum), and the Schirmer 1 test (ST1, mm) were examined and compared between each FBUP. RESULTS: In each FBUP, eye dryness and fatigue were the severest symptoms. Characteristic symptoms were sensitivity to light, heavy eyelids, pain, foreign body sensation, difficulty opening the eye, and discharge for AB, heavy eyelids for SB, and foreign-body sensation for LB. Statistically significant differences were found in TMR (AB-SB, -DB, and -RB; LB-RB), IG (AB-all other FBUP; LB-SB and -DB), and SG (AB-all other FBUPs), FBUT (AB-LB, -DB, and -RB; SB-DB and -RB; LB-RB; DB-RB), and NIBUT (AB-all other FBUPs; SB-DB and-RB, and LB-RB), CED (AB-all other FBUPs; LB-SB, -DB, and -RB) and OSED (AB-SB, -LB, and -DB; LB-SB, -DB, and -RB), and ST1 (AB-SB, -DB, and -LB) (P < .05 in each comparison). CONCLUSIONS: The 5 different FBUPs constituted different groups, reflecting different pathophysi-ologies.

Corneal permeability changes in dry eye disease: an observational study.

BACKGROUND: Diagnostic tests for dry eye disease (DED), including ocular surface disease index (OSDI), tear breakup time (TBUT), corneal fluorescein staining, and lissamine staining, have great deal of variability. We investigated whether fluorophotometry correlated with previously established DED diagnostic tests and whether it could serve as a novel objective metric to evaluate DED. METHODS: Dry eye patients who have had established signs or symptoms for at least 6 months were included in this observational study. Normal subjects with no symptoms of dry eyes served as controls. Each eye had a baseline fluorescein scan prior to any fluorescein dye. Fluorescein dye was then placed into both eyes, rinsed with saline solution, and scanned at 5, 10, 15, and 30 min. Patients were administered the following diagnostic tests to correlate with fluorophotometry: OSDI, TBUT, fluorescein, and lissamine. Standard protocols were used. P < 0.05 was considered significant. RESULTS: Fifty eyes from 25 patients (DED = 22 eyes, 11 patients; Normal = 28 eyes, 14 patients) were included. Baseline scans of the dry eye and control groups did not show any statistical difference (p = 0.84). Fluorescein concentration of DED and normal patients showed statistical significance at all time intervals (p < 10(-5), 0.001, 0.002, 0.049 for 5, 10, 15, & 30 min respectively). Fluorophotometry values converged towards baseline as time elapsed, but both groups were still statistically different at 30 min (p < 0.01). We used four fluorophotometry scoring methods and correlated them with OSDI, TBUT, fluorescein, and lissamine along with adjusted and aggregate scores. The four scoring schemes did not show any significant correlations with the other tests, except for correlations seen with lissamine and 10 (p = 0.045, 0.034) and 15 min (p = 0.013, 0.012), and with aggregate scores and 15 min (p = 0.042, 0.017). CONCLUSIONS: Fluorophotometry generally did not correlate with any other DED tests, even though it showed capability of differentiating between DED and normal eyes up to 30 min after fluorescein dye instillation. There may be an aspect of DED that is missed in the current regimen of DED tests and only captured with fluorophotometry. Adding fluorophotometry may be useful in screening, diagnosing, and monitoring patients with DED.

Regional Differences in Tear Film Stability and Meibomian Glands in Patients With Aqueous-Deficient Dry Eye.

OBJECTIVES: To noninvasively investigate regional differences in tear film stability and meibomian glands in patients with aqueous-deficient dry eye. METHODS: Forty-nine dry eyes and 31 normal eyes were analyzed. A corneal topographer with a tear film scanning function was used for noninvasive tear film break-up time (NI-TFBUT) measurements and meibomian gland observations. The NI-TFBUT values and location of the first tear film break-up point were recorded in four quadrants. Meibomian gland loss was graded for each eyelid using meiboscores. Lid margin abnormality was scored from zero to four according to the number of existing abnormalities. The NI-TFBUT values and meiboscores were compared between two groups, and regional differences in NI-TFBUT values and meiboscores were analyzed. Also, the correlation between the NI-TFBUT and ocular surface examination results were investigated. RESULTS: The NI-TFBUT values and meiboscores were significantly lower and higher, respectively, for the dry eye group than for the normal group. In the dry eye group, the occurrence rate for first tear film break-up was the highest in the inferior nasal quadrant, and the mean meiboscore was significantly higher for the upper eyelids than for the lower eyelids. The NI-TFBUT and lid margin abnormality scores showed a weak negative correlation, and the NI-TFBUT values and meiboscores showed no correlation. CONCLUSIONS: Compared to normal eyes, aqueous-deficient dry eyes show significant regional differences in tear film stability and meibomian glands. Considering these regional differences, the overall observation of the ocular surface, including both upper and lower eyelids, will aid clinicians in understanding this condition better.

Fluorophotometric Analysis of the Ocular Surface Glycocalyx in Soft Contact Lens Wearers.

PURPOSE: Unstable tear film characterized by shorting of tear break-up time (BUT) is associated with discomfort and dryness in contact lens wearers. The glycocalyx is thought to be crucial in maintaining the wettability and lubrication of the ocular surface. We evaluated the ocular surface glycocalyx in soft contact lens (SCL) wearers using a fluorescein-labeled wheat germ agglutinin (F-WGA) as a marker to demonstrate the ocular surface glycoconjugates in vivo. METHODS: Twenty experienced SCL wearers and 20 healthy volunteers with no history of CL wearing (controls) were enrolled in the study. After applying a 5% F-WGA solution to the eyes of study individuals, fluorescent intensities in their respective central corneas were measured by fluorophotometry. The relationship between F-WGA intensity in the corneal surface and clinical parameters associated with contact lens wear were analyzed. RESULTS: F-WGA fluorescence intensity in the SCL group was 418.5 ± 103.3, which was significantly lower than that of the controls (825.0 ± 179.8; p < 0.0001, Mann-Whitney test). F-WGA fluorescence intensity was not correlated with Schirmer's test values or age, whereas a statistically significant correlation between F-WGA fluorescence intensity and tear film BUT was observed (r = 0.77, p < 0.0001). The decrease in F-WGA fluorescence intensity could be reversed by discontinuation of SCL use. CONCLUSION: Reduction and/or compositional alteration of ocular surface glycocalyx may be one of the causative factors of SCL-induced eye dryness.

A fluorometric microarray with ZnO substrate-enhanced fluorescence and suppressed "coffee-ring" effects for fluorescence immunoassays.

A glass slide was first patterned with hydrophobic hexadecyltrimethoxysilane (HDS) and then microspotted with hydrophilic ZnO nanoparticles in an aminopropyltriethoxysilane (APS) matrix. The resulting HDS-ZnO-APS microarray could present the capability of suppressing the undesirable "coffee-ring" effects through its hydrophobic pattern so as to allow the fabrication of ZnO-APS testing microspots with a highly dense and uniform distribution. The lotus-like "self-cleaning" function could also be expected to effectively curb the cross contamination of multiple sample droplets. More importantly, the introduction of ZnO nanoparticles could endow the testing microspots with substrate-enhanced fluorescence leading to signal-amplification microarray fluorometry. The practical application of the developed HDS-ZnO-APS microarray was investigated by the sandwiched fluorometric immunoassays of human IgG, showing a linear detection range from 0.010 to 10.0 ng mL(-1). Such a throughput-improved fluorometric microarray could be tailored for probing multiple biomarkers in complicated media like serum or blood.

A novel analysis method for lactate dehydrogenase activity in serum samples based on fluorescence capillary analysis.

Based on fluorescence capillary analysis technology, a method for quantitating lactate dehydrogenase (LDH) activity in a micro-volume sample was developed. Sample and reagent consumptions were merely 2 and 16 µL per time, respectively. The optimized test conditions were as follows. The reaction reagent consisted of 0.10 M phosphate buffer (pH 6.5), 0.30 mM NADH and 1.20 mM pyruvate. NADH standard was prepared with a phosphate buffer of pH 8.0, and its linear response was controlled in 0.05 - 0.30 mM. LDH standards containing 2.0 mM PEG could exhibit long-term stability. Under the optimized conditions, a linear response for LDH from 50 to 1200 U L(-1) and a detection limit of 31 U L(-1) were obtained with good precision (RSD: 2.1 - 2.2%, n = 10) and better recovery of 96 - 105%. The method's characteristics was high sensitivity, low consumptions, simple operations, good precision and reliability, lending itself to the miniaturization of fluorophotometer which transformed into a bedside instrument in the hospital.

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 180 s subjectively and 15 to 105 s in captured images. CONCLUSION: Although the stability of fluorescein intensity can start to decline in as little as 45 s post fluorescein instillation, the diagnostic pattern of alignment, steep or flat fit is seen in each meridian by subjective observation from about 30s to 3 min indicating this is the most appropriate time window to evaluate GP lenses in clinical practice.

New method for viewing Krehbiel flow by polymethylmethacrylate particles suspended in fluorescein solution.

PURPOSE: To investigate the changes in the tear flow velocities caused by ageing. METHODS: Ninety-nine subjects (41 men, mean age 48.3 ± 20.7 years) were recruited from the Department of Ophthalmology of the Ehime University Hospital. None of the subjects had serious abnormalities of the external surface of the eye. The Krehbiel flow of tears was determined by 40-µm polymethylmethacrylate (PMMA) beads suspended in a fluorescein sodium solution (PPF). The movement of the beads was video recorded through a slit-lamp during normal blinking. The flow of the beads was determined with a Motion ANALYZER(®) software (KEYENCE Co., Osaka, Japan). The velocity of the beads in young age, 20-40 years, middle age, 41-60 years and old age, ≥61 years, groups was determined. RESULTS: The equation describing the velocity (mm/second) of the PMMA particles as a function of age in the lower tear meniscus measured in the direction of the lacrimal punctum was Y = 2.49-0.04X, where Y = velocity and X = age (r(2) = 0.214; p < 0.0001). For the upper meniscus, the equation was Y = 4.83-0.05X (r(2) = 0.195, p < 0.0001). The average velocity was 0.70 ± 1.66 mm/second in the lower and 2.16 ± 1.93 mm/second in the upper tear meniscus (p < 0.0001). The particle velocity decreased significantly with increasing age, but no significant difference between the male and female groups except for the lower tear meniscus when all subjects were analysed. CONCLUSION: The PPF technique is a simple method of examining Krehbiel flow of tears and may be used for evaluating functional nasolacrimal duct obstruction quantitatively.

Rose bengal staining as an assessment of ocular surface damage and recovery in dry eye disease-a review.

PURPOSE: To review the use of ophthalmic rose bengal to assess the ocular surface, especially considering the grading scales used. METHODS: A literature search was undertaken to identify reports where either rose bengal or lissamine green had been used as a 'vital' stain, with a special interest in identifying studies that provided data before and after treatment for dry eye and also considered the mechanism of action of these two chemicals. RESULTS: Between 1985 and 2006, numerous clinical studies used a consistent grading scheme (that attributed to Van Bijsterveld) to assess the outcome of artificial tear treatments on dry eye patients. With such consistency, including the adoption of a treatment period of 1 month, comparisons can be made to indicate the efficacy of rose bengal staining to assess reduction in ocular surface desiccation. However, in the following years, several alternative grading schemes have been used for both rose bengal and lissamine green and assessment periods have been variable so making inter-study comparisons considerably more difficult to undertake. An attribute of rose bengal appears to be its ability to stain the nuclei of cells, but whether this also occurs for lissamine green is unclear. CONCLUSIONS: Ophthalmic rose bengal has been successfully adopted for use to assess the ocular surface over many years as a vital stain. More research is needed to assess whether lissamine green ocular surface staining can simply be substituted for rose bengal in evaluation of dry eye treatments.