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PubMed1984

Fluorescein-diacetate, a fluorescent dye compound stain for rapid evaluation of the viability of mammalian oocytes prior to in vitro studies.

A method for a quantitative measurement of the fluorescence activity of porcine egg cells is described. The non-polar fluorescein-diacetate molecules enter the cell, are hydrolyzed by cell esterases, and fluorescein is produced. This polar compound can not leave the cell because it is unable to pass through the intact cell membrane, and it therefore accumulates in the cytoplasm of the cell. Damaged cells however show a distinct loss of fluorescein through the cell membrane. With the aid of a fluorescence microscope, a photometer and a recorder, the amount of radiated light can be measured. The advantages of this method in oocyte research are briefly discussed.

Animals↗
PubMed1980

Nuclear uptake of a 17 beta-estradiol-fluorescein derivative as a marker of estrogen dependence.

17-Fluorescein-labeled estradiol was prepared by linkage of fluorescein-isothiocyanate through a succinamide-ethyl-amine chain to the 17 position of 17 beta-estradiol. This compound, N-fluoresceino-N'[17 beta-(estradiol-hemisuccinamido)-ethyl]-thiourea, displaced tritiated estradiol from purified estrogen-receptor protein and readily crossed intact cellular membranes. In female rats, in-vivo injection was followed by nuclear labeling of endometrium, whereas nuclear labeling of non-target tissue was absent. Temperature-dependent nuclear transfer could be visualized directly by fluorescent microscopy when human estradiol target tissue (mammary and proliferative endometrium) was incubated in vitro with the compound. Temperature-dependent nuclear labeling was not present in human non-target tissue, including skeletal muscle, skin, stomach, colon, appendix, and lung, after in-vitro incubation. Temperature-dependent nuclear labeling with the compound was inhibited by estradiol and mixtures of natural and synthetic estrogens. A significant positive correlation of the presence of nuclear labeling in human mammary cancer was obtained with standard dextran-coated charcoal radioligand assay for estradiol receptor. Unlike radiologand assay and cytochemical assays, nuclear uptake of the fluorescein-labeled estradiol appears to require an intact estradiol-receptor mechanism.

Adult↗
PubMed2004

Transglutaminase-mediated N- and C-terminal fluorescein labeling of a protein can support the native activity of the modified protein.

Fluorescein and its analogs are among the best fluorophores to label proteins and the labeling generally involves chemical modification of a translated protein. Using this methodology, labeling at a specific position remains difficult. It is known that the guinea pig liver transglutaminase (TGase)-catalyzed enzymatic modification method can allow terminal-specific fluorophore labeling of a protein by monodansylcadaverine. However, native activity of the fluorescent protein has not been investigated so far, nor has direct comparison between the chemical modification and the TGase-catalyzed modification been attempted. Therefore, we compared the possibility of fluorescein labeling via chemical labeling and via TGase-catalyzed modification. The latter method was found to be very practical and overcame some of the problems associated with the specificity of the former; fluorescein was covalently attached only to the N- or C-terminal site of glutathione S-transferase when the reaction was catalyzed by TGase and the resulting labeled protein completely retained its native activity. The TGase-mediated labeling occurred not only at room temperature but also at 4 degrees C to the same extent, which is more desirable for preventing the inactivation of proteins.

Base Sequence↗
PubMed2000

Observation of corneal epithelial disturbance through soft contact lens using fluorescein dextran.

PURPOSE: To confirm the usefulness of fluorescein dextran as a vital dye for observing corneal epithelial disturbance through a medical-use soft contact lens. METHODS: Ten percent fluorescein isothiocyanate-dextran (FITC-dextran) was used as the staining dye. Patients with corneal epithelial disturbance who were wearing therapeutic soft contact lenses were observed using FITC-dextran. RESULTS: Through the soft contact lens, FITC-dextran enabled corneal disturbance observation similar to that seen with sodium fluorescein. In a case of corneal perforation, as well, positive Seidel's test was confirmed through the soft contact lens using this technique. CONCLUSION: FITC-dextran is considered a useful dye for examining the corneal epithelial condition without contact lens removal.

Adult↗
PubMed1996

Fluorescein angiography in the progressive outer retinal necrosis syndrome.

PURPOSE: Progressive outer retinal necrosis syndrome is a devastating retinopathy seen primarily in patients with acquired immune deficiency syndrome. To provide additional details of the pathogenesis of this disease, the authors describe the evolution of clinical and fluorescein angiographic changes during the course of progressive outer retinal necrosis syndrome. METHODS: The authors performed serial clinical examinations, fundus photography, and fluorescein angiography in a patient with acquired immune deficiency syndrome with progressive outer retinal necrosis syndrome. Clinical and fluorescein angiographic findings were correlated to provide detailed sequential analysis of the pathologic changes occurring during the course of this disorder. RESULTS: The angiographic changes seen during the various stages of the disease consisted of zonal microvascular alterations, retinal pigment epithelium (RPE) destruction, and choroidal leakage. Retinal damage was correlated closely with regions of choroidal leakage and was clinically evident as outer retinal whitening. Disease reactivation occurred as a prominent brush-fire border of intense leakage involving the retina, RPE, and choroid. Extensive damage to the retinal vasculature and RPE was noted in the wake of clinical infection. CONCLUSIONS: The angiographic findings in our patient demonstrate that the progressive outer retinal necrosis syndrome is a retinochoroiditis that involves the full thickness of retina as well as the RPE and choroid. The inflammatory changes seen throughout the course of this disease correlate with the histopathologic patterns reported to date.

Acquired Immunodeficiency Syndrome↗
PubMed1999

Fluorescein angiography of extreme peripheral retina and rubeosis iridis in proliferative diabetic retinopathy.

PURPOSE: To study the fluorescein angiographic appearance of the extreme peripheral retina in patients with proliferative diabetic retinopathy with rubeosis iridis. METHODS: Using a modified endoscope, fluorescein angiography was performed prospectively during pars plana vitrectomy on 14 eyes of 12 patients with proliferative diabetic retinopathy who had or developed postoperative rubeosis. These findings were compared with those obtained from 41 eyes of 35 patients without rubeosis. RESULTS: A fibrovascular ridge was observed at the ora serrata in 11 of 12 eyes (92%) with rubeosis but in only 15 of 35 (43%) eyes without rubeosis. Seven of 12 (58%) eyes with rubeosis showed the continuous type of fibrovascular ridge, whereas only 5 of 35 (14%) eyes without rubeosis showed the continuous type. A broom-shaped continuous fibrovascular ridge was observed only in eyes with rubeosis. CONCLUSIONS: Many eyes with proliferative diabetic retinopathy showed unusual findings in the extreme retinal periphery, which were more frequent in the eyes with rubeosis iridis. Fluorescein angiography of this region may provide valuable information for possible anterior neovascularization.

Adult↗
PubMed2000

Choroidopathy in patients with sarcoidosis observed by simultaneous indocyanine green and fluorescein angiography.

PURPOSE: To examine choroidopathy in patients with sarcoidosis. METHODS: In a prospective clinical study, 10 consecutive patients (20 eyes) with sarcoidosis underwent simultaneous indocyanine green (ICG) and fluorescein angiography (FA) with a double detector of scanning laser ophthalmoscopy. Angiographic findings recorded on videotapes were evaluated, and the relation of ICG angiographic findings with systemic activity and the extent of retinal inflammation was analyzed. RESULTS: Indocyanine green angiographic findings, not evident with fluorescein, were multiple lobular hypofluorescent spots in the posterior pole in 10 eyes of 5 patients, isolated hypofluorescent plaques in 2 eyes of 1 patient, hyperfluorescent spots in 3 eyes of 2 patients, and segmental choroidal vascular wall staining in 6 eyes of 4 patients. The multiple lobular hypofluorescent spots with ICG were observed at a significantly higher rate in eyes with extensive retinal vascular leakage of fluorescein than in eyes with minimal leakage (chi-square test, P = 0.0007). CONCLUSIONS: The patients with sarcoidosis showed choroidal abnormalities that could be revealed by ICG angiography, but not by funduscopy or FA. Simultaneous ICG and FA angiography may be useful for examining choroidal lesions in sarcoidosis.

Adult↗
PubMed2001

Hemodynamic changes in two patients with retinal circulatory disturbances shown by fluorescein angiography using a scanning laser ophthalmoscope.

PURPOSE: To assess hemodynamic changes in two patients with severely affected retinal circulation. METHODS: A 62-year-old man with central retinal artery occlusion and a 46-year-old woman with branch retinal vein occlusion were studied by fluorescein angiography with a scanning laser ophthalmoscope (SLO). Fluorescein angiography with SLO revealed hypofluorescent clumps of different sizes and hyperfluorescent dots in large retinal vessels. The velocities of the hypofluorescent clumps were calculated between two points on the same vessel, and movements of the hypofluorescent clumps and the hyperfluorescent dots were investigated. RESULTS: The velocities of the hypofluorescent clumps were slow and varied in the same vessel. The velocities of the hypofluorescent clumps increased at the sites with narrow calibers. The hypofluorescent clumps occasionally changed size in the bloodstream. The hypofluorescent clumps flowed along the walls of retinal vessels. Distance between consecutive hypofluorescent clumps was wide. Some vessels filled with hypofluorescent clumps were also detected. Rolling hyperfluorescent dots were seen in fluorescent plasma. CONCLUSIONS: The hypofluorescent clumps were concluded to be packed erythrocytes and the hyperfluorescent dots corresponded to leukocytes and platelets moving in the vessels. Fluorescein angiography with SLO is a useful method for evaluating hemodynamic changes using the hypofluorescent clumps in severely affected retinal circulation.

Blood Flow Velocity↗
PubMed2002

Indocyanine green and fluorescein angiography in nonarteritic anterior ischemic optic neuropathy.

PURPOSE: To assess the value of indocyanine green angiography (ICGA) for demonstrating choroidal vascular abnormalities in patients with nonarteritic anterior ischemic optic neuropathy (NAION). METHODS: The authors compared the ICGA and fluorescein fundus angiography characteristics of peripapillary circulation in 11 patients with acute NAION. There were 7 men and 4 women; the age range for the patients was 36 years to 72 years (mean +/- SD, 47.7 +/- 10.76 years). The angiographic factors considered significant were delay of peripapillary choroidal filling in the vertical watershed zone, leakage from the optic disk, and absolute filling defects on the disk. The authors compared the incidence of a vertical peripapillary watershed zone in the eyes of the 11 patients with that in the normal eyes of 50 controls (age range, 44-79 years) who had unilateral age-related macular degeneration. RESULTS: Indocyanine green angiography revealed a peripapillary watershed zone in 8 of 11 patient eyes and 23 of 50 control eyes. There was no statistical difference in the number of eyes affected in each group (chi2 = 0.53; P = 0.47). Fluorescein fundus angiography showed leakage from the disk in 10 of 11 patients, whereas ICGA highlighted this problem in only 7 of the patients. The choroidal filling time of the watershed zones was significantly longer with ICGA (t = 3.13; P = 0.011). CONCLUSION: Although ICGA allows better visualization of the choroidal watershed zones associated with NAION, it did not reveal any significantly different incidence of vertical choroidal watershed zone encompassing the optic disk for patients with NAION and controls. Fluorescein fundus angiography better visualized leakage from the disk in the patient group. These findings indicate that ICGA offers no significant advantage in terms of clinical diagnosis and management of NAION.

Acute Disease↗
PubMed2005

Digital simultaneous fluorescein and indocyanine green angiography, autofluorescence, and red-free imaging with a solid-state laser-based confocal scanning laser ophthalmoscope.

PURPOSE: To describe and to evaluate a novel confocal scanning laser ophthalmoscope (cSLO) for fluorescence angiography, fundus autofluorescence (FAF), and red-free imaging. METHODS: Digital infrared, red-free, FAF, fluorescein, and indocyanine green (ICG) angiography images were obtained with a cSLO in 766 patients. An optically pumped solid-state laser generates the excitation wavelength (488 nm) required for red-free, FAF, and fluorescein angiography images. For ICG angiography and infrared imaging, diode laser sources at 790 and 820 nm are used. Further features include an internal fixation control and a focus range of -24 to +30 diopters. RESULTS: High-image quality is achieved with a resolution of up to 5 microm per pixel in 30- x 30-degree images and allows for accurate delineation of normal and pathologic features. Simultaneous angiography offers high-contrast images. Corresponding display of quasi-simultaneous frames facilitates interpretation. A small focus difference between fluorescein and ICG scans occurs because of chromatic aberrations. Automated alignment and generation of mean images from several single frames allow for acquisition of high-resolution FAF images. CONCLUSION: Various laser-source related, optical, and electronic innovations improve cSLO fundus imaging for routine clinical application. A solid-state laser has advantages compared to argon gas laser sources, including less space occupation, heat emission, and noise production.

Adolescent↗
PubMed2006

Topography and fluorescein angiography of the optic nerve head in primary open-angle and chronic primary angle closure glaucoma.

PURPOSE: The purpose of this study is to correlate optic nerve head topography with fluorescein angiography of the optic nerve head in patients with primary open-angle glaucoma (POAG), chronic primary angle closure glaucoma (CPACG), and normal controls. METHODS: This was an institution-based, cross-sectional, case-control study of 30 consecutive patients each with POAG or CPACG, which were compared with 30 age- and sex-matched controls. The fluorescein angiograms undertaken in one eye of each of the 90 subjects were then analyzed both qualitatively and quantitatively. RESULTS: The mean age of controls (group 1) was 51.73 +/- 9.6 years, patients with CPACG (group II) was 53.26 +/- 9.5 years, and patients with POAG (group III) was 54.5 +/- 10.4 years. The mean deviation and corrected pattern standard deviation on Humphrey visual field analyzer, respectively, were -1.51 +/- 2.01 dB and 2.09 +/- 1.04 dB the in control group, -9.4 +/- 9.3 dB and 5.32 +/- 4.02 dB in the CPACG group, and -11.27 +/- 7.7 dB and 7.57 +/- 5.34 dB in the POAG group. There was no significant difference in the disc areas between the three groups (analysis of variance [ANOVA], p = 0.157). All circulatory parameters were delayed in both glaucoma groups compared with controls with the disc filling time (ANOVA, p = 0.001) and the choroidal filling time being significantly delayed (ANOVA, p = 0.006). The Moorfield regression analysis showed good correlation with the pattern of disc fluorescence in all quadrants in cases of CPACG and POAG. CONCLUSION: The optic nerve head and choroidal circulation was delayed in both patients with POAG and those with CPACG, which correlates with loss of neuroretinal rim and retinal nerve fiber layer on the Heidelberg Retina Tomograph II (HRT). Patients with POAG showed diffuse damage with significant rim loss, whereas patients with CPACG showed marked sectorial abnormalities (superotemporal and the inferior-temporal) on fluorescein angiography and HRT. One possible reason for this discrepancy could be sectorial ischemia occurring in cases of CPACG as a result of a sudden rise of intraocular pressure causing disc and visual field damage.

Blood Flow Velocity↗
PubMed2004

Comparison of colour Doppler imaging and retinal scanning laser fluorescein angiography in healthy volunteers and normal pressure glaucoma patients.

PURPOSE: To correlate retinal circulatory measurements using scanning laser fluorescein angiography and flow velocities of retrobulbar vessels measured by means of colour Doppler imaging. METHODS: Fifteen patients with normal pressure glaucoma (NPG) and 15 healthy volunteers underwent colour Doppler imaging and fluorescein angiographic studies. Peak systolic velocities (PSVs), end-diastolic velocities (EDVs) and resistive indices (RIs) of the ophthalmic artery (OA) and central retinal artery were obtained. In the fluorescein angiograms arteriovenous passage time (AVP) was quantified by means of digital dye dilution curve analysis. RESULTS: Arteriovenous passage time was significantly prolonged in NPG patients compared to healthy subjects (p = 0.0026). In the central retinal artery PSV (p = 0.023) and EDV (p < 0.0001) were significantly decreased and RI was increased (p < 0.0001) in patients with NPG. The EDV of the central retinal artery showed a significant correlation with AVP (EDV: r = - 0.53, p = 0.0023). The RI of the central retinal artery correlated significantly to AVP (RI: r = 0.63, p < 0.0001). The AVP did not correlate to EDV or PSV, nor to the RI measured in the ophthalmic artery. CONCLUSION: Arteriovenous passage time, which represents blood flow in a vascular segment of artery, capillary bed and corresponding vein, was found to be correlated to the EDV and the RI of the central retinal artery. The combination of different techniques allows further interpretation of ocular circulatory responses.

Adult↗
PubMed1993

Horse plasma gelsolin labelled with fluorescein isothiocyanate responds to calcium and actin.

Reaction between horse plasma gelsolin and fluorescein-5-isothiocyanate (FITC) resulted in incorporation of 4.8 +/- 0.6 fluorescein groups/gelsolin molecule. The sites of modification were not clustered in any one portion of the gelsolin polypeptide chain; all major peptides produced by proteolytic digestion with alpha-chymotrypsin exhibited a fluorescence characteristic of fluorescein. FITC-gelsolin has a peptide-backbone circular dichroism spectrum at 20 degrees C that is indistinguishable from that of native gelsolin, but FITC-gelsolin is considerably more resistant than native gelsolin to thermally induced precipitation. FITC-gelsolin is fully able to carry out severing of F-actin filaments, the prime function of gelsolin in plasma. An opening up of the structure of gelsolin on binding Ca2+ is evident from an increased susceptibility of FITC-gelsolin to quenching by I-. Ca2+ dependence of the interaction between gelsolin and actin is evident in titrations both of intensity and polarization of the fluorescence of FITC-gelsolin solutions. A Ca(2+)-sensitive interaction between gelsolin and tropomyosin also is observed.

Actins↗
PubMed1980

Continual monitoring of fluorescein angiography.

A system has been developed which enables continuous monitoring and recording of circulating fluorescein in the retinal blood vessels. This is achieved with minimal modification of a Zeiss fundus camera. A night vision television camera (lkegami) is used with illumination provided by the standard observation light of the fundus camera. This method offers advantages over standard fluorescein angiography in patient comfort, information obtained and the immediate evaluation of the fluorescein angiograms being available.

Fluorescein Angiography↗
PubMed2006

Optical coherence tomographic pattern of fluorescein angiographic leakage site in acute central serous chorioretinopathy.

PURPOSE: To study the optical coherence tomography (OCT) pattern of the fluorescein angiographic leakage in acute central serous chorioretinopathy (CSCR). METHODS: This is a non-interventional pilot case study. OCT line scan was performed over the fluorescein angiographic leak site in eyes clinically diagnosed acute CSCR. Clinical fundus photograph, site of leakage on fundus fluorescein angiography and corresponding OCT were analysed. RESULTS: The mean age of 10 consecutive patients was 38.8 +/- 6.9 years. Six patients were male and the mean duration of symptom was 7 days. Six eyes (60%) showed a characteristic dipping pattern of neurosensory retina with intervening hyper-reflective echoes suggestive of fibrin over the leakage site. All these eyes had ink-blot leak and subretinal fibrin. CONCLUSIONS: Ink-blot leak in acute CSCR with subretinal fibrin generates a dipping morphological pattern on OCT.

Acute Disease↗
PubMed1991

Skin perfusion patterns (fluorescein perfusography) during reactive hyperaemia in peripheral arterial occlusive disease.

From the pathophysiological point of view the regional distribution of blood flow is of special importance in ischaemic tissues. Within this study foot sole skin perfusion was investigated by means of fluorescein perfusography at rest and during reactive hyperaemia in patients with peripheral arterial occlusive disease confined to one limb (Fontaine stage II). Ambient temperatures were maintained around 21 degrees C. Mean fluorescein appearance times on the one side and their standard deviations (SD) and coefficients of variation (CV) on the other side were taken as measures of overall blood supply and homogeneity of flow, respectively. At rest no differences in these parameters could be detected between diseased legs and controls. After a 3-min supra-systolic circulatory arrest at the thigh, a significant reduction of fluorescein appearance times was observed for both groups but was statistically more pronounced in the controls. Furthermore, during reactive hyperaemia standard deviations as well as coefficients of variation decreased significantly only in normal limbs whereas they either remained constant (SD) or even increased (CV) in those with arterial obstructions. All effects associated with reactive hyperaemia showed statistically significant correlations with systolic ankle pressure indices. From these results it is concluded that haemodynamically effective arterial obstructions are followed by not only a restriction of overall hyperaemic blood supply but also a failure to homogenize microcirculatory perfusion in the case of increased flow requirements.

Arterial Occlusive Diseases↗
PubMed1993

Changes in skin blood flow in ischaemic limbs after vascular reconstruction measured by fluorescein flowmetry and laser Doppler flowmetry.

It is not known to what extent the skin blood flow increases after vascular reconstruction in ischaemic limbs. Thus, a study was undertaken to measure skin blood flow, using the two techniques laser Doppler flowmetry and fluorescein flowmetry, before and after vascular reconstruction. The skin temperature was measured also. The plantar circulation was assessed in 14 patients (9 non-diabetics and 5 diabetics; mean age 65 years; range 47-80), with the patient in the supine position, 1-2 days before and 8-10 days after surgery. After vascular reconstruction the skin blood flow increased by 240% (P < 0.01) when measured by fluorescein flowmetry and by 148% (P < 0.01) when measured by laser Doppler flowmetry, and the skin temperature rose by 3.2 degrees C (P < 0.01). In the contralateral non-operated limb there was no significant change in skin blood flow as measured by the two methods. The results imply that after vascular reconstruction in ischaemic limbs the skin blood flow increases both in the superficial layer as determined by fluorescein flowmetry (which measures the blood flow down to 0.6 mm) and in deeper layers as determined by laser Doppler flowmetry (which measures flux down to 6 mm).

Adult↗
PubMed1996

Precise correlation of histopathological and fluorescein angiographic morphology using retinal vascular casting.

The histopathology of three eyes obtained post mortem from 2 patients with age-related macular degeneration was correlated with the pre mortem fluorescein angiographic morphology. A precise point-by-point correlation between histopathology and the corresponding angiographic appearance was ensured by using the cast retinal vascular system as a pattern of reference. The study showed that both the photoreceptors, the pigment epithelium, and substances accumulated between the retinal and the choroidal vascular systems, may have a blocking effect on choroidal background fluorescence as seen on fluorescein angiograms. Furthermore, it is confirmed that fluorescein angiographic hyperfluorescence may be due to a lack of blocking of the choroidal fluorescence because of a window defect in the retinal photoreceptor layer and/or the pigment epithelium.

Aged↗