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1.  Multi-modality imaging on multiple evanescent white dot syndrome-A Spectralis Study 
AIM
To present retinal microstructure, metabolism and function abnormalities in the course of multiple evanescent white dot syndrome (MEWDS) by Heidelberg spectralis modality imaging platform and observe its outcome by EDI-SD-OCT and two wavelength autofluorescence.
METHODS
A case of multiple evanescent white dot syndrome in a 23-year-old female presented initially with a 15-day history of floaters and a central scotoma in the right eye. To establish the diagnosis, multimodality imaging was performed, namely, blue light-fundus autofluorescence (BL-FAF, excitation 488nm, emission >500nm), near-infrared fundus autofluorescence (NIR-FAF, excitation 787nm, emission >800nm) using a confocal scanning laser ophthalmoscope, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA), spectrum-domain enhance depth imaging optical coherence tomography (SD-EDI-OCT), multifocal electroretinography (mf-ERG) and fundus photogragh were performed and followed up at the eighth month after initially visiting.
RESULTS
Optical coherence tomography (OCT) showed a transient disruption of the foveal photoreceptor outer segments in correspondence to foveal granularity. NIR-FAF showed hypoautofluorescent areas, ≤40µm in size, mostly concentrated around the posterior pole and its temporal side less than that in BL-FAF. Mf-ERG show pinnacle disappeared in fovea and macula and responses decreased markedly compared with the follow eye. At the eighth month follow up, hyperfluorescence in BL-FAF were disappear, while, NIR-FAF Hypofluorescent spots in early stage of such lesion were reduced. But OCT demonstrated the structure was recovered in residual Hypofluorescent area in NIR-FAF. The subfoveal choroidal thickness was decreased from 372µm to 307µm slightly and cost line was recovered.
CONCLUSION
MEWDS is a benign self-healing disease and there is no pathological evidence to investigate the natural course of such disease. SD-OCT allows highly detailed images approaching histopathology to certify the microstructural changes. Two-wave length FAF and mf-ERG provide more information about metabolism in outer retina especial RPE and photoreceptor. Spectralis OCT combined with two-wavelength FAF and mf-ERG provide a new way to analyze this disease and offer more details for therapy and follow-up.
doi:10.3980/j.issn.2222-3959.2012.05.21
PMCID: PMC3484713  PMID: 23166880
MEWDS; Spectralis OCT; NIR-FAF; BL-FAF; mf-ERG
2.  Fundus autofluorescence imaging compared with different confocal scanning laser ophthalmoscopes 
The British Journal of Ophthalmology  2003;87(11):1381-1386.
Background: With the advent of confocal scanning laser ophthalmoscopes (cSLO), fundus autofluorescence (FAF) resulting mainly from lipofuscin accumulation on the level of the retinal pigment epithelium can be visualised in vivo. Various cSLOs are available to document FAF. The authors analysed and compared results of FAF using three different instruments.
Methods: Eight eyes of eight normal volunteers and 18 eyes of 12 patients with different retinal diseases (age related macular degeneration, macular dystrophy, central serous retinopathy) were examined. FAF images were recorded from each subject with the Heidelberg retina angiograph (HRA), the Rodenstock cSLO (RcSLO) and the Zeiss Prototype SM 30-4024 (ZcSLO). For excitation an argon laser (488 nm) was used (barrier filter: HRA 500 nm; RcSLO 515 nm; ZcSLO 521 nm). 32 FAF images were aligned and averaged using the same software for all cSLOs. FAF distribution was measured and grey scale values as well as root mean square (RMS) contrast were compared.
Results: Mean age of all subjects was 55.5 (SD 21.4) years. The maximum grey scale value averaged across all eyes was 76.19 (39.34) for the HRA, 61.44 (22.12) for the ZcSLO and 37.0 (9.97) for the RcSLO. The RMS contrast was 0.46 (0.20) for the ZcSLO, 0.40 (0.12) for the HRA, and 0.13 (0.05) for the RcSLO. The differences between the cSLOs were statistically significant with higher grey scale levels and more contrast for the HRA and ZcSLO than the RcSLO (repeated measures ANOVA; p<0.0001). The differences between the HRA and the ZcSLO were not significant (post hoc comparisons; p<0.05).
Conclusions: All cSLOs allow clinically useful FAF imaging in retinal diseases. However, grey scale levels and contrast were much lower on the RcSLO. Therefore, RcSLO images appear much darker than HRA or ZcSLO images. Furthermore, not all cSLOs have a fixed photodetector gain and a standardised value for the argon laser amplification, which is mandatory for an absolute comparison of FAF imaging results.
PMCID: PMC1771908  PMID: 14609839
scanning laser ophthalmoscope; fundus autofluorescence; lipofuscin; retinal pigment epithelium; macular degeneration
3.  The Minnesota Grading System Using Fundus Autofluorescence of Eye Bank Eyes: A Correlation To Age-Related Macular Degeneration (An AOS Thesis) 
Purpose
To establish a grading system of eye bank eyes using fundus autofluorescence (FAF) and identify a methodology that correlates FAF to age-related macular degeneration (AMD) with clinical correlation to the Age-Related Eye Disease Study (AREDS).
Methods
Two hundred sixty-two eye bank eyes were evaluated using a standardized analysis of FAF. Measurements were taken with the confocal scanning laser ophthalmoscope (cSLO). First, high-resolution, digital, stereoscopic, color images were obtained and graded according to AREDS criteria. With the neurosensory retina removed, mean FAF values were obtained from cSLO images using software analysis that excludes areas of atrophy and other artifact, generating an FAF value from a grading template. Age and AMD grade were compared to FAF values. An internal fluorescence reference standard was tested.
Results
Standardization of the cSLO machine demonstrated that reliable data could be acquired after a 1-hour warm-up. Images obtained prior to 1 hour had falsely elevated levels of FAF. In this initial analysis, there was no statistical correlation of age to mean FAF. There was a statistically significant decrease in FAF from AREDS grade 1, 2 to 3, 4 (P < .0001). An internal fluorescent standard may serve as a quantitative reference.
Conclusions
The Minnesota Grading System (MGS) of FAF (MGS-FAF) establishes a standardized methodology for grading eye bank tissue to quantify FAF compounds in the retinal pigment epithelium and correlate these findings to the AREDS. Future studies could then correlate specific FAF to the aging process, histopathology AMD phenotypes, and other maculopathies, as well as to analyze the biochemistry of autofluorescent fluorophores.
PMCID: PMC2646441  PMID: 19277247
4.  Fundus Autofluorescence Patterns in Primary Intraocular Lymphoma 
Retina (Philadelphia, Pa.)  2010;30(1):23-32.
Purpose
To evaluate fundus autofluorescence (FAF) patterns in patients with primary intraocular (vitreoretinal) lymphoma (PIOL/PVRL).
Methods
Records of all PIOL patients who underwent FAF imaging at the National Eye Institute (NEI) were reviewed. FAF patterns were evaluated with respect to clinical disease status and the findings on fluorescein angiography (FA) and spectral-domain optical coherence tomography (SD-OCT) images.
Results
There were 18 eyes (10 patients) with PIOL who underwent FAF imaging. Abnormal autofluorescence in the form of granular hyper- and hypoautofluorescence was seen in 11 eyes (61%) and blockage by mass lesion was seen in 2 eyes (11%). All eyes with granular pattern on FAF had active PIOL at the time of imaging, but there were 5 eyes with unremarkable FAF which were found to have active lymphoma. The most common pattern on FA was hypofluorescent round spots with a “leopard-spot” appearance (43%). These hypofluorescent spots on FA correlated to hyperautofluorescent spots on FAF in 5 (36%) eyes (inversion of FAF). Nodular hyperreflective spots at the level of RPE on OCT were noted in 43% of eyes. The hyperautofluorescent spots on FAF correlated with nodular hyperreflective spots on OCT in 6 eyes (43%).
Conclusion
Granularity on fundus autofluorescence was associated with active lymphoma in majority of cases. An inversion of FAF (hyperautofluorescent spots on FAF corresponding to hypofluorescent spots on FA) was observed in less than half of the eyes.
doi:10.1097/IAE.0b013e3181b408a2
PMCID: PMC3707141  PMID: 19834356
Cytokines; diagnosis; fundus autofluorescence; primary intraocular lymphoma; primary vitreoretinal lymphoma; uveitis
5.  Prevalence of Peripheral Abnormalities on Ultra-Widefield Greenlight (532 nm) Autofluorescence Imaging at a Tertiary Care Center 
Purpose.
To assess the prevalence of peripheral fundus autofluorescence (FAF) abnormalities in a variety of diseases seen at a tertiary retina clinic.
Methods.
We conducted a retrospective review of cases seen at the Doheny Eye Institute between November 2009 and May 2011, who had ultra-widefield FAF and pseudocolor imaging performed on new models of scanning laser ophthalmoscopes. Patients with a history of previous therapies that could alter the FAF findings, including vitrectomy, cryotherapy, laser photocoagulation, or photodynamic therapy, were excluded from the analysis. Based on their primary diagnosis the eyes were grouped into nine disease categories: age-related macular degeneration, central serous retinopathy, dystrophy, inflammatory disorders, ocular tumor, retinal vascular disorders, other, normal, and unknown. All FAF and accompanying pseudocolor images were reviewed independently by two reading center–certified graders.
Results.
A total of 470 eyes of 248 patients were included for analysis of which 461 eyes had images of sufficient quality for grading. The prevalence of peripheral findings was 65.5% (n = 302) for FAF images and 68.5% (n = 316) for the pseudocolor images (P < 0.001). The prevalence of peripheral abnormalities differed significantly between the disease categories ranging from 18.5% to 82.2% for FAF and 18.5% to 82.4% for pseudocolor images.
Conclusions.
Peripheral FAF abnormalities are frequent and readily revealed by FAF imaging. Interestingly, even cases with presumably macular disease demonstrated a high prevalence of peripheral findings. Further investigation in prospective studies is warranted.
Peripheral abnormalities on ultra-widefield autofluorescence images are common in a large variety of diseases and reliably identifiable. There is a high correspondence between changes seen on autofluorescence and pseudocolor images. Peripheral changes in ultra-widefield imaging are a common finding in a variety of different diseases.
doi:10.1167/iovs.12-9909
PMCID: PMC3450919  PMID: 22871828
6.  Role of wide-field autofluorescence imaging and scanning laser ophthalmoscopy in differentiation of choroidal pigmented lesions 
AIM
To evaluate the diagnostic properties of wide-field fundus autofluorescence (FAF) scanning laser ophthalmoscope (SLO) imaging for differentiating choroidal pigmented lesions.
METHODS
A consecutive series of 139 patients were included, 101 had established choroidal melanoma with 13 untreated lesions and 98 treated with radiotherapy. Thirty-eight had choroidal nevi. All patients underwent a full ophthalmological examination, undilated wide-field imaging, FAF and standardized US examination. FAF images and imaging characteristics from SLO were correlated with the structural findings in the two patient groups.
RESULTS
Mean FAF intensity of melanomas was significantly lower than the FAF of choroidal nevi. Only 1 out of 38 included eyes with nevi touched the optic disc compared to 31 out of 101 eyes with melanomas. In 18 out of 101 melanomas subretinal fluid was seen at the pigmented lesion compared to none seen in eyes with confirmed choroidal nevi. In “green laser separation”, a trend towards more mixed FAF appearance of melanomas compared to nevi was observed. The mean maximal and minimal transverse and longitudinal diameters of melanomas were significantly higher than those of nevi.
CONCLUSION
Wide-field SLO and FAF imaging may be an appropriate non-invasive diagnostic screening tool to differentiate benign from malign pigmented choroidal lesions.
doi:10.3980/j.issn.2222-3959.2014.04.21
PMCID: PMC4137210  PMID: 25161946
imaging; autofluorescence; scanning laser ophthalmoscopy; choroidal lesion; melanoma
7.  A Comparison of Fundus Autofluorescence and Retinal Structure in Patients with Stargardt Disease 
Purpose
To improve our understanding of Stargardt disease by comparing structural changes seen on spectral domain optical coherence tomography (SD-OCT) to those visible on fundus autofluorescence (FAF).
Methods
FAF and SD-OCT were obtained on 22 eyes of 11 patients with Stargardt disease. SD-OCT images were obtained at the fovea and at the eccentric preferred retinal locus (PRL). The diameters of “absent” (hypo-autofluorescent) and “abnormal” FAF areas were measured. The extent of the transverse defect of the junction between the inner and outer segments of the photoreceptors (IS-OS) was measured in the foveal area. The PRL was evaluated with fundus photography and microperimetry.
Results
Twenty-one of 22 eyes showed defective FAF. For 17 eyes, FAF was absent in the fovea and for 4 eyes the FAF was abnormal. All eyes showed disorganization and/or loss of the IS-OS junction in the foveal area on SD-OCT. The diameter of the absent FAF area was smaller than the measurement of the IS-OS junction loss; the latter was closer to the diameter of the abnormal FAF area. Seventeen eyes had an eccentric PRL associated with a retinal area with no defects on FAF.
Conclusions
For the majority of eyes changes on SD-OCT correlated well with changes on FAF. However for 3 patients, photoreceptor abnormalities were seen in the fovea on SD-OCT without an equivalent abnormality on FAF. This suggests that for these patients, the structural integrity of the photoreceptors may be affected earlier than changes in the RPE at least as detected by FAF.
doi:10.1167/iovs.08-2657
PMCID: PMC2749553  PMID: 19324865
8.  Microperimetry and fundus autofluorescence in patients with early age‐related macular degeneration 
The British Journal of Ophthalmology  2007;91(11):1499-1503.
Background
Early age‐related macular degeneration (AMD) has been correlated with different functional alterations, but the exact relationship between fundus lesions and overlying sensitivity is not well known. The aim of this study was to compare fundus‐related sensitivity (microperimetry) and fundus autofluorescence (FAF) of the macular area with drusen and pigment abnormalities in early AMD.
Methods
13 consecutive patients with early AMD and visual acuity of 20/20 were studied by means of microperimetry, which automatically analyses macular light differential threshold and fixation patterns. Fundus colour photo and FAF of the macular area were recorded on the same day. Microperimetry was exactly (topographically) superimposed over FAF images.
Results
Macular sensitivity significantly decreased over large drusen (11.2 ± 5.6 dB, p<0.0001) and over pigment abnormalities (13.1 ± 3.6 dB, p<0.0001). When both characteristics were present the reduction was greater if compared with its absence (9.6 ± 4.3 versus 15.0 ± 4.5 dB, p<0.0001). Sensitivitity reduction was significant in areas with altered FAF when compared with areas with normal FAF (p<0.0001).
Conclusions
Increased FAF in early AMD has a functional correlate exactly quantified by microperimetry. In retinal areas affected by early AMD retinal sensitivity deteriorates, despite good visual acuity. Microperimetry may allow the early detection of functional impairment caused by these lesions. Both microperimetry and FAF may be useful to monitor AMD progression.
doi:10.1136/bjo.2007.119685
PMCID: PMC2095427  PMID: 17504849
9.  Green-light fundus autofluorescence in diabetic macular edema 
AIM
To evaluate the role of central green-light fundus autofluorescence (FAF) in diabetic macular edema (DME).
METHODS
A consecutive series of 92 study eyes with diabetic retinopathy were included. Out of those, 51 diabetic eyes had DME and were compared to 41 diabetic eyes without DME. In all subjects, green-light FAF images were obtained, quantified and classified into various FAF patterns. Cross-sectional optical coherence tomography (OCT) scans were obtained for evaluation of Inner/Outer segment (IS/OS) layer integrity, measurements of central RPE-IS/OS layer thickness as well as classification of DME into various subtypes.
RESULTS
Mean central green-light FAF intensity of eyes with DME (1.289±0.140)log did not significantly differ from diabetic patients without DME (1.317±0.137)log. Most classifiable FAF patterns were seen in patients with cystoid DME. Mean central retinal thickness (CRT) of all study eyes with DME was (501.9±112.4)µm compared to (328.2±27.0)µm in diabetic patients without DME. Patients with DME had significantly more disrupted photoreceptor IS/OS layers than diabetic patients without DME (28/51 vs 5/41, P<0.001). Mean RPE-IS/OS thickness of patients with DME (60.7±14.1)µm was significantly (P<0.001) lower than in diabetic eyes without DME (73.5±9.4)µm. Correlation analys1s revealed non-significant correlations of green-light FAF intensity and OCT parameters in all subtypes of DME.
CONCLUSION
Our results indicate a poor correlation of central green-light FAF intensity with CRT, IS/OS layer integrity or RPE-IS/OS layer thickness in diabetic patients with or without DME and its various subtypes. Thus, central green-light FAF is not suitable for detection of retinal thickening in DME.
doi:10.3980/j.issn.2222-3959.2013.01.16
PMCID: PMC3580255  PMID: 23549658
diabetic macular edema; fundus autofluorescence; optical coherence tomography
10.  Fundus Autofluorescence and Spectral Domain OCT in Central Serous Chorioretinopathy 
Journal of Ophthalmology  2011;2011:706849.
Background. To describe the standard autofluorescence (FAF), the near infrared autofluorescence (NIA) and optical coherence tomography (OCT) patterns in central serous chorioretinopathy, correlating them with fluorescein angiography. Methods. Cross-sectional observational study, in which patients with at least seven months of CSC underwent ophthalmologic examination, fundus photography, FAF, NIA, fluorescein angiography (FA), and spectral-domain OCT. Results. Seventeen eyes of thirteen patients were included. The presentation features were a mottled hyperFAF in the detached area and areas with pigment mottling. NIA images showed areas of hyperNIA similar to FAF and localized areas of hypoNIA, which correlated with the points of leakage in the FA. OCT showed pigment epithelium detachment at the location of these hypoNIA spots. Discussion. FAF showed increased presence of fluorophores in the area of retinal detachment, which is believed to appear secondary to lipofuscin accumulation in the RPE or the presence of debris in the subretinal fluid. NIA has been related to the choroidal melanin content and there were areas of both increased and decreased NIA, which could be explained by damage ahead the retina, basically RPE and choroid. These findings, along with the PEDs found in the areas of hypoNIA, support the notion of a primary choroidal disease in CSC.
doi:10.1155/2011/706849
PMCID: PMC3153919  PMID: 21845214
11.  Fundus Autofluorescence in Type 2 Idiopathic Macular Telangiectasia: Correlation with Optical Coherence Tomography and Microperimetry 
American journal of ophthalmology  2009;148(4):573-583.
Purpose
To employ multiple modality imaging to described patients with type 2 idiopathic macular telangiectasia (IMT) at different disease severity stages so as to characterize and categorize disease progression through the full spectrum of disease phenotypes.
Design
Observational case series.
Methods
Twelve patients with type 2 IMT (22 eyes) with type 2 IMT examined with fundus photography, angiography, optical coherence tomography (OCT) imaging, fundus autofluorescence (FAF), and microperimetry (MP) testing in an institutional setting.
Results
Eyes examined by multiple modality imaging were classified into five proposed categories (0–4): Category 0 (fellow) eyes were normal on all imaging modalities. Category 1 eyes had increased foveal autofluorescence on FAF imaging as the only imaging abnormality. Category 2 eyes had increased foveal autofluorescence together with funduscopic and angiographic features typical of type 2 IMT. Category 3 had additional evidence of foveal atrophy on OCT and while category 4 has all the above features plus clinically evident pigment clumping. FAF signal increased in intensity in the foveal region from category 0 through category 3, while category 4 eyes demonstrated a mixed pattern of increased and decreases FAF signal.
Conclusions
The findings here outline a sequence of progressive changes seen with multiple imaging modalities in advancing stages of disease. Increases in foveal autofluorescence is an early anatomical change in type 2 IMT that may precede typical clinical and angiographic changes. Loss of macular pigment density in the fovea and a changing composition of flurophores in the retinal pigment epithelium may underlie these changes in FAF in the fundus.
doi:10.1016/j.ajo.2009.04.030
PMCID: PMC2753760  PMID: 19573860
12.  Fundus Autofluorescence and Spectral-Domain Optical Coherence Tomography Characteristics in a Rapidly Progressing Form of Geographic Atrophy 
A rapidly progressing geographic atrophy phenotype shows characteristic fundus autofluorescence and SD-OCT features.
Purpose.
To further characterize a previously described phenotypic variant of geographic atrophy (GA) associated with rapid progression and a diffuse-trickling appearance on fundus autofluorescence (FAF).
Methods.
Thirty-six patients (60 eyes; 72.2% women; mean age, 69.4 ± 10.7 years) with this distinct phenotype were examined by simultaneous confocal scanning laser ophthalmoscopy (cSLO) and spectral-domain optical coherence tomography (SD-OCT) imaging. Images were qualitatively and quantitatively analyzed and compared with 60 eyes (38 patients) with non diffuse-trickling GA.
Results.
The atrophic area in the diffuse-trickling phenotype showed a grayish FAF signal and characteristic coalescent lobular configuration at the lesion boundaries. SD-OCT revealed a marked splitting of band 4 (the presumptive retinal pigment epithelium (RPE)/Bruch's membrane (BM) complex) in all 240 analyzed border sections of diffuse-trickling GA eyes (four borders/eye) with a mean distance between the inner and outer parts of band 4 of 23.2 ± 7.5μm. This finding was present in only 13.8% (33/240) of analyzed border sections in non diffuse-trickling GA.
Conclusions.
Patients with the rapidly progressing diffuse-trickling GA phenotype exhibited a characteristic marked separation within the RPE/BM complex on SD-OCT-imaging. The presumed histopathologic correlates are basal laminar deposits. Such deposits may promote RPE cell death and, thus, contribute to rapid GA progression. The persistence of these deposits within the atrophic lesion may account for the distinct grayish FAF appearance, which differs from the markedly reduced signal in other forms of GA. Identification of such alterations based on FAF and SD-OCT imaging may be helpful in future interventional trials directed toward slowing GA progression. (ClinicalTrials.gov number, NCT00393692.)
doi:10.1167/iovs.10-7021
PMCID: PMC3109052  PMID: 21310912
13.  Predictive Value of Fundus Autofluorescence for Development of Geographic Atrophy in Age-Related Macular Degeneration 
Purpose
It has been suggested that lipofuscin accumulation, as measured by increased fundus autofluorescence (FAF), precedes progression or development of junctional zone geographic atrophy (GA) in age-related macular degeneration (AMD). The tools of biomedical image analysis were used to measure the probabilistic relationship of GA progression to increased FAF.
Methods
Serial AF images of eight eyes of six patients with AMD with GA were registered on computer. The images were leveled with a 12-zone quadratic polynomial mathematical model to minimize background variability. Semiautomated segmentation of GA was performed on the leveled images. Increased FAF was defined as a gray level greater than 2 standard deviations above the leveled image mean, identified on the initial image with automated segmentation, and measured as a fraction of the 250-μm border zone surrounding the initial GA lesion. Areas of GA lesions were identified on the final image. The positive predictive value (PPV) of increased FAF was determined as the probability that any pixel with increased FAF in the initial image would become part of new GA in the final image. Relative PPV was determined relative to the total quantity of new GA. The NPV (NPV) of increased FAF was calculated as the probability that any pixels without increased FAF would not become atrophic. The relative NPV was determined similarly. A similar analysis was also conducted with a 500-μm border zone to determine the predictive value of proximity to the original GA lesion (“proximity”) for GA progression.
Results
As a fraction of the geographic atrophy border zone, the mean new GA was 0.44 ± 0.20, and the mean increased FAF was 0.06 ± 0.06. The mean PPV of increased FAF for new GA formation was 0.50 ± 0.26. Compared with the relative PPV of chance of 1.0, the mean relative PPV of increased FAF was 1.15 ± 0.28. The mean NPV of increased FAF was 0.57 ± 0.20. The mean relative NPV of increased FAF was 1.00 ± 0.02. In the 500-μm border zone, the mean relative PPV of FAF and of proximity were essentially equal (1.56 ± 0.70 and 1.52 ± 0.26, respectively), whereas the mean relative NPV of proximity was significantly greater than that of FAF (1.26 ± 0.19 and 1.01 ± 0.01, respectively, P = 0.02)
Conclusions
The results of digital image analysis suggest that although increased FAF may have a modest PPV for new GA development, the relative PPV is generally no greater than chance. Similarly, the relative NPV demonstrates negligible difference from chance and is also lower than the relative NPV of proximity. This suggests that increased FAF, though a disease manifestation, is not a strong risk factor for development or extension of GA.
doi:10.1167/iovs.05-1027
PMCID: PMC2754747  PMID: 16723483
14.  Assesment of fundus autofluorescence in serpiginous and serpiginous-like choroidopathy 
Eye  2012;26(9):1232-1236.
Aims
The aim of this study was to correlate the activity status disclosed in fluorescein angiography (FA) and fundus autofluorescence (FAF) imaging, and the variations of FAF images in the evolution of serpiginous choroidopathy (SC) and serpiginous-like choroidopathy (SLC).
Methods
Prospective consecutive case series. Patients with SC or SLC were included from July 2009 to December 2010. All patients underwent FAF imaging (Spaide Autofluorescence Filters, Topcon TRC 50IX) and FA (Topcon TRC 50IX).
Results
Twelve patients (eight males, mean age 51.2 years) were included. Bilateral involvement in nine cases. Three different patterns of FAF images were present: active inflammation, transitional, and inactive inflammation.
Conclusion
FAF may be a useful tool for following patients with SC and SLC. It is possible to reserve other invasive techniques, such as FA, for cases with suspicious activity disclosed by FAF imaging.
doi:10.1038/eye.2012.121
PMCID: PMC3443827  PMID: 22744387
serpiginous choroidopathy; serpiginous-like choroidopathy; choroiditis; posterior uveitis; autofluorescence
15.  Multimodality Diagnostic Imaging in Unilateral Acute Idiopathic Maculopathy 
Archives of ophthalmology  2012;130(1):50-56.
Objective
To describe the clinical features and imaging characteristics in unilateral acute idiopathic maculopathy (UAIM).
Methods
This is a retrospective review of four patients diagnosed with UAIM. Clinical characteristics (age, symptoms, Snellen visual acuity (VA), and funduscopic features) and images from spectral-domain optical coherence tomography (SD-OCT), fundus autofluorescence (FAF), fluorescein (FA), and indocyanine green (ICG) angiography were analyzed.
Results
The median age at presentation was 31 years (range 27–52 years). The median interval between symptom onset and presentation was four weeks (range 1–20 weeks). Associated systemic findings included a viral prodrome (50%), orchitis (50%), hand-foot-mouth disease (25%), and positive Coxsackie virus titers (50%). The median presenting VA was 20/400 (range 20/70–1/400), which improved to 20/30 (range 20/20–20/60) at final follow-up. The median follow-up time was 6 weeks (range 0–8 weeks). Early in the disease course, the central macula developed irregular, circular areas of white-grey discoloration. Following recovery, the macula had a stippled retinal pigment epithelium characterized by rarefaction and hyperplasia. FA demonstrated irregular early hyperfluorescence and late subretinal hyperfluorescence. SD-OCT showed a partially reversible disruption of the outer photoreceptor layer. FAF initially revealed stippled autofluorescence that eventually became more hypoautofluorescent. ICG showed “moth-eaten” appearing choroidal vasculature, suggestive of choroidal inflammation.
Conclusions
The imaging characteristics highlight the structural changes during the active and resolution phases of UAIM. The visual recovery correlates with structural changes and suggests that the pathogenesis involves inflammation of the inner choroid, retinal pigment epithelium, and outer photoreceptor complex that is partially reversible.
doi:10.1001/archophthalmol.2011.359
PMCID: PMC4116109  PMID: 22232475
16.  Clinicopathologic findings in Best vitelliform macular dystrophy 
Purpose
To correlate the clinical and histopathologic features of Best vitelliform macular dystrophy (BVMD).
Methods
Two eyes were obtained postmortem from a patient with BVMD. The patient’s clinical information was reviewed. Series sections of the globes were performed and sequentially stained with hematoxylin-eosin, periodic acid-Schiff or Masson trichrome. A section of the left macula was submitted for electron microscopic processing. Histopathologic findings were reconstructed in a scaled two-dimensional map and compared with fundus photography, fundus autofluorescence (FAF), fundus fluorescein angiography (FFA) and optical coherence tomography (OCT) images.
Results
The macular lesion of the right eye was identified as a well-demarcated region with pigment, elevated submacular yellow material and subretinal fluid. This corresponded histopathologically to a well-circumscribed area of RPE hyperplasia, accumulation of lipofuscin in the RPE, deposition of granular material in the photoreceptors, macrophages and drusen. The left eye displayed a 1 disc diameter chorioretinal scar with surrounding shallow fluid and submacular pigment. This corresponded to RPE changes and a fibrocellular proliferation in the choriocapillaris.
Conclusion
Histopathologic mapping revealed retinal edema, RPE abnormalities, drusen and a chorioretinal scar in BVMD that correlated with the fundus, FFA, FAF and OCT findings.
doi:10.1007/s00417-010-1587-3
PMCID: PMC3425361  PMID: 21136072
Best vitelliform macular dystrophy; Optical coherence tomography; Fundus autofluorescence; Fundus fluorescein angiography; Two-dimensional reconstruction; Clinicopathologic correlation
17.  Correlation between peripapillary retinal nerve fiber layer thickness and fundus autofluorescence in primary open-angle glaucoma 
Purpose
To investigate the relationship between retinal nerve fiber layer (RNFL) thickness and retinal pigment epithelium alterations in patients with advanced glaucomatous visual field defects.
Methods
A consecutive, prospective series of 82 study eyes with primary open-angle glaucoma and advanced glaucomatous visual field defects were included in this study. All study participants underwent a full ophthalmic examination followed by visual field testing with standard automated perimetry as well as spectral-domain optical coherence tomography (SD-OCT) for peripapillary RNFL thickness and Optos wide-field fundus autofluorescence (FAF) images. A pattern grid with corresponding locations between functional visual field sectors and structural peripapillary RNFL thickness was aligned to the FAF images at corresponding location. Mean FAF intensity (range: 0 = black and 255 = white) of each evaluated sector (superotemporal, temporal, inferotemporal, inferonasal, nasal, superonasal) was correlated with the corresponding peripapillary RNFL thickness obtained with SD-OCT.
Results
Correlation analyses between sectoral RNFL thickness and standardized FAF intensity in the corresponding topographic retina segments revealed partly significant correlations with correlation coefficients ranging between 0.004 and 0.376 and were statistically significant in the temporal inferior central field (r = 0.324, P = 0.036) and the nasal field (r = 0.376, P = 0.014).
Conclusion
Retinal pigment epithelium abnormalities correlate with corresponding peripapillary RNFL damage, especially in the temporal inferior sector of patients with advanced glaucomatous visual field defects. A further evaluation of FAF as a potential predictive parameter for glaucomatous damage is necessary.
doi:10.2147/OPTH.S49112
PMCID: PMC3788816  PMID: 24092967
glaucoma; fundus autofluorescence; FAF; retinal nerve fiber layer; RNFL; optical coherence tomography; OCT; imaging
18.  FUNDUS AUTOFLUORESCENCE OF CHOROIDAL MELANOCYTIC LESIONS AND THE EFFECT OF TREATMENT 
Purpose
To correlate fundus autofluorescence (FAF) patterns in choroidal melanocytic lesions with changes present on the surface of such lesions, including lipofuscin, hyperpigmentation, drusen, and fibrous metaplasia, and to describe the effect of treatment on FAF.
Methods
Retrospective chart review of 23 consecutive patients with choroidal nevi and melanoma who underwent FAF photography. The correlation between increased FAF patterns and foci of lipofuscin, hyperpigmentation, drusen, or fibrous metaplasia was defined as a complete correlation, partial correlation, or no correlation. The posttreatment FAF photographs of 6 patients with choroidal melanoma who were managed with plaque radiotherapy or plaque radiotherapy and transpupillary thermotherapy were also analyzed.
Results
Lipofuscin was present in 13 tumors, hyperpigmentation in 9 tumors, drusen in 6 tumors, and fibrous metaplasia in 4 tumors. A complete correlation between increased FAF and lipofuscin was found in 8 tumors (61.5%), a partial correlation in 3 tumors (23.1%), and no correlation in 2 tumors (15.4%). A complete correlation between hyperpigmentation and increased FAF was found in 5 tumors (55.6%), a partial correlation in 3 tumors (33.3%), and no correlation in 1 tumor (11.1%). A partial correlation was found between drusen and increased FAF in all 4 tumors. A partial correlation was found between fibrous metaplasia and increased FAF in all 3 tumors. Following treatment, increased FAF was observed in 6 choroidal melanomas owing to an increase in lipofuscin and hyperpigmentation.
Conclusions
Choroidal melanocytic lesions with overlying lipofuscin and hyperpigmentation are associated with increased FAF in about 90% of cases. Fundus autofluorescence photography may be helpful in evaluating small melanocytic tumors, since lipofuscin is a risk factor for growth. Following treatment, choroidal melanomas may show increased FAF.
PMCID: PMC2258096  PMID: 18427607
19.  Classification of abnormal fundus autofluorescence patterns in the junctional zone of geographic atrophy in patients with age related macular degeneration 
Aim: To describe and classify patterns of abnormal fundus autofluorescence (FAF) in the junctional zone of geographic atrophy (GA) in patients with age related macular degeneration.
Methods: Digital FAF images were recorded in 164 eyes of 107 patients using a confocal scanning laser ophthalmoscope (cSLO; excitation 488 nm, detection above 500 nm) as part of a prospective multicentre natural history study (FAM Study). FAF images were obtained in accordance with a standardised protocol for digital image acquisition and generation of mean images after automated alignment.
Results: Image quality was sufficient for classification of FAF patterns in 149 eyes (90.9%) with lens opacities being the most common reason for insufficient image quality. Abnormal FAF outside GA in 149 eyes was classified into four patterns: focal (12.1%), banded (12.8%), patchy (2.0%), and diffuse (57.0%), whereby 12.1% had normal background FAF in the junctional zone. In 4% there was no predominant pattern. The diffuse pattern was subdivided into four groups including reticular (4.7%), branching (27.5%), fine granular (18.1%), and fine granular with peripheral punctate spots (6.7%).
Conclusions: Different phenotypic patterns of abnormal FAF in the junctional zone of GA can be identified with cSLO FAF imaging. These distinct patterns may reflect heterogeneity at a cellular and molecular level in contrast with a non-specific ageing process. A refined phenotypic classification may be helpful to identify prognostic determinants for the spread of atrophy and visual loss, for identification of genetic risk factors as well as for the design of future interventional trials.
doi:10.1136/bjo.2004.057794
PMCID: PMC1772741  PMID: 15965170
fundus autofluorescence; confocal scanning laser ophthalmoscopy; age related macular disease; geographic atrophy; retinal imaging
20.  COMPARISON OF COLOR FUNDUS PHOTOGRAPHS AND FUNDUS AUTOFLUORESCENCE IMAGES IN MEASURING GEOGRAPHIC ATROPHY AREA 
Retina (Philadelphia, Pa.)  2012;32(9):1884-1891.
Purpose
To assess the agreement between color fundus photographs (CFP) and fundus autofluorescence (FAF) images when measuring geographic atrophy (GA) area and reproducibility of measurements between graders. Frequency and disagreement types were also determined.
Methods
Eyes with GA secondary to age-related macular degeneration had CFP and FAF imaging on the same day. Seventy-two eyes from 72 patients were included in the analysis. Three graders calculated GA area using digital imaging software. Main outcome measures included agreement between graders for GA area on both FAF and CFP and agreement between both imaging modalities.
Results
The intraclass correlation for the 3 graders for FAF images was 0.99 (95% confidence interval, 0.98–0.99). For CFP, it was 0.96 (95% confidence interval, 0.94–0.97). The intraclass correlation between imaging modalities for Graders 1, 2, and 3 were 0.93, 0.85, and 0.87, respectively. Sensitivities to detect involvement of fovea (CFP, 86–97%; FAF, 72–93%) and specificities to detect sparing of fovea (CFP, 74–76%; FAF, 59–88%) overlapped between imaging modalities.
Conclusion
Both CFP and FAF imaging are reliable for measuring GA area. Interobserver agreement was slightly higher for FAF images. Although the high agreement between modalities suggests that either would be appropriate for measuring GA area, using both may be the best approach for following GA progression.
doi:10.1097/IAE.0b013e3182509778
PMCID: PMC3697023  PMID: 22547167
age-related macular degeneration; color fundus photographs; fundus autofluorescence; geographic atrophy
21.  Centrifugal Expansion of Fundus Autofluorescence Patterns in Stargardt Disease Over Time 
Archives of ophthalmology  2011;130(2):171-179.
Objective
Changing lipofuscin and melanin content in RPE cells has been hypothesized to contribute to Stargardt disease pathogenesis. Longitudinal study of autofluorescence in Stargardt disease which reflect changing fluorophore compositions can reveal aspects of disease progression not previously evident.
Method
We examined the temporal-spatial patterns of fundus autofluorescence with excitation at both 488 nm (standard fundus autofluorescence, FAF) and 795nm (near infrared autofluorescence, NIA) in a longitudinal case series involving 8 eyes of 4 patients (range of follow-up = 11 to 57 months; mean = 39 months). Image processing was performed to analyze spatial and temporal cross-modality associations.
Results
Longitudinal FAF imaging of fleck lesions revealed hyperautofluorescent lesions that extended in a centrifugal direction from the fovea with time. Patterns of spread were non-random and followed a radial path that leaves behind a trail of diminishing autofluorescence. Longitudinal NIA imaging also demonstrated centrifugal lesion spread, but with fewer hyperautofluorescent lesions, suggestive of more transient hyperautofluorescence and more rapid decay at longer wavelengths. FAF and NIA abnormalities were spatially correlated to each other, and together reflect systematic progressions in fleck distribution and fluorophore composition occurring during the natural history of the disease.
Conclusion
Stargardt disease fleck lesions do not evolve randomly in location but instead follow consistent patterns of radial expansion and a systematic decay of autofluorescence that reflect changing lipofuscin and melanin compositions in RPE cells. These progressive foveal-to-peripheral changes are helpful in elucidating molecular and cellular mechanisms underlying Stargardt disease and may constitute potential outcome measures in clinical trials.
doi:10.1001/archophthalmol.2011.332
PMCID: PMC3768260  PMID: 21987580
22.  Atypical vitelliform macular dystrophy misdiagnosed as chronic central serous chorioretinopathy: case reports 
BMC Ophthalmology  2012;12:25.
Background
To report two cases of atypical vitelliform macular dystrophy misdiagnosed as chronic central serous chorioretinopathy.
Case presentation
Two patients with incidentally discovered abnormalities of the retina without specific symptoms were referred to our hospital for consultation. Bilateral macula atrophic lesions were observed and optical coherence tomography revealed serous retinal detachment in the macula. Fluorescein angiography showed multiple leakages around the central hypofluorescent area and indocyanine green angiography showed partially dilated choroidal vessels. Fundus autofluorescence (FAF) showed a decreasing pattern of autofluorescence in the subretinal fluid area, and increasing autofluorescence at the border of the serous retinal detachment. Both patients were diagnosed with chronic central serous chorioretinopathy. Photodynamic therapy and intravitreal bevacizumab injection were administered for engorged choroidal vessels during follow-up, but neither patient showed improvement in symptoms or ophthalmologic findings. Based on re-evaluation by fundus photography, optical coherence tomography, fluorescein angiography, and comparison of the results of FAF with the first visit, vitelliform macular dystrophy was suspected and a definite diagnosis was made by electrooculography and genetic testing.
Conclusion
In patients with continuous serous retinal detachment without response to photodynamic therapy or intravitreal bevacizumab injection, careful fundus exam and FAF can be used to diagnose atypical vitelliform macular dystrophy.
doi:10.1186/1471-2415-12-25
PMCID: PMC3549736  PMID: 22817759
23.  SLO-infrared imaging of the macula and its correlation with functional loss and structural changes in patients with Stargardt disease 
Retina (Philadelphia, Pa.)  2011;31(5):949-958.
Purpose
To correlate the degree of functional loss with structural changes in patients with Stargardt disease.
Methods
Eighteen eyes of 10 Stargardt patients were studied. Scanning laser ophthalmoscope (SLO) infrared images were compared to corresponding spectral domain optical coherence tomography (SD-OCT) scans. Additionally, SLO microperimetry was performed and results were superimposed on SLO infrared images and in selected cases on fundus autofluorescence (FAF) images.
Results
Seventeen of 18 eyes showed a distinct hypo-reflective foveal and/or perifoveal area with distinct borders on SLO-infrared images which was less evident on funduscopy and incompletely depicted in FAF images. This hypo-reflective zone corresponded to areas of significantly elevated psychophysical thresholds on microperimetry testing, in addition to thinning of the retinal pigment epithelium (RPE), disorganization or loss of the photoreceptor cell inner-outer segment (IS-OS) junction and external limiting membrane (ELM) on SD-OCT.
Conclusion
SLO-infrared fundus images are useful for depicting retinal structural changes in Stargardt patients. An SD-OCT/SLO microperimetry device allows for a direct correlation of structural abnormalities with functional defects that will likely be applicable for the determination of retinal areas for potential improvement of retinal function in these patients during future clinical trials and for the monitoring of the diseases' natural history.
doi:10.1097/IAE.0b013e3181f441f6
PMCID: PMC3116073  PMID: 21293320
microperimetry; SLO infrared imaging; Stargardt disease; fundus autofluorescence imaging
24.  Fundus autofluorescence features in the inflammatory maculopathies 
Purpose
To describe the fundus autofluorescence (FAF) features of the inflammatory maculopathies and develop a quantification method for FAF analysis.
Methods
This is a retrospective, consecutive case series of patients with inflammatory maculopathies from two tertiary centers. The clinical findings, demographics, and FAF imaging characteristics were reviewed. Foveal autofluorescence (AF) was analyzed. Median and standard deviation (SD) of foveal AF intensity were measured.
Results
Thirty eyes of 15 patients were evaluated with both qualitative and quantitative FAF analysis. In acute macular neuroretinopathy, the active phase showed foveal hypoautofluorescence, which became hypoautofluorescent with resolution. In acute posterior multifocal placoid pigment epitheliopathy, multiple lesions with hypoautofluorescent centers with hyperautofluorescent borders were observed in active disease and became hypoautofluorescent with disease convalescence. In multifocal choroiditis and punctate inner choroiditis, the active hyperautofluorescent lesions progressed to inactive, hypoautofluorescent scars. Active serpiginous choroiditis showed hyperautofluorescent borders adjacent to a helicoid-shaped, hypoautofluorescent scar. Active unilateral acute idiopathic maculopathy (UAIM) showed a complex pattern of hypo- and hyperautoflourescence in the macula. The median foveal AF was the greatest in acute macular neuroretinopathy and UAIM among the maculopathies, while the greatest SD of foveal AF intensity was observed in UAIM.
Conclusion
The active phase of the majority of inflammatory maculopathies was characterized by hyperautofluorescent lesions. Increased SD of foveal AF correlated with a mixture of hypo-and hyperautoflourescence. Median and SD may be useful metrics in foveal AF and quantifiable values that may be assessed over time as a disease process evolves. Improvements in quantification methods of FAF imaging may allow us to objectively evaluate posterior uveitis.
doi:10.2147/OPTH.S68446
PMCID: PMC4189704  PMID: 25302012
posterior uveitis; foveal autofluorescence; quantification; fundus autofluorescence imaging
25.  Wide-field fundus autofluorescence corresponds to visual fields in chorioretinitis patients 
Background and objectives
Detection of peripheral fundus autofluorescence (FAF) using conventional scanning laser ophthalmoscopes (SLOs) is difficult and requires pupil dilation. Here we evaluated the diagnostic properties of wide-field FAF detected by a two-laser wavelength wide-field SLO in uveitis patients.
Study design/materials and methods
Observational case series of four patients suffering from different types of posterior uveitis/chorioretinitis. Wide-field FAF images were compared to visual fields. Panretinal FAF was detected by a newly developed SLO, which allows FAF imaging of up to 200° of the retina in one scan without the need for pupil dilation. Visual fields were obtained by Goldmann manual perimetry.
Results
Findings from wide-field FAF imaging showed correspondence to visual field defects in all cases.
Conclusion
Wide-field FAF allowed the detection of visual field defect-related alterations of the retinal pigment epithelium in all four uveitis cases.
doi:10.2147/OPTH.S26224
PMCID: PMC3236712  PMID: 22174575
fundus autofluorescence (FAF); Optomap; wide-field scanning laser ophthalmoscopy; imaging; uveitis; visual field

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