Evaluation Of Autofluorescence Research Articles

Panel Design and Optimization for Full Spectrum Flow Cytometry.

Technological advancements in fluorescence flow cytometry and an ever-expanding understanding of the complexity of the immune system have led to the development of large flow cytometry panels, reaching up to 40 markers at the single-cell level. Full spectrum flow cytometry, which measures the full emission range of all the fluorophores present in the panel instead of only the emission peaks, is now routinely used in laboratories around the world, and the demand for this technology is rapidly increasing. With the ability to use larger and more complex staining panels, optimized protocols are vital for achieving the best panel design, panel optimization, and high-dimensional data analysis outcomes. In addition, a better understanding of how to fully characterize the autofluorescence of the sample, coupled with an intelligent panel design approach, allows improved marker resolution on highly autofluorescent tissues or cells. Here, we provide optimized step-by-step protocols for full spectrum flow cytometry, covering panel design and optimization, autofluorescence evaluation and strategy selection, and methods for performing longitudinal studies.

Clinical Evaluation of the Optical Filter for Autofluorescence Glasses for Oral Cancer Curing Light Exposed (GOCCLES®) in the Management of Potentially Premalignant Disorders: A Retrospective Study.

Background: Any oral potentially malignant disorders (OPMDs) must be regularly monitored through clinical examination to detect any possible malignant transformation. Conventional intraoral exams, however, can be difficult because these conditions may resemble benign lesions. For this reason, several non-invasive diagnostic technologies have been developed to help the clinician in detecting and distinguishing between cancerous and benign lesions. Epithelial dysplasia can be considered the most important predictor of malignant evolution. Therefore, in this study we aim to evaluate the ability of an optical filter for autofluorescence Glasses for Oral Cancer Curing Light Exposed (GOCCLES®) and of toluidine blue staining in identifying dysplastic areas in patients with OPMDs. Methods: In this retrospective study, medical records, photographs and videos of 25 patients with oral lesions were analyzed. Forty-two biopsy samples in 25 patients with OPMDs and at least one suspicious oral mucosa lesion that were evaluated in white light, autofluorescence with optical filter GOCCLES®, toluidine blue staining and then biopsied with histopathological analysis were analyzed. Results: The sensitivity and specificity for the autofluorescence evaluation with GOCCLES® for identifying dysplasia or carcinoma were 66% and 48%, respectively. The positive and negative predictive values were 34% and 77%, respectively, and the accuracy was 53%. The sensitivity and specificity for toluidine blue staining were 91% and 68%, respectively. The positive and negative predictive values were 55% and 95%, respectively, and the accuracy was 75%. Conclusions: The optical filter for autofluorescence (GOCCLES®) and toluidine blue staining are simple, inexpensive, rapid and non-invasive procedures that can assist the clinician in distinguishing OPMDs from healthy mucosa but they are not able to distinguish benign and malignant lesions.

Evaluation of Autofluorescence in Identifying Parathyroid Glands by Measuring Parathyroid Hormone in Fine-Needle Biopsy Washings

BackgroundNear-infrared autofluorescence imaging has potentially great value for assisting endocrine surgeons in identifying parathyroid glands and may dramatically change the surgical strategy of endocrine surgeons in thyroid surgery. This study is designed to objectively evaluate the role of near-infrared autofluorescence imaging in identifying parathyroid glands during thyroid surgery by measuring intraoperative parathyroid hormone in fine-needle aspiration biopsy washings.MethodsThis study was conducted at a tertiary referral teaching hospital in China from February 2020 to June 2020. Patients undergoing total thyroidectomy with or without neck lymph node dissection were consecutively included. The surgeon used near-infrared autofluorescence imaging to identify parathyroid glands during thyroid surgery and confirmed suspicious parathyroid tissues by measuring their intraoperative parathyroid hormone. Nanocarbon was injected into the thyroid gland if the thyroid autofluorescence intensity was too strong. The sensitivity and accuracy of near-infrared autofluorescence imaging and vision for identifying parathyroid glands, and the difference in autofluorescence intensity in various tissues were the main outcomes.ResultsOverall, 238 patients completed the trial. Based on the pathological and aIOPTH results, the sensitivity of near-infrared autofluorescence imaging for detecting parathyroid glands (568 of 596 parathyroid glands; 95.30%)was significantly higher than that of vision (517 of 596 parathyroid glands; 86.74%, P<.001). The accuracy of near-infrared autofluorescence imaging (764 of 841 tissues; 90.84%) was significantly higher than that of vision (567 of 841 tissues; 67.42%, P<.001) when the evaluations of certain tissues were inconsistent. There was a significant difference between the autofluorescence intensity of the parathyroid glands and that of the lymph nodes (74.19 ± 17.82 vs 33.97 ± 10.64, P<.001).ConclusionThe use of near-infrared autofluorescence imaging, along with intraoperative parathyroid hormone and nanocarbon for the identification of parathyroid glands in thyroid surgery may increase the number of confirmed parathyroid glands. Using near-infrared autofluorescence imaging can effectively distinguish lymph nodes and parathyroid glands during lymph node dissection.

Intraoperative detection of parathyroid glands by autofluorescence identification using image-based system: report of 15 cases

BackgroundA detection of parathyroid glands by the evaluation of their autofluorescence in the near-infrared spectrum is considered as a promising tool in addition to their visual verification. The aim of this study was to evaluate the role of near-infrared autofluorescence application by using two different image-based systems for the identification of parathyroid glands during surgery of thyroid and parathyroid benign and malignant lesions.MethodsEvaluation of near-infrared autofluorescence was performed in 15 patients by using two different image-based systems equipped with a near-infrared laser camera. Intravenous injection of fluorophore indocyanine green was used for the enhancement of near-infrared autofluorescence signal.ResultsNormal parathyroid glands were identified and mobilized after visual inspection in 12 (80%) patients, which was confirmed by near-infrared autofluorescence evaluation. Confident recognition of parathyroid glands by near-infrared autofluorescence signal and their subsequent distinction from lymph nodes was achieved in two (13%) patients with prior surgery for papillary thyroid carcinoma. In one (7%) case, parathyroid gland was identified as fragments of tissue within the postoperative scarring area by near-infrared autofluorescence evaluation, but not by visual inspection. A less intensive near-infrared autofluorescence signal was detected in the parathyroid gland owing to unintentional excision in one (7%) case. Better signal intensity from parathyroid glands was noticed after changes of the near-infrared camera in Fluobeam 800 image-based system in position to an angle of approximately 45–65° in relation to area of interest in all cases as compared with holding straight on the parathyroid gland. Fluobeam LX demonstrated a good near-infrared autofluorescence signal without any specific changes in the camera angle. Thyroid carcinoma demonstrated low-intensity signal in the case of invasion to thyroid capsule. No fluorescent signal was identified from metastatic, or from normal, lymph nodes.ConclusionsThe application of near-infrared autofluorescence imaging is considered as a useful, but additional, tool for the visual assessment of parathyroid gland in the case of primary neck exploration. The utility of near-infrared autofluorescence imaging for parathyroid detection is increased in the case of repeated surgical intervention owing to increased risk of unintentional parathyroid removal as well as for discrimination of parathyroids from the lymph nodes in cases of thyroid malignancy.

Evaluation of autofluorescence quenching techniques on formalin- fixed chicken tissues

Autofluorescence (AF) in formalin-fixed and paraffin-embedded tissues limit their use in immunofluorescence staining techniques. Various methods have been used to reduce AF in human and animal tissues but no protocol has been optimized for avian tissues. The present study was undertaken to evaluate different treatment methods including ammonium chloride, glycine, Trypan blue, sodium borohydride, Sudan Black B, potassium permanganate, LED light, cupric sulphate combined with glycine, ammonium chloride and cupric sulphate in reducing AF in FFPE chicken tissues for the detection of FITC labelled antibodies against immune cell markers. Chicken tissues including conjunctiva, trachea and Harderian gland presented intense non-homogenous AF in cells resembling erythrocytes, connective cells and melanocytes. Only Sudan Black B effectively reduced AF in FFPE tissues; however, no specific fluorescent signal was observed for six FITC labelled antibodies against immune cell markers. Specific fluorescent signal from the FITC-labelled antibodies was observed in frozen chicken tissue sections with minimal AF, suggesting that the AF in FFPE tissues is related to the use of formaldehyde fixatives. In conclusion, this study demonstrates for the first time that AF quenching methods commonly used for other animal species are not appropriate for use in avian tissues and that frozen tissue sections are recommended for immunofluorescence staining techniques in poultry.

CORRELATION STUDY BETWEEN DRUSEN MORPHOLOGY AND FUNDUS AUTOFLUORESCENCE.

To correlate drusen morphology and outer retinal status with autofluorescence (AF) imaging in patients with intermediate age-related macular degeneration. Drusen type and morphology were analyzed using color fundus photography and spectral-domain optic coherence tomography, whereas fundus AF was used for drusen AF evaluation. Additional structural changes on spectral-domain optic coherence tomography, such as disruption of external limiting membrane, ellipsoid zone, and retinal pigment epithelium/Bruch membrane complex, as well as the presence of choroidal hypertransmission at correspondent locations were also evaluated and correlated with fundus AF findings. Spearman's correlation coefficient was used to analyze the correlation between spectral-domain optic coherence tomography morphological characteristics of drusen and AF appearance of the corresponding drusen. Strength of correlation was calculated (r), and a P value < 0.05 was considered statistically significant. Two hundred and twenty-eight drusen from 53 eyes of 53 patients were analyzed, 130 soft drusen (57.02%) and 98 cuticular drusen (42.98%). Sixty percent of the drusen were isoautofluorescent (n = 136), 35% hyperautofluorescent (n = 80), and 5% hypoautofluorescent (n = 12). We found positive correlation between drusen AF and hyperreflective foci (r = 0.4). Outer retinal layers morphology (external limiting membrane and ellipsoid zone status and hypertransmission) also correlates with autofluorescent findings (r = 0.3). Multimodal imaging reveals a broad spectrum of ultrastructural changes, which may reflect different stages in the evolution of drusen. Our results suggest that drusen morphological characteristics and autofluorescent findings are correlated but other factors or cofactors may be involved. The described correlations will help us understand new progression biomarkers of nonexudative age-related macular degeneration.

Evaluation of autofluorescence and photodynamic diagnosis in assessment of bladder lesions.

The ability to detect and diagnose bladder cancer early and precisely is crucial for effective treatment. The aim of this study is to assess the utility of optical biopsy performed with autofluorescence cystoscopy (AFC) using the Onco-LIFE system with numerical color values (NCVs) and by ALA/PDD. Histopathological examination of material obtained during TURBT and/or biopsy of the bladder was carried out in 251 patients. In the case of 35 patients, the selection of the specimen collected for histopathological examination was based using ALA/PDD. In the remaining 216 patients, tissue was collected based on the findings of AFC with NCV. Using AFC, the observed NCV ranged from 0 to 3.86; the highest mean NCV was observed in neoplastic muscle invasive lesions and was equal to 3.18. Furthermore, non-muscle invasive tumors were characterized by a mean NCV equal to 1.54. Tissue with inflammation, metaplasia, and healthy tissue demonstrated significantly lower mean NCV values. The presence of a muscle-invasive tumor increased the NCV by approximately 2.86 compared to healthy tissue. The rates of postoperative complications depend on the examining operator and are observed more often, as much as 65.7 % during ALA/PDD. AFC with NCV using the Onco-LIFE system, as well as ALA/PDD are helpful tools for early diagnosis of bladder precancerous and cancer lesions and for performing targeted biopsies. A significant correlation was found between lesion NCV index and the grade of dysplasia or tumor malignancy. Tissue with inflammation, metaplasia, and healthy tissue demonstrated significantly lower mean NCV values. AFE with NCV have a significantly higher sensitivity than specificity. Low rates of postoperative complications are correlated to the experience of the endoscopist and with AFE/NCV in comparison of ALA/PDD.

Eight-year multimodal follow-up of recurrent idiopathic acute exudative polymorphous vitelliform maculopathy

To describe and analyze clinical findings in a patient with recurrent idiopathic acute exudative polymorphous vitelliform maculopathy (AEPVM), followed in detail, and to propose the diagnostic and follow-up algorithm. Retrospective observational analysis. A young adult male patient diagnosed with idiopathic AEPVM who developed two relapses in a 12-month period eight years after the initial onset. Review of clinical charts, multimodal imaging, and electrophysiology findings. The patient repeatedly underwent complete ophthalmic examinations, including best-corrected visual acuity testing (BCVA), slit-lamp and fundus examinations; digital fundus photography, time-domain optical coherence tomography (OCT) in 2009 (Stratus OCT, Carl Zeiss Meditec, USA) and spectral-domain OCT in 2017-2018 (Spectralis-OCT, Heidelberg Engineering, Germany), together with fundus autofluorescence (FAF), fluorescein angiography (FA), and indocyanine green angiography (ICGA), all with HRA2 (Heidelberg Engineering, Germany); microperimetry (MP-1 microperimeter, Nidek, Japan). Laser flare photometry (Kowa FM-600, Japan) and electrophysiology testing were also performed. Clinical features of long-lasting recurrent idiopathic AEPVM, and diagnostic and follow-up algorithm in such rare cases. Case report of a 25-year-old male Caucasian patient with typical features of AEPVM, including serous neuroepithelial detachment with irregular retinal elevations, ophthalmoscopically resembling retinal folds, with subsequent subretinal accumulation of characteristic yellow-white vitelliform deposits. Features in this case rarely described, or even not yet reported, include indocyanine- and fluorescein-negative intraretinal cystic changes, optic disc hyperfluorescence on FA, serous retinal elevations mimicking retinal folds, increased choroidal thickness, lack of rapid visual recovery, and very slow anatomical improvement of the relapses. Bimonthly fundus autofluorescence evaluation together with SD-OCT were the most informative diagnostic methods, demonstrating the evolution of pathological signs. AEPVM may be a recurrent or even chronic condition with uncertain long-term visual outcomes. It may have variable clinical presentations depending on the stage of the disease, and both clinical manifestations and imaging features of different stages of the pathologic process may overlap. Patients should be made aware that visual improvement occurs very slowly, if at all. Bimonthly fundus autofluorescence evaluation together with SD-OCT should be recommended in such cases.

Can fluorescence and autofluorescence imaging be useful in diagnosis of basal cell cancer? Proposition of algorithms.

The most common non-melanoma skin carcinomas (NMSC) are basal cell carcinomas (BCC). Autofluorescence study may allow a non-invasive distinction of neoplastic lesions from benign lesions, whereas fluorescence spectroscopy enables qualification of the patient for possible photodynamic or surgical treatment. The aim of the study is to assess the suitability of autofluorescence and fluorescence imaging in the diagnosis of the stage of advancement, and the morphological type of changes in basal cell carcinoma of the skin. A group of 382 patients with 430 skin lesions of basal cell carcinoma were subjected to an autofluorescence study and a spectral imaging using a spectral camera. The acquired spectra and fluorescence images were analyzed using the Image Pro-Plus 5.0.2 software. An analysis of fluorescence intensity profiles in the long and short axis of the studied changes was carried out. The fluorescence emission observed in BCC lesions with an area of ​​up to 3 cm2 showed a statistically significantly higher level of intensity in comparison to changes in lesions exceeding 3 cm2 (#p < 0.05, ## p < 0.01). In the autofluorescence evaluation, we observed significant differences in the mean values of the Numerical Colour Value (NCV) depending on the size of the area of skin occupied by skin lesions. Based on the spatial and profile assessment of the types of BCC lesions a proposed algorithm for identifying changes was developed. Analysis of green fluorescence intensity profiles, analogous to fluorescence profiles analysis, allowed to propose an algorithm of autofluorescence evaluation in the diagnosis of BCC. Autofluorescence and fluorescence imaging allow assessment of the extent of neoplastic infiltration and distinguish types of skin lesions with BCC. The determined diagnostic algorithms may be an effective diagnostic solution in the diagnosis of skin lesions of this nature.

Anatomical and functional changes in neovascular AMD in remission: comparison of fibrocellular and fibrovascular phenotypes

PurposeTo investigate the anatomical changes and the macular function in neovascular age-related macular degeneration (AMD) eyes, according to the recognition of either fibrocellular or fibrovascular phenotype.MethodsWe enrolled eyes with previously...

Influence of darkness on pigments of Tetraselmis indica (Chlorodendrophyceae, Chlorophyta)

In the photic zone, phytoplankton experience diurnal variation in light intensity. However, prolonged exposure to aphotic condition influences their physiological state. Pigment composition is a useful biomarker to decipher cells physiological state and adaptive response to changing environmental conditions. Chlorophyll, a natural pigment, is biosynthesised even in darkness and studies have shown this ability is determined by genetic characteristics of an organism. The purpose of this study was to examine the influence of darkness on pigments and chlorophyll autofluorescence of Tetraselmis indica. Dark exposure (up to 6 months) had no significant impact on chlorophyll a and b concentration, whereas carotenoids were enhanced. Upon re-illumination pigments gradually recovered to pre-dark phase condition. These adaptive survival strategies of T. indica by altering pigment concentration in response to prolonged darkness are interesting. The absence of loroxanthin and loroxanthin esters in T. indica is reported in a first Tetraselmis species so far. In addition, the evaluation of autofluorescence and cellular chlorophyll concentration pointed out that they are not interdependent in this species. Hence, careful consideration of these two factors is needed when either of them is used as a proxy for other. The results obtained encourage a thorough study of pigment analysis, especially when subjected to darkness, to elucidate potential role in the evolution, chemotaxonomy, and survivability of species.

A new model of fundus autofluorescence time evolution in multiple evanescent white dot syndrome

PurposeFundus Autofluorescence proved to be a valuable non‐invasive tool for Multiple Evanescent White Dot Syndrome evaluation. Until now several models describing time evolution of the Fundus Autofluorescence in MEWDS have been proposed.The aim of our study is to present a new model of the Fundus Autofluorescence evaluation based on four examinations in a 56‐year‐old patient during a 3 months follow‐up period.MethodsFundus Autofluorescence images were captured by HRA Spectralis Heidelberg Engineering Imaging System. The ImageJ image analysis software was used for image denoising by median filtering. We have applied Otsu's multilevel tresholding method for image segmentation to distinguish and differentiate autofluorescent areas from normal fundus.ResultsWe have observed two levels of fluorescence intensity in the analyzed images. Bright fluorescence areas changed significantly with recovery time. Overall fluorescence areas changed to a lesser extent ‐ the areas were shrinking centripetally with functional improvement.ConclusionsCombination of Fundus Autofluorescence with image analysis methods brings us quantitative data, facilitating the trend analysis.

Optical Coherence Tomography and Fundus Autofluorescence evaluation in an animal model of Retinal Degeneration

PurposeTo evaluate fundus autofluorescence (FAF) and OCT changes comparing with immunohistochemical (ICC) analysis in long term degeneration of P23H rat and to investigate retinal and choroidal vascularization using fluorescein and indocianin green angiography.MethodsTwenty‐albino homozygous P23H line 1 rats aging from 18 postnatal days (P18) to 27 months and wild‐type albino Sprague‐Dawley (SD rats) (2 and 15 months old) were used for this study. Normal pigmented Long Evans (LE) 2 months old rats were used to compare FAF findings. ICC was performed to correlate with the findings of OCT and AF changes.ResultsFAF pattern varied from not findings at P18 to a mosaic of hyperfluorescent dots in the rats of 6 months or older. Retinal thicknesses diminished during the time: 205.2–183.18 μm in SD rats and 189.88–58.15 μm in P23H rats. In long term degeneration, OCT showed severe changes at the retinal layers; ICC helped to identify the cell loss and remodeling changes.ConclusionsAutofluorescent ophthalmoscopy is a non‐invasive method that can detect changes in metabolic activity at the RPE in animal models of retinal degeneration in vivo. Retinal vascular plexus changed with aging. OCT showed a diminution of retinal thickness and retinal layer changes with the degeneration. ICC shows a good correlation.

Autofluorescence imaging in recurrent oral squamous cell carcinoma.

The survival of patients with oral cancer is decreased by locoregional recurrence after an initial multimodal treatment. In order to identify lesions in the oral cavity for a possible recurrence, clinical evaluation as well as MRI or CT scanning is advised. The evaluation of mucosa lesions is hampered by changes related to radio- and chemotherapy as well as reconstruction with tissue flaps. Several techniques for easier identification of tissue abnormalities in the oral cavity have been advocated as adjuncts in order to facilitate identification. Especially methods using altered tissue fluorescence have gained much interest during the last decade. The aim of our prospective study was to evaluate fluorescence properties of undiagnosed mucosa lesions with the VELscope device in patients with multimodal treated oral cancer prior to histological confirmation. In total, 41 patients with a history of oral squamous cell carcinomas (OSCC) (19 females and 22 males) with undiagnosed mucosa lesions where included in the study. After clinical evaluation, examination and documentation using the VELscope® device were performed. Then, an incisional biopsy was performed. An autofluorescence loss indicating a malignant or dysplastic mucosa condition could be detected in six patients (14.6%); however, only one OSCC and one SIN revealed a complete autofluorescence loss. In four patients, OSCC was present in lesions with retained autofluorescence. Sensitivity and specificity for the VELscope® examination to identify malignant oral lesions by autofluorescence were 33.3 and 88.6%, respectively. The positive and negative predictive values were 33.3 and 88.6%, respectively. No statistical correlation between gender and lesion appearance versus autofluorescence loss could be detected. In contrast to mucosa lesions in patients with no prior treatment, the autofluorescence evaluation with the VELscope reveals no additional information in our analysis. Accordingly, invasive biopsies as gold standard are still needed to get sufficient evidence regarding potential malignancy in patients after multimodal treatment for oral cancer.

Evaluation of autofluorescence and toluidine blue in the differentiation of oral dysplastic and neoplastic lesions from non dysplastic and neoplastic lesions: a cross-sectional study.

The objective was to compare toluidine blue (TB) and autofluorescence (AF) for the detection of oral dysplasia and squamous cell carcinoma (OSCC) in clinically suspicious lesions according to conventional examination. Fifty-six clinically suspicious lesions were subjected to AF and TB examination. Data were compared using two different scenarios: in the first, mild dysplasia was considered as positive, while in the second, it was considered as negative. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV), accuracy, and concordance were calculated. AF sensitivity and specificity were 70.0 and 57.7%, respectively, while TB showed a sensitivity of 80% and a specificity of 61.5%. The sensitivity increased in the second scenario in both AF (76.5%) and TB (88.2%). The specificity decreased in AF and TB, showing the same value (51.3%). PPV was higher in TB than in AF (70.6 versus 65.6%) and similarly for NPV (72.7 versus 62.5%). In the second scenario, TB PPV was 44.1% and NPV was 90.9%; AF PPV was 40.6% and NPV was 83.3%. TB showed greater accuracy than AF in the first scenario (62.5 versus 58.9%). AF and TB are both sensitive but not specific in OSCC and dysplasia diagnosis.

Autofluorescence Spectroscopy Augmented by Multivariate Analysis as a Potential Noninvasive Tool for Early Diagnosis of Oral Cavity Disorders

Oral leukoplakia is one of the common potentially malignant lesions encountered worldwide. We report the results of an in vivo clinical evaluation of autofluorescence (AF) spectroscopy for differential diagnosis of oral leukoplakia. Multivariate analysis of spectral data has been incorporated to improve the efficacy of the technique. The results of this noninvasive study are expected to provide potential for extending the technique to other disorders. A total of 18 patients and 30 normal volunteers participated in this study. AF spectra were acquired from affected sites of patients and from right and left buccal mucosa of normal volunteers. Diagnostic performance was analyzed using spectral intensity ratio (SIR), and principal component analysis followed by linear discriminant analysis (PCA-LDA). AF spectra of leukoplakic patients showed characteristic emissions from flavin adenine dinucleotide (FAD) and porphyrin at 500 and 630 nm, respectively. But the emission from porphyrin is not very prominent in the case of healthy volunteers. Also, significant decrease in spectral intensity is observed for leukoplakia compared with normal volunteers in the unprocessed spectra. Method of SIR yielded 96% sensitivity and 100% specificity and an overall 100% for PCA-LDA respectively for efficient differentiation of the lesions. The result of this preliminary study shows that PCA-LDA or SIR applied to AF spectroscopy is a useful tool for the differential diagnosis of oral cavity disorders. This has been demonstrated in leukoplakia in a clinical setting, and it is expected that the technique can be extended to other oral cavity disorders as well.

Foveal Duplication Artifact with Spectral-Domain Optical Coherence Tomography

The authors describe a rare artifact on two different spectral-domain optical coherence tomography (OCT) systems and explain its origin in patient- rather than hardware-related errors. During evaluation on spectral-domain OCT, four patients registered twin foveal depressions on the retinal thickness map display and were initially unable to fixate on the given target. Repeat scans on the same OCT system without changing scan protocol demonstrated no artifact after the patients began to fixate well, except in one patient who had nystagmus. Fundus and autofluorescence evaluation revealed no true duplication of fovea. Performing a repeat scan with greater focus on patient compliance is necessary when such artifactual anomalies are encountered.

Indocyanine green angiography in chronic central serous chorioretinopathy

SummaryBackgroundCentral serous chorioretinopathy (CSC) is a condition that originates from alterations of the choroidal circulation. The aim of this paper was to evaluate the use of indocyanine green angiography (ICGA) in patients with chronic CSC.Material/MethodsThe analysis included 17 patients (34 eyes) with chronic CSC in at least 1 eye. The eye examination included: distance and near visual acuity, biomicroscopy, applanation tonometry, fundus examination, colored and red-free fundus photography, evaluation of autofluorescence, optical coherence tomography, and fluorescein and indocyanine green angiography.ResultsIn 34 eyes (100%) involved in the ICGA study the results revealed zones of transient increased choroidal vessels permeability. In 18 eyes (52.9%) choroidal changes were accompanied by a focal serous pigment epithelial detachment. In 4 eyes (11.8%) of 3 patients’ the ICGA examination confirmed the presence of occult choroidal neovascularization (CNV). In the patient with bilateral diffuse retinal pigment epitheliopathy, CNV was present in 1 eye, in the patient with unilateral chronic CSC it was also present in 1 eye, and in the third patient with bilateral chronic CSC it was detected in both eyes.ConclusionsICGA is a very useful examination that enables ophthalmologists to visualize choroidal changes due to chronic CSC, as well as to diagnose occult CNV in chronic CSC.

Novel KCNV2 Mutations in Cone Dystrophy with Supernormal Rod Electroretinogram

To describe patients with cone dystrophy and supernormal rod electroretinogram (ERG) and search for mutations in the recently described KCNV2 gene. Clinical and molecular study. Patients from three families originating from France, Morocco, and Algeria had standard ophthalmologic examination and color vision analysis, Goldmann perimetry, International Society for Clinical Electrophysiology of Vision (ISCEV) protocol in accordance with ERG testing, autofluorescence evaluation, and optical coherence tomography 3 scanning. The two coding exons of KCNV2 were polymerase chain reaction amplified and sequenced. All patients had the characteristic features of supernormal, delayed rod ERG responses at the highest levels of stimulation and markedly reduced cone responses. In the French family, two affected sisters were compound heterozygotes for the recurrent c.1381G>A (Gly461Arg) mutation and for a novel c.442G>T (Glu148Stop) mutation. In the Moroccan family, affected members were homozygotes for the novel c.1404delC mutation (His468fsX503) and in the Algerian family, the proband was homozygote for the novel c.1001delC mutation (Ala334fsX453). In the three families, parents were unaffected heterozygote carriers. None of the mutations were present in 50 control chromosomes. The three novel truncative mutations are likely to be null mutations leading to loss of function, with no difference in the phenotype presentation. Amino acid changes are found exclusively in the N-terminal fragment of the protein and in the P-loop, indicating the importance of those regions for the function of the KCNV2 protein.

DAFE autofluorescence assessment of oral cavity, larynx and bronchus in head and neck cancer patients

Head and neck (H&N) cancer patients have a high incidence of local field change as well as second primary lung tumours. We have applied the Wolf Diagnostic Autofluorescence Endoscopy (DAFE) system in a novel way, combining autofluorescence evaluation of both H&N region and bronchial. New H&N cases as well as "old" cases with symptoms were included. The DAFE procedure was done separate to panendoscopy. The H&N region was examined first; images were recorded of the known primary with reference to subsequent resection margins, as well as of adjacent mucosa inspecting for additional abnormal sites. Then autofluorescence bronchoscopy was performed. Changes in management were only on the basis of histology taken because of abnormal autofluorescence. One hundred and seven cases were referred, including 96 new cases and 11 old cases. Autofluorescence examination of H&N detected sites which led to change of management in 11 patients. This included additional sites in nine patients (which then either had extra surgery or radiotherapy as a result) and wider resection margins were made possible in two patients. In the bronchus there were 21 significant lesions in 16 patients. Immediate management change occurred in one invasive cancer, two microinvasive carcinomas and three carcinomas in situ (in four patients). There were 15 sites of severe and moderate dysplasia in 12 patients. None of these bronchoscopic lesions were detectable on CT chest. Therefore, overall an immediate change in management occurred in 15 of 107 patients (14% of patients). This combined procedure yielded a significant number of lesions particularly as a simple addition to preoperative work-up in new H&N cancer cases.