Near-infrared Autofluorescence Research Articles

Simultaneous Expression of ABCA4 and GPR143 Mutations: A Complex Phenotypic Manifestation

PurposeTo describe the complex, overlapping phenotype expressed in a two generation family harboring pathogenic mutations in the ABCA4 and GPR143 genes.MethodsClinical evaluation of a two generation family included quantitative autofluorescence imaging (qAF, 488-nm excitation) using a modified confocal scanning laser ophthalmoscope equipped with an internal fluorescent reference to account for varying laser power detector sensitivity, spectral-domain optical coherence tomography, and full-field ERG testing. Complete sequencing of the ABCA4 and GPR143 genes was carried out in each individual.ResultsAffected individuals presented with bull's eye lesions and qAF levels above the 95% confidence interval for healthy eyes; full-field ERG revealed no generalized rod dysfunction but mild implicit time delays in cone responses. Complete sequencing of the ABCA4 gene revealed two disease-causing mutations, p.L541P and p.G1961E; and mutational phase was confirmed in each unaffected parent. Further examination in the affected patients revealed a peripheral “mud-splattered” pattern of hypopigmented RPE after which sequencing of GPR143 revealed a novel missense variant, p.Y157C. The GPR143 variant segregated from the father who did not exhibit any indications of retinal disease with the exception of an abnormal near-infrared autofluorescence (NIR-AF) signal distribution in the macula.ConclusionsAn individual carrying both ABCA4 and GPR143 disease-causing mutations can express a complex, overlapping phenotype associated with both Stargardt disease and X-linked ocular albinism (OA1). The absence of OA1-related disease changes (with the exception of NIR-AF changes associated with melanin distribution) in the father may be indicative of mild expressivity or variable gene penetrance.

Intraoperative Near-infrared Imaging for Parathyroid Gland Identification by Auto-fluorescence: A Feasibility Study.

Parathyroid glands (PGs) can be particularly hard to distinguish from surrounding tissue and thus can be damaged or removed during thyroidectomy. Postoperative hypoparathyroidism is the most common complication after thyroidectomy. Very recently, it has been found that the parathyroid tissue shows near-infrared (NIR) auto-fluorescence which could be used for intraoperative detection, without any use of contrast agents. The work described here presents a histological validation ex vivo of the NIR imaging procedure and evaluates intraoperative PG detection by NIR auto-fluorescence using for the first time to our knowledge a commercially available clinical NIR imaging device. Ex vivo study on resected operative specimens combined with a prospective in vivo study of consecutive patients who underwent total or partial thyroid, or parathyroid surgery at a comprehensive cancer center. During surgery, any tissue suspected to be a potential PG by the surgeon was imaged with the Fluobeam 800 (®) system. NIR imaging was compared to conventional histology (ex vivo) and/or visual identification by the surgeon (in vivo). We have validated NIR auto-fluorescence with an ex vivo study including 28 specimens. Sensitivity and specificity were 94.1 and 80%, respectively. Intraoperative NIR imaging was performed in 35 patients and 81 parathyroids were identified. In 80/81 cases, the fluorescence signal was subjectively obvious on real-time visualization. We determined that PG fluorescence is 2.93±1.59 times greater than thyroid fluorescence in vivo. Real-time NIR imaging based on parathyroid auto-fluorescence is fast, safe, and non-invasive and shows very encouraging results, for intraoperative parathyroid identification.

Clinical Characterization of Coronary Atherosclerosis With Dual-Modality OCT and Near-Infrared Autofluorescence Imaging

ObjectivesThe authors present the clinical imaging of human coronary arteries in vivo using a multimodality optical coherence tomography (OCT) and near-infrared autofluorescence (NIRAF) intravascular imaging system and catheter. BackgroundAlthough intravascular OCT is capable of providing microstructural images of coronary atherosclerotic lesions, it is limited in its capability to ascertain the compositional/molecular features of plaque. A recent study in cadaver coronary plaque showed that endogenous NIRAF is elevated in necrotic core lesions. The combination of these 2 technologies in 1 device may therefore provide synergistic data to aid in the diagnosis of coronary pathology in vivo. MethodsWe developed a dual-modality intravascular imaging system and 2.6-F catheter that can simultaneously acquire OCT and NIRAF data from the same location on the artery wall. This technology was used to obtain volumetric OCT-NIRAF images from 12 patients with coronary artery disease undergoing percutaneous coronary intervention. Images were acquired during a brief, nonocclusive 3- to 4-ml/s contrast purge at a speed of 100 frames/s and a pullback rate of 20 or 40 mm/s. OCT-NIRAF data were analyzed to determine the distribution of the NIRAF signal with respect to OCT-delineated plaque morphological features. ResultsHigh-quality intracoronary OCT and NIRAF image data (>50-mm pullback length) were successfully acquired without complication in all patients (17 coronary arteries). The maximum NIRAF signal intensity of each plaque was compared with OCT-defined type, showing a statistically significant difference between plaque types (1-way analysis of variance, p < 0.0001). Interestingly, coronary arterial NIRAF intensity was elevated only focally in plaques with a high-risk morphological phenotype (p < 0.05), including OCT fibroatheroma, plaque rupture, and fibroatheroma associated with in-stent restenosis. ConclusionsThis OCT-NIRAF study demonstrates that dual-modality microstructural and fluorescence intracoronary imaging can be safely and effectively conducted in human patients. Our findings show that NIRAF is associated with a high-risk morphological plaque phenotype. The focal distribution of NIRAF in these lesions furthermore suggests that this endogenous imaging biomarker may provide complementary information to that obtained by structural imaging alone.

Iris autofluorescence in Fuchs' heterochromic uveitis.

To explore the characteristic autofluorescence patterns of iris depigmentation in eyes diagnosed with Fuchs' heterochromic uveitis (FHU). Near-infrared autofluorescence images and colour images of iris were taken in 21 eyes of 21 patients with FHU, 30 eyes of 15 normal subjects, 30 eyes of 15 normal age-related iris atrophy and 33 eyes of 20 patients with uveitis other than FHU. The confocal scanning laser ophthalmoscope (Heidelberg Retina Angiograph 2, HRA2) was used for melanin-related autofluorescence imaging. The indocyanine green angiography mode of HRA2 was applied for near-infrared laser imaging, and the wavelength of the excitation laser was 795 nm. Iris colour images were also taken with the slit lamp. In normal iris, moderately intense autofluorescence was noted for the pigment ruff at the pupillary border, the crests in the pupillary zone and the collarette; and there was mild autofluorescence in the ciliary zone. In eyes with age-related iris atrophy and uveitis, much less autofluorescence was seen than the healthy normal irides. In eyes with FHU, there was moderate but discontinuous autofluorescence in the pigment ruff, a petaloid pattern of autofluorescence in the pupillary zone, moderate autofluorescence in the collarette and reticular pattern of autofluorescence in the ciliary zone. Characteristic autofluorescence patterns appeared in eyes diagnosed with FHU. Near-infrared autofluorescence is a promising objective technique to document the iris changes in FHU.

Analysing the Progression Rates of Macular Lesions with Autofluorescence Imaging Modes in Dry Age-Related Macular Degeneration.

Objectives:In this study we aimed to compare the sensitivity of blue-light fundus autofluorescence (FAF) and near-infrared autofluorescence (NI-AF) imaging for determining the progression rates of macular lesions in dry age-related macular degeneration (AMD).Materials and Methods:The study was designed retrospectively and included patients diagnosed with intermediate and advanced stage dry AMD. Best corrected visual acuities and FAF and NI-AF images were recorded in 46 eyes of 33 patients. Lesion borders were drawn manually on the images using Heidelberg Eye Explorer software and lesion areas were calculated using Microsoft Excel software. BCVA and lesion areas were compared with each other.Results:Patients’ mean follow-up time was 30.98±13.30 months. The lesion area progression rates were 0.85±0.93 mm2/y in FAF and 0.93±1.01 mm2/y in NI-AF, showing statistically significant correlation with each other (r=0.883; p<0.01). Both imaging methods are moderately correlated with visual acuity impairment (r=0.362; p<0.05 and r=0.311; p<0.05, respectively). In addition, larger lesions showed higher progression rates than smaller ones in both imaging methods.Conclusion:NI-AF imaging is as important and effective as FAF imaging for follow-up of dry AMD patients.

Near-infrared autofluorescence spectroscopy of in vivo soft tissue sarcomas

Soft tissue sarcomas (STS) are a rare and heterogeneous group of malignant tumors that are often treated via surgical resection. Inadequate resection can lead to local recurrence and decreased survival rates. In this study, we investigate the hypothesis that near-infrared (NIR) autofluorescence can be utilized for tumor margin analysis by differentiating STS from the surrounding normal tissue. Intraoperative in vivo measurements were acquired from 30 patients undergoing STS resection and were characterized to differentiate between normal tissue and STS. Overall, normal muscle and fat were observed to have the highest and lowest autofluorescence intensities, respectively, with STS falling in between. With the exclusion of well-differentiated liposarcomas, the algorithm's accuracy for classifying muscle, fat, and STS was 93%, 92%, and 88%, respectively. These findings suggest that NIR autofluorescence spectroscopy has potential as a rapid and nondestructive surgical guidance tool that can inform surgeons of suspicious margins in need of immediate re-excision.

Lessons talk by imaging about atrophy in retinal disease: Fundus autofluorescence

SummaryFundus autofluorescence (AF) and near‐infrared autofluorescence (NIA) have been used for many years to assess clinically, in a non‐invasive manner, the status of the retinal pigment epithelium (RPE). Their value in the evaluation of patients with posterior segment disorders has expanded; both are currently used in ophthalmic clinics throughout the world for the assessment of patients with degenerative, inflammatory and neoplastic disorders, among others. This talk will provide an overview on how AF and NIA have contributed to the characterisation of atrophy in retinal diseases, from lessons learnt using these imaging technologies on the understanding of mechanisms underlying this pathology, to identification of people at risk of visual loss as well as providing enhanced disease phenotyping and assessment of potential, unwanted, treatment effects. Recent advances, such as wide angle autofluorescence imaging, will likely contribute furthering the understanding of atrophy in retinal disease.

Predicting Progression of ABCA4-Associated Retinal Degenerations Based on Longitudinal Measurements of the Leading Disease Front.

To evaluate the progression of the earliest stage of disease in ABCA4-associated retinal degenerations (RDs). Near-infrared excited reduced-illuminance autofluorescence imaging was acquired across the retina up to 80 degrees eccentricity in 44 patients with two ABCA4 alleles. The eccentricity of the leading disease front (LDF) corresponding to the earliest stage of disease was measured along the four meridians. A mathematical model describing the expansion of the LDF was developed based on 6 years of longitudinal follow-up. The extent of LDF along the superior, inferior, and temporal meridians showed a wide spectrum from 3.5 to 70 degrees. In patients with longitudinal data, the average centrifugal expansion rate was 2 degrees per year. The nasal extent of LDF between the fovea and ONH ranged from 4.3 to 16.5 degrees and expanded at 0.35 degrees per year. The extent of LDF beyond ONH ranged from 19 to 75 degrees and expanded on average at 2 degrees per year. A mathematical model fit well to the longitudinal data describing the expansion of the LDF. The eccentricity of the LDF in ABCA4-RD shows a continuum from parafovea to far periphery along all four meridians consistent with a wide spectrum of severity observed clinically. The model of progression may provide a quantitative prediction of the LDF expansion based on the age and eccentricity of the LDF at a baseline visit, and thus contribute significantly to the enrollment of candidates appropriate for clinical trials planning specific interventions, efficacy outcomes, and durations.

Flecks in Recessive Stargardt Disease: Short-Wavelength Autofluorescence, Near-Infrared Autofluorescence, and Optical Coherence Tomography.

We evaluated the incongruous observation whereby flecks in recessive Stargardt disease (STGD1) can exhibit increased short-wavelength autofluorescence (SW-AF) that originates from retinal pigment epithelium (RPE) lipofuscin, while near-infrared AF (NIR-AF), emitted primarily from RPE melanin, is usually reduced or absent at fleck positions. Flecks in SW- and NIR-AF images and spectral-domain optical coherence tomography (SD-OCT) scans were studied in 19 STGD1 patients carrying disease-causing ABCA4 mutations. Fleck spatial distribution and progression were recorded in serial AF images. Flecks observed in SW-AF images typically colocalized with darkened foci in NIR-AF images; the NIR-AF profiles were larger. The decreased NIR-AF signal from flecks preceded apparent changes in SW-AF. Spatiotemporal changes in fleck distribution usually progressed centrifugally, but in one case centripetal expansion was observed. Flecks in SW-AF images corresponded to hyperreflective deposits that progressively traversed photoreceptor-attributable bands in SD-OCT images. Outer nuclear layer (ONL) thickness negatively correlated with expansion of flecks from outer to inner retina. In the healthy retina, RPE lipofuscin fluorophores form in photoreceptor cells but are transferred to RPE; thus the SW-AF signal from photoreceptor cells is negligible. In STGD1, NIR-AF imaging reveals that flecks are predominantly hypofluorescent and larger and that NIR-AF darkening occurs prior to heightened SW-AF signal. These observations indicate that RPE cells associated with flecks in STGD1 are considerably changed or lost. Spectral-domain OCT findings are indicative of ongoing photoreceptor cell degeneration. The bright SW-AF signal of flecks likely originates from augmented lipofuscin formation in degenerating photoreceptor cells impaired by the failure of RPE.

Non-Invasive In Vivo Imaging and Quantification of Tumor Growth and Metastasis in Rats Using Cells Expressing Far-Red Fluorescence Protein.

Non-invasive in vivo imaging is emerging as an important tool for basic and preclinical research. Near-infrared (NIR) fluorescence dyes and probes have been used for non-invasive optical imaging since in the NIR region absorption and auto fluorescence by body tissue is low, thus permitting for greater penetration depths and high signal to noise ratio. Currently, cell tracking systems rely on labeling cells prior to injection or administering probes targeting the cell population of choice right before imaging. These approaches do not enable imaging of tumor growth, as the cell label is diluted during cell division. In this study we have developed cell lines stably expressing the far-red fluorescence protein E2-Crimson, thus enabling continuous detection and quantification of tumor growth. In a xenograft rat model, we show that E2-Crimson expressing cells can be detected over a 5 week period using optical imaging. Fluorescence intensities correlated with tumor volume and weight and allowed for a reliable and robust quantification of the entire tumor compartment. Using a novel injection regime, the seeding of MDA-MB-231 breast cancer cells in the lungs in a rat model was established and verified.

Reticular Pseudodrusen in Sorsby Fundus Dystrophy

To investigate the association of reticular pseudodrusen (RPD) with Sorsby fundus dystrophy (SFD). Prospective, monocenter, cross-sectional case series. Sixteen patients of 4 unrelated families with SFD caused by mutations in TIMP3. All subjects underwent multimodal imaging including near-infrared (NIR) reflectance and fundus autofluorescence with a confocal scanning laser ophthalmoscope and spectral-domain optical coherence tomography (SD OCT). Prevalence, topographic distribution, and phenotype of RPD. Mean age of the investigated patients was 56.8 years (range, 23-78 years). Reticular pseudodrusen were identified frequently in SFD patients in the sixth decade of life (5 of 7 [71%]) and were absent in younger (n = 3) or older (n = 6) patients. They were most abundant in the superior quadrant and spared the foveal region. Reticular pseudodrusen appeared as yellowish round to oval (dot subtype; n = 5) or confluent, wriggled (ribbon subtype; n = 3) lesions, sometimes forming irregular networks. Reticular pseudodrusen were hyporeflective on NIR reflectance and hypofluorescent on fundus autofluorescence imaging. They appeared as subretinal deposits on SD OCT imaging. Other lesions, such as peripheral pseudodrusen and soft drusen, were present less frequently. Reticular pseudodrusen are a frequent finding in patients with SFD. Although SFD patients with RPD are younger, distribution and phenotype of RPD are similar to those observed in patients with age-related macular degeneration. The association of RPD with SFD implicates a role of Bruch's membrane, the Bruch's membrane-retinal pigment epithelium interface, or both in the pathogenesis of RPD.

Observation on imaging features of two types of fundus autofluorescence and spectral-domain optical coherence tomography in acute central serous chorioretinopathy

Objective To observe the imaging features of short-wave length fundus autofluorescence (SW-AF), near-infrared fundus autofluorescence (NIR-AF)and spectral-domain optical coherence tomograph (SD-OCT) in acute central serous chorioretinopathy (CSC). Methods This is a retrospective observational consecutive case series study. Eighty-two eyes with acute CSC from 76 patients were enrolled in this study.There were 50 males (56 eyes) and 26 females (26 eyes), average age was (41.32±7.37) years old, average course of the disease was (0.47±1.73) months. The routine clinical examinations included best corrected visual acuity, slit lamp microscope, indirect ophthalmoscope, fundus fluorescein angiography (FFA), SW-AF, NIR-AF and SD-OCT. The imaging features of NIR-AF, SW-AF, SD-OCT and FFA in same eye with acute CSC were analyzed comparatively. Results SD-OCT showed retinal pigment epithelium (RPE) detachment corresponding to RPE leakage point on FFA images in all CSC eyes. RPE leakage points in all eyes corresponding to local hypo-NIR-AF, whereas corresponding to normal SW-AF. NIR-AF demonstrated the area of decreased AF corresponding to serous retinal detachment in all eyes, of which, the area of hypo-SW-AF was consistent with that of sensory retinal detachment in 75 eyes. Except for RPE leaking fluorescein in 82 eyes, FFA showed window defects corresponding to clusters of hypo-NIR-AF in 45 eyes, of which, 27 eyes showed hypo-SW-AF appearing concurrently with the locations of window defects of FFA. In 18 eyes, the extent and amount of hypo-SW-AF were less than that of hypo-NIR-AF. Conclusions There was local RPE detachment in all eyes with acute CSC. The locations of sensory retinal detachment, local RPE detachment and RPE depigmentation showed hypo-NIR-AF. The locations of sensory retinal detachment and RPE depigmentation showed hypo-SW-AF in most of acute CSC eyes. Key words: Central serous chorioretinopathy; Tomography, optical coherence; Fluorescein angiography; Diagnostic imaging

Near-infrared autofluorescence: its relationship to short-wavelength autofluorescence and optical coherence tomography in recessive stargardt disease.

We compared hypoautofluorescent (hypoAF) areas detected with near-infrared (NIR-AF) and short-wavelength autofluorescence (SW-AF) in patients with recessive Stargardt disease (STGD1) to retinal structure using spectral domain optical coherence tomography (SD-OCT). The SD-OCT volume scans, and SW-AF and NIR-AF images were obtained from 15 eyes of 15 patients with STGD1 and registered to each other. Thickness maps of the total retina, receptor-plus layer (R+, from distal border of the RPE to outer plexiform/inner nuclear layer boundary), and outer segment-plus layer (OS+, from distal border of the RPE to ellipsoid zone [EZ] band) were created from SD-OCT scans. These were compared qualitatively and quantitatively to the hypoAF areas in SW-AF and NIR-AF images. All eyes showed a hypoAF area in the central macula and loss of the EZ band in SD-OCT scans. The hypoAF area was larger in NIR than SW-AF images and it exceeded the area of EZ band loss for 12 eyes. The thickness maps showed progressive thinning towards the central macula, with the OS+ layer showing the most extensive and severe thinning. The central hypoAF areas on NIR corresponded to the OS+ thinned areas, while the hypoAF areas on SW-AF corresponded to the R+ thinned areas. Since the larger hypoAF area on NIR-AF exceeded the region of EZ band loss, and corresponded to the OS+ thinned area, RPE cell loss occurred before photoreceptor cell loss. The NIR-AF imaging may be an effective tool for following progression and predicting loss of photoreceptors in STGD1.

Morpho-functional correlation of fundus autofluorescence in Stargardt disease

BackgroundTo correlate patterns in short-wavelength (SW) and near-infrared (NIR) fundus autofluorescence (FAF) with morpho-functional outcomes in eyes affected by Stargardt disease.MethodsFifty-four eyes of 27 patients were prospectively enrolled. All patients...

Inherited retinal or optic nerve disorders – five steps to diagnosis

An early diagnosis of inherited retinal or optic nerve disorders is often delayed due to unspecific clinical signs, multiple clinical manifestations and striking genetic heterogeneity of the underlying molecular defects. This study represents a retrospective analysis of findings in 4,021 patients with inherited retinal or optic nerve disorders seen between 1986 and 2014 (1,171 with follow-up). In addition to the basic ophthalmological examination, electrophysiological tests (ERG, n = 2,088, since 1986; EOG, n = 381, since 1986; VEP n = 595, since 1986; mfERG, n = 819, since 1998) and non-invasive retinal imaging (fundus autofluorescence (FAF, n = 1,784, since 2002), near-infrared autofluorescence (NIA, n = 1,091, since 2006), spectral domain OCT (SD-OCT, n = 848, since 2008) and three-wavelengths multicolour spectral reflection imaging (MC, n = 366, since 2013) were performed at least once. Molecular DNA testing was done in 383 patients between 2006 and 2014. Based on these data an efficient diagnostic strategy is suggested: 1) inclusion of inherited retinal and optic nerve disorders into the differential diagnosis of visual loss or visual field defects with undefined causes; 2) non-invasive retinal imaging; 3) electrophysiological tests; 4) DNA testing to confirm the initial clinical diagnosis; 5) examination in specialised centres, therapy and follow-up. In recent years, the spectrum of diagnostic techniques has continuously expanded. Importantly, non-invasive retinal imaging has become the primary diagnostic tool and DNA testing based on state-of-the-art high throughput techniques increases the identification of associated gene mutations. In conclusion, a structured process in the diagnostic procedure of inherited retinal and optic nerve disorders greatly reduces a diagnostic delay, enables an earlier counselling and therapy and avoids further unnecessary diagnostic tests.

Ex vivo catheter-based imaging of coronary atherosclerosis using multimodality OCT and NIRAF excited at 633 nm.

While optical coherence tomography (OCT) has been shown to be capable of imaging coronary plaque microstructure, additional chemical/molecular information may be needed in order to determine which lesions are at risk of causing an acute coronary event. In this study, we used a recently developed imaging system and double-clad fiber (DCF) catheter capable of simultaneously acquiring both OCT and red excited near-infrared autofluorescence (NIRAF) images (excitation: 633 nm, emission: 680nm to 900nm). We found that NIRAF is elevated in lesions that contain necrotic core - a feature that is critical for vulnerable plaque diagnosis and that is not readily discriminated by OCT alone. We first utilized a DCF ball lens probe and a bench top setup to acquire en face NIRAF images of aortic plaques ex vivo (n = 20). In addition, we used the OCT-NIRAF system and fully assembled catheters to acquire multimodality images from human coronary arteries (n = 15) prosected from human cadaver hearts (n = 5). Comparison of these images with corresponding histology demonstrated that necrotic core plaques exhibited significantly higher NIRAF intensity than other plaque types. These results suggest that multimodality intracoronary OCT-NIRAF imaging technology may be used in the future to provide improved characterization of coronary artery disease in human patients.

Long-term efficacy of half-dose photodynamic therapy on chronic central serous chorioretinopathy

BackgroundTo evaluate the long-term efficacy and complications of half-dose photodynamic therapy (PDT) for treating chronic central serous chorioretinopathy (CSC).MethodsA retrospective, interventional case series of patients with chronic CSC (symptoms ≧3...

Near-Infrared Autofluorescence Imaging in Geographic Atrophy Using Spectralis Single and Combined Wavelength Modes.

The Spectralis Heidelberg retina angiography + optical coherence tomography (OCT) device often fails to acquire near-infrared autofluorescence (NI-AF) due to poor signal because of interference by the beam splitter used for aligning the laser source with the OCT diode light source. We report the rates of successful NI-AF image acquisition using indocyanine green angiography (ICGA) mode (without dye) compared with combined fluorescein angiography (FA) + ICGA mode (without dye) in patients with geographic atrophy (GA). This was a retrospective review. Two hundred images from 50 eyes of 25 patients with GA were included. All patients underwent NI-AF imaging using ICGA (787 nm) and combined FA + ICGA (488 + 787 nm) modes in both eyes. Each eye had macula- and disc-centered imaging. The rates of successful image acquisition were compared using McNemar test. Successful NI-AF acquisition using ICGA was 48% to 52% in macula-centered views and 36% to 56% in disc-centered views. Only 36% of patients had successful bilateral NI-AF imaging. This improved to 100% in all views in both eyes after switching to combined FA + ICGA mode (P < 0.001). Logistic regression showed no patient or ocular factors were predictive of NI-AF acquisition failure in ICGA mode. Acquisition of NI-AF images in GA using ICGA mode has a high failure rate, which can be eliminated by using combined FA + ICGA mode. This finding has implications on the design of imaging protocols for routine assessment and clinical trials of geographic atrophy.

Label-free intraoperative parathyroid localization with near-infrared autofluorescence imaging.

The inability to accurately localize the parathyroid glands during parathyroidectomy and thyroidectomy procedures can prevent patients from achieving postoperative normocalcemia. There is a critical need for an improved intraoperative method for real-time parathyroid identification. The objective of the study was to test the accuracy of a real-time, label-free technique that uses near-infrared (NIR) autofluorescence imaging to localize the parathyroid. The study was conducted at the Vanderbilt University endocrine surgery center. Patients undergoing parathyroidectomy and/or thyroidectomy were included in this study. To validate the intrinsic fluorescence signal in parathyroid, point measurements from 110 patients were collected using NIR fluorescence spectroscopy. Fluorescence imaging was performed on 6 patients. Imaging contrast is based on a previously unreported intrinsic NIR fluorophore in the parathyroid gland. The accuracy of fluorescence imaging was analyzed in comparison with visual assessment and histological findings. The detection rate of parathyroid glands was measured. The parathyroid glands in 100% of patients measured with fluorescence imaging were successfully detected in real time. Fluorescence images consistently showed 2.4 to 8.5 times higher emission intensity from the parathyroid than surrounding tissue. Histological validation confirmed that the high intrinsic fluorescence signal in the parathyroid gland can be used to localize the parathyroid gland regardless of disease state. NIR fluorescence imaging represents a highly sensitive, real-time, label-free tool for parathyroid localization during surgery. The elegance and effectiveness of NIR autofluorescence imaging of the parathyroid gland makes it highly attractive for clinical application in endocrine surgery.

Use digital subtraction images of blue-light and near-infrared autofluorescence for the assessment of irregular foveal contour.

The aims of this study are to generate subtraction images of blue-light autofluorescence (BL-AF) and near-infrared autofluorescence (NIR-AF) from normal eyes, eyes with full thickness macular holes, and eyes with irregular foveal contour, and to compare their autofluorescence patterns. This retrospective study included 44 normal eyes of 22 health individuals, 32 eyes with full thickness macular holes of 32 patients, and 36 eyes with irregular foveal contour of 36 patients. BL-AF and NIR-AF were obtained from all patients and used to generate subtraction images using the Image J software. The decreased signal of central patch was recorded. The central foveal thickness (CFT) and outer nucleus layer (ONL) thickness of fovea were measured to calculate the ONL thickness/CFT ratio. The subtraction images showed regularly increased signal in the central macula of all normal eyes. In contrast, decreased signal of central patch was detected in all full thickness macular holes eyes and 26 out of 36 eyes with irregular foveal contour. No significant difference of the ONL thickness/CFT ratio (F = 2.32, P = 0.113) was observed between normal and irregular foveal contour eyes with or without decreased signal of central patch. Both regularly increased signal and decreased signal of central patch were detected in the eyes with irregular foveal contour. Our results suggest that subtraction images are useful for the assessment of certain macular conditions by providing supplementary information to the green-light autofluorescence and BL-AF.