Retinal examinations were performed using a Heidelberg Engineering Spectralis SD-OCT (Heidelberg Engineering, Heidelberg, Germany) with automatic real-time (ART) function. Scan quality was checked using the OSCAR-IB Criteria (Schippling et al., 2015 (link)) with the Advised Protocol for OCT Study Terminology and Elements APOSTEL recommendations (Cruz-Herranz et al., 2016 (link)). 3.4 mm ring scans around the optic nerve head were used to measure the peripapillary retinal nerve fibre layer (12°, 1536 A-scans 16 ≤automated real time tracking (ART) ≤ 100). The ganglion cell inner plexiform (GCIP) layer volume was measured using a 6 mm diameter cylinder around the fovea from macular volume scans (25° × 30°, 61 vertical B-scans, 768 A-scans per B-scan, ART = 15). Layer segmentation was performed using Eye Explorer 1.9.10.0, viewing module 6.3.4.0 (Heidelberg Engineering, Heidelberg, Germany).
Eye explorer 1
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Eye Explorer 1.9.10.0 is a software application developed by Heidelberg Engineering for analyzing and visualizing ophthalmic data. The core function of the software is to provide a platform for healthcare professionals to view, manage, and assess patient eye data from various diagnostic devices.
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9 protocols using eye explorer 1
1
Retinal Nerve Fiber Layer and Ganglion Cell Measurements
2
Retinal Nerve Fiber Layer Evaluation in Neuroinflammatory Disorders
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Optical coherence tomography (OCT) investigations were performed in all CIS/RRMS-NON patients, in 17 out of 18 CIS/RRMS-ON patients, in 22 out of 23 NMOSD-ON patients and in 21 out of 26 HC using a Heidelberg Engineering Spectralis spectral domain OCT (Heidelberg Engineering, Heidelberg, Germany) with automatic real-time (ART) function for image averaging. The peripapillary retinal nerve fiber layer (pRNFL) was measured with activated eye tracker using 3.4-mm ring scans around the optic nerve head (12°, 1536 A-scans 16 ≤ ART ≤ 100). Segmentation of global RNFL was performed semiautomatically using software provided by the OCT manufacturer (Eye Explorer 1.9.10.0 with viewing module 6.0.9.0; Heidelberg Engineering). Visual acuity tests were performed by either using ETDRS charts or the Traditional Snellen Eye Chart in all CIS/RRMS-NON, in 17 out of 18 CIS/RRMS-ON patients, in 21 out of 23 NMOSD-ON patients and in 21 out of 26 HC. Visual testing outcomes were converted in decimals.
3
Optical Coherence Tomography for Neurodegeneration
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Optical coherence tomography examination was performed using a SD-OCT (Spectralis, Heidelberg Engineering, Heidelberg, Germany) with automatic real-time (ART) function for image averaging. Data are reported for peripapillary retinal nerve fiber layer thickness (pRNFL) to assess axonal degeneration. Total macular volume (TMV), volumes of combined ganglion cell and inner plexiform layer (GCIPL = GCL + IPL), and inner retinal layer (IRL = GCL + IPL + mRNFL) were assessed as markers for neuronal degeneration. Data for inner nuclear layer (INL) were evaluated to detect edema-related retinal changes. Calculation of macular layers is given for a 3 mm diameter cylinder around the fovea from a macular volume scan (20° × 20°, 25 vertical B-scans, ART ≤ 49). The pRNFL was measured with an activated eye tracker using 3.4 mm ring scans around the optic nerve (12°, 1,536 A-scans, ART ≤ 100). Segmentation of all the layers was performed semi-automatically using software provided by the OCT manufacturer (Eye Explorer 1.9.10.0 with viewing module 6.3.4.0, Heidelberg Engineering, Heidelberg, Germany). All the scans were checked for sufficient quality and segmentation errors and corrected, if necessary. OCT data are reported according to the APOSTEL and OSCAR-Ib recommendations (47 (link)–49 (link)). Data were analyzed separately for the patients up to 17 years of age and adults.
4
Comprehensive Optic Nerve Assessment Using SD-OCT
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OCT examination was performed using an SD-OCT (Spectralis, Heidelberg Engineering, Heidelberg, Germany) with automatic real-time (ART) function for image averaging. We acquired peripapillary retinal nerve fiber layer thickness (pRNFL), macular RNFL (mRNFL), and volumes of combined ganglion cell and inner plexiform layer (GCIP), and combined outer plexiform layer and outer nuclear layer (OPONL). All macular layers were calculated as a 3 mm-diameter cylinder around the fovea from a macular volume scan (20° × 20°, 25 vertical B-scans, ART ≤ 49). The peripapillary RNFL (pRNFL) was measured with activated eye tracker using 3.4 mm ring scans around the optic nerve (12°, 1536 A-scans, ART ≤ 100). Segmentation of all layers was performed semi-automatically using software provided by the OCT manufacturer (Eye Explorer 1.9.10.0 with viewing module 6.3.4.0, Heidelberg Engineering, Heidelberg, Germany). One experienced rater (JH), blinded for the diagnosis, carefully checked all scans for sufficient quality and segmentation errors and corrected if necessary. OCT data in this study are reported and analyzed according to the APOSTEL and OSCAR-IB recommendations [18 , 19 (link)]. The data were correlated with VOG measures and clinical scores.
5
Spectral Domain OCT for Neurological Disorders
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Patients underwent spectral domain OCT (Spectralis SD-OCT; Heidelberg Engineering, Heidelberg, Germany) with the Eye Explorer 1.9.10.0 and automatic real-time (ART) image averaging. pRNFL was calculated from a standard ring scan around the optic nerve head (12°, 1536 A-scans, 16 ≤ ART ≤ 100) using segmentation by the device's software with viewing module 6.0.14.0. A macular volume scan (25° × 30°, 61 B-scans, 768 A-scans per B-scan, 12 ≤ ART ≤ 15) was acquired for intraretinal segmentation of GCIP and INL. Segmentation of macular scans was performed with SAMIRIX (52 (link)). All OCT scans were revised for retinal changes unrelated to MS, sufficient quality (53 (link), 54 (link)), segmentation errors and were manually corrected by a blinded experienced grader if necessary. OCT methods are reported in line with the APOSTEL criteria (55 (link)).
6
Quantitative Retinal Imaging Protocol
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All retinal examinations were performed using a Heidelberg Engineering Spectralis spectral domain OCT (Heidelberg Engineering, Heidelberg, Germany) with automatic real-time (ART) function for image averaging. The peripapillary retinal nerve fiber layer (pRNFL) was measured with activated eye tracker using 3.4-mm ring scans around the optic nerve head (12°, 1,536 A-scans 16 ≤ ART ≤ 100). The combined ganglion cell and inner plexiform layer (GCIPL) volume was measured using a 6-mm diameter cylinder around the fovea from a macular volume scan (25° × 30°, 61 vertical B-scans, 768 A-scans per B-scan, ART = 15).13 (link) Segmentation of pRNFL and GCIPL was performed semiautomatically using software provided by the OCT manufacturer (Eye Explorer 1.9.10.0 with viewing module 6.0.9.0; Heidelberg Engineering). All measurements were checked for segmentation errors and corrected if necessary by an experienced rater. Foveal thickness (FT) was measured as the mean thickness of a 1-mm diameter cylinder around the fovea from each collected macular scan. We report our quantitative OCT data in line with the APOSTEL recommendations.14 (link)
7
Automated Retinal Layer Analysis
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All OCT data were collected as part of routine clinical practice in our outpatient clinic using a spectral-domain OCT with automatic real-time function (SD-OCT) (SPECTRALIS, Heidelberg Engineering, Heidelberg, Germany), layer segmentation was performed by the OCT manufacturer software (Eye Explorer 1.9.10.0 Heidelberg Engineering, Heidelberg, Germany). Data analysis followed the international APOSTEL 2.0 and OSCAR-IB criteria [Aytulun et al., 2021 (link), Schippling et al., 2015 (link)]. A 3.5 mm ring scan around the optic nerve (12°, 1536 A-scans, ART ≤ 100) was used to measure the peripapillary retinal nerve fiber layer thickness (pRNFL). Total macular volume (TMV) as well as thickness of the ganglion cell and inner plexiform layer (GCIPL) and inner nuclear layer (INL) were measured on a macular volume scan (20° × 20°, 25 B-scans, ART ≤ 49) using a ø 3 mm ring fixed to the fovea.
8
Comprehensive Retinal Imaging Protocol for Neurodegeneration Assessment
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OCT examination was performed using a SD-OCT (Spectralis, Heidelberg Engineering, Heidelberg, Germany) with automatic real time (ART) function for image averaging as described before (35 (link)). Data are reported for peripapillary retinal nerve fiber layer thickness (pRNFL) to assess axonal degeneration, volume of combined ganglion cell and inner plexiform layer (GCIPL = GCL + IPL) as marker for neuronal degeneration, and for inner nuclear layer (INL). Macular layers were calculated for a 3 mm diameter cylinder around the fovea from a macular volume scan (20° x 20°, 25 vertical B-scans, ART ≤ 49). The pRNFL was measured with activated eye tracker using 3.4 mm ring scans around the optic nerve (12°, 1536 A-scans, ART ≤100). Segmentation of all layers was performed semi-automatically using software provided by the OCT manufacturer (Eye Explorer 1.9.10.0 with viewing module 6.3.4.0, Heidelberg Engineering, Heidelberg, Germany). All scans were checked for sufficient quality and segmentation errors and corrected, if necessary. OCT data are reported according to the APOSTEL (2.0) and OSCAR-Ib recommendations (51 (link)–53 (link)). Both eyes of each subject were included in subsequent analysis as statistically dependent duplicates.
9
Retinal Imaging Protocol for NMOSD Study
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All OCT data were acquired on a spectral domain OCT device (Spectralis, Heidelberg Engineering) with automated real-time function. No pupil dilatation was used. We report the OCT acquisition settings and scanning protocol according to the APOSTEL recommendations [35 (link)]: The pRNFL thickness was measured using 3.4-mm ring scans around the optic nerve head (12°, 1536 A-scans, 9 ≤ ART ≤ 100). The GCIPL volume was measured using a 6-mm diameter cylinder around the fovea from a macular volume scan (25°×30°, 61 vertical B-scans, 768 A-scans per B-scan, ART = 15). Segmentation of the pRNFL and the intraretinal layers in the macular scan was performed semi-automatically using software provided by the optical coherence tomography manufacturer (Eye Explorer 1.9.10.0 with viewing module 6.0.9.0; Heidelberg Engineering). Quality was evaluated according to the OSCAR-IB criteria [36 (link), 37 (link)].
Two patients did not have OCT data. Eight eyes from six NMOSD-ON had to be excluded due to incidental findings or quality reasons. Only the macular scan from two additional NMOSD-ON eyes was excluded due to quality reasons.
Two patients did not have OCT data. Eight eyes from six NMOSD-ON had to be excluded due to incidental findings or quality reasons. Only the macular scan from two additional NMOSD-ON eyes was excluded due to quality reasons.
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