This study has been transferred to the Human Research Ethics Committee of St Vincent's Hospital Melbourne.

This study investigates a novel, non-invasive imaging technique called hyperspectral retinal imaging to improve the identification and characterisation of retinal and choroidal structures in both healthy and diseased eyes. Hyperspectral imaging captures retinal images across multiple wavelengths of light, generating detailed spectral information that may reveal biological and structural features not visible with conventional retinal photography.

Approximately 1000 participants will undergo retinal imaging at specialist eye clinics in Melbourne, Australia. The study aims to determine whether hyperspectral imaging can detect spectral signatures associated with retinal and optic nerve diseases such as diabetic retinopathy, glaucoma, and age-related macular degeneration, and whether these signatures correlate with disease severity.

This investigator-initiated imaging study aims to evaluate the utility of hyperspectral retinal imaging (HSI) for the identification and characterisation of retinal and choroidal structures in both normal and diseased eyes.

Hyperspectral imaging is a non-invasive retinal imaging modality that captures sequential images across multiple wavelengths of light (typically >25 spectral bands and up to approximately 90 wavelengths). This produces a three-dimensional dataset ("hypercube") containing spatial and spectral information for each pixel, enabling analysis of tissue-specific spectral reflectance properties. The technique differs from conventional retinal photography, which typically uses three colour channels (red, green, and blue), by providing significantly enhanced spectral resolution.

The study population will include approximately 1000 participants recruited from multiple ophthalmic clinics in Melbourne, Australia. Participants will be recruited across a range of normal ocular health and retinal disease states, including but not limited to diabetic retinopathy, glaucoma, and age-related macular degeneration.

Imaging will be performed at the Centre for Eye Research Australia (CERA) using two hyperspectral imaging systems: the Optina Diagnostics Metabolic Hyperspectral Retinal Camera and a prototype hyperspectral camera developed at CERA. Both systems acquire rapid sequential retinal images across visible to near-infrared wavelengths. The Optina device provides a field of view of approximately 26 degrees and uses a tunable supercontinuum light source, while the CERA prototype uses LED-based illumination with optical filtering and provides a field of view of approximately 35 degrees.

Participants may undergo pharmacological pupil dilation prior to imaging to improve image quality. Each study visit will last approximately 60 minutes, including dilation and imaging procedures.

Image data will undergo registration and processing to correct for eye movement and system response, enabling extraction of pixel-level spectral signatures. Computational analysis methods will be developed to identify and quantify retinal and choroidal features and to explore associations between spectral signatures and structural or functional measures of disease.

The primary objectives are to optimise hyperspectral imaging acquisition protocols for retinal and optic nerve structures, to determine whether spectral signatures associated with retinal pathology can be detected using hyperspectral imaging, and to assess whether these spectral changes correlate with clinical measures of disease severity.

Inclusion Criteria:

• Adults aged 18 years and older
• Able to provide informed consent
• Willing and able to attend study imaging sessions
• Participants with healthy eyes or diagnosed retinal/optic nerve disease (including but not limited to diabetic retinopathy, glaucoma, and age-related macular degeneration)
• Adequate media clarity to permit retinal imaging as determined by investigator

Exclusion Criteria:

• Inability to provide informed consent
• Known history of narrow anterior chamber angles or risk of acute angle closure where pupil dilation is contraindicated
• Ocular media opacities preventing adequate retinal imaging (e.g., dense cataract, severe corneal opacity, vitreous hemorrhage)
• Any condition that, in the opinion of the investigator, would make participation unsafe or compromise study data quality
• Allergy or contraindication to pharmacologic mydriatic agents used for pupil dilation

The plan for sharing individual participant data (IPD) has not yet been finalised. Given the exploratory and image-based nature of this study, which involves large hyperspectral retinal imaging datasets and ongoing development of analysis methods, decisions regarding data sharing will depend on the completion of primary analyses, ethical approvals, and institutional data governance requirements. The feasibility of de-identification and appropriate data access frameworks is currently under review.