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In a latest research printed within the Journal of Biomedical Optics, researchers reveal the multimodal performance of focused ocular fluorescence spectroscopy in vitro and in vivo

Research: Focused spectroscopy within the eye fundus. Picture Credit score: – Yuri A /


Some typical structural and useful adjustments happen within the eyes, particularly the attention fundus, as a consequence of ocular illnesses like diabetic retinopathy (DR), age-related macular degeneration (AMD), and glaucoma. Neurological illnesses like Alzheimer’s illness (AD) and Parkinson’s illness (PD) also can result in retinal adjustments, akin to thinning of the retinal nerve fiber layer (RNFL) and adjustments in hemodynamics.

Given the extremely heterogeneous options and composition of the attention fundus, biomarkers both get broadly dispersed all through this tissue or localize to particular areas. For instance, β-amyloid plaques unfold all through the retina of AD sufferers, whereas sufferers with DR have localized hemorrhages. 

Typical imaging strategies don’t present sufficient knowledge on retinal adjustments induced by these illnesses as in comparison with focused ocular diffuse reflectance spectroscopy (DRS). Ocular DRS strategies allow spectral evaluation of particular components of the attention fundus, together with the optic disc, peripheral retina, and fovea between 500 and 800 nanometers (nm).

Diffuse reflectance and fluorescence spectroscopy also can elucidate the affect of things like lipofuscin accumulation, RNFL structural adjustments, blood absorption spectrum, and melanin spectral profile, all of which affect the optical properties of retinal tissues.

Concerning the research

Within the present research, researchers establish the important thing options of the focused ocular spectroscopy know-how in vitro utilizing a reference goal and mannequin eye. The reference goal was an ultrahigh-definition display screen with a grid of eight completely different colours, with the fundus digicam positioned in entrance of it and solely gathering gentle emitted by the display screen. The OEMI-7 eye mannequin, a seven mm pupil precisely simulating the human eye, helped validate these DRS acquisitions.

Subsequently, in vivo imaging and DRS have been used to evaluate blood oxygen saturation (StO2) within the optic nerve head and parafovea of eight wholesome research members who offered knowledgeable consent earlier than the research. These people have been between 27 and 35 years of age, had no systemic illnesses or treatment, and had regular outcomes following eye examinations.

The pointing gentle emitting diode (LED) illuminated the precise place of the particular area of spectral acquisition (ROSA), which allowed the digicam to seize its location. A two-step acquisition sequence was used, adopted by mixed imaging and focused spectroscopy.

The placement of the DRS acquisition space was decided primarily based on the ROSA picture segmentation. Spectra have been acquired by shifting the ROSA to 6 completely different areas throughout the subject of view of the reference goal for spectral evaluation. 

Bandpass filters isolate the excitation illumination for inexperienced fluorescence imaging. Comparatively, long-pass filters enabled unique imaging and spectral acquisition of sunshine emitted by fluorescence.

Spectral evaluation concerned three processing steps, whereby ambient gentle spectral contributions from the spectrum have been eliminated, and the impact of the illumination supply spectrum was subsequently decided. The sunshine spectrum was then normalized to appropriate for variations in sign depth.

Research findings

The mannequin eye acquired reflectance spectra from blood vessels, the retina close to the optic nerve head, the optic nerve head, and the retina removed from the optic nerve head (D). The blood vessels and optic nerve exhibited clearly completely different reflectance spectra. Equally, the mannequin eye helped carry out fluorescence evaluation for 4 areas, with solely the blood vessels and optic nerve head emitting fluorescence alerts.

5-second DRS acquisitions corresponded to 13 acquired spectra and have been made on the optic nerve head and parafovea for all eight members. Common absorbance spectra for each areas confirmed interindividual variability.

All earlier strategies to evaluate blood oxygen saturation within the eye had restricted sensitivity and, consequently, allowed for the relative evaluation of StO2 just for massive blood vessels of the attention fundus. Within the present research, blood oxygen saturation measurements made in several areas led to completely different values of StO2

Decrease oxygen saturation and extra interindividual variability in StO2 have been noticed within the parafovea than within the optic nerve head, starting from 30.4-58.4% and 62.1-69.7%, respectively.

An ocular oximetry algorithm was applied for spectra acquired in vivo and demonstrated the potential to evaluate the presence of various fluorophores/chromophores that can be utilized to diagnose completely different retinal pathologies. Extra particularly, this strategy focused particular areas of curiosity that have been recognized by way of wide-field fluorescence and purchased a complete emission spectral profile of those molecules.


The multimodal system offered on this research enabled concurrent and steady imaging and focused spectroscopy within the eye fundus. Furthermore, it confirmed excessive sensitivity, spectral decision, and quick acquisition pace for retinal biomarker detection. It’s noteworthy as a result of different programs, akin to hyperspectral imaging, compromise between spectral decision and acquisition pace.

Additional, this know-how acquired distinct spectral profiles within the completely different areas examined throughout in vitro and through in vivo testing. To conclude, focused ocular spectroscopy may open novel methods to diagnose and deal with eye illnesses over time.

Journal reference:

  • Lapointe, N., Akitegetse, C., Poirier, J., et al. (2023). Focused spectroscopy within the eye fundus. Journal of Biomedical Optocs 28(12).doi:10.1117/1.JBO.28.12.126004


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