title: Autofluorescence imaging with near-infrared excitation:normalization by reflectance to reduce signal from choroidal fluorophores.
abstract: OBJECTIVE
  We previously developed reduced-illuminance autofluorescence imaging (RAFI) methods involving near-infrared (NIR) excitation to image melanin-based fluorophores and short-wavelength (SW) excitation to image lipofuscin-based flurophores. Here, we propose to normalize NIR-RAFI in order to increase the relative contribution of retinal pigment epithelium (RPE) fluorophores.
  
  
  METHODS
  Retinal imaging was performed with a standard protocol holding system parameters invariant in healthy subjects and in patients. Normalized NIR-RAFI was derived by dividing NIR-RAFI signal by NIR reflectance point-by-point after image registration.
  
  
  RESULTS
  Regions of RPE atrophy in Stargardt disease, AMD, retinitis pigmentosa, choroideremia, and Leber congenital amaurosis as defined by low signal on SW-RAFI could correspond to a wide range of signal on NIR-RAFI depending on the contribution from the choroidal component. Retinal pigment epithelium atrophy tended to always correspond to high signal on NIR reflectance. Normalizing NIR-RAFI reduced the choroidal component of the signal in regions of atrophy. Quantitative evaluation of RPE atrophy area showed no significant differences between SW-RAFI and normalized NIR-RAFI.
  
  
  CONCLUSIONS
  Imaging of RPE atrophy using lipofuscin-based AF imaging has become the gold standard. However, this technique involves bright SW lights that are uncomfortable and may accelerate the rate of disease progression in vulnerable retinas. The NIR-RAFI method developed here is a melanin-based alternative that is not absorbed by opsins and bisretinoid moieties, and is comfortable to view. Further development of this method may result in a nonmydriatic and comfortable imaging method to quantify RPE atrophy extent and its expansion rate.
authors.forename: Artur V
authors.initials: AV
authors.lastname: Cideciyan
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Swider
authors.forename: Samuel G
authors.initials: SG
authors.lastname: Jacobson
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002829:Choroid / Q000378:metabolism*; D005260:Female; D005451:Fluorescein Angiography / Q000379:methods*; D006801:Humans; D008062:Lipofuscin / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009887:Ophthalmoscopy / Q000379:methods; D061848:Optical Imaging / Q000379:methods*; D012164:Retinal Diseases / Q000175:diagnosis*; D055213:Retinal Pigment Epithelium / Q000378:metabolism; D019265:Spectroscopy, Near-Infrared / Q000379:methods; D055815:Young Adult
chemical_list: D008062:Lipofuscin