Methods for tissue preparation

  • Eyes were obtained from the Alabama Eye Bank within 4 hr of death (>900 pairs, 1995-2009) and were preserved by immersion in 2% glutaraldehyde and 1% paraformaldehyde following anterior segment removal.
  • Eye health history was available for 33% of these eyes.
  • All donor eyes remaining in this collection in 2010 and having a gross macular appearance consistent with any stage of AMD were sectioned and evaluated, along with many eyes with unremarkable maculas from age-matched donors.
  • Tissue was post-fixed by osmium tannic acid paraphenylenediamine to highlight neutral lipids in extracellular AMD-associated lesions.
  • Macula-wide, high-resolution sections were generated from 8 mm diameter full-thickness punches, and 0.8 µm-thick sections were stained with toluidine blue.