Abstract
Purpose: :
To obtain in vivo laser scanning confocal images of fresh and cryopreserved human amniotic membrane (AM).
Methods: :
Fresh and cryopreserved AMs from 3 different donors were examined using a cornea–specific in vivo laser scanning confocal microscope (Heidelberg Retina Tomograph II Rostock Cornea Module, HRT II–RCM). Fresh AMs were kept at room temperature, and were examined within 2 hours after harvesting, while cryopreserved AMs were examined after being thawed at room temperature for at least 1 hour. Tomo–cap was mounted on the holder to cover the microscope lens. Then, the epithelial side of the AM samples was attached to the front surface of the Tomo–cap, and each AM sample was examined layer by layer with HRT II–RCM
Results: :
Comparable with histologic sections, the HRT II–RCM resolved five distinct layers: the epithelium, a basement membrane, a compact stromal layer, a cell–rich stromal layer, and a spongy stromal layer, in both fresh and cryopreserved AMs. In the compact stromal layer, microfolds were more prominent in fresh AMs than in cryopreserved AMs. For the cell–rich stromal layer, mesenchymal cells were observed more distinctively in fresh AMs than in cryopreserved AMs.
Conclusions: :
These baseline confocal images visualized by in vivo HRT II–RCM can be used for future investigation of in vivo remodeling after AM transplantation.
Keywords: imaging/image analysis: clinical • cornea: clinical science • anatomy