Staining characteristics of preserved human amniotic membrane.
OBJECTIVE
Amniotic membrane is an ultra-thin cellophane-like membrane that is used in ocular surface reconstruction. We evaluated the staining characteristics of commonly available dyes on preserved human amniotic membrane to aid in handling of amniotic membrane during transplantation.


METHODS
Five dyes, indocyanine green (2.5%, 1.0%, and 0.5%), fluorescein (0.25%), rose bengal (1%), lissamine green B (1%), and trypan blue (0.5%), were used to stain amniotic membrane. After staining, the specimens were observed under a dissecting microscope to evaluate for the uptake of the stains. Positively stained membranes were evaluated for the persistence of staining by placing them in 2 to 3 mL of balanced saline solution that was changed every 30 minutes over 6 hours.


RESULTS
Preserved human amniotic membrane is stained by indocyanine green, rose bengal, lissamine green B, and trypan blue. Of these four dyes, only the membrane stained with 1% lissamine green B was free of stain after 120 minutes. Indocyanine green, rose bengal, and trypan blue continued to strongly stain the membrane after 24 hours.


CONCLUSIONS
Indocyanine green, rose bengal, trypan blue, and lissamine green B all stain amniotic membrane. Lissamine green B appears to have advantages over the other dyes in that it will stain the membrane well, and in our model, dissipate in 120 minutes. Intraoperative staining with lissamine green B may be a simple and effective way to assist surgeons in the proper handling of amniotic membrane.
Hu
Daniel J
DJ
Department of Ophthalmology, Northwestern University, Feinberg School of Medicine, Chicago, IL 60611, USA.
Basti
Surendra
S
Bryar
Paul J
PJ
D000650:Amnion / Q000187:drug effects* / Q000378:metabolism; D004396:Coloring Agents / Q000494:pharmacology*; D003287:Contrast Media / Q000494:pharmacology; D015925:Cryopreservation; D003890:Desiccation; D019793:Fluorescein / Q000494:pharmacology; D006801:Humans; D007208:Indocyanine Green / Q000494:pharmacology; D008086:Lissamine Green Dyes / Q000494:pharmacology; D012395:Rose Bengal / Q000494:pharmacology; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors; D014021:Tissue Preservation; D014343:Trypan Blue / Q000494:pharmacology
D004396:Coloring Agents; D003287:Contrast Media; D008086:Lissamine Green Dyes; D012395:Rose Bengal; C024537:green 5; D014343:Trypan Blue; D007208:Indocyanine Green; D019793:Fluorescein