Human conjunctival tissue samples were from patients with SJS/TEN undergoing ocular surface reconstruction. The controls were nearly normal conjunctival tissue samples from operated patients with conjunctivochalasis.11–14 (link)
Conjunctival tissue sections mounted on slides were fixed for 10 min at 4°C with 4% PFA/0.1M phosphate-buffered saline (PBS), incubated overnight in a moist chamber at 4°C with sheep anti-human teneurin-2 polyclonal antibody (AF4578; R&D, MN, USA) or isotype control sheep IgG (R&D), and then washed in PBS without Ca and Mg [PBS(−)]. Alexa Fluor 488 donkey anti-sheep IgG (H+L) (Molecular Probes, Eugene, OR) was applied for 1 hour at room temperature. After washing the slides, antifade mounting medium with 4′,6-diamidino-2-phenylindole was applied (Vectashield; Vector Laboratories, Burlingame, CA).
Conjunctival tissue sections mounted on slides were fixed for 10 min at 4°C with 4% PFA/0.1M phosphate-buffered saline (PBS), incubated overnight in a moist chamber at 4°C with sheep anti-human teneurin-2 polyclonal antibody (AF4578; R&D, MN, USA) or isotype control sheep IgG (R&D), and then washed in PBS without Ca and Mg [PBS(−)]. Alexa Fluor 488 donkey anti-sheep IgG (H+L) (Molecular Probes, Eugene, OR) was applied for 1 hour at room temperature. After washing the slides, antifade mounting medium with 4′,6-diamidino-2-phenylindole was applied (Vectashield; Vector Laboratories, Burlingame, CA).
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