Fixation and embedding of human cerebral organoids (hCOs)

MD Martina Donadoni
SC Senem Cakir
AB Anna Bellizzi
MS Michael Swingler
IS Ilker K Sariyer
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Before immunoanalytical assays, hCOs were collected and washed three times with D-PBSfor 10 min. hCOs were fixed with 4% PFA overnight at 4˚C for 16 h, washed with 0.1% PBS-T, and kept in D-PBS for 1 to 7 days at 4˚C. The 30% Sucrose in D-PBS solution was used for dehydration for 1–2 days until the hCOs were submerged. hCOs were then embedded in Optimal Cutting Temperature Embedding Medium (OCT) to obtain 10 μm thick sections at cryostat and air-dried for 20 min at RT before staining or storing at -80˚C.

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