Henrik Renner Author Answered Jul 14, 2021
Max Planck Institute for Molecular Biomedicine Münster
Dear Anu Sebin,
Thank you very much for your question and interest in our protocol. Using our standard workflow for the generation of organoids each well does, indeed, contain exactly one organoid. Please also refer to our previous publications (https://elifesciences.org/articles/52904 and https://bio-protocol.org/e4049) for more information on organoid generation and culture. If your culture system generates several organoids per well, this should also be compatible with the procedure we outlined here. However, we have not optimized this approach.
In order to prevent the loss of samples during the various washing steps, careful liquid handling is crucial. To facilitate the aspiration of liquids, we recommend slightly tilting the plate and performing all liquid handling by pipetting (not by vacuum pumps or similar devices). Leave a small amount of liquid (≤ 10 μl) in the well so as to not damage the organoids by extensive direct contact with a pipet tip.
Best regards,
Henrik Renner & Jan Bruder