In the lab I work, we're using the free-floating technique to label BrdU in brain sections. We are using 12-well plates, but still wondering what's the adequate number of sections per volume proportion or number of sections per well.
Dear Daniel,
I use 24 well plates and incubate single brain slices per well with 200 microliter antibody solution per well. Having multiple sections per well is not a good idea as the sections can cover over/seal and prevent antibody access. Also you need to make sure that the sections you have are not too large to fold and stick to itself and again prevent access. I seal the top of the wells with parafilm to prevent evaporation, and incubate at 4 degrees Celsius.
best wishes, Refik
Technical Report Research Methods and Technical Considerations: Answers to ov...
How about covering a large petri dish with parafilm, 100 drops of buffer on top of the parafilm. Each drop containing one section. The buffer can easily be exchanged with a pipet, without touching or disturbing the section. For long time incubations I would advise to have the petri dish moist with e.g. wet paper.
Thank you both, Mr. Kanjhan and Mr. Kramer. We will try those ways.
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