Optimized Protocol of Methanol Treatment for Immunofluorescent Staining in Fixed Brain Slices

We optimized methanol treatment in paraformaldehyde-fixed slices for immunofluorescent staining of ependymal basal bodies in brain ventricles. As 100% methanol induced severe deformations to the slices (including rolling and folding over), we tried to decrease methanol concentration. We found that 33.3% to 75% methanol could result in ideal immunostaining of basal bodies without inducing obvious deformations. Instead of treating slices at −20°C (without proper cryoprotection measurements) as suggested in previous studies, we carried out methanol treatment at room temperature. Our modified protocol can not only raise immunostaining efficiency in tissue slices, it may also prevent potential freezing damages to the samples.

• Publication year

  2017

• Venue

  Applied immunohistochemistry & molecular morphology (Print)

• Publication date

  2017-03-01

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1097/PAI.0000000000000293PMID 26509907PMCID 4848166
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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