Quantitative validation of immunofluorescence and lectin staining using reduced CLARITY acrylamide formulations.

ABSTRACT: The CLARITY technique enables three-dimensional visualization of fluorescent-labeled biomolecules in clarified intact brain samples, affording a unique view of molecular neuroanatomy and neurocircuitry. It is therefore, essential to find the ideal combination for clearing tissue and detecting the fluorescent-labeled signal. This method requires the formation of a formaldehyde-acrylamide fixative-generated hydrogel mesh through which cellular lipid is removed with sodium dodecyl sulfate. Several laboratories have used differential acrylamide and detergent concentrations to achieve better tissue clearing and antibody penetration, but the potential effects upon fluorescent signal retention is largely unknown. In an effort to optimize CLARITY processing procedures we performed quantitative par

SUBMITTER: Krolewski DM 

PROVIDER: S-EPMC5828880 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature