Traditional histological approaches using tissue sections of conventional thickness (7–100 μm) fail to fully capture the intricate three-dimensional architecture of neuronal and glial cells. Visualization of larger tissue blocks is hindered by tissue opacity and limited antibody penetration, where opacity arises from light scattering at interfaces with mismatched refractive indices (RIs). Objective: Сompare two RI-matching methods, CLARITY and SWITCH, for processing archival human fetal brain tissue blocks fixed long-term in 4% buffered paraformaldehyde at room temperature. Results: Cerebral cortex blocks from four human fetuses (25–39 gestational weeks) were examined. Confocal microscopy revealed imaging depths of up to 2 mm for both methods. However, CLARITY exhibited shallow antibody penetration, with staining predominantly restricted to superficial layers (150–180 μm), while SWITCH enabled uniform antibody penetration beyond 300 μm. Our findings demonstrate that
both CLARITY and SWITCH are applicable to archival human fetal brain tissue with prolonged fixation (up to 3.5 years). Conclusion: SWITCH outperformed CLARITY in tissue processing time, antibody penetration depth, and low post-treatment sample deformation.
Original languageEnglish
Pages (from-to)S186-S191
Number of pages5
JournalCell and Tissue Biology
Volume19
Issue numberSuppl 1
DOIs
StatePublished - 1 Aug 2025

    Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Neuroscience(all)

    Research areas

  • optical clearing, immunofluorescence, CLARITY, SWITCH, human, renatal brain development, prenatal brain development

ID: 140020726