hFRUIT: An optimized agent for optical clearing of DiI-stained adult human brain tissue
Sven Hildebrand, Anna Schueth, Klaus von Wangenheim, C. Mattheyer, Francesco Pampaloni, H. Bratzke, Alard Roebroeck, Ralf A. W. Galuske
Abstract
Here, we describe a new immersion-based clearing method suitable for optical clearing of thick adult human brain samples while preserving its lipids and lipophilic labels such as 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). This clearing procedure is simple, easy to implement, and allowed for clearing of 5 mm thick human brain tissue samples within 12 days. Furthermore, we show for the first time the advantageous effect of the Periodate-Lysine-Paraformaldehyde (PLP) fixation as compared to the more commonly used 4% paraformaldehyde (PFA) on clearing performance.
Topics & Concepts
ParaformaldehydeClearingHuman brainFixation (population genetics)Brain tissueChemistryBiomedical engineeringBiologyBiochemistryPathologyMedicineNeuroscienceEconomicsFinanceGeneAdvanced Fluorescence Microscopy TechniquesCell Image Analysis TechniquesOptical Coherence Tomography Applications