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  • hFRUIT: An optimized agent for optical clearing of DiI-stained adult human brain tissue.

hFRUIT: An optimized agent for optical clearing of DiI-stained adult human brain tissue.

Scientific reports (2020-06-21)
Sven Hildebrand, Anna Schueth, Klaus von Wangenheim, Christian Mattheyer, Francesco Pampaloni, Hansjürgen Bratzke, Alard F 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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Silicone oil, high temperature
Sigma-Aldrich
1-Thioglycerol, ≥97%
Sigma-Aldrich
Mineral oil, light oil, (neat)