Arezoo Khoradmehr
Fahime Mazaheri - Research and Clinical Center for Infertility, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Morteza Anvari - Department of Biology and Anatomical Sciences, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Amin Tamadon - The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran

The whole brain vasculature three-dimensional (3D) imaging can be done by the Fast Free-of-Acrylamide Clearing Tissue (FACT) protocol. The FACT protocol removes lipids from the brain tissue using sodium dodecyl sulfate (SDS) to make the tissue transparent for imaging. This study was done using FACT protocol in combination with imaging of auto-fluorescency of red blood cells in vessels to image the vasculature of a translucent mouse brain. In this experimental study, brain of adult female mice were dissected out without the perfusion. Mice brains were sliced for vasculature imaging before the clearing. Brain slices were cleared by the FACT protocol. After 1 mm of the brain slice clearing, the blood vessels containing auto-fluorescent red blood cells were imaged by a z-stack motorized epi-fluorescent microscope. The 3D structures of the brain vessels were reconstructed by Imaris software. Auto-fluorescent blood vessels were 3D segmented by the FACT in mouse brain cortex. The FACT method is a simple technique which might be appropriate for laboratories in developing countries lacking advanced microscopes such as confocal microscopes. Successful labeling the vessels in murine species after clearing by the FACT approach resulted in 3D imaging of brain cortex vessels for the first time. Imaging the vasculature was performed without any staining; rather accomplished by a simple auto-fluorescence imaging of the RBC.

FACT, Three-dimensional imaging, Tissue, Vasculature, Mice