Abstract
In recent years, three-dimensional mesoscopic imaging has gained significant importance in life sciences for fundamental studies at the whole-organ level. In this manuscript, we present an optical projection tomography (OPT) method designed for imaging of the intact mouse brain. The system features an isotropic resolution of ~50 µm and an acquisition time of four to eight minutes, using a 3-day optimized clearing protocol. Imaging of the brain autofluorescence in 3D reveals details of the neuroanatomy, while the use of fluorescent labels displays the vascular network and amyloid deposition in 5xFAD mice, an important model of Alzheimer's disease (AD). Finally, the OPT images are compared with histological slices.
Highlights
Since its invention in the 19th century, histopathology has been and continues to be a widely used method for tissue-based medical diagnosis
Standard benzyl alcohol and benzyl benzoate (BABB) clearing techniques require tissue immersion in increasing series of methanol concentration for dehydration to prevent cellular distortions and excessive shrinkage of the tissue. We found that it is not needed for optical projection tomography (OPT) as the agarose embedding acts as a protective coating
We present a novel OPT method for whole mouse brain imaging
Summary
Since its invention in the 19th century, histopathology has been and continues to be a widely used method for tissue-based medical diagnosis. While it excels at producing two-dimensional images of tissue sections, extracting information from whole three-dimensional tissues remains challenging. Micro-optical sectioning tomography (MOST) [2] is a block-face imaging method where the process of histology is automatized via simultaneous imaging and sectioning of the tissue. This technique has been successfully applied to image a whole mouse brain with a submicrometric resolution [3]. The acquisition time of approximately two weeks represents an experimental limitation for studies involving numerous animals
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