Wide field block face imaging using deep ultraviolet induced autofluorescence of the human brain.

Imaging large volume human brains at cellular resolution involve histological methods that cause structural changes. A reference point prior to sectioning is needed to quantify these changes and is achieved by serial block face imaging (BFI) methods that have been applied to small volume tissue (∼1 cm3). We have developed a BFI uniquely designed for large volume tissues (∼1300 cm3) with a very large field of view (20 × 20 cm) at a resolution of 70 µm/pixel under deep ultraviolet (UV-C) illumination which highlights key features. The UV-C imaging ensures high contrast imaging of the brain tissue and highlights salient features of the brain. The system is designed to provide uniform and stable illumination across the entire surface area of the tissue and to work at low temperatures, which are required during cryosectioning. Most importantly, it has been designed to maintain its optical focus over the large depth of tissue and over long periods of time, without readjustments. The BFI was installed within a cryomacrotome, and was used to image a large cryoblock of an adult human cerebellum and brainstem (∼6 cm depth resulting in 2995 serial images) with precise optical focus and no loss during continuous serial acquisition. The deep UV-C induced BFI highlights several large fibre tracts within the brain including the cerebellar peduncles, and the corticospinal tract providing important advantage over white light BFI. The 3D reconstructed serial BFI images can assist in the registration and alignment of the microscopic high-resolution histological tissue sections. • Large field of view block face imaging of human brain. • Deep ultraviolet induced fluorescence of the brain highlights important structures prior to cryosectioning. • Robust reference volume for high throughput high resolution histology pipeline. [ABSTRACT FROM AUTHOR]