Karthik, Srinivasa, Joseph, Jayaraj, Jayakumar, Jaikishan, Manoj, Rahul, Shetty, Mahesh, Bota, Mihail, Verma, Richa, Mitra, Partha, Sivaprakasam, Mohanasankar (September 2023) Wide field block face imaging using deep ultraviolet induced autofluorescence of the human brain. Journal of Neuroscience Methods, 397. p. 109921. ISSN 0165-0270
Abstract
BACKGROUND: 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). NEW METHOD: 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. RESULTS: 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. COMPARISON WITH EXISTING METHOD(S): 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. CONCLUSIONS: The 3D reconstructed serial BFI images can assist in the registration and alignment of the microscopic high-resolution histological tissue sections.
| Item Type: | Paper |
|---|---|
| Subjects: | Investigative techniques and equipment organism description > animal organs, tissues, organelles, cell types and functions > organs types and functions > brain organism description > animal > mammal > primates > hominids organism description > animal > mammal > primates > hominids > human organism description > animal > mammal organs, tissues, organelles, cell types and functions > organs types and functions organs, tissues, organelles, cell types and functions organism description > animal > mammal > primates |
| CSHL Authors: | |
| Communities: | CSHL labs > Mitra lab |
| SWORD Depositor: | CSHL Elements |
| Depositing User: | CSHL Elements |
| Date: | 1 September 2023 |
| Date Deposited: | 22 Sep 2023 15:38 |
| Last Modified: | 10 Jan 2024 19:49 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/40978 |
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