Dear Collegues,
Translucence Biosystems was honored to present at the AD/PD™ 2022, Alzheimer's & Parkinson's Diseases Conference. Please see our poster presentation entitled, "Brain-Wide Cellular Resolution Measurement of Neuronal Activity, Microglial Activation and β-Amyloid Deposition" below.
Abstract
Recent advances in optical clearing and light sheet imaging have opened an exciting new avenue for brain-wide, cellular resolution immunostaining at the forefront of a dimensional shift from 2D to 3D histology. With access to the intricate anatomy of the whole intact brain, we are developing unbiased and complete pictures of brain pathology for pre-clinical studies.
Methods: With our optimized iDISCO-based clearing methods and our Mesoscale Imaging System for the ZEISS Lightsheet Z.1 microscope, we can image entire mouse brains in ~10 min. Further, our machine learning-enabled 3TK software identifies individual cells or protein aggregates throughout the brain and aligns them to the Allen Reference Atlas to produce a regionalized read-out of patterns across 100’s of brain areas.
Results: We have applied this technology to generate cellular resolution maps of neuronal activity by measuring the immediate-early gene (IEG) products Npas4 and cFos in response to both a light exposure task and in response to contextual fear conditioning. Further, we have optimized methods to identify and count microglia throughout the brain and perform regional analyses of microglial activation.
Importantly, we have used Alzheimer’s Disease model 5xFAD mice to develop assays to generate brain-wide signatures of β-Amyloid deposition. Our automated analysis of microglia and β-Amyloid co-staining allows for brain-wide quantification of plaque-associated microglia. Conclusions: These new methods for whole-brain, next generation 3D immunohistochemistry and anatomics are ideally suited to pre-clinical studies for unbiased, complete and anatomically precise mapping of the efficacy of CNS therapeutics. We thank NIMH for Grants R43MH119989 and R43MH122070.
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