Tissue clearing is a powerful technique that renders large biological samples optically transparent and allows 3Dvisualization of wide-brain molecular anatomy. To help disseminate this technology, Translucence Biosystemsdeveloped a Mesoscale Imaging System⢠and, using previous SBIR NIH support, an analytical softwarepackage for advanced analysis of brain samples, 3TKTM. While complete solutions exist for imaging and analysisof cleared samples, no complete solution exists for the highly labor intensive and error prone tissue clearingprocess. Here Translucence Biosystems introduce Clearbotâ¢, an automated system for sample clearing andimmunostaining that will complement previous Translucence devices and complete a pipeline for reliable brain-wide molecular anatomy of intact rodent and human tissues. In this Phase I proposal, Translucence Biosystemswill build a prototype of Clearbot⢠and demonstrate automated brain tissue clearing for whole-brain analysis.Specific aims: Aim 1: Assemble and evaluate a Clearbot prototype: the basic functionality of the prototypedeveloped by an engineering consultant will be evaluated for functionality, robustness, energy consumption, andoverall reliability. Aim 2: Evaluate the utility of Clearbot for automated clearing and immunostaining ofintact brains: quality and repeatability of tissue clearing will be evaluated using the prototype assembled in Aim1. Multiple mouse brains will be cleared and immunostained with Clearbot and benchmarked to manualprocessing done in parallel. This Phase I project will demonstrate the feasibility of developing an automateddevice that is comparable to or better than manual clearing. Following success of this Phase I feasibility testing,future developments will advance the prototype to a flexible, commercial-grade system with a robust and simpleuser-interface, helping researchers apply this game-changing technology to their research questions.
Public Health Relevance Statement: NARRATIVE
Translucence is developing Clearbotâ¢, an automated system for high throughput tissue clearing and
immunostaining. Clearbot will promote the implementation of tissue clearing and intact brain histological analysis,
an emerging field that has been identified by the NIH BRAIN Initiative's Brain 2025 Report as one of its key
targets for development. In this project, Translucence will demonstrate the feasibility and functionality of Clearbot
with validation tests that compare the quality of clearing and staining between manual and automated
processing.
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