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A Tissue Clearing Method for Neuronal Imaging from Mesoscopic to Microscopic Scales
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Optimized Protocol for Imaging Cleared Neural Tissues Using Light Microscopy.

Yoh Isogai1,2, Douglas S Richardson3, Catherine Dulac1

  • 1Department of Molecular and Cellular Biology, Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 13, 2016
PubMed
Summary
This summary is machine-generated.

This study presents a streamlined protocol for rapidly imaging intact brain tissue at high resolution. The method integrates tissue clearing, imaging, and analysis to facilitate mesoscale biological investigations.

Keywords:
CLARITYConfocalLightsheetOptiViewTissue clearingWhole brain imaging

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Area of Science:

  • Biology
  • Neuroscience
  • Biotechnology

Background:

  • Understanding organismal-scale biological processes, especially in the nervous system, is challenging despite advances in molecular and cellular biology.
  • Bridging the gap between molecular/cellular functions and organismal-scale phenomena requires new methodologies.
  • Large-scale tissue analysis has been hindered by limitations in preserving molecular and cellular detail in intact samples.

Purpose of the Study:

  • To outline a protocol for rapid, high-resolution imaging of intact brain tissue up to thousands of cubic millimeters.
  • To address logistical considerations in analyzing cellular processes within intact tissues.
  • To provide an integrated solution for researchers entering the field of large-scale tissue imaging.

Main Methods:

  • The protocol involves a three-step pipeline: tissue clearing, tissue imaging, and data analysis.
  • Tissue clearing techniques are employed to enable imaging of large, intact biological samples.
  • An integrated solution is described to streamline the imaging and analysis process.

Main Results:

  • The developed protocol enables rapid imaging of intact brain tissue at cellular resolution.
  • The method facilitates the study of mesoscale biological questions.
  • The optimized pipeline aims to lower the barrier for implementing high-resolution tissue imaging.

Conclusions:

  • This optimized protocol facilitates high-resolution imaging of intact biological tissues.
  • It enables the investigation of mesoscale questions in biology and neuroscience.
  • The streamlined approach supports researchers in exploring complex biological systems at an unprecedented scale.