Considerations for Cleared Tissue Imaging with Light-Sheet Fluorescence Microscopy in Neuroscience Research

Conventional neuroimaging methods often rely on destructive and technically-complex sample preparation and optical reconstruction techniques to overcome the limitations in penetration depth caused by light-scattering and light-absorbing components in biological tissue samples.

Conversely, Light-sheet fluorescence microscopy (LSFM) can image large, optically cleared (ex vivo) samples and — when combined with tissue clearing for non-transparent samples — can penetrate many millimeters through a specimen, unrestricted by scattering and absorption, enabling the acquisition of whole tissues, brains, and even organisms.

Readers can expect to learn more about:

• The unique capabilities of light-sheet fluorescence microscopy for neuronal to whole-brain imaging;
• The advantages and limitations of light-sheet fluorescence microscopy in neuroscience applications;
• Important considerations for neuroscientists adopting LSFM techniques for cleared sample imaging; and
• The innovative neuroscience research LSFM now supports.

This brief includes a case study demonstrating mouse brain imaging with LSFM cleared by CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging / Immunostaining / in situ-hybridization-compatible Tissue hYdrogel).

_{KEYWORDS: Neuroscience Research; Case Study; Light-Sheet Microscopy; Cleared Sample Imaging; Tissue Clearing; CLARITY}