Low-cost multimodal light sheet microscopy for optically cleared tissues and living specimens

J Biomed Opt. 2016 Dec 1;21(12):126008. doi: 10.1117/1.JBO.21.12.126008.

Abstract

Light sheet microscopy techniques have expanded with designs to address many new applications. Due to rapid advancements in computing power, camera/detector technologies, and tissue clearing techniques, light sheet methods are becoming increasingly popular for biomedical imaging applications at the cellular and tissue levels. Light sheet imaging modalities couple rapid imaging rates, low-levels of phototoxicity, and excellent signal to noise ratios, contributing to their popularity for experimental biology. However, the current major limitation of light sheet microscopy arises from optical aberrations, with the main drawback being the defocusing introduced by refractive index variations that accompany clearing techniques. Here, we propose an inexpensive and easy to build light sheet based instrumentation to overcome this limitation by optomechanically decoupling the sample scanning movement from the detection step. Our solution is relatively simple to implement and also provides increased modularity by using a swappable excitation arm. This expands the range of samples we can image on a single system, from high resolution for single cells at ? m spatial resolution, to tissues with mm spatial resolution. We demonstrate our approach, using the system to image iDISCO cleared embryos and sciatic nerves, and provide the full three-dimensional reconstruction of these objects in minutes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Embryo, Mammalian / diagnostic imaging
  • Equipment Design
  • Female
  • Ganglia, Spinal / diagnostic imaging
  • HEK293 Cells
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Lasers
  • Mice
  • Microscopy, Fluorescence / methods*
  • Multimodal Imaging / methods*
  • Neurobiology
  • Single-Cell Analysis

Grants and funding