In Vivo Mammalian Brain Imaging Using One- and Two-Photon Fluorescence Microendoscopy

TLDR

One of the major limitations in current neuroscience techniques is the lack of methods to image individual cells deep within the brains of live animals. The authors developed two minimally invasive fluorescence microendoscopy methods and tested their ability to image individual cells in vivo. Both one‑ and two‑photon fluorescence microendoscopy use compound GRIN lenses (350–1,000 µm) to achieve micron‑scale resolution; one‑photon provides full‑frame imaging suitable for fast frame rates, while two‑photon offers laser‑scanning optical sectioning. The technique yielded video‑rate movies of thalamic and CA1 hippocampal red‑blood‑cell dynamics and still‑frame images of CA1 neurons and dendrites in anesthetized rats and mice, positioning it to support the growing demand for in vivo cellular imaging with new fluorophores.

Abstract

One of the major limitations in the current set of techniques available to neuroscientists is a dearth of methods for imaging individual cells deep within the brains of live animals. To overcome this limitation, we developed two forms of minimally invasive fluorescence microendoscopy and tested their abilities to image cells in vivo. Both one- and two-photon fluorescence microendoscopy are based on compound gradient refractive index (GRIN) lenses that are 350-1,000 microm in diameter and provide micron-scale resolution. One-photon microendoscopy allows full-frame images to be viewed by eye or with a camera, and is well suited to fast frame-rate imaging. Two-photon microendoscopy is a laser-scanning modality that provides optical sectioning deep within tissue. Using in vivo microendoscopy we acquired video-rate movies of thalamic and CA1 hippocampal red blood cell dynamics and still-frame images of CA1 neurons and dendrites in anesthetized rats and mice. Microendoscopy will help meet the growing demand for in vivo cellular imaging created by the rapid emergence of new synthetic and genetically encoded fluorophores that can be used to label specific brain areas or cell classes.