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J Neurophysiol 92: 3121-3133, 2004. First published May 5, 2004; doi:10.1152/jn.00234.2004
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INNOVATIVE METHODOLOGY

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

Juergen C. Jung1,2,3, Amit D. Mehta1,3, Emre Aksay3,4, Raymond Stepnoski3 and Mark J. Schnitzer1,3

1Department of Biological Sciences and Department of Applied Physics, Stanford University, Stanford, California 94305; 2Department of Pharmacology, Oxford University, Oxford OX1 3QT, United Kingdom; 3Bell Laboratories, Lucent Technologies, Murray Hill, New Jersey 07974; and 4Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544

Submitted 8 March 2004; accepted in final form 22 April 2004

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 µm 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.

Address for reprint requests and other correspondence: M. J. Schnitzer, Stanford Univ., James H. Clark Center, Stanford CA 94305-5435 (E-mail: mschnitz{at}stanford.edu).




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