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J Neurophysiol (May 5, 2004). doi:10.1152/jn.00234.2004
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Submitted on March 8, 2004
Accepted on April 22, 2004

In vivo mammalian brain imaging using one- and two-photon fluorescence microendoscopy

Juergen C Jung1, Amit D Mehta2, Emre Aksay3, Raymond Stepnoski4, and Mark J Schnitzer2*

1 Department of Biological Sciences and Department of Applied Physics, Stanford University, Stanford, CA, USA; Department of Pharmacology, Oxford University, Oxford, United Kingdom; Bell Laboratories, Lucent Technologies, Murray Hill, NJ, USA
2 Department of Biological Sciences and Department of Applied Physics, Stanford University, Stanford, CA, USA; Bell Laboratories, Lucent Technologies, Murray Hill, NJ, USA
3 Bell Laboratories, Lucent Technologies, Murray Hill, NJ, USA; Department of Molecular Biology, Princeton University, Princeton, NJ, USA
4 Bell Laboratories, Lucent Technologies, Murray Hill, NJ, USA

* To whom correspondence should be addressed. E-mail: mschnitz{at}stanford.edu.

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 350-1000 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.




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