In vivo mammalian brain imaging using one- and two-photon fluorescence microendoscopy.
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.
Jung
Juergen C
JC
Department of Biological Sciences, Stanford University, Stanford, CA 94305-5435, USA.
Mehta
Amit D
AD
Aksay
Emre
E
Stepnoski
Raymond
R
Schnitzer
Mark J
MJ
D000818:Animals; D003712:Dendrites / Q000648:ultrastructure*; D004724:Endoscopy; D004912:Erythrocytes / Q000166:cytology; D005260:Female; D005456:Fluorescent Dyes; D008322:Mammals; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D017785:Photons; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D014741:Video Recording
D005456:Fluorescent Dyes