JN Fuel your research with LabChart
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

J Neurophysiol (March 3, 2004). doi:10.1152/jn.00087.2004
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
92/1/609    most recent
00087.2004v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Roorda, R. D.
Right arrow Articles by Miesenbock, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Roorda, R. D.
Right arrow Articles by Miesenbock, G.
Submitted on January 29, 2004
Accepted on February 25, 2004

Video-rate Nonlinear Microscopy of Neuronal Membrane Dynamics with Genetically Encoded Probes

Robert D. Roorda1, Tobias M. Hohl1, Ricardo Toledo-Crow1, and Gero Miesenbock1*

1 Laboratory of Neural Systems, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

* To whom correspondence should be addressed. E-mail: g-miesenboeck{at}ski.mskcc.org.

Biological membranes decorated with suitable contrast agents give rise to nonlinear optical signals such as two-photon fluorescence and harmonic up-conversion when illuminated with ultra-short, high-intensity pulses of infrared laser light. Microscopic images based on these nonlinear contrasts were acquired at video or higher frame rates by scanning a focused illuminating spot rapidly across neural tissues. The scan engine relied on an acousto-optic deflector (AOD) to produce a fast horizontal raster and on corrective prisms to offset the AOD-induced dispersion of the ultra-short excitation light pulses in space and time. Two membrane-bound derivatives of the green fluorescent protein (GFP) were tested as nonlinear contrast agents. Synapto-pHluorin, a pH-sensitive GFP variant fused to a synaptic vesicle membrane protein, provided a time-resolved fluorescent read-out of neurotransmitter release at genetically specified synaptic terminals in the intact brain. Arrays of dually lipidated GFP molecules at the plasma membrane generated intense two-photon fluorescence but no detectable second-harmonic power. Comparison with second-harmonic generation by membranes stained with a synthetic styryl dye suggested that the genetically encoded chromophore arrangement lacked the orientational anisotropy and/or dipole density required for efficient coherent scattering of the incident optical field.




This article has been cited by other articles:


Home page
 J. Neurosci.Home page
L. Sjulson and G. Miesenbock
Rational Optimization and Imaging In Vivo of a Genetically Encoded Optical Voltage Reporter
J. Neurosci., May 21, 2008; 28(21): 5582 - 5593.
[Abstract] [Full Text] [PDF]

Home page
 PhysiologyHome page
L. Sjulson and G. Miesenbock
Optical Recording of Action Potentials and Other Discrete Physiological Events: A Perspective from Signal Detection Theory
Physiology, February 1, 2007; 22(1): 47 - 55.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurophysiol.Home page
V. Iyer, T. M. Hoogland, and P. Saggau
Fast Functional Imaging of Single Neurons Using Random-Access Multiphoton (RAMP) Microscopy
J Neurophysiol, January 1, 2006; 95(1): 535 - 545.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurophysiol.Home page
D. A. Dombeck, L. Sacconi, M. Blanchard-Desce, and W. W. Webb
Optical Recording of Fast Neuronal Membrane Potential Transients in Acute Mammalian Brain Slices by Second-Harmonic Generation Microscopy
J Neurophysiol, November 1, 2005; 94(5): 3628 - 3636.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Visit Other APS Journals Online
Copyright © 2004 by the The American Physiological Society.