期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 148, 期 1, 页码 94-102出版社
ELSEVIER
DOI: 10.1016/j.jneumeth.2005.06.027
关键词
voltage-sensitive dyes; potentiometric dyes; two-photon microscopy; cross-sections; styryl dyes; nonlinear optical properties; chimeric dyes
资金
- NINDS NIH HHS [NS16824, NS40966] Funding Source: Medline
Microscopy based on voltage-sensitive dyes has proven effective for revealing spatio-temporal patterns of neuronal activity in vivo and in vitro. Two-photon microscopy using voltage-sensitive dyes offers the possibility of wide-field visualization of membrane potential on sub-cellular length scales, hundreds of microns below the tissue surface. Very little information is available, however, about the utility of voltage-sensitive dyes for two-photon imaging purposes. Here we report on measurements of two-photon fluorescence excitation cross-sections for nine voltage-sensitive dyes in a solvent, octanol, intended to simulate the membrane environment. Ultrashort light pulses from a Ti: sapphire laser were used for excitation from 790 to 960 nm, and fluorescein dye was used as a calibration standard. Overall, dyes RH795, RH421, RH414, di-8-ANEPPS, and di-8-ANEPPDHQ had the largest two-photon excitation cross-sections (similar to 15 x 10(-50) cm(4) s photon(-1)) in this wavelength region and are therefore potentially useful for two-photon microscopy. Interestingly, di-8-ANEPPDHQ, a chimera constructed from the potentiometric dyes RH795 and di-8-ANEPPS, exhibited larger cross-sections than either of its constituents. (c) 2005 Elsevier B.V. All rights reserved.
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