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Neurophotonics • Open Access

Computer-generated holography enhances voltage dye fluorescence discrimination in adjacent neuronal structures
Author(s): Amanda J. Foust; Valeria Zampini; Dimitrii Tanese; Eirini Papagiakoumou; Valentina Emiliani

Paper Abstract

Voltage-sensitive fluorescence indicators enable tracking neuronal electrical signals simultaneously in multiple neurons or neuronal subcompartments difficult to access with patch electrodes. However, efficient widefield epifluorescence detection of rapid voltage fluorescence transients necessitates that imaged cells and structures lie sufficiently far from other labeled structures to avoid contamination from out of focal plane and scattered light. We overcame this limitation by exciting dye fluorescence with one-photon computer-generated holography shapes contoured to axons or dendrites of interest, enabling widefield detection of voltage fluorescence with high spatial specificity. By shaping light onto neighboring axons and dendrites, we observed that dendritic back-propagating action potentials were broader and slowly rising compared with axonal action potentials, differences not measured in the same structures illuminated with a large “pseudowidefield” (pWF) spot of the same excitation density. Shaped illumination trials showed reduced baseline fluorescence, higher baseline noise, and fractional fluorescence transient amplitudes two times greater than trials acquired with pWF illumination of the same regions.

Paper Details

Date Published: 7 January 2015
PDF: 8 pages
Neurophoton. 2(2) 021007 doi: 10.1117/1.NPh.2.2.021007
Published in: Neurophotonics Volume 2, Issue 2
Show Author Affiliations
Amanda J. Foust, Univ. Paris Descartes (France)
Valeria Zampini, Univ. Paris Descartes (France)
Dimitrii Tanese, Univ. Paris Descartes (France)
Eirini Papagiakoumou, Univ. Paris Descartes (France)
INSERM (France)
Valentina Emiliani, Univ. Paris Descartes (France)

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