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Neurophotonics • Open Access

Superresolving dendritic spine morphology with STED microscopy under holographic photostimulation
Author(s): Marcel A. Lauterbach; Marc Guillon; Claire Desnos; Dany Khamsing; Zahra Jaffal; François Darchen; Valentina Emiliani

Paper Abstract

Emerging all-optical methods provide unique possibilities for noninvasive studies of physiological processes at the cellular and subcellular scale. On the one hand, superresolution microscopy enables observation of living samples with nanometer resolution. On the other hand, light can be used to stimulate cells due to the advent of optogenetics and photolyzable neurotransmitters. To exploit the full potential of optical stimulation, light must be delivered to specific cells or even parts of cells such as dendritic spines. This can be achieved with computer generated holography (CGH), which shapes light to arbitrary patterns by phase-only modulation. We demonstrate here in detail how CGH can be incorporated into a stimulated emission depletion (STED) microscope for photostimulation of neurons and monitoring of nanoscale morphological changes. We implement an original optical system to allow simultaneous holographic photostimulation and superresolution STED imaging. We present how synapses can be clearly visualized in live cells using membrane stains either with lipophilic organic dyes or with fluorescent proteins. We demonstrate the capabilities of this microscope to precisely monitor morphological changes of dendritic spines after stimulation. These all-optical methods for cell stimulation and monitoring are expected to spread to various fields of biological research in neuroscience and beyond.

Paper Details

Date Published: 22 June 2016
PDF: 8 pages
Neurophoton. 3(4) 041806 doi: 10.1117/1.NPh.3.4.041806
Published in: Neurophotonics Volume 3, Issue 4
Show Author Affiliations
Marcel A. Lauterbach, Univ. Paris Descartes (France)
Marc Guillon, Univ. Paris Descartes (France)
Claire Desnos, Univ. Paris Descartes (France)
Dany Khamsing, Univ. Paris Descartes (France)
Zahra Jaffal, Univ. Paris Descartes (France)
François Darchen, Univ. Paris Descartes (France)
Valentina Emiliani, Univ. Paris Descartes (France)

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