ABSTRACT Stimulation of light-sensitive chemical probes has become a powerful tool for the study of dynamic signaling processes in living tissue. Classically, this approach has been

constrained by limitations of lens-based and point-scanning illumination systems. Here we describe a microscope configuration that incorporates a nematic liquid-crystal spatial light

modulator to generate holographic patterns of illumination. This microscope can produce illumination spots of variable size and number, and in patterns shaped to precisely match user-defined

elements in a specimen. Using holographic illumination to photolyze caged glutamate in brain slices, we show that shaped excitation on segments of neuronal dendrites and simultaneous,

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ULTRAFAST LIGHT TARGETING FOR HIGH-THROUGHPUT PRECISE CONTROL OF NEURONAL NETWORKS Article Open access 05 April 2023 FAST OPTICAL RECORDING OF NEURONAL ACTIVITY BY THREE-DIMENSIONAL

CUSTOM-ACCESS SERIAL HOLOGRAPHY Article 23 December 2021 CONSTRUCTION AND USE OF AN ADAPTIVE OPTICS TWO-PHOTON MICROSCOPE WITH DIRECT WAVEFRONT SENSING Article 01 November 2023 REFERENCES *

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Article  Google Scholar  Download references ACKNOWLEDGEMENTS We thank D. Cojoc, E. Ferrari and E. di Fabrizio for collaborating in the development of the software; E. Papagiakoumou for

helping to set up the double microscope system; D. Oron (Weizmann Institute of Science) for stimulating discussions and providing the coumarin samples; D. Ogden for stimulating discussions,

comments on the manuscript and suggesting the use of 405-nm laser as a source; and A. Silver and A. Marty for critical reading of the manuscript. This work was supported by grants from the

European Science Foundation and the Centre Nationale de la Recherche Scientifique through the European Young Investigators program (to V.E.), the European Commission FP6 Specific Targeted

Project “Photolysis” LSHM-CT-2007-037765 (to V.E., S.C. and D.A.D.), Fondation Fyssen, the Actions Thématiques et Incitatives sur Programme Jeune Chercheurs, Federation pour la Recherche sur

le Cerveau (to D.A.D.), Foundation Bettencourt-Schueller (to V.E. and S.C.), US National Institutes of Health grant NS49238 (to T.S.O.), the Network of European Science Institutes (to

D.A.D. and V.E.) and the Alexander von Humboldt Foundation (to C.L.). AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Neurophysiology and New Microscopy Laboratory, Centre National de la

Recherche Scientifique, Unité Mixte de Recherche 8154, Institut National de la Santé et de la Recherche Médicale, U603, Université Paris Descartes, 45 Rue des Saints Pères, Paris, 75270,

Cedex 06, France Christoph Lutz, Vincent DeSars, Serge Charpak & Valentina Emiliani * Laboratory of Brain Physiology, Centre National de la Recherche Scientifique, Unité Mixte de

Recherche 8118, Université Paris Descartes, 45 Rue des Saints Pères, Paris, 75270, Cedex 06, France Thomas S Otis & David A DiGregorio * Department of Neurobiology, David Geffen School

of Medicine, University of California Los Angeles, 10833 LeConte Avenue, Los Angeles, 90095, California, USA Thomas S Otis Authors * Christoph Lutz View author publications You can also

search for this author inPubMed Google Scholar * Thomas S Otis View author publications You can also search for this author inPubMed Google Scholar * Vincent DeSars View author publications

You can also search for this author inPubMed Google Scholar * Serge Charpak View author publications You can also search for this author inPubMed Google Scholar * David A DiGregorio View

author publications You can also search for this author inPubMed Google Scholar * Valentina Emiliani View author publications You can also search for this author inPubMed Google Scholar

CONTRIBUTIONS C.L. set up the holographic optical microscope and performed the theoretical simulation for axial beam propagation; C.L., T.S.O. and D.A.D. performed and analyzed the

cerebellar brain slice experiments; C.L. and S.C. performed and analyzed the hippocampal brain slice experiments; T.S.O., S.C. and D.A.D. contributed to writing the manuscript. V.D.

developed the software. V.E. conceived and supervised the project, and prepared the manuscript. CORRESPONDING AUTHOR Correspondence to Valentina Emiliani. SUPPLEMENTARY INFORMATION

SUPPLEMENTARY TEXT AND FIGURES Supplementary Notes 1–3, Supplementary Methods (PDF 274 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Lutz, C.,

Otis, T., DeSars, V. _et al._ Holographic photolysis of caged neurotransmitters. _Nat Methods_ 5, 821–827 (2008). https://doi.org/10.1038/nmeth.1241 Download citation * Received: 28 March

2008 * Accepted: 08 July 2008 * Published: 10 August 2008 * Issue Date: September 2008 * DOI: https://doi.org/10.1038/nmeth.1241 SHARE THIS ARTICLE Anyone you share the following link with

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