Arbitrary two-dimensional multiphoton excitation patterns with temporally focused digital holograms

Dan Oron; Eirini Papagiakoumou; Vincent de-Sars; Valentina Emiliani

doi:10.1117/12.808489

3 March 2009 Arbitrary two-dimensional multiphoton excitation patterns with temporally focused digital holograms

Dan Oron, Eirini Papagiakoumou, Vincent de-Sars, Valentina Emiliani

Proceedings Volume 7184, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVI; 71840S (2009) https://doi.org/10.1117/12.808489
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

Multiphoton excitation has recently found application in the fields of bioimaging, uncaging and lithography. In order to fully exploit the advantages of nonlinear excitation, in particular the axial resolution due to nonlinearity, most systems to date operate with point or multipoint excitation, while scanning either the laser beam or the sample to generate the illumination pattern. Here we combine the recently introduced technique of scanningless multiphoton excitation by temporal focusing with recent advances in digital holography to generate arbitrarily shaped, depth resolved, two-dimensional excitation patterns completely without scanning. This is of particular importance in applications requiring uniform excitation of large areas over short time scales, such as neuronal activation by multiphoton uncaging of neurotransmitters. We present an experimental and theoretical analysis of the effect of spatial patterning on the depth resolution achieved in temporal focusing microscopy. It is shown that the depth resolution for holographic excitation is somewhat worse than that achieved for uniform illumination. This is also accompanied by the appearance of a speckle pattern at the temporal focal plane. The origin of the two effects, as well as means to overcome them, are discussed.

Citation Download Citation

Dan Oron, Eirini Papagiakoumou, Vincent de-Sars, and Valentina Emiliani "Arbitrary two-dimensional multiphoton excitation patterns with temporally focused digital holograms", Proc. SPIE 7184, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVI, 71840S (3 March 2009); https://doi.org/10.1117/12.808489

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KEYWORDS

Objectives

Digital holography

Speckle pattern

Holography

Holograms

Multiphoton microscopy

Diffraction gratings

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