Three-dimensional spatiotemporal focusing of holographic patterns.
Two-photon excitation with temporally focused pulses can be combined with phase-modulation approaches, such as computer-generated holography and generalized phase contrast, to efficiently distribute light into two-dimensional, axially confined, user-defined shapes. Adding lens-phase modulations to 2D-phase holograms enables remote axial pattern displacement as well as simultaneous pattern generation in multiple distinct planes. However, the axial confinement linearly degrades with lateral shape area in previous reports where axially shifted holographic shapes were not temporally focused. Here we report an optical system using two spatial light modulators to independently control transverse- and axial-target light distribution. This approach enables simultaneous axial translation of single or multiple spatiotemporally focused patterns across the sample volume while achieving the axial confinement of temporal focusing. We use the system's capability to photoconvert tens of Kaede-expressing neurons with single-cell resolution in live zebrafish larvae.
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, CNRS UMR 8250, Paris Descartes University, UFR Biomédicale, 45 rue des Saints-Pères, 75270 Paris Cedex 06, France.
Oscar
O
Hernandez
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, CNRS UMR 8250, Paris Descartes University, UFR Biomédicale, 45 rue des Saints-Pères, 75270 Paris Cedex 06, France.|Institut national de la santé et de la recherche médicale (Inserm), France.
Eirini
E
Papagiakoumou
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, CNRS UMR 8250, Paris Descartes University, UFR Biomédicale, 45 rue des Saints-Pères, 75270 Paris Cedex 06, France.
Dimitrii
D
Tanese
Institut du Cerveau et de la Moelle Épinière, UPMC, Inserm UMR S975, CNRS UMR 7225, Campus Hospitalier Pitié Salpêtrière, 47 building de l'Hôpital, 75013 Paris, France.
Kevin
K
Fidelin
Institut du Cerveau et de la Moelle Épinière, UPMC, Inserm UMR S975, CNRS UMR 7225, Campus Hospitalier Pitié Salpêtrière, 47 building de l'Hôpital, 75013 Paris, France.
Claire
C
Wyart
Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, CNRS UMR 8250, Paris Descartes University, UFR Biomédicale, 45 rue des Saints-Pères, 75270 Paris Cedex 06, France.
Valentina
V
Emiliani
D000818:Animals; D006696:Holography / Q000295:instrumentation / Q000379:methods*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007814:Larva / Q000254:growth & development / Q000648:ultrastructure; D008027:Light; D008858:Microscopy, Phase-Contrast / Q000295:instrumentation / Q000379:methods*; D009474:Neurons / Q000648:ultrastructure*; D055096:Optical Devices; D015027:Zebrafish / Q000254:growth & development