Theses – Optical System for Two-Photon Excitation with Temporal Focusing

Master Thesis Phy/PoL/ET/MT

Motivation

Background:
Many different optical methods are used for studying biological tissue. The penetration depths of these methods is usually limited to several tens of micrometers by the strongly wavelength-dependent scattering in tissue. Using much less scattered near infrared light, powerful ultrashort pulsed lasers allow for a much deeper tissue penetration and simultaneous micrometer-scale optical sectioning similar to confocal microscopy e.g. by the use of two-photon fluorescence processes.

Scope:
In this work, a femtosecond laser will be used to set up an optical system to realize the cutting edge two-photon excitation method of temporal focusing. Here, femtosecond light pulses are spread out temporally by a grating such that all components only overlap temporally in the desired measurement volume to reach the field strengths necessary for two-photon processes. Adaptive optics will be used to shape illumination patterns in the sample plane. The setup will be characterized optically as well as in its ability to stimulate light-sensitized biological tissue in exemplary measurements.

Left: Exemplary setup for the two-phtoton stimulation of light-sensitive tissue with temporal focusing. Right: Time-averaged
imaging volume of Ca2+ signaling of neurons in mouse hippocampus [Prevedel, R. et al. Fast volumetric calcium imaging across multiple
cortical layers using sculpted light. Nat Methods 13, 1021–1028 (2016).]

Range of Tasks

  • Design and setting up of an optical system for two photon stimulation with temporal focusing
  • Optical characterization of the system
  • Assessment off suitability for cell stimulation with exemplary measurements

Related Topics

Optics, two-photon microscopy, temporal focusing, ultrashort pulsed laser

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