First Advisor
Erik Sánchez
Date of Award
Spring 6-2024
Document Type
Thesis
Degree Name
Bachelor of Science (B.S.) in Physics and University Honors
Department
Physics
Language
English
Subjects
2-photon, fluorescence, far-field, nonlinear optics, bioimaging
Abstract
The objective of this project was to convert a Sarastro 2000 confocal laser scanning microscope (CLSM) into a system capable of far-field two-photon excitation (TPE) imaging for the use of the PSU Biology department. TPE microscopy operates on the ability of fluorophores to accept two photons each with half the energy of a desired transition in a single quantum event via a virtual energy state and then emit a higher energy photon upon relaxation. This is preferable to single-photon excitation (SPE) imaging due to lower photon imaging, causing less damage to delicate biological samples, as well as the inherent localization due to the low excitation volume. The adaptation process included physically altering the CLSM by replacing the onboard argon laser with a Ti:sapphire laser, replacing optical components for compatibility with the new laser, and implementing a new scan mechanism. The new scan mechanism, a custom analog two-mirror galvanometer head, interfaces with an analog data acquisition board and software designed for analog motor control and data analysis. Components were successfully replaced, and future work includes software compatibility and laser alignment and enclosure.
Recommended Citation
Hollenbaugh, Jesseca, "Development of a Two-Photon Imaging System" (2024). University Honors Theses. Paper 1497.