Presentation Type
Poster
Start Date
5-8-2024 11:00 AM
End Date
5-8-2024 1:00 PM
Subjects
Fluorescence microscopy, Laser science and technology
Advisor
Erik Sánchez
Student Level
Undergraduate
Abstract
The objective of this project was to convert a Sarastro 2000 confocal laser scanning microscope into a system capable of imaging using two-photon excitation (TPE) fluorescence 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) due to the lower energy photons causing less damage to delicate biological samples. The adaptation process included physically altering the system by replacing the on-board argon laser with a Ti:sapphire laser, replacing optical components for optimization with the new laser, and implementing a new scan mechanism. The new scan mechanism, a custom all-analog two-mirror galvo head, interfaces with an analog data acquisition board and a computer with software designed for analog motor control and data analysis.
Creative Commons License or Rights Statement
This work is licensed under a Creative Commons Attribution 4.0 License.
Included in
Development of a Two-Photon Laser Scanning Microscope
The objective of this project was to convert a Sarastro 2000 confocal laser scanning microscope into a system capable of imaging using two-photon excitation (TPE) fluorescence 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) due to the lower energy photons causing less damage to delicate biological samples. The adaptation process included physically altering the system by replacing the on-board argon laser with a Ti:sapphire laser, replacing optical components for optimization with the new laser, and implementing a new scan mechanism. The new scan mechanism, a custom all-analog two-mirror galvo head, interfaces with an analog data acquisition board and a computer with software designed for analog motor control and data analysis.