Dan Fu
University of Washington, Department of Chemistry, USA
Title: Quantitative multiphoton microscopy based on multiplex and ratiometric techniques
Biography
Biography: Dan Fu
Abstract
Multiphoton microscopy is a powerful technique for understanding cellular structure and function in tissue and in living organisms. Currently fluorescence is the most popular contrast mechanism, but there is also a surge in interests of using absorption and Raman as chemical contrasts. Using a high frequency modulation transfer technique, two-photon absorption and stimulated Raman scattering can be detected with the same microscope configuration for multiphoton fluorescence. However, a common challenge in multiphoton microscopy is to quantitatively determine molecular concentration in tissue samples. Light scatters in tissue, resulting in signal loss as it travels deeper into tissue. We developed multiplex and ratiometric techniques that allows us to correct for scattering induced signal loss and recover quantitative molecular information. We further demonstrated quantitative stimulated Raman scattering and two-photon imaging of cells and tissue. We believe these developments will be essential for future biomedical applications of multiphoton microscopy.