Adaptive optics two-photon fluorescence microscopy

Yaopeng Zhou; Thomas Bifano; Charles Lin

doi:10.1117/12.705045

9 February 2007 Adaptive optics two-photon fluorescence microscopy

Yaopeng Zhou, Thomas Bifano, Charles Lin

Proceedings Volume 6467, MEMS Adaptive Optics; 646705 (2007) https://doi.org/10.1117/12.705045
Event: MOEMS-MEMS 2007 Micro and Nanofabrication, 2007, San Jose, California, United States

Abstract

Non-linear imaging is widely used in biological imaging, primarily because of its ability to image through tissue to depth of a few hundred micrometers. Because two photons need to be absorbed to excite a fluorophore in this instrument, the probability of fluorescence emission of a detectable photon scales with the intensity squared of the beam. As a result, aberrations in the beam path that reduce the peak intensity of the focused, scanned laser spot have a significant effect on the instrument performance. Methods for reducing those aberrations should allow higher resolution and detection sensitivity, and deeper tissue imaging. In this paper, I will describe a non-linear imaging microscope that has an adaptive optics (AO) subsystem to compensate for beam path aberrations. The AO system relies on a 140 actuator deformable mirror, controlled using a stochastic gradient descent algorithm with feedback from a fluorescence sensor. The controlled instrument will be used for in vivo imaging of mouse skin, lymph nodes, and skull bone marrow at depths up to 500 &mgr;m.

Citation Download Citation

Yaopeng Zhou, Thomas Bifano, and Charles Lin "Adaptive optics two-photon fluorescence microscopy", Proc. SPIE 6467, MEMS Adaptive Optics, 646705 (9 February 2007); https://doi.org/10.1117/12.705045

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