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Notes on two-photon microscopy

Yoshida, Shawn (2022) Notes on two-photon microscopy. [Teaching Resource] (Unpublished)

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Two-photon microscopy allows for the imaging of both fixed and live tissues orders of magnitude deeper than what is possible with standard widefield microscopy techniques and does so at higher resolutions than those conventional techniques. There are numerous structures that were previously unable to be imaged in live specimens until the development of two-photon microscopy due to their depth in the tissue. While confocal microscopy can also image deeper into tissues than conventional widefield techniques, the penetration depth is still insufficient for many biological structures of interest. Before the development of two-photon microscopy, the primary methods for imaging such structures were tissue sectioning and scaling, i.e. making the tissues transparent via soaking in chemical solvents. Both of these methods are incompatible with living samples, and while they can also achieve two-photon-like resolutions, they cannot visualize any dynamics. Two-photon microscopy exploits the nonlinearity of its namesake effect, and by only exciting near the waist of the beam in the imaging plane, is able to achieve millimeter-depth imaging in tissues at higher resolutions.

Item Type:Teaching Resource
Yoshida, Shawn0000-0002-0866-2741
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Alternate Title:Two-photon microscopy enables sub-diffraction limit characterizations of millimeter-depth features in living specimens
Subject Keywords:Two-photon microscopy, two-photon excitation, confocal microscopy, fluorescence, nonlinear optics, biological imaging
Record Number:CaltechAUTHORS:20220603-000125134
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:115012
Deposited By: Shawn Yoshida
Deposited On:03 Jun 2022 16:50
Last Modified:03 Jun 2022 16:50

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