Simultaneous multiple-excitation multiphoton microscopy yields increased imaging sensitivity and specificity |
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| Authors: | Margaret T Butko Mikhail Drobizhev Nikolay S Makarov Aleksander Rebane Brendan C Brinkman Joseph G Gleeson |
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| Affiliation: | (1) Departments of Neuroscience and Biomedical Graduate Program, Howard Hughes Medical Institute, University of California, 92093 San Diego, La Jolla, CA, USA;(2) Department of Physics, Montana State University, 59717 Bozeman, MT, USA |
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| Abstract: | Background Multiphoton microscopy (MPM) offers many advantages over conventional wide-field and confocal laser scanning microscopy (CLSM) for imaging biological samples such as 3D resolution of excitation, reduced phototoxicity, and deeper tissue imaging. However, adapting MPM for critical multi-color measurements presents a challenge because of the largely overlapping two-photon absorption (TPA) peaks of common biological fluorophores. Currently, most multi-color MPM relies on the absorbance at one intermediate wavelength of multiple dyes, which introduces problems such as decreased and unequal excitation efficiency across the set of dyes. |
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