Real time imaging of the excitation volume of a multiphoton microscope

Knowledge of the excitation profile in a confocal or multiphoton microscope can improve the image resolution, e.g. by using deconvolution, pixel reassignment or adaptive optics strategies. Here we demonstrate a method by which the scanning beam can be used to place a stationary, virtual ‘guide star’...

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Bibliographic Details
Published in:Journal of optics (2010) 2022-06, Vol.24 (6), p.64012
Main Authors: Kumar Maity, Barun, Roy, Debsankar Saha, Maiti, Sudipta
Format: Article
Language:English
Subjects:
deconvolution
guide star
point spread function (PSF)
probe volume spread function (PVSF)
PSF
super-resolution
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Summary:Knowledge of the excitation profile in a confocal or multiphoton microscope can improve the image resolution, e.g. by using deconvolution, pixel reassignment or adaptive optics strategies. Here we demonstrate a method by which the scanning beam can be used to place a stationary, virtual ‘guide star’ at any chosen location in the sample, during imaging. This can then be used to directly image the excitation profile. The major advantage of our easy-to-install method, compared to competing methods, is that it can work for non-descanned multiphoton microscopy, the method of choice for deep tissue or ultraviolet imaging. Our experimental results reproduce diffraction theory based calculations in a minimally-scattering sample, and provide detailed information about the aberrated excitation profile in a highly scattering sample.
ISSN:2040-8978
2040-8986
DOI:10.1088/2040-8986/ac69f5