High-speed amplitude modulation of femtosecond laser pulses for frequency-multiplexed two photon imaging

Dmitri Tsyboulski; Natalia Orlova; Peter Saggau

doi:10.1117/12.2252627

21 February 2017 High-speed amplitude modulation of femtosecond laser pulses for frequency-multiplexed two photon imaging

Dmitri Tsyboulski, Natalia Orlova, Peter Saggau

Author Affiliations +

Proceedings Volume 10069, Multiphoton Microscopy in the Biomedical Sciences XVII; 100691T (2017) https://doi.org/10.1117/12.2252627
Event: SPIE BiOS, 2017, San Francisco, California, United States

Abstract

We present a frequency-multiplexed multi-site two-photon imaging method utilizing amplitude modulation of femtosecond laser pulses in the MHz range to tag each excitation beam and the corresponding fluorescence signals with specific frequencies. The frequency tags are generated with an interferometric setup employing acousto-optic deflectors (AODs) to achieve precise spatial overlap of femtosecond pulses with acoustically-shifted frequencies. Creating matching excitation beam patterns in each interferometer arm using multiple AOD driving frequencies and subsequently overlapping these matching patterns results in multiple excitation beams with unique beat frequencies available for scanning. As a proof-of-concept, we demonstrate multiplexed two-photon image acquisition using test samples, and compare the performance of this approach to conventional two-photon laser scanning microscopy.

Citation Download Citation

Dmitri Tsyboulski, Natalia Orlova, and Peter Saggau "High-speed amplitude modulation of femtosecond laser pulses for frequency-multiplexed two photon imaging", Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 100691T (21 February 2017); https://doi.org/10.1117/12.2252627

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available