Data and image processing for a simultaneous time-and spectrum-resolved multifocal multiphoton microscopy

Zhe Fu; Junle Qu D.D.S.; Ziyang Lin; Lixin Liu; Baoping Guo; Hanben Niu

doi:10.1117/12.741586

1 May 2007 Data and image processing for a simultaneous time-and spectrum-resolved multifocal multiphoton microscopy

Zhe Fu, Junle Qu D.D.S., Ziyang Lin, Lixin Liu, Baoping Guo, Hanben Niu

Proceedings Volume 6534, Fifth International Conference on Photonics and Imaging in Biology and Medicine; 65341S (2007) https://doi.org/10.1117/12.741586
Event: Fifth International Conference on Photonics and Imaging in Biology and Medicine, 2006, Wuhan, China

Abstract

A Simultaneous Time- and Spectrum-Resolved Multifocal Multiphoton Microscopy (STSR-MMM) system has been developed for the acquisition of fluorescence lifetime information of any spectrum of the biological samples in only one measurement. The near-infrared light from mode-locked titanium: sapphire femtosecond laser is split with a microlens array into an array of beams that are transformed into an array of high-aperture foci at the sample for parallel multiphoton excitation. The Gaussian shape of the excitation beam results in non-uniformity and asymmetry of the excitation foci array and also of the reconstructed fluorescence image. A coefficient matrix has been presented for the correction of the non-uniformity and asymmetry in the fluorescence image. Data acquisition and image processing methods have also been investigated to increase the image acquisition speed and improve the reconstruction of time- and spectrum-resolved fluorescence image. The methods for obtaining fluorescence intensity image, spectrally resolved intensity image and spectrally resolved lifetime image are also presented.

Citation Download Citation

Zhe Fu, Junle Qu D.D.S., Ziyang Lin, Lixin Liu, Baoping Guo, and Hanben Niu "Data and image processing for a simultaneous time-and spectrum-resolved multifocal multiphoton microscopy", Proc. SPIE 6534, Fifth International Conference on Photonics and Imaging in Biology and Medicine, 65341S (1 May 2007); https://doi.org/10.1117/12.741586

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