Multifocal multiphoton microscopy with adaptive optical correction

Coelho, Simon, Poland, Simon, Krstajic, Nikola, Li, David, Moneypenny, James, Walker, Richard, Tyndall, David, Ng, Tony, Henderson, Robert and Ameer-Beg, Simon (2013) Multifocal multiphoton microscopy with adaptive optical correction. In: SPIE BiOS.

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Abstract

Fluorescence lifetime imaging microscopy (FLIM) is a well established approach for measuring dynamic signalling events inside living cells, including detection of protein-protein interactions. The improvement in optical penetration of infrared light compared with linear excitation due to Rayleigh scattering and low absorption have provided imaging depths of up to 1mm in brain tissue but significant image degradation occurs as samples distort (aberrate) the infrared excitation beam. Multiphoton time-correlated single photon counting (TCSPC) FLIM is a method for obtaining functional, high resolution images of biological structures. In order to achieve good statistical accuracy TCSPC typically requires long acquisition times. We report the development of a multifocal multiphoton microscope (MMM), titled MegaFLI. Beam parallelization performed via a 3D Gerchberg–Saxton (GS) algorithm using a Spatial Light Modulator (SLM), increases TCSPC count rate proportional to the number of beamlets produced. A weighted 3D GS algorithm is employed to improve homogeneity. An added benefit is the implementation of flexible and adaptive optical correction. Adaptive optics performed by means of Zernike polynomials are used to correct for system induced aberrations. Here we present results with significant improvement in throughput obtained using a novel complementary metal-oxide-semiconductor (CMOS) 1024 pixel single-photon avalanche diode (SPAD) array, opening the way to truly high-throughput FLIM. © (2013) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.

Item Type: Conference or Workshop Item (Paper)
Additional Information: Publisher: International Society for Optics and Photonics
Schools and Departments: School of Engineering and Informatics > Engineering and Design
Subjects: Q Science > QC Physics > QC0350 Optics. Light > QC0450 Spectroscopy
T Technology > TK Electrical engineering. Electronics Nuclear engineering > TK0452 Electric apparatus and materials. Electric circuits. Electric networks
Depositing User: David Day-Uei Li
Date Deposited: 26 Feb 2013 15:14
Last Modified: 18 Apr 2013 06:09
URI: http://sro.sussex.ac.uk/id/eprint/43831
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