Using adaptive optics for deep in-vivo Multiphoton FLIM

TY - CHAP

T1 - Using adaptive optics for deep in-vivo Multiphoton FLIM

AU - Poland, Simon

AU - Fruhwirth, Gilbert

AU - Ng, Tony

AU - Ameer-beg, Simon

PY - 2011/2/10

Y1 - 2011/2/10

N2 - Multiphoton microscopy (MPM) is a high resolution (sub-mu m) 3D optical imaging technique that has seen widespread use for microscopy at moderate depth within biological tissue (similar to 1 mm). MPM combined with Fluorescence lifetime imaging (FLIM) and Fluorescent resonant energy transfer (FRET) provides the ability to image protein-protein interactions. When applied in-vivo at depth, it will be a key component to identifying and evaluating drug interaction in tumours.Unfortunately as one images more deeply into biological tissue, depth is restricted due to the specimen induced aberrations, which result in deterioration in both the image quality and resolution. Adaptive optics (AO), a technique first developed for astronomy, has been shown to be successful in overcoming problems associated with imaging in depth in confocal, multiphoton, CARS and SHG microscopy. The principle relies on shaping the wavefront with a wavefront modulator to compensate for the distortions introduced by the biological tissue sample. The success of such a technique relies on being able to correctly determine the mirror shape requiredIn this paper we will discuss the development a dedicated MPM FLIM-FRET microscope incorporating an AO for use in-vivo applications. Using a deformable membrane mirror as a wavefront modulator, a strategy for implementation will be discussed.

AB - Multiphoton microscopy (MPM) is a high resolution (sub-mu m) 3D optical imaging technique that has seen widespread use for microscopy at moderate depth within biological tissue (similar to 1 mm). MPM combined with Fluorescence lifetime imaging (FLIM) and Fluorescent resonant energy transfer (FRET) provides the ability to image protein-protein interactions. When applied in-vivo at depth, it will be a key component to identifying and evaluating drug interaction in tumours.Unfortunately as one images more deeply into biological tissue, depth is restricted due to the specimen induced aberrations, which result in deterioration in both the image quality and resolution. Adaptive optics (AO), a technique first developed for astronomy, has been shown to be successful in overcoming problems associated with imaging in depth in confocal, multiphoton, CARS and SHG microscopy. The principle relies on shaping the wavefront with a wavefront modulator to compensate for the distortions introduced by the biological tissue sample. The success of such a technique relies on being able to correctly determine the mirror shape requiredIn this paper we will discuss the development a dedicated MPM FLIM-FRET microscope incorporating an AO for use in-vivo applications. Using a deformable membrane mirror as a wavefront modulator, a strategy for implementation will be discussed.

KW - Microscopy

KW - multiphoton

KW - in-vivo imaging

KW - adaptive optics

KW - FLIM

KW - FRET

KW - ABERRATION CORRECTION

KW - CONFOCAL MICROSCOPY

KW - CELL MOTILITY

KW - ENERGY-TRANSFER

KW - IMPLEMENTATION

KW - ASSOCIATION

U2 - 10.1117/12.875102

DO - 10.1117/12.875102

M3 - Conference paper

SN - 978-0-8194-8440-6

VL - 7903

BT - Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79032C

A2 - Periasamy, Ammasi

A2 - König, Karsten

A2 - T C So, Peter

PB - SPIE - INT SOC OPTICAL ENGINEERING

T2 - Conference on Multiphoton Microscopy in the Biomedical Sciences XI

Y2 - 23 January 2011 through 25 January 2011

ER -