Multiphoton Imaging - Biomedical Optical Imaging Technologies: Design and Applications - page 230

Chapter 7
Multiphoton Imaging

Shakil Rehman and Colin J.R. Sheppard

The nonlinear process of multiphoton imaging used in an optical microscope has
inherent out-of-focus rejection of light and hence gives superior optical sectioning
without a pinhole when compared with a confocal microscope. The lateral resolution
remains close to the diffraction limit of the imaging optics. The flexibility of
combining the various nonlinear contrast mechanisms, multiphoton microscopy is
likely to become a major imaging modality in biomedical fields. With the current
development of femtosecond pulsed laser technology, it is to be expected that
multiphoton optical microscopes will continue to be advanced for high-resolution
imaging in a variety of biomedical applications. This chapter will first discuss the
principle of multiphoton process and then multiphoton imaging modes. This chapter
will also provide a detailed discussion on the design of a multiphoton imaging
system.

7.1 Introduction

Multiphoton imaging is fundamentally different from single photon imaging in
the sense that at very high energy density of photons at the focus of an imaging
system, nonlinear absorption or scattering dominates. This results in the observation

S. Rehman ( )
Division of BioEngineering, National University of Singapore, Singapore 117576, Singapore
Singapore-MIT Alliance for Research and Technology, 1 CREATE Way,
#10-01 CREATE Tower, Singapore 138602
e-mail: [e-mail]
C.J.R. Sheppard
Division of BioEngineering, National University of Singapore, Singapore 117576, Singapore
Department of Biological Sciences, National University of Singapore, Singapore 117543,
Singapore